ISSN 2087-3940 (PRINT) | ISSN 2087-3956 ... - Biodiversitas

Jatropha curcas photo by A. Abdurrahman 

| Nus Biosci | vol. 3 | no. 1 | pp. 1‐58 | March 2011 | 

ISSN 2087‐3940 (PRINT) | ISSN 2087‐3956 (ELECTRONIC)

THIS PAGE INTENTIONALLY LEFT BLANK


ISSN 2087‐3940 (PRINT) | ISSN 2087‐3956 (ELECTRONIC) 

I S E A Journal o f B i o l o g i c a l S c i e n c e s 

FIRST PUBLISHED: 

2009 

ISSN: 

2087-3940 (printed edition), 2087-3956 (electronic edition) 

EDITORIAL BOARD: 

Abdulaziz M. Assaeed (King Saud University, Riyadh, Saudi Arabia), Alfiono (Sebelas Maret University, Surakarta), Edwi Mahajoeno 

(Sebelas Maret University, Surakarta), Ehsan Kamrani (Hormozgan University, IR Iran), Eko Handayanto (Brawijaya University, 

Malang), Endang Sutariningsih (Gadjah Mada University, Yogyakarta), Faturochman (Gadjah Mada University, Yogyakarta), Iwan 

Yahya (Sebelas Maret University, Surakarta), Jamaluddin (R.D. University, Jabalpur, India), Lien A. Sutasurya (Bandung Institute of 

Technology, Bandung), Magdy Ibrahim El-Bana (Suez Canal University, Al-Arish, Egypt), Mahendra K. Rai (Amravati University, 

India), Marsetyawan H.N. Ekandaru (Gadjah Mada University, Yogyakarta), Oemar Sri Hartanto (Sebelas Maret University, Surakarta), 

R. Wasito (Gadjah Mada University, Yogyakarta), Rugayah (Indonesian Institute of Science, Cibinong-Bogor), Sameer A. Masoud 

(Philadelphia University, Amman, Jordan), Supriyadi (Balitbiogen, Bogor), Sri Margana (Gadjah Mada University, Yogyakarta), Suranto 

(Sebelas Maret University, Surakarta), Sutarno (Sebelas Maret University, Surakarta), Sutiman B. Sumitro (Brawijaya University, 

Malang), Taufikurrahman (Bandung Institut of Technology, Bandung), Wayan T. Artama (Gadjah Mada University, Yogyakarta) 

EDITOR-IN-CHIEF: 

Sugiyarto (sugiyarto_ys@yahoo.com) 

EDITORIAL STAFF: 

Yansen M. Toha (lingkungan_global@yahoo.com), Ari Pitoyo (aripitoyo@yahoo.co.id) 

MANAGING EDITORS: 

Ahmad Dwi Setyawan (unsjournals@gmail.com) 

PUBLISHER: 

“Bioscience Community”, School of Graduates, Sebelas Maret University, Surakarta 

ADDRESS: 

Bioscience Program, School of Graduates, Sebelas Maret University 

Jl. Ir. Sutami 36A Surakarta 57126. Tel. & Fax.: +62-271-663375, Email: nusbioscience@yahoo.com 

ONLINE: 

www.unsjournals.com/nusbioscience 

EXPERTATION OF THE EDITORIAL BOARD: 

AGRICULTURAL SCIENCES: Eko Handayanto (ehn_fp@brawijaya.ac.id), ANTHROPOLOGY: Sri Margana (margo15id@yahoo.com), APPLIED 

BIOLOGICAL SCIENCES: Suranto (surantouns@gmail.com), BIOCHEMISTRY: Wayan T. Artama (artama@ugm.ac.id), NATURAL PRODUCT 

BIOCHEMISTRY: MAHENDRA K. RAI, BIOPHYSICS AND COMPUTATIONAL BIOLOGY: Iwan Yahya (iyahya@uns.ac.id), CELL BIOLOGY: 

Sutiman B. Sumitro (sutiman@brawijaya.ac.id), DEVELOPMENTAL BIOLOGY: Lien A. Sutasurya (lien@bi.itb.ac.id), ECOLOGY: Magdy 

Ibrahim El-Bana (magdy.el-bana@ua.ac.be), ENVIRONMENTAL SCIENCES: Abdulaziz M. Assaeed (assaeed@ksu.edu.sa), EVOLUTION: 

Taufikurrahman (taufik@bi.itb.ac.id), GENETICS: Sutarno (nnsutarno@yahoo.com), IMMUNOLOGY: Marsetyawan H.N. Ekandaru 

(marsetyawanhnes@yahoo.com), MEDICAL SCIENCES: Alfiono (afieagp@yahoo.com), ANIMAL AND VETERINARY SCIENCES: R. Wasito 

(wasito@ugm.ac.id), MICROBIOLOGY: Endang Sutariningsih (annisah-endang@ugm.ac.id), NEUROSCIENCE: Oemar Sri Hartanto 

(oemarsrihartanto@yahoo.com), PHARMACOLOGY: Supriyadi (supriyadi@cbn.net.id), PHYSIOLOGY: Sameer A. Masoud 

(smasoud@philadelphia.edu), PLANT BIOLOGY: Rugayah (titikrugayah@yahoo.com), POPULATION BIOLOGY: Ehsan Kamrani 

(kamrani@hormozgan.ac.ir), PSYCHOLOGICAL AND COGNITIVE SCIENCES: Faturochman (fatur@cpps.or.id), SUSTAINABILITY SCIENCE: 

Jamaluddin (jamaluddin_123@hotmail.com), SYSTEMS BIOLOGY: Edwi Mahajoeno (edmasich@yahoo.com)

GUIDANCE FOR AUTHORS 

NUSANTARA BIOSCIENCES, the ISEA Journal of Biological 

Sciences publishes scientific articles, namely original full research and 

review in all Biological Sciences, including: Agricultural Sciences, 

Anthropology, Applied Biological Sciences, Biochemistry, Natural 

Product Biochemistry, Biophysics and Computational Biology, Cell 

Biology, Developmental Biology, Ecology, Environmental Sciences, 

Evolution, Genetics, Immunology, Medical Sciences, Microbiology, 

Neuroscience, Pharmacology, Physiology, Plant Biology, Population 

Biology, Psychological and Cognitive Sciences, Sustainability Science, 

and Systems Biology. Scientific feedback (short communication) is only 

received for manuscript, which criticize published article before. 

Manuscripts will be reviewed by managing editor, editorial board and 

invited peer review according to their disciplines. The only articles written in 

English (U.S. English) and Bahasa Indonesia are accepted for publication. 

This journal periodically publishes in March, July and November. In order to 

support reduction of global warming and forest degradation, editor prefers 

receiving manuscripts via e-mail rather than in hard copy. Manuscript and its 

communications can only be addressed to the managing editor; better to 

forward to one of the editorial board member for accelerating evaluation. A 

letter of statement expressing that the author (s) is responsible for the 

original content of manuscript, the result of author(s)’s research and never 

been published must be declared. Manuscript of original research should be 

written in no more than 25 pages (including tables and figures), each page 

contain 700-800 word, or proportional with article in this publication number. 

Invited review articles will be accommodated. Avoid expressing idea with 

complicated sentence and verbiage, and used efficient and effective sentence. 

Manuscript is typed at one side of white paper of A4 (210x297 mm 2 ) 

size, in a single column, double space, 12-point Times New Roman font, 

with 2 cm distance step aside in all side. Smaller letter size and space can be 

applied in presenting table. Word processing program or additional software 

can be used, however, it must be PC compatible and Microsoft Word based. 

Names of sub-species until phylum should be written in italic, except for 

italic sentence. Scientific name (genera, species, author), and cultivar or 

strain should be mentioned completely at the first time mentioning it, 

especially for taxonomic manuscripts. Name of genera can be shortened after 

first mentioning, except generating confusion. Name of author can be 

eliminated after first mentioning. For example, Rhizopus oryzae L. UICC 

524, hereinafter can be written as R. oryzae UICC 524. Using trivial name 

should be avoided, otherwise generating confusion. Mentioning of scientific 

name completely can be repeated at Materials and Methods. Biochemical 

and chemical nomenclature should follow the order of IUPAC-IUB, while 

its translation to Indonesian-English refers to Glossarium Istilah Asing- 

Indonesia (2006). For DNA sequence, it is better used Courier New font. 

Symbols of standard chemical and abbreviation of chemistry name can 

be applied for common and clear used, for example, completely written 

butilic hydroxytoluene to be BHT hereinafter. Metric measurement use IS 

denomination, usage other system should follow the value of equivalent with 

the denomination of IS first mentioning. Abbreviation set of, like g, mg, mL, 

etc. do not follow by dot. Minus index (m -2 , L -1 , h -1 ) suggested to be used, 

except in things like “per-plant” or “per-plot”. Equation of mathematics 

can be written separately. Number one to ten are expressed with words, 

except if it relates to measurement, while values above them written in 

number, except in early sentence. Fraction should be expressed in decimal. 

In text, it should be used “%” rather than “gratuity”. 

Title of article should be written in compact, clear, and informative 

sentence preferably not more than 20 words (generally 135 characters 

including spaces). Name of author(s) should be completely written. Running 

title is about five words, refelcting the idea of the manuscript. Name and 

institution address should be also completely written with street name and 

number (location), zip code, telephone number, facsimile number, and e-mail 

address. Manuscript written by a group, author for correspondence along 

with address is required. First page of the manuscript is used for writing 

above information. 

Abstract should not be more than 250 words, written in English, on 

page two of the manuscript. Keywords is about five words, covering 

scientific and local name (if any), research theme, and special methods 

which used. Introduction is about 400-600 words, covering background and 

aims of the research. Materials and Methods should emphasize on the 

procedures and data analysis. Results and Discussion should be written as a 

series of connecting sentences, however, for manuscript with long discussion 

should be divided into sub titles. Thorough discussion represents the causal 

effect mainly explains for why and how the results of the research were 

taken place, and do not only re-express the mentioned results in the form of 

sentences. Conclusion should preferably be given at the end of the 

discussion. Acknowledgments list and funding sources are expressed in a 

brief. Dedications are rarely allowed. 

Figures and Tables of maximum of three pages should be clearly 

presented. Title of a picture is written down below the picture, while title of a 

table is written in the above the table. Colored picture and photo can be 

accepted if information in manuscript can lose without those images. Photos 

and pictures are preferably presented in a digital file. JPEG format should be 

sent in the final (accepted) article. Author could consign any picture or photo 

for front cover, although it does not print in the manuscript. There is no 

appendix, all data or data analysis are incorporated into Results and 

Discussions. For broad data, it can be displayed in website as Supplement. 

Citation in manuscript is written in “name and year” system; and is 

arranged from oldest to newest and from A to Z. The sentence sourced from 

many authors, should be structured based on the year of recently. In citing an 

article written by two authors, both of them should be mentioned, however, 

for three and more authors only the family (last) name of the first author is 

mentioned followed by et al., for example: Saharjo and Nurhayati (2006) or 

(Boonkerd 2003a, b, c; Sugiyarto 2004; El-Bana and Nijs 2005; Balagadde et 

al. 2008; Webb et al. 2008). Extent citation as shown with word “cit” should 

be avoided, and suggested to refer an original reference. 

References. APA style in double space is used in the journal reference. 

Only published or in-press papers and books may be cited in the reference 

list. Unpublished abstracts of papers presented at meetings or references to 

"data not shown" are not permitted. References should be cited in alphabetic 

order. All authors should be named in the citation (unless there are more than 

five). If there are more than five, list the first author's name followed by et al. 

Include the full title for each cited article. Authors must translate foreign 

language titles into English, with a notation of the original language (except 

for Spanish, France, and Germany). For Indonesian manuscript, translation 

of Indonesian title to English is not necessary. For correct abbreviations of 

journal titles, refer to Chemical Abstracts Service Source Index (CASSI). 

Provide inclusive volume, number, and page ranges for journal articles, but 

not for book or book chapters. 

Journal: 

Saharjo BH, Nurhayati AD. 2006. Domination and composition structure 

change at hemic peat natural regeneration following burning; a case 

study in Pelalawan, Riau Province. Biodiversitas 7: 154-158. 

Book: 

Rai MK, Carpinella C. 2006. Naturally occurring bioactive compounds. 

Elsevier, Amsterdam. 

Chapter in book: 

Webb CO, Cannon CH, Davies SJ. 2008. Ecological organization, biogeography, 

and the phylogenetic structure of rainforest tree communities. In: Carson 

W, Schnitzer S (eds) Tropical forest community ecology. Wiley- 

Blackwell, New York. 

Abstract: 

Assaeed AM. 2007. Seed production and dispersal of Rhazya stricta. 50 th 

annual symposium of the International Association for Vegetation 

Science, Swansea, UK, 23-27 July 2007. 

Proceeding: 

Alikodra HS. 2000. Biodiversity for development of local autonomous 

government. In: Setyawan AD, Sutarno (eds) Toward mount Lawu 

national park; proceeding of national seminary and workshop on 

biodiversity conservation to protect and save germplasm in Java island. 

Sebelas Maret University, Surakarta, 17-20 July 2000. [Indonesia] 

Thesis, Dissertation: 

Sugiyarto. 2004. Soil macro-invertebrates diversity and inter-cropping plants 

productivity in agroforestry system based on sengon. [Dissertation]. 

Brawijaya University, Malang. [Indonesia] 

Information from internet: 

Balagadde FK, Song H, Ozaki J, Collins CH, Barnet M, Arnold FH, Quake 

SR, You L. 2008. A synthetic Escherichia coli predator-prey ecosystem. 

Mol Syst Biol 4: 187. www.molecularsystemsbiology.com 

Progress of manuscript. Notification of manuscript whether it is accepted 

or refused will be notified in about three months since the manuscript 

received. Manuscript is refused if the content does not in line with the 

journal mission, low quality, inappropriate format, complicated language 

style, dishonesty of research authenticity, or no answer of correspondence in 

a certain period. Author or first authors at a group manuscript will get one 

original copy of journal containing manuscript submitted not more than a 

month after publication. Offprint or reprint is only available with special request. 

NOTE: Author(s) agree to transfer copy right of published paper to 

NUSANTARA BIOSCIENCES, the ISEA Journal of Biological Sciences. 

Authors shall no longer be allowed to publish manuscript completely without 

publisher permission. Authors or others allowed multiplying article in this 

journal as long as not for commercial purposes. For the new invention, 

authors suggested to manage its patent before publishing in this journal. 

NOTIFICATION: All communications are strongly recommended to be undertaken through email.

Vol. 3, No. 1, Pp. 1-6 

March 2011 

ISSN: 2087-3940 (print) 

ISSN: 2087-3956 (electronic) 

Optimization of DNA extraction of physic nut (Jatropha curcas) by 

selecting the appropriate leaf 

EDI PRAYITNO, EINSTIVINA NURYANDANI ♥ 

Open University, UPBJJ Semarang. Jl. Semarang-Kendal, Mangkang Wetan, Semarang 50156, Central Java, Indonesia, Tel. +62-24-8666044, Fax. +62- 

24-8666045; ♥ email: vina_ut@yahoo.co.id 

Manuscript received: 11 November 2010. Revision accepted: 24 February 2011 (stay empty) 

Abstract. Prayitno E, Nuryandani E. 2011. Optimization of DNA extraction of physic nut (Jatropha curcas) by selecting the appropriate 

leaf. Nusantara Bioscience 3: 1-6. Jatropha curcas L. has important roles as renewable source of bioenergy. The problem occurs on 

difficult of DNA extraction for its molecular breeding programs. The objectives of this research were to study which leaf best as source 

of DNA extraction. Four accession were used, namely J1 and J2 (Jawa Tengah), S1 (South Sumatra), and S2 (Bengkulu). First, third, 

fifth, seventh, and yellow leaves for each accession were extracted using modification of Doyle and Doyle (1987) method. Visualization 

and comparation with Lambda DNA, Spectrophotometer UV-Vis and cutting DNA with EcoRI enzyme were show quality and quantity 

of DNA. The result showed that third leaves have sufficient quality and quantity as source of DNA. Third leaves DNA quantity for J1 

(19.33 µg/mL), J2 (26.21 µg/mL), S1 (31.20 µg/mL), dan S2 (61.03 µg/mL), and quality for each accession were 1.9063 (J1), 2.0162 

(J2), 2.0116 (S1), and 2.0856 (S2). 

Key words: Jatropha curcas, DNA extraction, appropriate, leaf. 

Abstrak. Prayitno E, Nuryandani E. 2011. Optimalisasi ekstraksi DNA jarak pagar (Jatropha curcas) melalui pemilihan daun yang 

sesuai. Nusantara Bioscience. Nusantara Bioscience 3: 1-6. Jarak pagar (Jatropha curcas L.) mempunyai peran penting sebagai sumber 

bahan bakar nabati. Usaha pemuliaan tanaman ini secara molekuler sering terkendala sulitnya ekstraksi DNA. Penelitian ini bertujuan 

untuk mengetahui daun yang sesuai untuk digunakan sebagai sumber DNA. Penelitian ini dilakukan pada empat aksesi jarak pagar yaitu 

J1 dan J2 (Jawa Tengah), S1 (Sumatera Selatan), dan S2 (Bengkulu). Ekstraksi dilakukan pada daun pertama, ketiga, kelima, ketujuh, 

dan daun kuning dari setiap aksesi dengan metode Doyle and Doyle (1987) yang dimodifikasi. Kualitas dan kuantitas DNA hasil 

ekstraksi diketahui melalui visualisasi dengan pembanding DNA lambda, spektrofotometer UV-Vis pada panjang gelombang 260/280, 

dan pemotongan menggunakan enzim EcoRI. Hasil penelitian menunjukkan bahwa daun ketiga memadai untuk digunakan sebagai 

sumber DNA. Kuantitas DNA daun ketiga J1 (19,33 µg/mL), J2 (26,21 µg/mL), S1 (31,20 µg/mL), dan S2 (61,03 µg/mL). Sedangkan 

kemurniannya masing-masing yaitu 1,9063 (J1), 2,0162 (J2), 2,0116 (S1), dan 2,0856 (S2). 

Kata kunci: Jatropha curcas, ekstraksi DNA, daun, sesuai. 

INTRODUCTION 

Increased economic growth and spur the growth of 

population of high energy consumption. Energy source the 

world today is still dominated by fossil fuel that cannot be 

renewed (unrenewable). Various efforts have been made to 

solve energy problems (Raharjo 2007). Fuel from the plant 

has several advantages such as ease of storage and 

environmentally friendly, therefore biofuels were given 

priority for development. On January 25, 2006, the 

President of Indonesia issued Presidential Regulation No. 

5/2006 regarding the national energy policy and 

Presidential Instruction No. 1/2006 concerning the 

provision and use of biofuels as alternative fules. Then on 

July 1, 2006, presidential and state officials conducting a 

retreat in the village of Losari, Grabag subdistric, 

Magelang district, and decided to develop a bioenergy or 

biofuel as an alternative energy. 

Biofuel can be divided into two major categories, 

namely bioethanol and biodiesel. Bioethanol is ethanol 

derived from fermentation of raw materials that contain 

starch or sugar such as molasses and cassava. This fuel can 

be used to replace regular gasoline (gasoline). Ethanol can 

be used is alcohol-free pure water (anhydrous alcohol) and 

levels of more than 99.5%, or called with a fuel grade 

ethanol (FGE). Blend of premium and FGE is called 

gasohol. In Indonesia, Pertamina give biopremium 

trademark for the product. Biodiesel is a popular name for 

FAME (fatty acid methyl ester), is a biofuel that is used to 

power diesel engines as an alternative to diesel. This fuel 

derived from vegetable oils are converted through chemical 

and physical reactions, so that the nature of the chemical 

has changed from its original nature. Currently, Pertamina 

has issued such a product with trade name which is a 

blending FAME biodiesel with regular diesel (petrosolar) 

(Prihandana et al. 2007).

2 

3 (1): 1-6, March 2011 

Jatropha curcas is a native plant of Central America 

(Fairless 2007) and has been naturalized in tropical and 

subtropical regions, including Indonesia. This species is 

drought resistant and is commonly planted as a garden 

fence, but is also useful as an ornamental plant shrubs and 

herbs. Oil from the seeds is useful for medicine, 

insecticides, making soap and candles, as well as raw 

material for biodiesel (Gubitz et al. 1999). The use of 

castor oil as biodiesel ingredient is an ideal alternative, 

because it is a renewable oil resources (renewable fuels) 

and non-edible oil so it does not compete with human 

consumption requirements, such as palm oil, corn, 

soybeans and others (Dwimahyani 2005). In addition, 

Jatropha also contains secondary metabolites which are 

useful as protectant for plants and as an ingredient for 

human medicine (Debnath and Bisen 2008) 

Some of the obstacles encountered in developing castor oil, 

among others, lack of information about varieties that have 

beneficial properties such as high production, fast 

multiplication, high oil yield in seeds, as well as resistance 

to pests and diseases. This happens because so far the 

Jatropha plant is only regarded as hedgerows that have low 

economic value so that research and development of this 

plant is rarely done. To overcome this, plant breeding has a 

significant role. 

Characterization of jatropha plant in Indonesia is 

carried out simply and not be universal. Often, the mention 

of Jatropha plant species is based solely on phenotypic 

appearance or region of origin. Characterization using 

morphological or phenotypic description has limitations 

because it is very influenced by the environment. Different 

morphological features can be caused by environmental 

stress, whereas the same genotype, whereas the same 

morphological features do not necessarily indicate that both 

types of plants are closely related, because the outer shape 

of a plant is the result of cooperation between the genotype 

by environment (Joshi et al. 1999; Karsinah 1999 .) 

Therefore, it is necessary to develop universal genetic 

information. Molecular markers can provide information 

universally because it is not influenced by the environment 

(Azrai 2005), so that they can answer the problem in the 

characterization of physic nut plants. 

Jatropha curcas is one of the many plants that contain 

latex, which is a true plant secondary metabolites. The 

presence of secondary metabolites such as polyphenols, 

tannins, and polysaccharides can inhibit the action of the 

enzyme (Porebski 1997; Pirtilla et al. 2001). Isolation of 

plant DNA at a distance often experienced problems due to 

high levels of secondary metabolites in the form of 

polysaccharides and polyphenols. According to Sharma et 

al. (2002) the presence of metabolites in several crops 

affect DNA isolation procedure, he was using a modified 

CTAB to isolate DNA from plant tissue containing high 

polysaccharide. In line with this Kiefer et al. (2000), Pirtilla 

et al. (2001) and Sanchez-Hernandes, C. and J.C. Gaytan- 

Oyarzun (2006), states that the extraction of DNA and 

RNA from plants containing polysaccharides, polyphenols 

as well as sap and difficult. 

Proper techniques of DNA extraction is needed in the 

plant breeding process to obtain DNA with a high quality 

and quantity. To obtain pure DNA from plant sap, 

generally carried out repeated purification and modification 

of procedures (Kiefer et al. 2000), thus requiring additional 

cost and effort. For that, you can use parts of plants that 

contain little secondary metabolites. The content of 

secondary metabolites in plant tissues fluctuate in line with 

its development. Secondary metabolites may vary because 

of differences in age and plant part (Cirak et al. 2007a, b, 

2008; Achakzai et al. 2009). Therefore, to simplify the 

DNA extraction process jatropha, have done research to 

learn the parts of plants containing secondary metabolites 

in small amounts and produce DNA with high quality and 

quantity. 

This research aims to study the jatropha plant leaves at 

different levels of development that have the potential to 

produce the best quality and quantity of DNA in the DNA 

extraction process. 

MATERIALS AND METHODS 

Time and place of study 

Research was conducted at the Open University UPBJJ- 

Semarang, Central Research Laboratory Tropical Fruit IPB, 

Bogor, West Java, and Laboratory of Structure and 

Function of Plant Diponegoro University in March to 

November 2009. 

Plant material 

Jatropha plant materials used in this study are the three 

accessions of jatropha plants originated from areas of 

Klaten (Central Java) with the code J1 and J2, Palembang 

(South Sumatra) with codes S1, and Bengkulu, with the 

code S2. 

Procedures 

Isolation of DNA. About 0.5 g of leaves from the first, 

third, fifth, seventh and yellow leaves from each sample 

was crushed in porcelain bowls by adding 0.1 grams of 

silica sand to be easily crushed. To prevent network 

browning by oxidation, polivinilpolipirilidon (PVPP) as 

much as 40 mg and added extraction buffer (2% CTAB, 

100 mM Tris-HCl pH 8, 1.4 M NaCl, 20 mM EDTA) as 

much as 1 mL is added into a cup containing the sample 

which has added 1% merkaptoetanol. Samples that have 

been incorporated into the fine volume of 1.5 mL 

Eppendorf tube. Subsequently the mixture incubated at 

65oC for 30 minutes while inverted, and then added 1 mL 

solution of chloroform: isoamilalchohol (24:1 = v/v) and 

divortek for 5 seconds. This solution was then separated 

using a centrifuge with a speed of 11,000 rpm for 10 

minutes at a room temperature. Supernatant was separated 

from the pellet by putting it into a new Ependorf tube. 

DNA in the supernatant was purified by adding 1 mL 

solution of chloroform: isoamilalkohol (24:1 = v/v) and 

disentrifuse at a speed of 11,000 rpm for 10 minutes at 

room temperature. Supernatant was transferred into a tube 

and added with 1 mL of cold isopropanol, shaken gently 

until white threads arise, which is DNA. Subsequently 

DNA was precipitated by incubation for 30 minutes at a

PRAYITNO & NURYANDANI – Optimization of DNA extraction of Jatropha curcas 3 

temperature of -20ºC. Solution containing the DNA that 

has been purified disentrifuse with speed 11 000 rpm for 10 

minutes at room temperature and then the supernatant was 

discarded. DNA precipitate was washed with 70% alcohol 

and dried at room temperature. Further the DNA samples 

that was obtained was dissolved in 100 mL TE buffer (10 

mM Tris-HCl pH 7.5, 10 mM EDTA) and incubated at 37° 

C for one hour and then mixed until uniform to further test 

its quality. 

Test the quality and quantity of DNA. The quantity 

(concentration) and quality of DNA determined by UV-Vis 

spectrophotometer at wavelength 260 and 280 nm. 

Determination of the total DNA quantity was calculated 

based on the value of absorbance at a wavelength of 260 

nm. A at 260 = 1.0 equivalent amount of DNA is 50 

ug/mL. λ DNA quality is considered good if the value of 

A260/280 approaching 1.8 to 2. To determine the 

concentration and quality of DNA, electrophoresis results 

were soaked in a solution of 1% EtBr and then observed 

under UV transluminator. The quantity of DNA is based on 

the thickness of the electrophoresis results of DNA samples 

are compared with the amount of lambda DNA of known 

concentration, ie 250 ug/mL. This study also tested the 

quality of DNA by cutting genomic DNA using EcoRI 

enzyme are visualized by electrophoresis on agarose gel. 

RESULTS AND DISCUSSION 

Visualization of the extracted DNA 

The success of the isolation and extraction process of 

genomic DNA can be marked with resultant large DNA 

(high molecular weight DNA), that is not degraded during 

extraction and purification process, and can be cut by 

restriction enzymes that has been used (Herison 2003). 

Results of isolation and extraction of jatropha’s DNS 

employed Doyle and Doyle method (1987) which has been 

modified to produce the desired genomic DNA bands, 

although relatively small quantity when compared to 

lambda DNA. Genomic DNA was seen as a ribbon that 

lights up at the top sinks electrophoresis results. 

In general, smear on DNA extracted from young leaves 

(code J11, J21, S11, S21) and concentrated look taller than 

the smear on DNA extracted from the older leaves, then 

gradually decreasing concentration smear on leaves more 

old (leaves the third, fifth, and seventh), and smear the least 

present in yellow leaves, except on J1 where J1k (yellow) 

has a thicker ribbon smears compared J15 and J17 (Figure 

1). 

In genomic DNA extracted from young leaves, which 

are visible smear on the bottom of genomic DNA. Ribbon 

smear is a molecule with varying weights that can be 

derived from degraded DNA or other follow-up material 

that is not known (Herison 2003). Smears indicated that the 

isolated genomic DNA was not intact anymore, probably 

dismembered during the extraction takes place (Sisharmini 

et al. 2001). Genomic DNA damage can be caused by 

degradation of secondary compounds that are released 

when the cells were destroyed or damaged due to physical 

handling. The decline is likely influenced by the smear of 

secondary metabolites of plants and physical handling. In 

this case the physical handling for each sample the same 

can be said for using the same standard procedure, 

therefore, the greatest influences that cause differences in 

high and low smear is a secondary metabolite from the 

leaves of plants (Milligan 1992). 

In certain plants, plant metabolites will be seen visually 

in the form of sap. Jatropha curcas is a plant sap, with pink 

latex (de Padua et al. 1999) or nodes in the young gradually 

turns cloudy/older if left in free air or dark brown when 

taken from the older plants (Heyne 1987). Young leaves 

contain more secondary metabolites than older leaves 

(Badawi 2006; Mulyani 2006). Young leaves generally 

contain secondary metabolites and enzymes that high 

because it requires in the process of growth, development, 

and division of cells’ leaf. In the development of plant 

secondary metabolite concentrations will gradually decline 

as the decline in leaf growth activity, and the leaves have 

yellowed, the concentration of enzymes and secondary 

metabolites in the leaves decreased significantly due to the 

ongoing process of senesensi (Salisbury and Ross 1995). 

At this stage the plant will attract substances and enzymes 

that are still useful to the plant from old leaves for use in 

the process of development of the younger plants, so the 

possibility of plant secondary metabolites present in a very 

low level so that the DNA is not much degraded by the 

follow-up compound (Salisbury and Ross 1995; Herison 

2003). Although the smear on the older leaves less and less, 

but the quantity of genomic DNA was also decreased, 

which lights up genomic DNA bands at the top of the wells 

that are running low on older leaves. 

L J11 J13 J15 J17 J1K J21 J23 J25 J27 J2K S11 S13 S15 S17 S1K L 

L S21 S23 S25 S27 S2K L 

Figure 1. Visualization of the extracted DNA from four accessions of Jatropha curcas Klaten (J1, J2), Palembang (S1) and Bengkulu 

(S2). L = LAMDA (ladder)

4 

3 (1): 1-6, March 2011 

The young leaves have a high cleavage activity. In the 

division process, DNA replication will experience, so the 

amount of DNA will double itself, thus DNA concentration 

is relatively high in young leaves. On older leaves, the 

division process could decrease, until finally stopped 

altogether. On the leaves that have yellowed, in addition to 

the absence of the division process, it also exacerbated the 

death of cells that were old, so the amount of DNA was 

also decreased dramatically (Salisbury and Ross 1995). 

Test the quality and quantity of DNA with UV-Vis 

spectrophotometer 

The quantity (concentration) and quality of DNA 

determined by UV-Vis spectrophotometer at wavelength 

260 and 280 nm. Determination of the total DNA quantity 

was calculated based on the value of absorbance at a 

wavelength of 260 nm. The highest DNA purity can be 

seen in the A260/280 ratio that produces the value of 1.8 to 

2. According to Sambrook et al. (1989) DNA with a ratio 

in the range of figures have met the requirements of purity 

required in molecular analysis. Spectrophotometer results 

show relatively good purity DNA that has yet to reach 

100% purity in some accessions. The concentration and 

purity of genomic DNA was analyzed using UV-Vis 

spectrophotometer can be seen in Table 1. 

Genomic DNA which has a purity of 100% contained in 

the accession J1 was extracted from the third leaf with 

value ratio of 1.9063. Genomic DNA from the first leaf 

accession J1 has a value ratios approaching 100% purity 

with ratio of 2.0131. While the three other leaves, that 

leaves the fifth, seventh, and yellow leaves have a value 

ratio of less than 1.8 respectively, 1.7417, 1.2578, and 

1.2356. Results DNA extraction leaves first, third, and fifth 

of the accession J2 has a value closer to purity ratio, 

respectively 2.0697, 2.0162, 2.0914, while the seventh 

leaves and yellow leaves have a ratio value that is still far 

from purity, namely 1.5873 and 1, 1940. 

On the accession of S1, almost all of the extracted DNA 

purity approaching leaves, each leaf of the first, third, fifth, 

and seventh ratio is 2.0768, 2.0116, 2.0792, 2.0225, while 

the yellow leaves have value ratio far from the purity of 

1.4434. DNA extracted first and third leaf from the 

accession of S2 close to the purity of the value ratio of 

2.0611 and 2.0856. While leaf fifth, seventh, and yellow 

leaves have a ratio that is far from the purity of the 

respective ratios 2.2187, 2.1782, and 1.5177. 

Besides the purity of genomic DNA samples, another 

consideration that must be considered is the quantity of 

genomic DNA was generated from the DNA extraction 

process. Readings A260 = 1 means the concentration of 

DNA obtained at 50 ug/mL (Herison 2003). The 

concentration of genomic DNA was extracted was 

calculated by the formula: DNA concentration (ug/mL) = 

A260 x dilution factor x 50 ug/mL. 

DNA concentration resulting from the extraction 

process represents the amount of DNA contained in the leaf 

tissue used for the sample and treatment methods used in 

each sample is the same. Table 1 below is the concentration 

of DNA from samples of twenty leaves from four 

accessions of jatropha plant that is used. From Table 1, 

note the concentration ratio of genomic DNA from leaf 

tissue of each first, third, fifth, seventh, and yellow leaves, 

and comparison of genomic DNA concentration between 

sections. In general, genomic DNA concentration 

decreased with increasing age of leaves used as a sample. 

Samples from the first leaf shows the quantity of 

genomic DNA is much larger than the sample leaves the 

third, fifth, seventh, and yellow leaves. Measurement of the 

quantity of genomic DNA samples from accessions J1 

genomic DNA in Klaten produces relatively little 

compared to the accession of J2, S1, and S2, which is 27.69 

ug/mL for the first leaf, 19.33 ug/mL for the third leaf, 3.68 

tg/mL for the fifth leaf, 2.03 g/mL for the seventh leaf, and 

4.51 ug/mL for yellow leaves. This is due to a smaller 

sample size compared to other accessions due to spill some 

of the samples by laboratory staff who worked on, so that 

DNA samples that were tested got reduced. While the 

accession J2, where accession was also derived from the 

same home with the accession of J1, which was from 

Klaten, Central Java, and comes from the same parent, the 

quantity of genomic DNA generated greater than J1, which 

is 62.06 ug/mL for the extraction of the first leaf, 26.21 

ug/mL for the third leaf, 27.69 ug/mL for the fifth leaf, 

5.37 g/mL for the seventh leaf, and 4.37 ug/mL for yellow 

leaves. 

The concentration of genomic DNA for S1 accession on 

the first leaves produced 67.61 g/mL DNA, whereas the 

third leaf, the concentration of genomic DNA was 31.20 

ug/mL, on the fifth leaves of 46.71 ug/mL, on the seventh 

leaf, 22, 90 ug/mL, and the yellow leaves of 7.59 g/mL. 

Accession S2 on the first leaves produced 101.35 g/mL 

genomic DNA, while the third leaf, the concentration of 

genomic DNA was 61.03 ug/mL, on the fifth leaves of 

44.18 ug/mL, leaves the seventh, 26.27 ug/mL , and the 

yellow leaves of 5.37 g/mL. The Figure 1 shows the 

concentration of the extracted genomic DNA of 

Table 1. Test the quality (purity) and quantity (concentration) of DNA using UV-Vis spectrophotometer in four accessions of jatropha 

from Klaten (J1, J2), Palembang (S1) dan Bengkulu (S2). 

Leaves 

DNA purity 

DNA concentration (µg/mL) 

J1 J2 S1 S2 J1 J2 S1 S2 

First 2.0131 2.0697 2.0768 2.0611 27.69 62.06 67.61 101.35 

Third 1.9063 2.0162 2.0116 2.0856 19.33 26.21 31.20 61.03 

Fifth 1.7417 2.0914 2.0792 2.2187 3.68 27.69 46.71 44.18 

Seventh 1.2578 1.5873 2.0225 2.1782 2.03 5.37 22.90 26.27 

Yellow 1.2356 1.1940 1.4434 1.5177 4.51 4.37 7.59 5.37

PRAYITNO & NURYANDANI – Optimization of DNA extraction of Jatropha curcas 5 

diminishing. This is related to the phase of leaf 

development that has been outlined above. 

Results spectrophotometer for quantity of genomic 

DNA of the above shows that the largest quantity of 

genomic DNA from four accessions were found in the 

extraction of the first leaf. But considering the quality of 

the resulting DNA, the highest purity approaching 100% 

are found in the sample using the third leaf as a source of 

genomic DNA, although in terms of quantity, the number is 

lower than the samples originated from the first leaf. 

Comparison of DNA extracted from five types of leaf 

samples from accessions used in J1 and J2 from Klaten, 

from the same parent tree can be seen in Table 1. From 

Table 1 it can be seen that the DNA genome of the first and 

second leaf (accession J1) and leaves the first, third, and 

fifth (accession J2) approached the purity, but purity is 

closest to the third leaf (accession J1 on the ratio of 1, 9063 

(purity 100%) and the accession to the ratio of 2.0162 A2). 

But in terms of quantity, J1 and J2 are not comparable 

although originating from the same parent because of the 

sample is not the same J1 J2 terms of number of samples 

tested for spill samples by the laboratory. 

Some researches indicate that generally young leaves 

are used in DNA extraction because of the ease in getting 

the DNA with a high quantity. Mansyah et al. (2003) who 

conducted research on mangosteen states that extraction of 

DNA from old leaves is more difficult when compared 

with young leaves, so as to obtain DNA from old leaves 

with a sufficient quantity is required special treatment, 

namely with the addition of the extracted leaves up to 2 g 

and DNA purification with the addition of RNase. While 

Prana (2003) who perform DNA extraction on taro plants 

also use the young leaves (in this case the leaf shoots) as 

the source of DNA. 

Test the quality of DNA by using the enzyme EcoRI 

cuts 

The purity of DNA can be seen from the absence of a 

DNA sample can be cut by restriction enzyme such as 

EcoR1 (Figure 2). If a DNA sample has high purity, this 

DNA would be easy to cut by restriction enzymes. But if 

this is still contain DNA samples follow-up materials such 

as secondary metabolites, carbohydrates, proteins, and 

others, will hinder the work restriction enzymes. 

Whether DNA can be cut with restriction enzymes is 

visible from at least smear results of electrophoresis bands 

after DNA cut with EcoRI enzyme (Herison 2003). EcoRI 

produce DNA bands when smears were electrophoresed 

because this restriction enzyme included in the frequent 

cutter (Vos et al. 1995). The result of cutting with EcoRI 

enzyme produces DNA fragments that appear as a smear 

on some samples, but most other samples can not be cut by 

this enzyme because of the high follow-up compounds that 

inhibit enzymes work. Smear only be observed in J13 and 

J15, while the other samples have not seen a clear smear as 

a result of enzyme EcoRI. Visible is the presence of minor 

compounds in the lower section sinks. possible follow-up 

material that inhibits this enzyme EcoRI work so as not to 

cut the genomic DNA tested jatropha. 

The description above discussion shows that differences 

in leaf tissue age used influence the extraction of genomic 

DNA where the younger leaves will produce a quantity of 

genomic DNA was higher but also accompanied by the 

high follow-up material in the form of plant secondary 

metabolites that inhibit the work in the field of molecular 

further. Older leaves to produce the amount of genomic 

DNA are relatively fewer compared to young leaves, but 

the following secondary metabolites was also reduced in 

number. This study shows that the third leaf is better used 

as a source of genomic DNA since the DNA purity is better 

than the other leaves, and the quantity produced enough 

DNA to be used for further molecular analysis. 

CONCLUSION 

The third leaf physic nut plants suitable for use as a 

source of DNA for molecular analysis of genomes, as in 

quantity and quality sufficient to produce genomic DNA 

for further molecular analysis such as PCR. Genomic DNA 

extracted from the third leaf is generally close to 100% 

purity and quantity of DNA produced is also large enough 

to be used for further molecular analysis. 

M J11 J13 J15 J17 J1k J21 J23 J25 J27 J2K S11 S13 S15 S17 S1K S21 S23 S25 S27 S2k M 

Figure 2. Visualization of results by the enzyme EcoRI cuts at the four accessions of jatropha from Klaten (J1, J2), Palembang (S1) and 

Bengkulu (S2).

6 

3 (1): 1-6, March 2011 

REFERENCES 

Achakzai AKK, Achakzai P, Masood A, Kayan SA, Tareen RB. 2009. 

Response of plant parts and age on the distribution of secondary 

metabolites on plants found in Quetta. Pak J Bot 41 (5): 2129-2135. 

Azrai M. 2005. Pemanfaatan markah molekuler dalam proses seleksi 

pemuliaan tanaman. J Agrobiogen 1 (1): 26-37. [Indonesia] 

Badawi A. 2007. Pengaruh tingkat ketuaan daun dan dosis filtrat daun 

saga (Abrus precatorius) terhadap kadar billirubin serum 

darah tikus putih (Ratus novergicus) yang diinduksi 

dengan karbon tetraklorida (CCl 4 ) [Tesis S1]. Universitas 

Muhammadiyah Malang. Malang. [Indonesia] 

Cirak C, Radušienė J, Ivanauskas L, Janulis V. 2007b. Variation of 

bioactive secondary metabolites in Hypericum perfoliantum during its 

phonological cycle. Acta Physiol Plant 29: 197-203. 

Cirak C, Radušienė J, Janulis V, Ivanauskas L. 2007a. Secondary 

metabolites in Hypericum perfoliantum: variation among plant parts 

and phonological stages. Bot Helv 117: 29-36. 

Cirak C, Radušienė J, Janulis V, Ivanauskas L. 2008. Pseudohypercin and 

hyperforin in Hypericum perforatum from Northern Turkey; variation 

among populations, plant parts and phonological stages. J Integ Plant 

Biol 50: 575-580. 

De Padua LS, Bunyaprahatsara N, Lemmens RHMJ. 1999. Plant 

Resources of South East Asia No. 12 (1) Medicinal and poisonous 

plants. Backhuys. Leiden. 

Debnath M, Bisen PS. 2008. Jatropha curcas L., a multipurpose stress 

resistant plant with a potential for ethnomedicine and renewable 

energy. Curr Pharm Biotechnol 9 (4): 288-306. 

Doyle JJ, Doyle JL. 1987. A rapid DNA isolation procedure for small 

quantities of fresh leaf tissue. Phytochem Bull19: 11-15. 

Dwimahyani I. 2005. Pemuliaan mutasi tanaman jarak pagar (Jatropha 

curcas L.). P3TIR-BATAN. Jakarta. [Indonesia] 

Fairless D. 2007. Biofuel: The little shrub that could – maybe. Nature 

449: 652-655. 

Gübitz GM, Mittelbach M, Trabi M. 1999. Exploitation of the tropical oil 

seed plant Jatropha curcas L. Bioresour Technol 67: 73-82. 

Herison C, Rustikawati, Eliyanti. 2003. Penentuan protokol yang tepat 

untuk menyiapkan DNA genom cabai (Capsicum sp.). J Akta Agrosia 

6 (2): 38-43. [Indonesia] 

Heyne K. 1987. Tumbuhan berguna Indonesia II. Yayasan Sarana 

Wanajaya. Jakarta. [Indonesia] 

Instruksi Presiden No. 1 Tahun 2006 tanggal 25 Januari 2006 tentang 

penyediaan dan pemanfaatan bahan bakar nabati (biofuel) sebagai 

bahan bakar lain. [Indonesia] 

Joshi SP, Ranjekar PK, Gupta VS. 1999. Molecular markers in plant 

genome analysis. Curr Sci 77 (2): 230-240. 

Karsinah. 1999. Keragaman genetic plasma nutfah jeruk berdasarkan 

analisis penanda RAPD. [Tesis]. Institut Pertanian Bogor. Bogor. 

[Indonesia] 

Kiefer E, Heller W, dan Ernst D. 2000. A simple and efficient protocol for 

isolation of functional RNA from plant tissues rich in secondary 

metabolites. Plant Mol Biol Rep 18: 33-39. 

Mansyah E, Baihaki A, Setiamihardja R, Darsa JS, Sobir. 2003. Analisis 

variabilitas genetik manggis (Garcinia mangostana L.) di Jawa dan 

Sumatera Barat menggunakan teknik RAPD. Zuriat 14 (1): 36-44. 

[Indonesia] 

Milligan BG. 1992. Plant DNA isolation. In: Hoelzel AR (ed). Molecular 

genetic analysis of populations; a practical approach. Oxford 

University Press. New York. 

Mulyani A, Agus F, Allelorung D. 2006. Potensi sumber daya lahan untuk 

pengembangan jarak pagar (Jatropha curcas L.) di Indonesia. J 

Litbang Pertanian 25(4): 130-138. [Indonesia] 

Peraturan Presiden No. 5 Tahun 2006 tanggal 25 Januari 2006 tentang 

kebijakan energi nasional Porebski S, Bailey LG, Baum BR. 1997. 

Commentary modification of a CTAB DNA extraction protocol for 

plants containing high polysaccharide and polyphenol components. 

Plant Mol Biol Rep 15 (1): 8-15. 

Pirtilla AM, Hirsikorpi M, Kamarainen T, Zaakola L, and Hohtola A. 

2001. DNA isolation method for medicinal and aromatic plants. Plant 

Mol Biol Rep 19:273a-273f. 

Prana TK, Hartati NS. 2003. Identifikasi sidik jari DNA talas (Colocasia 

esculenta L. Schott) Indonesia dengan teknik RAPD (Random 

Amplified Polymorphic DNA): skrining primer dan optimalisasi 

kondisi PCR. J Natur Indonesia 5 (2): 107-112. [Indonesia] 

Prihandana R, Hambali E, Mujdalipah S, Hendroko R. 2007. Meraup 

untung dari jarak pagar. Agromedia Pustaka. Jakarta. [Indonesia] 

Raharjo S. 2007. Analisa performa mesin diesel dengan bahan bakar 

biodiesel dari minyak jarak pagar. Seminar Nasional Teknologi 2007 

(SNT 2007). Yogyakarta, 24 November 2007. [Indonesia] 

Sanchez-Hernandes C dan Gaytan-Oyarzun JC. 2006. Two minipreparation 

protocols to DNA extraction from plants with high 

polysaccharide and secondary metabolites. African J of Biotechnol 5 

(20): 1864-1867. 

Salisbury FB, Ross CW. 1995. Fisiologi tumbuhan. ITB Press. Bandung. 

Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning. 2nd ed. 

Cold Spring Harbor Lab Press. New York. 

Sisharmini A, Ambarwati AD, Santoso TJ, Utami, DW Herman. 2001. 

Teknik isolasi DNA dan analisis PCR gen pinII pada genom ubi jalar. 

Prosiding Seminar Hasil Penelitian Rintisan dan Bioteknologi 

Tanaman. Bogor, 26-27 Desember 2001. 

Sharma AD, Gill PK, dan Singh P. 2002. DNA isolation from dry and 

fresh samples of polysaccharide-rich plants. Plant Mol Biol Rep 20: 

415a-415f. 

Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, Frijters 

A, Pot J, Peleman J, Kuiper M, Zabeau M. 1995. AFLP: a new 

technique for DNA fingerprinting. Nucl Acids Res 233: 4407-4414.

Vol. 2, No. 1, Pp. 7-14 

March 2010 



Characterisation of taro (Colocasia esculenta) based on morphological and 

isozymic patterns markers 

TRIMANTO 1,♥ , SAJIDAN², SUGIYARTO² 

¹ SMP Negeri 2 Gemolong, Sragen, Jl. Citro Sancakan No. 249, Sragen 57274, Central Java, Indonesia; Tel.: +92-0818754378 

² Bioscience Program, School of Graduates, Sebelas Maret University, Surakarta 57126, Central Java, Indonesia 

Manuscript received: 25 October 2009. Revision accepted: 15 February 2010. 

Abstract. Trimanto, Sajidan, Sugiyarto. 2011. Characterization of taro (Colocasia esculenta) based on morphological and isozymic 

patterns markers. Nusantara Bioscience: 7-14. The aims of this research were to find out: (i) the variety of Colocasia esculenta based 

on the morphological characteristics; (ii) the variety of C. esculenta based on the isozymic banding pattern; and (iii) the correlation of 

genetic distance based on the morphological characteristics and isozymic banding pattern. Survey research conducted in the 

Karanganyar district, which include high, medium and low altitude. The sample was taken using random purposive sampling technique, 

including 9 sampling points. The morphological data was elaborated descriptively and then made dendogram. The data on isozymic 

banding pattern was analyzed quantitatively based on the presence or absence of bands appeared on the gel, and then made dendogram. 

The correlation based on the morphological characteristics and isozymic banding pattern were analyzed based on the product-moment 

correlation coefficient with goodness of fit criterion. The result showed : (i) in Karanganyar was founded 10 variety of C. esculenta; (ii) 

morphological characteristics are not affected by altitude; (iii) isozymic banding pattern of peroxides forms 14 banding patterns, esterase 

forms 11 banding patterns and shikimic dehydrogenase forms 15 banding patterns; (iv) the correlation of morphological data and the 

isozymic banding pattern of peroxidase has good correlation (0.893542288) while esterase and shikimic dehydrogenase isozymes have 

very good correlation (0.917557716 and 0.9121985446); (v) isozymic banding pattern of data supports the morphological character data. 

Key words: taro, Colocasia esculenta, morphology, isozyme. 

Abstrak. Trimanto, Sajidan, Sugiyarto. 2011. Karakterisasi talas (Colocasia esculenta) berdasarkan penanda morfologi dan pola pita 

isozim. Nusantara Bioscience: 7-14. Tujuan penelitian ini adalah untuk mengetahui: (i) keragaman Colocasia esculenta berdasarkan 

karakter morfologi; (ii) keragaman C. esculenta berdasarkan pola pita isozim, dan (iii) hubungan jarak genetik berdasarkan karakter 

morfologi dan pola pita isozim. Survei penelitian dilakukan di Kabupaten Karanganyar, di ketinggian tinggi, sedang dan rendah. Sampel 

diambil menggunakan teknik random sampling purposif, mencakup 9 titik cuplikan. Data morfologi diuraikan secara deskriptif dan 

kemudian dibuat dendogram kekerabatan. Data pola pita isozim dianalisis secara kuantitatif berdasarkan ada atau tidaknya pita di gel, 

kemudian dibuat dendogramnya. Korelasi berdasarkan karakter morfologi dan pola pita isozim dianalisis berdasarkan korelasi koefisien 

momen-produk kriteria goodness of fit. Hasil penelitian menunjukkan: (i) di Karanganyar terdapat 10 varietas C. esculenta; (ii) karakter 

morfologi tidak terpengaruh oleh ketinggian; (iii) peroksidase membentuk 14 pola pita isozim, esterse membentuk 11 pola pita dan 

shikimate dehidrogenase membentuk 15 pola pita; (iv) data morfologi dengan isozim peroksidase memiliki korelasi yang baik 

(0,893542288), sementara data morfologi dengan isozim esterse dan shikimate dehidrogenase memiliki korelasi yang sangat baik 

(0,917557716 dan 0,9121985446); (v) data pola pita isozim mendukung data karakter morfologi. 

Kata kunci: talas, Colocasia esculenta, morfologi, isozim 

INTRODUCTION 

The diversity of food crops in Indonesia can be 

developed to overcome the food problem. Types of tubers 

that can be utilized more optimally as a staple food rice 

substitutes include cassava, sweet potato, taro, purse, 

arrowroot and canna. These tubers have a lot of the preeminent, 

among them having a high content of 

carbohydrates as energy sources (Liu et al. 2006), not 

containing gluten (Rekha and Padmaja 2002), containing 

angiotensin (Lee et al. 2003), antioxidative ( Nagai et al. 

2006),which can be applied to various purposes (Aprianita 

2009), and produce more energy per hectWEREthan rice 

and wheat. Tubers can be grown on marginal areas 

(Louwagie et al. 2006), where other plants cannot grow and 

can be stored in the form of flour and starch (Aboubakar et 

al. 2008). 

Taro has a good variety of morphological characters 

such as tubers, leaves and flowers as well as chemicals 

such as flavor, aroma and others (Xu et al. 2001). 

Characterization of taro plants now has started to be 

developed through two approaches. The diversity among 

the varieties can be distinguished based on morphological 

and molecular markers. Diversity based on morphological 

marker has a weakness, because the morphological 

characteristics do not necessarily indicate genetic diversity. 

Morphological diversity is influenced by the environment, 

because every environment has different conditions, so the 

plants do adapt to their home range.

8 

2 (1): 7-14, March 2010 

Molecular marker is an effective technique in genetic 

analysis of a plant variety. Molecular markers have been 

applied widely in plant breeding programs. Molecular 

marker that is often used to distinguish plant diversity is a 

marker of isozyme and DNA (Asains et al. 1995; Setyo 

2001). Isozyme is a direct product of genes and relatively 

free from environmental factors. Isozyme can be used as a 

genetic trait to study and identify the diversity of 

individuals or a cultivar. Isozymes were enzymes that have 

active molecules and different chemical structure, but 

catalyze the same chemical reaction. Different forms of an 

enzyme molecule can be used as the basis of chemical 

separation, by electrophoresis method will result in banding 

patternsproduced by different distances (Purwanto et al. 2002). 

Information about the genetic diversity of taro 

(Colocasia esculenta L.) is needed for plant breeding and 

improvement for the offsprings to obtain superior varieties. 

Based on the background, the research was conducted on 

taro plants in different areas in a region that had high 

altitude, medium and low that included morphological 

characters and isozyme banding patter pita on different 

varieties of taro plants in Karanganyar, Central Java. 


The experiment was conducted in March 2009 to 

August 2009. Taro plants (Colocasia esculenta L.) were 

collected from Karanganyar District, Central Java 

differentiated by differences in altitude, namely: (i) the 

highlands (> 1000 m asl), (ii) plain medium (500-1000 m 

asl), and (iii) Lowland (

TRIMANTO et al. – Characterization of taro based on morphological and isozyme markers 9 

Tabel 1. Environmental conditions where the growth of taro in Karanganyar. 

Locations/ 

Subdistricts 

Type of taro 

Environmental factors 

Altitude Temp. Type of 

Shade 

(meter) (°C) soil 

In the dendrogram similarity coefficient of 60% was 

used to analyze the phylogenetic relationship of the 18 

samples found in different locations with 11 different 

varieties. According Cahyarini (2004) said the similarity 

distance away if less than 0.60 or 60%, so that separate 

groups at a distance of less than 0.60 still has a close 

resemblance. In this dendogram analysis, the number 1 or 

100% indicates that the group members have a perfect 

resemblance, while getting closer to the number 0 means 

the similarity distance farther. 

Benthul 

Dendogram analysis results showed that the Benthul 

taro of different height have the same morphological 

characteristics and have a high relationship. This is evident 

in the coefficient of 0.60 which was still in one group. But 

there was a tendency that Benthul of different heights 

showes different sizes, ranging from leaf size, plant height, 

stem and tuber. Benthul is commonly grown as a crop 

population between the rice fields and gardens, and 

allowed to grow without special treatment. Environmental 

factors such as temperature at any altitude, soil and 

availability of different light and water, thought to cause 

the size of the plants experience the difference. According 

to Park et al. (1997) and Djukri (2006) each deal with 

environmental stress of plants continues to do the 

adaptation, including changes in morphological 

characteristics and physiology. 

Benthul that grows in the highlands appear higher with 

habitus width, leaf midrib and stalk thin and big. This was 

observed in taro grown in ketinggianya more than 1500 m 

with high 22°C, and high rainfall reaches 2299 mm 

/±humidity, low temperature year. According to Basri 

Rainfall 

(mm/y) 

Cultivation 

Lowland 

Gondangrejo Benthul 

Mberek 

150 

150 

29 

29 

Grumosol 

Grumosol 

- 

- 

1537 

1537 

√ 

- 

Jaten Kladi 

Linjik 

98 

98 

29 

29 

Aluvial 

Aluvial 

- 

√ 

1680 

1680 

√ 

√ 

Karanganyar Lompongan 320 30 Mediteran - 2012 - 

Kebakkramat Plompong 95 29 Mediteran - 2012 √ 

Plain medium 

Karangpandan Benthul 

Lompongan 

Sarangan 

Matesih Jabon 

Laos 

Linjik 

Plateau 

Tawangmangu Kladitem 

Benthul 

Lompongan 

650 

600 

650 

700 

750 

700 

28 

28 

28 

28 

27 

28 

Mediteran 

Mediteran 

Mediteran 

Litosol 

Litosol 

Litosol 

- 

√ 

- 

- 

- 

√ 

2818 

2818 

2818 

2480 

2480 

2480 

1500 

1700 

1500 

23 

22 

23 

Andosol 

Andosol 

Andosol 

√ 

- 

√ 

3299 

3299 

3299 

√ 

√ 

- 

Ngargoyoso Laos 1000 26 Andosol √ 3182 √ 

Jatiyoso 

Sarangan 

Jepang 

1300 

1200 

26 

26 

Andosol 

Andosol 

- 

- 

3098 

3098 

√ 

- 

√ 

√ 

√ 

- 

√ 

√ 

(2002) plant growth is influenced by 

environmental factors. Altitude above 1500 

m cause gas and water vapor content 

(humidity) and the number of clouds 

blocking sunlight to the plants, so plants 

were capturing light by raising levels of 

chlorophyll and surface area. Taro plants 

tend to have broad leaves because of the 

availability of adequate water due to high 

rainfall in the area still support the optimum 

process in photosynthesis.±Low temperature 

22°C 

Benthul that grows in the lowlands tend 

to have narrower leaves and smaller and 

lighter bulbs. According to Menzel (1980) 

the temperature is too high may cause leaves 

to hinder the development of broad and 

narrow leaf photosynthetic rate high as a 

result reducing the weight of tuber. But 

when the temperature is too low to reach less 

than 10°C, the plant tissue can be damaged 

and an interruption of growth so the plants 

tend to be stunted. 

Lompongan 

Dendogram Lompongan relationship 

found in three different heights showed only 

the size difference. Broadly speaking taro 

from the highlands, medium and low still have the same 

morphological characteristics. Lompongan plants grow 

wildly around the edge of rice fields and waterways. 

Lompongan plants from the highlands have differences 

with the lowlands, such as: green leaf color is more 

concentrated, browner midrib color, and the size is larger. 

Unlike Lompongan plants in the highlands that were often 

found on the outskirts of the river with shade trees around 

it, the ones in the lowlands were found in around the edges 

of fields full of water. Environmental factors in the form of 

light, temperature and humidity cause the plants to have 

different adaptations. According to Taiz and Zeiger (1991), 

leaf surface area increased because of the shade, and color 

changes due to the increased levels of chlorophyll a and b. 

In the circumstances shaded light spectrum that is 

active in the process of photosynthesis (wavelength 400- 

700 nm) get decreased. Plants will make adjustments to 

streamline the capture of light energy that is by increasing 

leaf area, plant height and chlorophyll a and b (Lambers et 

al. 1998). 

Altitude causes humidity, light, temperature, and 

moisture content to vary. According to Fitter and Hay 

(1998) environmental factors were related one another so 

that the plant held a response to the environment. High 

water levels in the soil cause leaf’s cell turgor to increase 

which in turns causes leaf’s expansion. Reduced light 

causes the leaves to add the proportion of mesophyll tissue. 

Temperatures that were too high (> 40 ° C) cause defective 

enzyme and respiration is rapid, so the plants have stunted 

growth. The temperature is too low (

10 

2 (1): 7-14, March 2010 

Table 2. 18 samples of C. esculenta in Karanganyar district with 

characteristics 

Characteristics 

Varieties 

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 

A. Type of plant 

1. Rentang tanaman 

1. Sempit - - - - - √ √ √ √ √ - - - - √ - - - 

2. Sedang - √ √ √ √ - - - - - √ √ √ √ - - - √ 

3. Lebar √ - - - - - - - - - - - - - - √ √ - 

2. Tinggi tanaman 

1. Kerdil (< 50 cm) - - - - - √ √ √ - - - - - - √ - - - 

2. Sedang (< 50 cm) - √ √ √ √ - - - √ √ √ √ √ √ - - - - 

3. Tinggi (< 50 cm) √ - - - - - - - - - - - - - - √ √ √ 

3. Jumlah stolon 

1. 1-5 buah - - - - - - - - √ √ - - - √ - √ √ √ 

2. 6- 10 buah √ √ √ - - - - - - - √ √ √ - - - - - 

3. 11-20 buah - - - √ √ √ √ √ - - - - - - √ - - - 

4. Panjang stolon 

1. Pendek (15 cm) - - - √ √ - - - - - - - - √ - - √ - 

B. Cormus (umbi) 

1. Manifestasi cormus 

1. Ada √ √ √ - - - √ √ √ - - √ - √ √ √ - √ 

2. Tidak ada - - - √ √ √ - - - √ √ - √ - - - √ - 

2. Panjang Cormus 

1. Pendek (± 8 cm) - - √ - - - √ √ - - - - - - - - √ - 

2. Sedang (± 12 cm) √ √ - - √ √ - - - - - - √ √ √ √ - - 

3. Panjang (± 18 cm) - - - √ - - - - √ √ √ √ - - - - - √ 

3. Cabang cormus 

1. Bercabang - - - - - - √ √ - - - - - √ √ - - - 

2. Tidak bercabang √ √ √ √ √ √ - - √ √ √ √ √ - - √ √ √ 

4. Bentuk cormus 

1. Kerucut √ √ √ - - - - - - - - - √ - - √ √ - 

2. Membulat - - - - - - - - - - - - - - - - - √ 

3. Silindris - - - √ √ √ - - - - - - - - - - - - 

4. Memanjang - - - - - - - - √ √ √ √ - - - - - - 

5. Datar dan terbuka - - - - - - √ √ - - - - - √ √ - - - 

5. Berat cormus 

1. Ringan (± 0.5 kg) - - - - - - √ √ - - - - √ - - - √ - 

2. Sedang (± 2 kg) - - √ √ √ √ - - √ √ - - - √ √ √ - - 

3. Berat (± 4 kg) √ √ - - - - - - - - √ √ - - - - - √ 

6. Warna korteks cormus 

1. Putih - - - - - - √ √ √ √ - - - √ √ √ - √ 

2. Kuning-orange √ √ √ √ √ √ - - - - √ √ √ - - - √ - 

7. Warna daging tengah 

1. Putih - - - √ √ √ √ √ √ √ √ √ - √ √ √ √ √ 

2. Kuning √ √ √ - - - - - - - - - - - - - - - 

3. Orange - - - - - - - - - - - - √ - - - - - 

8. Warna serat daging 

1. Putih - - - √ √ √ - - - - - - - √ - - √ √ 

2. Kuning muda - - - - - - - - - - - - - - - √ - - 

3. Kuning-orange - - - - - - - - - - √ √ √ - √ - - - 

4. Merah √ √ √ - - - √ √ √ √ - - - - - - - - 

9. Permukaan kulit cormu 

1. Berserabut - - - √ √ √ - - - - √ √ - - - - √ - 

2. Bersisik - - - - - - √ √ √ √ - - √ - - √ - √ 

3. Berserabut dan bersisik√ √ √ - - - - - - - - - - √ √ - - - 

10. Ketebalan kulit 

1. Tebal √ √ √ - - - - - √ √ - - √ √ √ - - - 

2. Tipis - - - √ √ √ √ √ - - √ √ - - - √ √ √ 

11. Tingkat serabut 

1. Sedikit - - - √ √ √ √ √ - - - - - - - - √ - 

2. Banyak √ √ √ - - - - - √ √ √ √ √ √ √ √ - √ 

12. Warna tunas 

1. Kuning hijau - - - - - - √ √ - - √ √ - √ √ √ √ - 

2. Merah muda √ √ √ √ √ √ - - √ √ - - - - - - - √ 

3. Ungu - - - - - - - - - - - - √ - - - - - 

C. Daun 

1. Posisi daun dominan 

1. Mendatar - - - √ √ √ - - - - - - - - - - - - 

2. Mangkok √ √ √ - - - - - - - - - - - - - - √ 

3. Tegak keatas - - - - - - √ √ - - - - - √ √ - - - 

4. Tegak kebawah - - - - - - - - √ √ √ √ √ - - √ √ - 

2. Tepi daun 

1. Penuh - - - - - - - - - - - - - √ √ - - - 

2. Bergelombang - - - √ √ √ - - √ √ - - √ - - √ √ - 

3. Berlekok-lekok √ √ √ - - - √ √ - - √ √ - - - - - √ 

3. Warna Helai daun 

1. Hijau - - - √ √ √ √ √ √ √ √ √ - √ √ - - - 

2. Hijau tua √ √ √ - - - - - - - - - - - - √ √ √ 

3. Ungu - - - - - - - - - - - - √ - - - - - 

4. Warna tepi helai daun 

1. Keputihan - - - - - - √ √ - - √ √ - - - √ - - 

2. Hijau - - - - - - - - - - - - - √ - - √ - 

3. Merah muda - - - - √ √ - - - - - - - - - - - - 

4. Ungu √ √ √ - - - - - √ √ - - √ - √ - - √ 

5. Warna cairan ujung daun 

1. Keputihan - - - √ √ √ √ √ - - √ √ - - √ √ √ - 

2. Kuning - - - - - - - - - - - - - √ √ 

3. Merah muda √ √ √ - - - - - √ √ - - - - - - - - 

4. Merah tua - - - - - - - - - - - - √ - - - - - 

6. Warna utama tulang daun 

1. Kuning - - - - - - √ √ - - - - - - √ √ - - 

2. Hiaju - - - - - - - - √ √ √ √ - √ - - √ √ 

3. Merah muda √ √ √ - - - - - - - - - - - - - - - 

4. Ungu - - - √ √ √ - - - - - - √ - - - - - 

7. Pola utama tulang daun 

1. Bentuk Y √ √ √ √ √ √ √ √ - - √ √ - √ √ √ √ √ 

2. Bentuk Y meluas - - - - - - - - √ √ - - √ - - - - - 

8. Warna petiole 

Sepertiga atas 

1. Kuning - - - - - - √ √ - - - - - √ √ √ - - 

2. Hijau muda - - - - - - - - - - √ √ - - - - √ - 

3. Cokelat √ √ √ - - - - - √ √ - - - - - - - - 

4. Ungu - - - √ √ √ - - - - - - √ - - - - √ 

Sepwertiga bawah 

1. Kuning - - - - - - - - - - - - - √ - - - - 

2. Hijau muda - - - - - - √ √ - - √ √ - - √ √ √ 

3. Cokelat √ √ √ √ √ √ - - √ √ - - - - - - - √ 

4. Ungu - - - - - - - - - - - - √ - - - - - 

9. Warna garis petiole 

1. Hijau - - - - - - √ √ - - √ √ - √ √ - √ - 

2. Ungu √ √ √ √ √ √ - - √ √ - - √ - - √ - √ 

10 Irisan melintang bawa 

1. Terbuka √ √ √ √ √ √ - - √ √ √ √ √ √ √ √ - √ 

2. Tertutup - - - - - - √ √ - - - - - - - - √ - 

11. Warna cincin petiole 

1. Putih - - - √ √ √ - - - - √ √ - - - - - - 

2. Kuning kehijauan - - - - - - - - - - - - - √ √ √ √ - 

3. Merah muda √ √ √ - - - √ √ √ √ - - - - - - - √ 

4. Ungu - - - - - - - - - - - - √ - - - - - 

12. Warna pelepah daun 

1. Keputihan - - - - - - √ √ - - - - - - - - - - 

2. Hijau muda - - - - - - - - - - √ √ - √ √ √ √ 

3. Merah Keunguan √ √ √ √ √ √ - - √ √ - - √ - - - - √ 

13. Lapisan lilin 

1. Tidak ada - - - - - - - - √ √ - - √ - - - - - 

2. Rendah √ √ √ - - - √ √ - - √ √ - - - - √ - 

3. Sedang - - - √ √ √ - - - - - - - √ √ - - - 

4. Tinggi - - - - - - - - - - - - - - - √ - √


Note: 1. Benthul (plateau), 2. Benthul (plain medium), 3.Benthul 

(lowlands), 4. Lompongan (plateau), 5.Lompongan (plain medium), 

6. Lompongan (lowlands), 7. Laos (plateau), 8. Laos (plain 

medium), 9. Linjik (plain medium), 10. Linjik (lowland), 11. 

Sarangan (plateau), 12. Sarangan (plain medium), 13. Kladitem 

(plateau), 14. Plompong (lowland), 15. Kladi (lowland), 16. jabon 

(plain medium), 17. Mberek (lowland), 18. Japan (plateau). 

Figure 1. Dendogram relationship 18 samples of C. esculenta 

from three different heights based on morphological characters. 

Description: No. 1-18 same as Table 2. 

Characterization of isozyme taro 

Peroxidase 

Results with the dye peroxidase isozyme analysis, 

shikimate dehydrogenase and esterase can be seen in 

Figure 2. Peroxidase in 18 samples of C. esculenta tested to 

form 14 different banding pattern. Banding pattern I with 

migration distance (Rf) 0586, 0630, 0717, 0761 and 0804 

being owned by the sample 1. Banding pattern II with Rf 

0586, 0717, 0761 and 0804 were owned by the sample 2 

and 3. Banding pattern III is owned by sample 5 with Rf 

0630, 0739, 0782 and 0826. Banding pattern IV with Rf 

0630, 0739, 0782 owned by samples 4 and 6. Banding 

pattern V with Rf 0652, 0739, 0782 and 0874 were owned 

by the sample 7 and 8. Rf banding pattern VI 0652, 0739, 

0782 and 0869 were owned by the sample 9 and 10. 

Banding pattern VII with Rf 0565, 0717 0739 and 0847 

held by the sample 11. Banding pattern VIII with Rf 0565, 

0717 0739 owned by sample 12. IX banding pattern with a 

distance of 0630 and 0739 held by the sample 13. Banding 

pattern X with Rf 0630 and 0739 held by the sample 14. XI 

banding pattern with a distance of 0630, 0739, and 0804 is 

owned by the sample 15. XII banding pattern with a 

distance of 0630, 0739, and 0826 is owned by the sample 

16. XIII banding pattern with a distance of 0630, 0717 and 

0761 held by the sample 17. Banding pattern XIV with Rf 

0607, 0652 and 0.761 were owned by the sample 18. 

Shikimate dehydrogenase 

Isozyme analysis results with dye shikimate 

dehydrogenase (ShDH) on 18 samples of C. esculenta 

tested to form 15 different banding pattern. Banding pattern 

I with Rf 0523, 0568, and 0863 is owned by the sample 1. 

Banding pattern II with Rf 0523, 0568, 0614 and 0863 

were owned by the sample 2 and 3. Banding pattern III 

with Rf 0523, 0568, 0614 and 0840 were owned by the 

sample 4. Banding pattern IV with Rf 0500, 0523, 0568, 

0614 and 0840 were owned by samples 5 and 6. Banding 

pattern V with Rf 0568 and 0840 were owned by the 

sample 7 and 8. Banding pattern VI with Rf 0523 and 0840 

held by the sample 9. Banding pattern VII with Rf 0500, 

0523 and 0840 were owned by the sample 10. Banding 

pattern VIII with Rf 0523 and 0818 held by the sample 11. 

Banding pattern IX with Rf 0500, 0523 and 0818 were held 

by the sample 12. Banding pattern X with Rf 0416, 0432, 

0523, 0795 owned by sample 13. Banding pattern of Rf 

0500 XI, 0523, 0727 and 0750 were owned by the sample 

14. XII banding pattern of Rf 0523, 0546, 0581 and 0818 

held by the sample 15. Banding pattern XIII with Rf 0523, 

0546, 0568 and 0795 held by the sample 16. Banding 

pattern XIV with Rf 0500, 0546, and 0795 was owned by 

the sample 17. Banding pattern XV with Rf 0546, 0568 and 

0795 were held by the sample 18. 

Esterase 

Results with the dye esterase isozyme analysis on 18 

samples of C. esculenta were tested forming 11 different 

banding patterns. Banding pattern I with Rf the same but 

having different shapes, and shown at Rf 0.22, 12:26 and 

12:32 were owned by the sample 1, 2 and 3 (quantitative 

and qualitative). Banding pattern II with Rf 0.20, 0:28, 0:32 

and 0.36 were owned by the sample 4, 5 and 6 (quantitative 

and qualitative). Banding pattern III with Rf 0:30, 0:34, 

0:38, 0:40 was owned by the sample 7 and 8 (quantitative 

and qualitative). Banding pattern IV with Rf 0.20, 0:30, 

0:34, 0:38 and 0:44 is owned by the sample 9 and 10. 

Banding pattern V with Rf 0.20, 12:26 and 12:38 were 

owned by the sample 11 and 12 (quantitative and 

qualitative). VI banding pattern was owned by the sample 

13 with Rf 0.20, 0:28, 0:30, 0:46, 0:48. Banding pattern VII 

owned by the sample 14 with Rf 0.20, 0.26, 0:30, 0:34. 

VIII banding pattern VIII was owned by the sample 15 

with Rf 0.20, 0.26, 0:30, 0:36. Banding pattern IX was 

owned by the sample 16 with Rf 0.20, 0:22, 0:26, 0:32. 

Banding pattern X was owned by the sample 17 with Rf 

0.20, 0.24, 0:32 and banding pattern XI with Rf 0.20, 12:28 

and 12:32 were owned by the sample 18. 

Similarity on taro genetics based on isozyme markers 

Genetic similarity between samples can be tested using 

cluster analysis (group average analysis), which results in 

the form dendogram or tree diagram. The end result is a 

dendogram of relationship were tested by three different 

enzymes (peroxidase, shikimat dehydroginase, and 

esterase) (Figure 3). 

Election peroxidase has advantages including: a broad 

spectrum and has a very important role in the process of 

plant physiology. This enzyme can be isolated and 

scattered in the cell or plant tissue, especially in plant 

tissues that had been developed (Butt 1980; Hartati 2001). 

Shikimate dehydrogenase (ShDH) is an enzyme which 

spread to most living things. Shikimate dehydrogenase 

involved in oxidoreductase that catalyzes NADP + 

shikimate into three main products dehydroshikimate + 

NADPH+H + . At the plant, esterase is a hydrolytic enzyme 

that functions to withhold simple esters in organic acids, 

inorganic acids and phenols and alcohols have low 

molecular weight and easily soluble.

12 

2 (1): 7-14, March 2010 

A B C 

Figure 2. The variation of 18 isozyme banding pattern of sample C. esculenta from three different heights. Description: a. Banding 

pattern of peroxidase, b. Shikimate dehydrogenase banding pattern, c. Esterase banding pattern. No. 1-18 same as Table 2. 

A B C 

Figure 3. Relationship dendogram 18 samples of C. esculenta from three different heights based on isozyme banding pattern. A. 

peroxidase, B. shikimate dehydrogenase, c. Esterase. No. 1-18 same as Table 2. 

Results dendogram relationship between the use of 

peroxidase enzymes, shikimate dehydrogenase and esterase 

showed generally taro of the same variety have the same 

banding pattern, although from different locations 

ketinggianya, so that enzymatically still have a high 

relationship, since it is estimated the same parent. On a 

different taro varieties tend to have a different banding 

pattern. Formation of the group between the use of 

esterase, peroxidase and shikimate dehydrogenase gave 

different relationship relations, but in one variety is 

generally joined in one group at a distance of more than 

60% similarity, although originating from different 

locations’ height. 

Esterase formed seven groups which were of more than 

60% similarity between one another, where there were taro 

who joined another group. Jabon formed a group with 

Plompong which were of 0.80 similarity. Kladi formed a 

group with Plompong at a distance of 0.75 similarities. 

Lompongan joined with Japan at a distance of 0.70 

similarities. Laos and Linjik form one group at a distance 

of 0.67 similarity. Even when they are different taro 

varieties butwhen they form one group, they still have a 

high genetic relationship. 

Peroxidase also formed seven groups. In general, in a 

variety of taro is still present in one group even though 

planted in different locations altitude place. Peroxidase 

formed a different group variation taro with esterase. In 

peroxidase, Laos and Linjik in one group that were of 0.75 

similarity. Plompong and Kladi form one group, but Jabon 

joined at a distance of 0.70 similarity. Lompongan and 

Kladitem form one group at a distance of 0.75 similarity. 

Peroxidase added information that was not the presence of 

new groups formed on the use of esterase. 

Shikimate dehydrogenase provided the formation of 

different groups of taro with esterase and peroxidase. In 

shikimate dehydrogenase,formed three groups originating 

from different varieties, but it formed one group that was of 

more than 60% similarity. Lompongan and Benthul joined 

at 0.65 similarity distance. Kladi joined Sarangan at a 

distance of 0.62 similarity. Mberek and Japan formed one 

group that was of 0.67 similarity. 

The use of different enzymes gave results in different 

groups, although there is formation of the same group with


a different enzyme’s use. The use of different enzyme will 

complement the formation of groups of different taro 

varieties. The genetic pattern of bands that formed in the 

use of enzymes is the expression of taro varieties in 

question. With a specific enzyme that cannot afford some 

taro that express ribbon patterns, but with other enzymes 

can express ribbon patterns. So that more types of enzymes 

used then it will complete the formation of groups on 

varieties of taro. 

Results dendogram through morphological markers and 

isozyme banding pattern shows the difference. From the 

morphological marker of the 11 varieties, obtained taro 

formed 10 groups at a distance of 0.60 similarity. Different 

taro varieties, most will form a separate group means 

morphologically different taro varieties have different 

morphological characteristics. Talas who formed a group 

on the analysis of relationship is Kladi and Plompong. 

When the isozyme was used, more groups were 

formed, this means that between the different taro varieties 

there is still a high relationship. If the different varieties of 

taro belong to one group with a distance of close to 1 it is 

possible that the similarity comes from the older of taro. 

Environmental factors affect plant morphology, if the 

environmental factor is more dominant than genetic factors, 

the plant will experience a change in morphology (Suranto 

1999, 2001). In the long term it is possible crop genetic 

trait changes in her body. Plants that were stressed 

environment would be possible to have mutations, so that 

in the long term can happen speciation. 

New types were also possible as a result of 

hybridization, so having a close relationship with both of 

the parent species. The property of taro which has a close 

relationship is what can be used to search for a superior 

taro through crossbreeding. Some taros found in 

Karanganyar were a wild taro. Wild Taro and of likely no 

benefit are possibly to have genetic traits that superior, so 

that the hybridization process to obtain high yielding 

varieties can be applied. 

Generative breeding of taro is naturally difficult to 

occur because the male and female flowersg et mature at 

different times and a new flowering occurs after more than 

6 months of age. Many plants are not considered going 

through a flowering because the flowering process is too 

long. Many cultivated plants are harvested before 

adulthood, so many plants are difficult to perform in a 

generative breeding. 

Characterization of taro plants through morphological 

marker is more easily done, by observing external nature, 

taro plants can be assumed to have superior properties. But 

genetic markers also play an important role because it is 

more fundamental and is not influenced environment. Data 

morphology and isozyme banding pattern on taro plants in 

Karanganyar can be used in addition to the identification of 

the food plant breeding efforts. 

Characterization relations of morphology and isozyme 

The correlation between genetic distance based on 

morphological markers and similarity based on isozyme 

banding pattern were analyzed based on product-moment 

correlation coefficient with the criteria of goodness of fit 

according to Rohlf (1993). Result of calculation correlation 

between genetic distance based on morphological markers 

and genetic similarity based on isozyme banding pattern 

showed that between morphology and isozyme has a good 

correlation and a very good (Table 4). Correlation between 

morphological data and isozyme banding pattern of 

peroxidase, esterase, and shikimate dehydrogenase, 

respectively, also were on the value of 0.893542288, 

0.917557716, 0.9121985446. This shows the 

characterization of taro based on morphological markers 

consistent with isozyme banding pattern, so that the 

isozyme data support the morphological data. 

Diversity is difficult to observe the morphological 

marker would be more accurate if you have the genetic 

markers such as isozymes. Morphological characters that 

were equipped with the character of isozyme banding 

pattern adds accuracy of the data to identify plant diversity. 

Isozyme has advantages because it requires little sample of 

the plant, were not inhibited during plant dormancy, can be 

used to perform characterization of the plant in very much. 

Table 4. Relationships and morphological characterization 

characterization results based on isozyme banding pattern 

Characters that correlated Level Criteria 

Morphology and POD 0.893542288 good 

Morphology and EST 0.917557716 very good 

Morphology and ShDH 0.9121985446 very good 

The relationships of taro plants obtained from places of 

different heights can be made into a dendogram between 

morphology and marker pattern of the isozyme’s ribbon. 

Dendogram based on morphological markers and isozyme 

banding pattern of peroxidase, shikimate dehydrogenase, 

and esterase showed that taro with the same type from a 

different altitude did not show any difference at a distance 

of 60% similarity. Of the eighteen samples were divided 

into 10 groups. Each taro with the same type, although 

located in different places still reflect the height of high 

relationship. This proved that taro plants of the same type 

belonged to a single group. 

Figure 4. Dendogram relationship 18 samples of C. esculenta 

from three different heights based on morphological markers and 

isozyme banding pattern of peroxidase, esterase, and shikimate 

dehydrogenase. Description: No. 1-18 same as Table 2.

14 

2 (1): 7-14, March 2010 

Taro varieties which become one group is based on 

morphological markers and isozyme banding pattern, 

where the isozyme banding pattern supports the 

morphological data. This is evident in samples 1, 2, 3, ie 

Bentul from three different height locations which join one 

group. Other evidence were sample 4, 5 and 6, which were 

from three different altitude sites that also formed one 

group. This indicated that the isozyme data support the 

morphological data, so as to identify the plant in addition to 

morphological data, isozyme data is also needed to increase 

the accuracy of the data. There were varieties of taro which 

have a a close relationship that are Kladi and Plompong 

that have a high relationship when viewed from the merger 

with its isozyme morphological characteristics, both were 

at the coefficient of 0.68. Allegedly the two taro plants 

have elders who have a high kindship, because almost the 

same its relation of morphology and isozyme almost the 

same. From the characterization results obtained that has a 

relationship Kladi and Plompong highest compared with 

other varieties of taro. Taro with different varieties formed 

their own groups at a distance of 60% similarity. This 

means that at a distance of 60% of all varieties of taro had 

different characters. 

CONCLUSION 

There is a diversity of morphological characters in 18 

samples of taro plants (Colocasia esculenta L.) that grow in 

Karanganyar. Taro is still in one variety that is at different 

height diversity appears only on the size of the vegetative 

plant. The results showed isozyme banding pattern of the 

variability in isozyme banding pattern of peroxidase, 

esterase and shikimate dehydrogenase in taro varieties 

found in different locations. Characterization of taro based 

on morphological markers is consistent with the 

characterization based on isozymes. Isozyme data support 

the morphological character data. 

REFERENCES 

Aboubakar YN. Njintang, Scher J, Mbofung CMF. 2008. 

Physicochemical, thermal properties and microstructure of six 

varieties of taro (Colocasia esculenta L. Schott) flours and starches. J 

Food Engineer 86 (2): 294-305. 

Aprianita A, Purwandari U, Watson B, Vasiljevic T. 2009. Physicochemical 

properties of flours and starches from selected commercial 

tubers available in Australia. Intl Food Res 16: 507-520. 

Basri H. 2002. Agroecology; a physiological approach. Raja Grafindo 

Persada. Jakarta. [Indonesia] 

Butt VS. 1980. Direct oxidases and related enzymes. In: Stumpf PK, Cohn 

EE (eds). The biochemistry of plants. Vol. 2. Academic Press. New 

York. 

Cahyarini RD, Yunus A, Purwanto E. 2004. Identification of the genetic 

diversity of some local soybean varieties in Java based on isozyme 

analysis. [Thesis]. School of Graduates, Sebelas Maret University. 

Surakarta. [Indonesia] 

Djukri. 2006. The plant characters and corm production of taro as catch 

crop under the young rubber stands. Biodiversitas 7 (3): 256-259. 

[Indonesia] 

Fitter AH, Hay RKM. 1998. Environmental physiology of plants. Gadjah 

Mada University Press. Yogyakarta 

Hartati S, Prana T. 2001. Analysis of starch and crude fiber content of 

flour several taro cultivars (Colocasia esculenta L. Schott). J Natur 

Indonesia 6 (1): 29-33. [Indonesia] 

Kusumo S, Hasanah M, Moeljopawiro S, Thohari M, Subandriyo, 

Hardjamulia A, Nurhadi A, Kasim H. 2002. Panduan Karakterisasi 

dan Evaluasi Plasma Nutfah Talas. Komisi Nasional Plasma Nutfah, 

Badan Penelitian dan Pengembangan Pertanian, Departemen 

Pertanian. Jakarta. [Indonesia] 

Lee MH, Lin YS, Lin YH, Hsu FL and Hou WC. 2003. The mucilage of 

yam (Dioscorea batatas Decne) tuber exhibited angiotensin 

converting enzyme inhibitory activities. Bot Bull Acad Sinica 44: 

267-273. 

Liu Q, Donner E, Yin Y, Huang RL and Fan MZ. 2006. The 

physicochemical properties and in vitro digestibility of selected 

cereals, tubers, and legumes grown in China. Food Chem 99: 470- 

477. 

Louwagie G, Stevenson CM, Langohr R. 2006. The impact of moderate to 

marginal land suitability on prehistoric agricultural production and 

models of adaptive strategies for Easter Island (Rapa Nui, Chile) . J 

Anthropol Archaeol 25 (3): 290-317. 

Menzel CM. 1980. Tuberization in potato Solanum tuberosum cultivar 

Sebago at high temperatures: responses to gibberellins and growth 

inhibitors. Ann Bot 46: 259-266 

Nagai T, Suzuki N and Nagashima T. 2006. Antioxidative activity of 

water extracts from the yam (Dioscorea opposita Thunb.) tuber 

mucilage tororo. Eur J Lipid Sci Tech 108: 526-531. 

Purwanto E, Sukaya, Merdekawati P. 2002. Study on germplasm diversity 

of pummelo at magetan East Java based on isozyme markers). 

Agrosains 4 (2): 50-55. [Indonesia] 

Rekha MR, Padmaja G. 2002. Alpha-amylase inhibitor changes during 

processing of sweet potato and taro tubers. Plant Food Human Nutr 

52: 285-294. 

Rohlf FJ. 2005. NTSYS-pc: numerical taxonomy and multivariate 

analysis system, version 2.2. Exeter Software: Setauket, NY 

Suranto. 1991. Studies of population variation in species of Ranunculus. 

[Thesis]. Departement of Plant Science, University of Tasmania. 

Hobart, Australia. 

Suranto. 2001. Study on Ranunculus population: isozymic pattern. 

Biodiversitas 2 (1): 85-91. 

Taiz L, Zeiger E. 1991. Plant physiology. Benyamin/Cumming. Tokyo. 

Xu J, Yang Y, Pu Y, Ayad WG, Eyzaguirre PB. 2001. Genetic diversity in 

Taro (Colocasia esculenta Schott, Araceae) in China: an 

ethnobotanical and genetic approach. Econ Bot 55 (1): 14-31.

Vol. 3, No. 1, Pp. 15-22 

March 2011 



Study on floristic and plant species diversity in the Lebanon oak (Quercus 

libani) site, Chenareh, Marivan, Kordestan Province, western Iran 

HASSAN POURBABAEI ♥, SHIVA ZANDI NAVGRAN 

Determent of Forestry, Faculty of Natural Resources, University of Guilan, Somehsara, P.O.Box 1144, Tel.: +98-182-3220895, Fax.: +98-182-3223600, 

♥ E-mail: h_pourbabaei@guilan.ac.ir 

Manuscript received: 28 Augustus 2010. Revision accepted: 4 October 2010. 

Abstract. Pourbabaei H, Navgran SZ. 2011. Study on floristic and plant species diversity of the Lebanon oak site (Quercus libani) in 

Chenareh, Marivan, Kordestan Province, western Iran. Nusantara Bioscience 3: 15-22. In order to study floristic and plant species 

diversity, approximately 450 ha of oak forests were selected in Chenareh, Marivan of Kordestan province in western Iran. Inventory was 

selectively carried out in 50 m elevation range in the different aspects. Vegetation was surveyed in the four layers including: tree (dbh 

>5 cm), regeneration (dbh 5 cm), regenerasi (dbh

16 

3 (1): 15-22, March 2011 

affect on the natural regeneration of Pistacia species, that 

is, the seedlings of Pistacia species would be protected 

under the spiny bushes of Amygdalus species. 

The Zogros mountains are divided into two parts: 

northern and southern. The northern Zagros is consisted of 

the growing site of Quercus infectoria Oliv. and also 

Q.libani Oliv. and Q.persica J. & Sp. (Q.brantii Lindl.) 

species are found in this part. However, the southern 

Zagros is included Q.persica site which it extended to Fars 

province (i.e., 29º 5´ N). The northern Zagros is wetter and 

cooler than the southern one. The dispersion areas of 

Lebanon oak (Q.libani) are mostly restricted to central and 

eastern mountains of Tavrous and Amanous of Anatolia in 

Turkey, the mountains of northeastern of Iraq and 

northwestern of Syria and western part of Iran (i.e., 

Kordestan province) (Browicz 1994). In addition, this 

species is found over 1000 m asl elevation and the best 

conditions range from 1200 to 1600 even to 1800 m asl to 

growing it, and also this species is found higher than 2000 

m asl elevation in Ahir dagi and Herakol dagi mountains in 

southern Anatolia, Turkey. Western borderline of this 

species is located in the Goniah province in Anatolia and 

northern borderline restricted to latitude 40ºN in Erzincon 

province in Turkey (Davis 1982). In flora of Iraq, the 

distribution area of this species was cited in central regions 

of Iraq forests, north of Syria, Palestinian, Turkey and Iran 

on hillsides on the metamorphic and igneous rocks and on 

loam soils, and elevation ranges from 1800 to 2000 

(occasionally 2100) m asl (Townsend and Guest 1980) . In 

Iran, the distribution of this species is restricted to 

highlands of Sardasht in Kordestan and Euromiah 

provinces, and horizontal distribution is from north of 

Sardasht to south of Marivan in Kordestan province and 

vertical distribution is from 1400 to 2150 m asl elevation 

(Fattahi 1994). This species is situated in latitude from 25º 

to 36º N and longitude from 45º to 46º E and is grown cold 

humid, cold sub humid or humid climates. The pure type of 

this species is found in highlands and the mixed one mostly 

found associated with Q.infectoria and Q.brantii species. 

This species is in relation to soil and climatic conditions. 

The forests of this species are found as high and coppice 

forms, and the species covers 106316 ha area in western 

Iran of which 83844 is located in the Kordestsn province 

(Fattahi 1994). 

There are numerous studies in relation to floristic 

composition all over the world (e.g., Andel 2001; Nebel et 

al. 2001; Ipor et al. 2002; Blanckaert et al. 2004; Wardell- 

Johnson et al. 2004; Ramírez et al. 2007; Cayuela et al. 

2008; Gole et al. 2008; Macía 2008; El-Ghanim et al. , 

2010; Figueroa et al. , 2011). In addition, plant species 

diversity has been assessed in forest ecosystems in recent 

decades (e.g., Brockway 1998; Pitkänen 1998; Khera et al. 

2001; Ashton and Macintosh 2002; Aubert et al. 2003; 

Nagaike 2003; Jobidon et al. 2004; Chiarucci and Bonini 

2005; Pant and Samant 2007; Aparicio et al. 2008; Macía 

2008; Pe´rez-Ramos 2008; Hayat et al. , 2010). Whereas 

there is less studies about plant species diversity in Zagros 

forest ecosystems (Mirzaei et al. 2008; Pourbabaei et al. , 

2010). 

The aim of this study was to determine floristic 

composition and plant species diversity in the Lebanon oak 

site in Kordestan province of Iran. 


Study area 

The study area is located in Marivan city of Kordestan 

province in western Iran, and Chenareh is situated 25 km 

from northwestern Marivan city (35° 29′ to 35° 45′ N 

latitude, 46° 14′ to 46° 29′ W longitude). Mean annual 

precipitation is 909.5 mm, ranging from 590.8 to 1422.2 

mm (Figure 1). 

Kordestan 

Chenareh forest 

Marivan City 

Figure 1. Study site maps Chenareh’s forests (blank circle) in Marivan District, Kordestan Province, IR Iran.

POURBABAEI & NAVGRAN – Plant species diversity of Chenareh forest 17 

Mean annual temperature is 13.3º C, and the length of 

dry season is 4 month (based on embrothermic curve) from 

June to August. Type of climate is sub humid with cold 

winters in the basis of Emberger’s formula (Department of 

Forestry 2002). Edaphically, soils consist of developed 

brown (calciferous and eutroph), deep and semi deep, and 

young soils consisting of litho sol and colluvium which 

often are less deepness and shallow. Quercus brantii 

community are predominantly found on calcico brown 

soils, and Q.libani community often found on eutroph 

brown soils. 

The research was conducted in 450 ha of Chenareh’s 

forests where included Lebanon oak and altitude ranges 

from 1500 to 1800 m asl These forests are located steep 

areas, and slope is more than 50% in the most area. Main 

aspects of these forests are northern and southern. These 

forests have been under anthropogenic disturbances in the 

past, therefore they are considered as manipulated forests 

now. 

Sampling 

At first, oak site was quantified on the map with 

1:50000 scale with surveying forests. Inventory was 

selectively carried out in 50 m elevation range from 1500 

m to 1800 m asl in the different aspects in the basis of 

distribution of Lebanon oak population. Sampling plot area 

was 1000 m 2 in size and circular (Zobeiri 1994). In total, 

42 sampling plots were made. At each plot, diameter at 1.3 

m (DBH) of tree ≥ 5 cm was measured and identified (high 

and coppice origin), and crown diameters (i.e., large and 

small) of regeneration with DBH < 5 cm were measured. 

For shrub species, the number of individuals were recorded 

and identified. To collect herbaceous data, nested plot 

sampling was performed at center the plot (Muller- 

Dombois 1974), and minimal area ranged from 32 to 1000 

m 2 in the basis of different altitudes. Cover percentage was 

visually estimated, as accurately as possible, for each 

herbaceous species in the nested plots, and type of species 

was identified in the Herbarium of Faculty of Natural 

Resources, University of Guilan. 

Data analysis 

Species richness (total number of species present) and 

evenness (the manner in which abundance is distributed 

among species) are the two principal components of 

diversity. Species richness is frequently characterized by 

the number of species present (S), Margalef species 

richness (R 1 ) and Menhinick species richness (R 2 ) (Ludwig 

and Reynolds 1988). In this study, Smith and Wilson’s 

evenness index (E var ) was applied to calculate evenness 

measures (Krebs 1999). Diversity indices combine species 

richness and evenness components into a single numeric 

value. The most commonly used indices of diversity, 

Simpson (1-D) and Shannon-Wiener (H′) were used in this 

study (Magurran 2004). Moreover, Hill’s N 2 and 

McArthur’s N 1 were calculated in the basis of these indices 

(Krebs 1999). Vegetation data were analyzed in four layers 

(i.e., tree, regeneration, shrub and herb) using richness, 

evenness and diversity indices. In tree layer, DBH was 

converted to basal area (m 2 ) for each individual tree and 

summed for each species, and then substituted for the 

number of individuals in the diversity formula. 

Furthermore, crown cover area (m 2 ) was computed for each 

regeneration species and applied the formula. Data analyses 

were performed using Ecological Methodology and SPSS 

13.0 software (Krebs 1999; Kinnear 2001). 


Floristic composition 

A total of 82 plant species were found in the studied 

area, of which 12 woody species (9 trees, 3 shrubs) and 70 

herbaceous species existed (Table 1) while 4 trees, 3 

shrubs, one bush and 78 herbaceous species were identified 

in Ilam forests of Zagros (Pourbabaei et al. 2010). 

Therefore, it is concluded that tree richness is high in the 

studied area. Also, it can be deduced from Table 1 that 

Rosaceae and Fagaceae families play an important role in 

among woody species. Moreover, Asteraceae and Poaceae 

families were most abundant amongst herbaceous species. 

In addition, results were revealed that the Asteraceae 

family was dominant in Ilam forests of Zagros (Pourbabaei 

et al. 2010). 

The number of plant species was considerable in the 

studied area when compare with northern Zagros 

mountains since there is 165 woody species (tree and 

shrub) in Zagros and 182 bush and herbaceous species only 

in northern Zagros (Jazirehi and Rostaghi 2003). The 

highest richness of woody species belong to Fagaceae and 

Rosaceae and the highest richness of herbaceous species 

belong to Asteraceae and Poaceae families in the studied 

area, these results were confirmed in the Zagros zone 

(Jazirehi and Rostaghi 2003). 

Plant diversity based on different aspects 

Plant species diversity of four growth layers was 

obtained in terms of different aspects. The highest and 

lowest population of Lebanon oak was found in eastern 

(32%) and northwestern (24%) aspect, respectively. Figure 

1 displays mean tree (high and coppice forms) diversity in 

the basis of different aspects. 

The mean diversities were highest in southwestern and 

lowest in southeastern and northern aspects in the tree 

layer. The ANOVA test indicated that there were no 

significant differences amongst mean diversity measures in 

the different aspects (P > 0.05). Figure 3. displays mean 

tree richness, Margalef (R 1 ) and Menhinick (R 2 ) and 

evenness measures in the different aspects. 

The mean richness, R 1 and R 2 measures were highest in 

southwestern, and lowest in southeastern and northern 

aspects, respectively. The mean E var was the highest in 

southeastern and lowest in northeastern aspect. The 

Kruskal-Wallis test showed that there were no significant 

differences amongst mean richness values in the different 

aspects. Whereas, the ANOVA test indicated that there 

were significant differences amongst mean R 1 and R 2 in the 

different aspects (P < 0.05), and Tukey test showed that 

there was significant difference between southwestern and 

southeastern aspects in view of mean R 1 . Also, there was 

significant difference between southwestern and other 

aspects except southern in view of mean R 2 .

18 

3 (1): 15-22, March 2011 

Table 1. Plant species list based on growth layers 

Layer 

Tree 

Shrub 

Herbaceous 

Species 

Acer monspessulanum L. (Aceraceae), Amygdalus communis L. (Rosaceae), Cerasus mahaleb L. (Rosaceae), Crataegus 

pontica C.Koch. (Rosaceae), Pistacia atlantica (Anacardiaceae), Pyrus syriaca Boiss. (Rosaceae), Quercus brantii Lindl. 

(Fagaceae), Q.infectoria Oliv. (Fagaceae), Q.libani Oliv. (Fagaceae). 

Cerasus microcarpa (C.A.Mey) Boiss. (Rosaceae), Cotoneaster nummularia Fisch & Mey. (Rosaceae), Lonicera 

nummularifolia Jaub & Spach. (Caprifoliaceae). 

Acanthus dioscoridus L. (Acantaceae), Achillea filipendula L. (Asteraceae), A.millefolium L. (Asteraceae), Aegilops 

triuncialis L. (Poaceae), A.triuncialis L. (Poaceae), Alopecurus myosuroides Ovcz. (Poaceae), Antemis tinctoria L. 

(Asteraceae), Astragalus curvirstris Boiss. (Papilionaceae), A. michauxianus Boiss. (Papilionaceae), A. (tragacantha ) 

sp. (Papilionaceae), Aristolochia bottae Jaub & Spach. (Aristolochiaceae), Boissiera squarrosa Hochst. (Poaceae), 

Bromus tectorum L. (Poaceae), Buchingera axillaris Boiss. (Cruciferae), Bunium elegans (Fenzl.) Freyn. (Umbelliferae), 

Callipeltis cucularia Stev. (Rubiaceae), Centaurea virgata Lam. (Asteraceae), Cephalaria syriaca (L)Schrad. 

(Dipsaceae), Chaerophyllum macropodum Boiss. (Umbelliferae), Cornilla varia L. (Papilionaceae), Dactylis glomerata 

L. (Poaceae), Dianthus tabrizianus Adams. (Caryophylaceae), Echinops orientalis Trauth. (Asteraceae), E.ritrodes 

Bunge. (Asteraceae), Eremopoa persica (Trin.) Roshev (Poaceae), Eryngium thyrsoides F.Delaroche. (Umbelliferae), 

Euphorbia macroclada Boiss (Euphorbiaceae), Ferula orientalis L. (Umbelliferae), Fibijia macrocarpa Boiss. 

(Cruciferae), Galium aparine L. (Rubiaceae), Grammosciadium platycarpum Boiss. (Umbelliferae), Gundelia 

tournefortii L. (Asteraceae), Helianthemum ledifolium (L.) Miller. (Cistaceae), Heteranthelium piliferum (Banks & 

Soland) (Poaceae), Hordeum bulbosum L. (Poaceae), Hypericum scabrum L. (Hypericaceae), Inula britanica L. 

(Asteraceae), Lamium album L. (Labiatae), Marrubium vulgare L. (Labiatae), Mesostemma kotschyanum Wed. 

(Caryophylaceae), Milium pedicellare Bornm. (Poaceae), Onopordon kurdicum Bornm& Beauv (Asteraceae), Onosma 

elwendicum L. (Boraginaceae), O. microcarpa DC. (Boraginaceae), Phlomis olivieri Benth. (Labiatae), P.rigida Labill. 

(Labiatae), Picnomon acarna L. (Asteraceae), Poa bulbosa L. (Poaceae), Potentila kurdica Boiss & Hohen. (Rosaceae), 

Prangos ferulaceae L. (Umbelliferae), Rhaponticum insigne Boiss. (Asteraceae), Rhabdoscidium aucheri Boiss. 

(Umbelliferae), Salvia bracteata Banks & Soland. (Labiatae), Sanguisorba minor Scop. (Rosaceae), Scabiosa 

calocephala Boiss.(Dipsacaceae), S. leucactis Patzak. .(Dipsacaceae), Scutellaria pinnatifida A.Hamilt. (Labiatae), 

Serratula grandifolia Boiss. (Asteraceae), Smyrnium aucheri Boiss. (Umbelliferae), Stachys inflata Benth. (Labiatae), 

Taeniatherum crinitum (Schreb).Nevski (Poaceae), Teucrium polium L. (Labiatae), Trifolium campestre Schreb. 

(Papilionaceae), T.pratens L. (Papilionaceae), Turginia latifolia L. (Umbelliferae), Valerianella dactylophylla Boiss & 

Hohen. (Valerianaceae), Veronica kurdica Benth. (Scrophulariaceae), Vicia variabilis Freyn & Sint. (Papilionaceae), 

Xeranthemum inaepertum Boiss. (Asteraceae), Zoegea leptaurea L. (Asteraceae). 

The mean diversity measures were highest in 

northeastern (i.e., 1-D and H′) and southern (i.e., N 2 and 

N 1 ) and lowest in southwestern (i.e., 1-D, H′ and N 1 ) and 

southeastern (i.e., N 2 ) in the regeneration layer (Figure 4). 

The ANOVA test showed that there were significant 

differences amongst mean 1-D measures in the different 

aspects, but no significant differences amongst other 

diversity indices. In addition, Tukey test showed that there 

was significant difference between northeastern and 

southwestern aspects. 

The mean richness, R 1 and R 2 measures were highest in 

northern, southern and lowest in southwestern and 

southeastern aspects, and mean E var was the highest in 

southwestern and lowest in northern aspect (Figure 5). 

There were significant differences amongst mean richness, 

R 1 and R 2 measures. Tukey test showed that there was 

significant difference between northern and southwestern 

aspect in view of richness. 

The mean diversities were highest in northern (i.e. N 2 

and H′) and northwestern (i.e. 1-D and N 1 ) and lowest in 

northeastern aspect in the shrub layer (Figure 6). The 

ANOVA test showed that there were no significant differences 

amongst mean diversities measures in the different aspects. 

The mean richness and R 1 measures were highest in 

northern, while R 2 was the highest in eastern aspect. The 

mean richness was lowest in other aspects and also the 

mean of R 1 and R 2 were found the lowest in northwestern 

aspect, and the E var were found the highest in northwestern 

and lowest in northeastern (Figure 7). There were no 

significant differences amongst mean richness, R 1 , R 2 and 

E var measures in the different aspects. 

The mean diversities were highest in western and 

lowest in northeastern aspect in the herbaceous layer 

(Figure 8). The ANOVA test showed that there were 

significant differences amongst mean diversities measures 

in the different aspects. The differences among means were 

detected using Tukey’s test which are characterized by 

different letters on the histogram of Figure 8. 

The mean richness and E var measures were highest in 

western and lowest in northern aspect in the herbaceous 

layer and there were significant differences amongst mean 

measures in different aspects (Figure 9). 

The Lebanon oak was found in all aspects, but it had 

the most abundant in eastern and the least in northwestern 

aspect since it requires plenty of sunlight in eastern aspect 

(Maroufi 2000). This species is preferred northern and 

eastern aspects and ecological needs of Q.libani is higher 

than Q.infectoria and Q.brantii (Jazirehi and Rostaghi 2003). 

The tree species diversity was found the highest in 

southwestern and lowest in southeastern and northern 

aspects, because richness and richness indices had the 

highest and lowest values the mentioned aspects, and 

evenness had the highest and lowest values in southeastern 

and northeastern aspects, respectively.


Diversity indices 

4.5 

4 

3.5 

3 

2.5 

2 

1.5 

1 

0.5 

0 

1-D 

N 2 

H' 

N 1 

1 2 3 4 5 6 7 8 

Aspect code 

Figure 1. Mean diversity measures and their standard errors based 

on different aspects in the tree layer (1. northern, 2. northeastern, 

3. northwestern, 4. eastern, 5. southern, 6. southwestern, 7. 

southeastern, 8. western). 


2.5 

2 

1.5 

1 

0.5 

0 

1 2 3 4 

1-D 

N2 

H' 

N1 

Aspect code 


on different aspects in the shrub layer. 

Richness and evenness indices 

6 

5 

4 

3 

2 

1 

0 

S 

R1 

R2 

Evar 

1 2 3 4 5 6 7 8 

Aspect code 


3 

2.5 

2 

1.5 

1 

0.5 

0 

1 2 3 4 

S 

R1 

R2 

Evar 

Aspect code 

Figure 3. Mean richness, R 1 , R 2 and E var measures and their 

standard errors based on different aspects in the tree layer. 


standard errors based on different aspects in the shrub layer. 


3.5 

3 

2.5 

2 

1.5 

1 

0.5 

0 

1-D 

N2 

H' 

N1 

1 2 3 4 5 6 7 

Aspect code 


14 

12 

10 

8 

6 

4 

2 

0 

a 

a 

ab 

a 

abc 

abc 

ab 

a 

bc 

abcd 

abc 

abc 

cd 

cd d 

bcd 

a ab 

abc 

bc cd cd 

ab 

abc 

a abc abc ab bcd abc bcd cde 

1 2 3 4 5 6 7 8 

1-D 

N 2 

H' 

N 1 

Aspect code 


on different aspects in the regeneration layer. 


on different aspects in the herbaceous layer (The same letters on 

the histogram indicate that there are no significant differences 

amongst mean values). 


5 

4.5 

4 

3.5 

3 

2.5 

2 

1.5 

1 

0.5 

0 

S 

R1 

R2 

Evar 

1 2 3 4 5 6 7 


14 

12 

10 

8 

6 

4 

2 

0 

cd 

bc 

abc 

abc 

abc 

a a 

a 

a ab bc ab ab ab bc bc 

1 2 3 4 5 6 7 8 

S 

Evar 

Aspect cod 

Aspect code 


standard errors based on different aspects in the regeneration 

layer. 

Figure 9. Mean richness and E var measures and their standard 

errors based on different aspects in the herbaceous layer.

20 

3 (1): 15-22, March 2011 


3.5 

3 

2.5 

2 

1.5 

1 

0.5 

1-D 

N 2 

H' 

N 1 


2.5 

2 

1.5 

1 

0.5 

1-D 

N2 

H' 

N1 

0 

1500 1550 1600 1650 1700 1750 1800 

0 

1500 1550 1600 1650 1700 1750 1800 

Elevation (m a.s.l.) 


Figure 10. Mean diversity measures and their standard errors 

based on elevation classes in the tree layer. 


based on elevation classes in the shrub layer 


5 

4.5 

4 

3.5 

3 

2.5 

2 

1.5 

1 

0.5 

0 

S 

R1 

R2 

Evar 

1500 1550 1600 1650 1700 1750 1800 



3 

2.5 

2 

1.5 

1 

0.5 

0 

S 

R1 

R2 

Evar 

1500 1550 1600 1650 1700 1750 1800 



standard errors based on elevation classes in the tree layer. 


standard errors based on elevation classes in the shrub layer. 


3.5 

3 

2.5 

2 

1.5 

1 

0.5 

0 

1-D 

N2 

H' 

N1 

1500 1550 1600 1650 1700 1750 1800 


10 

9 

8 

7 

6 

5 

4 

3 

2 

1 

0 

1500 1550 1600 1650 1700 1750 1800 

1-D 

N 2 

H' 

N 1 




based on elevation classes in the regeneration layer. 


based on elevation classes in the herbaceous layer. 


5 

4.5 

4 

3.5 

3 

2.5 

2 

1.5 

1 

0.5 

0 

S 

R1 

R2 

Evar 

1500 1550 1600 1650 1700 1750 1800 



12 

10 

8 

6 

4 

2 

0 

S 

Evar 

1500 1550 1600 1650 1700 1750 1800 



standard errors based on elevation classes in the regeneration 

layer. 

Figure 17. Mean richness and E var measures and their standard 

errors based on elevation classes in the herbaceous layer.


These results are to be confirmed with obtained results 

from Zagros forests in Ilam (Mirzaei et al. 2008; 

Pourbabaei et al. 2010). The Lebanon oak trees have been 

overexploited in southwestern aspect and as a result, 

population of other species such as Amygdalus communis 

and Crataegus pontica have increased in this aspect and 

also caused to increase tree species diversity. 

The regeneration diversity of woody species was found 

the highest in northeastern (i.e., 1-D and H´) and southern 

(i.e., N 1 and N 2 ) and the lowest in southwestern (i.e., 1-D 

and H´) and southeastern (i.e., N 2 ). The highest value of 

richness, R1 and R2 were found in northern and southern 

aspects and the lowest in southwestern and southeastern 

aspects. The highest value of evenness was found in 

southwestern and the lowest in northern aspect. 

The shrub diversity was found the highest in northern 

(i.e., H´ and N 2 ) and northwestern (i.e., 1-D and N 1 ) and the 

lowest in northeastern aspect. The highest value of richness 

and R 1 was found in northern and R 2 in eastern aspect. The 

lowest value of richness was found in other aspects, and R 1 

and R 2 in northwestern aspect. The highest value of 

evenness was found in northwestern and the lowest in 

northeastern. 

The herbaceous diversity was highest in western and 

the lowest in northeastern aspect. The highest value of 

richness and evenness were found in western and the 

lowest were in northern aspect. The number of tree 

individuals per hectare and its crown cover were low in 

western aspect and as a result the herbaceous diversity was 

the highest in this aspect. The population of tree species 

was more in northeastern aspect and crown coverage was 

60 to 80 percent and as a result the herbaceous diversity 

was lower in this aspect. 

Plant diversity based on elevation classes 

The elevation distribution of Lebanon oak species stretch 

from 1500 to 1800 m asl in the studied area. The highest 

and lowest Lebanon oak population was found from 1600 

to 1750 m asl (18%) and from 1500 to 1600 m asl (8%), 

respectively. The mean diversities were found the highest 

in elevation 1500 m asl and lowest in elevation 1800 m asl in 

the tree layer (Figure 10). There were no significant 

differences amongst mean diversity measures in the 

different elevations (P > 0.05). These results are to be 

confirmed with gained results of Zagros forests in Ilam 

(Mizaei et al. 2008). 

The mean richness, R 1 and R 2 measures were found the 

highest and lowest in elevation 1500 and 1800 m asl, 

respectively in the tree layer, while the highest and lowest 

of mean E var was found in elevation 1650 and 1600 m asl, 

respectively (Figure 11). There were no significant differences 

amongst mean these parameters in the different elevations. 

The mean diversities were found the highest in 

elevation 1500 m asl and lowest in elevation 1800 m asl in 

the regeneration layer (Figure 12). The mean richness and 

R 1 measures were found the highest in elevation 1500 m 

asl, and the highest value of R 2 was in elevation 1700 m asl 

and these parameters were lowest in elevation 1650 and 

1800 m asl, respectively in the regeneration layer. The 

highest and lowest of E var were found in elevation 1700 and 

1800 m asl, respectively (Figure 13). There were no 

significant differences amongst mean diversity, richness 

and evenness measures in elevation classes in the 

regeneration layer. 



the shrub layer (Figure 14). The mean richness and R 1 

measures were also found the highest in elevation 1500 m 

asl, and the highest value of R 2 was in elevation 1750 m asl 

and these parameters were lowest in elevation 1600 m asl 

in the shrub layer. The highest and lowest of E var were 

found in elevation 1600 and 1750 m asl, respectively 

(Figure 15). There were no significant differences amongst 

mean diversity, richness and evenness measures in 

elevation classes in the shrub layer. 



the herbaceous layer (Figure 16). The mean richness was 

found the highest in elevation 1500 m asl, and lowest in 

elevation 1800 m asl, and the highest and lowest of E var 

were found in elevation 1600 and 1800 m asl, respectively, 

in the this layer (Figure 17). There were no significant 

differences amongst mean diversity, richness and evenness 

measures in elevation classes in the herbaceous layer. 

The most population of Lebanon oak was found from 

1600 to 1750 m asl elevation. Maroufi (2000) indicated that 

this tree was distributed upper 1400 m asl elevation and it 

formed pure stands in elevation from 1600 to 1700 m asl 

The Quercus brantii, Q.infectoria and Q.libani species 

were observed with each other in elevation from 1500 to 

1600 m asl, and in elevation of 1600 to 1650 m asl 

Q.infectoria and Q.libani species found with together, and 

from 1650 to 1800 m asl just Q.libani was found 

(Tabatabaei and Geisarani 1992).The Q.libani species is 

distributed from 1500 to 2100 m asl and the best 

elevational range of this species was characterized from 

1600 to 1800 m asl (Jazirehi and Rostaghi 2003). 

The herbaceous species of Vicia variabilis Fren & Sint. 

has more population in sites where Q.libani population is 

plentiful. With increasing elevation up to 1700 m asl, 

V.variabilis population is also increased. The Q.libani 

forms pure stands in higher elevations (1650 to 1800 m asl) 

and population of Mesostemma kotschyanum is increased 

in comparing with V.variabilis since ecological needs of 

Mesostemma kotschyanum lower than is Vicia variabilis. 

The herbaceous coverage is to be increased in western and 

eastern aspects due to decreasing crown cover of oak 

species, and Turginia latifolia L. species is formed the 

most coverage percent since it has less ecological needs 

and it also is a thorny species. 

CONCLUSION 

The Zogros are divided into two parts: northern and 

southern. Northern Zagros is determined in the basis of 

distribution of Quercus infectoria Oliv. and Q. libani Oliv. 

Southern Zagros is also determined based on distribution of 

Quercus brantii Lindl. The Lebanon oak was found in all 

aspects, but it had the most population in eastern aspect and 

also this species was preferred northern aspect due to high

22 

3 (1): 15-22, March 2011 

ecological needs. The most population of Lebanon oak was 

found from 1600 to 1750 m asl elevation because of 

suitable humidity and edaphically conditions. In fact, 

elevational distribution of Lebanon oak is as spindle shape, 

that is, population of this species is increasing when the 

elevation is increasing and the population is decreasing in 

higher elevation. The disturbance is approximately high in 

elevation of 1500 m asl, as a result herbaceous and other 

woody species have been dominated and Lebanon oak 

decreased. Therefore, in order to rehabilitate the northern 

Zagros it is recommended that plantation of Lebanon oak is 

greatly conducted in the mentioned aspects and elevations. 

Regarding that plant species diversity and richness are 

considerable in studied area, it is better that this site is 

considered as genetic reservoir. 

ACKNOWLEDGEMENTS 

We would like to thank to Hosein Maroufi who helped 

us in identification of plant species specimens. Also, we 

wish to acknowledge our field assistants that had helped us 

during the data collection. 

REFERENCES 

Andel TV (2001) Floristic composition and diversity of mixed primary 

and secondary forests in northwest Guyana. Biodiv Conserv 10: 1645- 

1682. 

Aparicio A, Albaladejo RG, Olalla-Ta´rraga M A' , Carrillo LF, 

Rodríguez M A' (2008) Dispersal potentials determine responses of 

woody plant species richness to environmental factors in fragmented 

Mediterranean landscapes. For Ecol Manag 255: 2894-2906. 

Ashton AC, Macintosh DJ (2002) Preliminary assessment of plant 

diversity community ecology of the sematan mangrove forest, 

Sarawak, Malaysia. For Ecol Manag 166: 111-129. 

Aubert M, Alard D, Bureau F (2003) Diversity of plant assemblages in 

managed temperate forests: a case study in Normandy (France). For 

Ecol Manag 175: 321-337. 

Blanckaert I, Swennen RL, Paredes FM, Rosas LR, Lira SR (2004) 

Floristic composition, plant uses and management practices in home 

gardens of San Rafael Coxcatlán, Valley of Tehuacan-Coxcatlán, 

Mexico. J Arid Environ 57:39-62. 

Brockway DG (1998) Forest plant diversity at local and landscape scales 

in the Cascade Mountains of southwestern Washington. For Ecol 

Manag 109: 323-341. 

Browicz K (1994) Chorology of trees and shrubs in south-west Asia. 

Pozznn 57: 39-62. 

Cayuela L, Benayas JMR, Maestre FT, Escudero A (2008) Early 

environments drive diversity and floristic composition in 

Mediterranean old fields: Insights from a long-term experiment. Acta 

Oecologica 34: 311-321. 

Chiarucci A, Bonini I (2005) Quantitative floristics as a tool for the 

assessment of plant diversity in Tuscan forests. For Ecol Manag 212: 

160-170. 

Davis PH (1982). Flora of Turkey and the East Aegean Islands. Vol. 7. 

Edinburgh University Press. Edinburg. 

Department of Forestry (2002) Forestry plan, Ghablianeh district, Dow 

viseh region, Marivan. University of Kordestan. Sanandaj. [Persian] 

El-Ghanim WM, Hassan LM, Galal TM, Badr A (2010) Floristic 

composition and vegetation analysis in Hail region north of central 

Saudi Arabia. Saudi J Biol Sci, 17: 119-128. 

Fattahi, M (1994) Study on Zagros oak forests and the most important 

their destruction causes. Institute of Forests and Rangelands Research 

press. Sanandaj. [Persian] 

Figueroa JA, Teillier S, Castro SA (2011) Diversity patterns and 

composition of native and exotic floras in central Chile. Acta 

Oecologica 37 (2): 103-109. 

Gole TW, Borsch T, Denich M., Teketay D (2008) Floristic composition 

and environmental factors characterizing coffee forests in southwest 

Ethiopia. For Ecol Manag 255: 2138-2150. 

Hayat MSA, Kudus KA, Faridah-Hanum I, Noor AGA, Nazre M (2010) 

Assessment of Plant Species Diversity at Pasir Tengkorak Forest 

Reserve, Langkawi Island, Malaysia. J Agric Sci 2(1): 31-38. 

Ipor I, Sani H, Tawan C (2002) Floristic composition of forest formation 

at Mahua, Crocker range national park, Sabah. ASEAN Review of 

Biodiversity and Environmental Conservation (ARBEC), July- 

September, 1-8. 

Jazirehi MH, Rostaghi EM (2003) Silviculture in Zagros. University of 

Tehran Press. Tehran. [Persian] 

Jobidon R, Cyr G, Thiffault N (2004) Plant species diversity and 

composition along an experimental gradient of northern hardwood 

abundance in Picea mariana plantations. For Ecol Manag 198: 209-221. 

Khera N, Kumar A, Ram J, Tewari A (2001) Plant biodiversity 

assessment in relation to disturbances in mid- elevational forest of 

Central Himalaya, India. Tropical Ecol 42 (1): 83-95 

Kinnear PR, Gray CO (2001) SPSS for windows made simple release 10 

(translated by Ardakani AF). Psychology Press. Tehran. [Persian] 

Krebs JC (1999) Ecological methodology. Harper and Row. New York. 

Ludwig JA, Reynolds JF (1988) Statistical ecology. John Wiley and Sons. 

New York. 

Macía MJ (2008) Woody plants diversity, Xoristic composition and land 

use history in the Amazonian rain forests of Madidi National Park, 

Bolivia. Biodiv Conserv 17: 2671-2690. 

Magurran AE (2004) Measuring biological diversity. Blackwell. New 

York. 

Maroufi H (2000) Study on site requirements of Lebanon oak (Quercus 

libani Oliv.) in Kordestan province. [Dissertation]. Institute of Higher 

Education of Imam Khomeini. Qom. [Persian] 

Mizaei J, Akbarinia M, Hosseini SM, Sohrabi H, Hosseinzade J (2008) 

Biodiversity of herbaceous species in related to physiographic factors 

in forest ecosystems in central Zagros. Iranian J Biol 20 (4): 375-382. 

[Persian] 

Mohadjer, MR (2005) Silviculture. University of Tehran Press. Tehran. 

[Persian] 

Mueller-Dombois, Ellenberg H (1974) Aims and methods of vegetation 

ecology. John Wiley and Sons. New York. 

Nagaike T, Hayashi A, Abe M, Arai N (2003) Differences in plant species 

diversity in Larix kaempferi plantations of different ages in central 

Japan. For Ecol Manag 183: 177-193. 

Nebel G, Kvist LP, Vanclay JK, Christensen H, Freitas L, Ruíz J (2001) 

Structure and floristic composition of flood plain forests in the 

Peruvian Amazon, I. Overstory. For Ecol Manag 150: 27-57. 

Pant S, Samant SS (2007) Assessment of plant diversity and prioritization 

of communities for conservation in Mornaula. Appl Ecol Environ Res 

5 (2): 123-138. 

Pérez-Ramos IM, Zavala MA, Marañón T, Dı´az-Villa MD, Valladares F 

(2008) Dynamics of understorey herbaceous plant diversity following 

shrub clearing of cork oak forests: A five-year study. For Ecol Manag 

255: 3242-3253. 

Pitkänen S (1998) The use of diversity indices to assess the diversity of 

vegetation in managed boreal forests. For Ecol Manag 112:121-137. 

Pourbabaei H, Heydari M, Najafifar A (2010) The relationship between 

plant diversity and physiographic factors in Ghalarang protected area. 

Ecol Environ Conserv 16 (4): 1-7. 

Ramírez N, Dezzeo N, Chacón N (2007) Floristic composition, plant 

species abundance, and soil properties of montane savannas in the 

Gran Sabana, Venezuela. Flora 202: 316-327. 

Sabeti H (1994) Forests, trees and shrubs in Iran. University of Yazd. 

Yazd, IR Iran. [Persian] 

Tabatabaei M, Geisarani F (1992) Natural resources of Kordestan. 

Jahadedaneshgahi publications. Tehran. [Persian] 

Townsend CC, Guest E (eds). (1980) Flora of Iraq, vol. 4. Ministry of 

Agriculture and Agrarian Reform. Baghdad. 

Wardell-Johnson GW, Williams MR, Mellican AE, Annells A (2004) 

Floristic patterns and disturbance history in karri forest, south-western 

Australia, 1. Environment and species richness. For Ecol Manag 199: 

449-460. 

Zobeiri M (1994) Forest inventory (measuring forest and tree). Tehran 

University. Tehran. [Persian]

Vol. 3, No. 1, Pp.: 23-27 

March 2011 



Evaluation structural diversity of Carpinus betulus stand in Golestan 

Province, North of Iran 

VAHAB SOHRABI 1,♥ , RAMIN RAHMANI 1 , SHAHROKH JABBARI 2 , HADI MOAYERI 1 

Faculty of Forestry, Gorgan University of Agricultural Science and Natural Resources. PO Box 386, Shahid Beheshti Street, Gorgan, Golestan, Islamic 

Republic of Iran, Tel. +98 (171) 222 0028, Fax. +98 (171) 222 598, ♥ Email: vahabsohrabi61@yahoo.com 

2 Super Council of Forests, Range, Watershade Management Organization, Islamic Republic of Iran. 

Manuscript received: 19 February 2011 Revision accepted: 3 March 2011. 

Abstract. Sohrabi V, Rahmani R, Jabbari S, Moayeri H. 2011. Evaluation structural diversity of Carpinus betulus stand in Golestan 

Province, Northern Iran. Nusantara Bioscience 3: 23-27. In order to investigate structural diversity of Carpinus betulus type in Golestan 

province 30 modified Whittaker plots by systematic random system were located. Per plot the characteristic of trees and shrubs species 

(Species name, diameter and height of trees) are recorded. The heterogenity indices of Simpson, Shannon–Wiener, Simpson’s reciprocal 

and number of equally common species were used for the quantitative data. Toward better understand from diversity condition in 

horizontal and vertical composition of stand, the diameter divided in 10 cm classes and Method of Mohajer and the height divided in 10 

m height classes and dominant height, then number of diversity of each class extracted by Ecological Methodology software V.7. The 

results showed with increase of diameter and height classes, decrease species diversity. Also regeneration layers diversity has significant 

difference with trees layers. Thus, the study of biodiversity changes in different diameter and height category cause ecologically precise 

perspective in management of forest stands. 

Key words: structure diversity, indices diversity, diameter and height classes. 

Abstrak. Sohrabi V, Rahmani R, Jabbari S, Moayeri H. 2011. Evaluasi keragaman struktur tegakan Carpinus betulus di Provinsi 

Golestan, Iran bagian utara. Nusantara Bioscience 3: 23-27. Dalam rangka untuk menyelidiki struktur keragaman tipe Carpinus betulus 

di provinsi Golestan, 30 plot Whittaker yang telah dimodifikasi dibuat secara sistem random sistematis. Pada setiap plot, karakteristik 

spesies pepohonan dan semak (nama spesies, diameter dan tinggi pohon) dicatat. Indeks heterogenitas dari beberapa macam indeks 

Simpson, Shannon-Wiener, Simpson’s reciprocal dan jumlah spesies yang umum ditemukan digunakan untuk data kuantitatif. Untuk 

lebih memahami kondisi keanekaragaman dalam tegakan horizontal dan vertikal, maka dikelompokkan ke dalam diameter dalam kelas 

10 cm, metode Mohajer, tinggi dalam kelas 10 m, dan ketinggian yang dominan, kemudian jumlah keragaman setiap kelas ditentukan 

dengan software Ecological Methodology v.7.0. Hasil penelitian menunjukkan bahwa peningkatan kelas diameter dan tinggi, 

menyebabkan penurunan keragaman spesies. Keragaman lapisan regenerasi memiliki perbedaan signifikan dengan lapisan pohon. Studi 

perubahan keanekaragaman hayati dengan kategori diameter dan tinggi yang berbeda memerlukan perspektif ekologis yang tepat dalam 

pengelolaan tegakan hutan. 

Kata kunci: keanekaragaman struktur, indeks diversitas, kelas diameter dan tinggi. 

INTRODUCTION 

Human knows the concept and the importance of 

biodiversity from earlier century. Plato frequently point out 

the diversity and believe that if there is more diversity in 

the world, the world will be better (Beasapour 2000, 

Ejtehadi et al. 2009). Today, the word of biodiversity 

applies by various science experts, such as ecologists. The 

convention of biodiversity of USA, describe biodiversity 

as: there is a difference in all the life type all sources such 

as marine, ground and ecological complex combination and 

include the diversity within species, between species and 

ecosystems (Markandya et al. 2008). One of the constant 

keys of management of uneven age forest is the true 

understanding about spatial structure of forest (Costanza et 

al. 2007). Forest structure is the important feature in 

management of forest ecosystems (Zenner and Hibbs 

2000). Structural features are used to determine the species 

neech heterogeneous experiment and plant dynamic time, 

management of regeneration patterns and fragmentation 

dynamic, description of microclimate diversity and 

predicting the wood production (Youngblooda et al. 2004). 

Management of forest stands performs by stands structure 

control (age, size and tree density) and forest structure (size 

and spatial order of tree) because the concept of forest 

structure is more important than species combination 

(Oheimb et al. 2005). The study of natural forests structure 

defined the way of desired structure that the use of 

appropriate silviculture operation and stimulation of natural 

structure in under management stands considered as the 

way to keep the biological diversity and forest dynamic and 

stability (Markandya et al. 2003). The study of forest

24 

3 (1): 23-27, March 2011 

structure especially in virgin forests is very important and 

gives us comprehensive information about the condition in 

forest for programming. The diversity of a forest stand may 

not be sufficiently described by tree species diversity alone. 

Structural diversity, resulting from recruitment of trees of 

different sizes into multilayered canopies, should also be 

taken into account (Liang et al. 2007). This characteristic, 

which can be approximated by the diversity of tree size, 

affects the amount of light and precipitation received by 

subordinate trees and understory plants (Anderson et al. 

1969), and may thus influence the productivity of forest 

ecosystems. Thus manipulating tree-size diversity is a 

practical tool for forestmanagers who strive for greater 

biodiversity and/or greater productivity (Varga et al. 

2005).Varus studing done about in forest structure. 

Ahani et al (2006) do the research about species 

diversity of tree based on the diameter class in Acer sites in 

Shafarud forests. So, rhombus plots in half hectare study in 

forest according to Acer (34 plots). First the feature within 

each plot, its slope, aspect, height from sea level and then 

total diameter of trees up to more than 10 cm 

measured.Biodiversity accounted in four diameter alasses 

(10-30, 35-50, 55-80, 80-120 cm). The result showed that 

the Shanon and N1 Mac Arthur indices in diameter class of 

35-50 cm, have greatest amount, while the index of 

Simpson and N2 hill shows the greatest amount in diameter 

class of 10-30 cm. The purpose of this paper is the 

evaluation of structural diversity in diameter and height 

classes and their changing process with changing of 

diameter classes and height category in Carpinus betulus 

(Persian: Mamarz) type in Golestan province, IR Iran. 


The regions of study 

Kohmian forestry plan is located in 98 wateshade 

domain which is limited in north is village of Kohmian, 

Fazel Abad, Khanduz Sadat and Marzbone, in south and 

west to Naeem s forestry plan and in east to vatan forestry 

plan. Its east longitude is 55-14-49 to 55-10-30 and its 

north width is 37-65-15 to 37-00-00 degrees (Figure 1). 

Golestan 

6 

4 

5 

1 

2 

3 

Figure 1. Map of the site study in in Golestan Province, North of Iran. 1. Shastkalateh, 2. Tavir, 3. Kohmian, 4. Takht, 5. Loveh, 6. 

Farsian.

SOHRABI et al. – Diversity of Carpinus betulus stand in Golestan, Iran 25 

Table 1. Indices used in this paper (Ejtehadi et al. 2009) 

Equation 

1− D = 1 −∑ ( p i 

) 

s 

Pi 

H ′ = ∑ ( Pi 

)( Log2 

) 

i = 1 

1 1 

= 

∑ 

2 

D p i 

H′ 

1 

= e 

N 

2 

Simpson 

Index 

Shannon–Wiener 

Simpson’s 

reciprocal 

Number of equally 

common species 

Description of equation 

(1-D) = Simson’s index of diversity 

p 1 = proportion of individual species I in the community 

H’ = information content of sample (bits/individual) = index of species diversity 

s = number of species 

p 1 = proportion of total sample belonging to i-th species 

1/D = Simson’s reciprocal index (= Hill’s N 2 ) 

p 1 = proportion of individual species i in the community 

H’ = information content of sample (bits/individual) = index of species diversity 

s = number of species 

p 1 = proportion of total sample belonging to i-th species 

Research method 

This research is basee on sampling by systematic 

random system and the center of plots in forest is 

determined. To study and investigation, 30 modified 

Whittaker plots in range of 850-950 m altitude from the sea 

level in north aspect were located. In this 20x50 meter 

frame, the characteristic of trees and shrubs species 

(species name, diameter and height of trees) are recorded. 

The heterogenity indices of Simpson, Shannon–Wiener, 

Simpson’s reciprocal and number of equally common 

species and evenness indices of Simpson, Camargo, Smith- 

Wilson and modified nee were used for the quantitative 

data (Table 1). Then aforesaid characteristics saved as 

information bank in Excell 2010. Then indices account by 

Ecological Methodology software v.7.0 (Krebs 1999). 

Analyze of data was done by analyze of variance 

(ANOVA) and Duncan’s multiple range test (DMRT). 


Next of survey recorded number of 10 trees species 

dependent of 8 families and 3 shrubs species dependent of 

2 families that show notable statistics (Table 2). 

Table 2. Composition of trees and shrubs species. 

Scientific name Family Trees/Shrubs 

Quercus castanefolia Fagaceae T 

Carpinus betulus Betulaceae T 

Parrotia persica Hamameliadaceae T 

Tilia begunda Tiliaceae T 

Acer insigne Acearaceae T 

Ulmus glabra Ulmaceae T 

Acer cappadocicum Acearaceae T 

Alnus glutinosa Betulaceae T 

Crataegus monogyna Rosaceae S 

Mespilus germanica Rosaceae S 

Prunus avium Rosaceae S 

Sorbus torminalis Rosaceae T 

Diospyros lotus Ebenaceae S 

Diversity indices in 10 cm diameter classes 

The under study diversity indices in this paper shows 

the decrease in the diameter classes of 10cm with increase 

of classes. The most diversity number is in diameter class 

of 0-10 cm and the least diversity number is in diameter 

class of 90-100 cm. other than Simpson diversity index that 

shows the least diversity number in diameter class of more 

than 100cm, the significant different is between diameter 

classes in 1% level (Figure 2). 

Diversity indices in diameter classes by method of 

Mohajer 

Diversity indices shows decrease process with the 

increase of diameter classes but it increase again in last 

class (dbh>80). The most diversity number is in the class of 

0-10 cm and the least diversity number is in class of 60_80 

cm. The diameter classas (20-30, 30-60, dbh>80) are not 

significant different. The significant different is between 

diameter classes in 1% level (Figure 3). 

Diversity indices in 10m height classes 

Diversity indices have orderly decrease process. The 

most diversity number is in height class of 0-10m and the 

least diversity number is in height class of 40-50m. , the 

significant different is between height classes in 1% level 

(Figure 4). 

Diversity indices in dominant height of height classes 

The most diversity number in all indices is for h

26 

3 (1): 23-27, March 2011 

Figure 2. The comparison of diversity indices in 10cm diameter 

classes. A. Simpson, B. Simpson’s reciprocal, C. Shannon– 

Wiener, D. Number of equally common species. 

Figure 5. The comparison of diversity indices in height classes by 

dominant height. A. Simpson, B. Simpson’s reciprocal, C. 

Shannon–Wiener, D. Number of equally common species. 

Figure 3. The comparison of diversity indices by method of 

Mohajer (2005). A. Simpson, B. Simpson’s reciprocal, C. 

Shannon–Wiener, D. Number of equally common species. 

Figure 4. The comparison of diversity indices in 10m height 

classes. A. Simpson, B. Simpson’s reciprocal, C. Shannon– 

Wiener, D. Number of equally common species 

forest stands. Forest stands have different structure in 

various sections (linear and phenomenal) like a building. 

For recognition, study and precise programming of forest 

stands, its features need to consider according to different 

sections. Various profiles (linear and phenomenal) could be 

dividing for forest stands. The study of forest stand profile 

especially in virgin forests is very important and gives us 

comprehensive information about structure of these forests 

(Mohajer 2005). For better understanding of the structure 

of forest stand, we analyzed it according to the vertical and 

horizontal structure. Species diversity of tree and shrub in 

this type have significant different in low diameter and 

height classes with up diameter and height classes classes. 

Diameter and height classes below of 10 cm, account as 10 

regeneration layer, so diversity of regeneration layer is 

more than the diversity of tree layers (Pourbabaei et al. 

2006; Sohrabi 2010). This is due to the decrease of canopy 

of small saplings and it need low light than higher age 

process in this classes. By the increase of diametrical and 

height classes, the diversity decrease. It is obvious that the 

structure diversity naturally in the virgin forest decrease 

depend on site condition and with increase of stand age and 

its move toward climax, because gradually increase of trees 

age dominant species dominant against the under species. 

Trees are the main elements in forest ecosystems that other 

living thing life of this ecosystem depends on the life of 

them. Therefore removing of the tree threatened the life of 

the existent in this ecosystem. The main role of forest 

engineer is the marketing of forest (Mohajer 2005). In this 

step choosing of trees perform by considering of target 

diameter from defined species and gradually the number of 

trees in defined diameter decreased and so the repeating act 

might remove some class of trees. It is threatened the 

structure diversity and the species diversity. Trees diversity 

in higher diametrical and altitudinal categories is part of the 

lower diametrical category diversity. Any changes in above 

level might change the ground cover. Tree dimension 

diversity has an effect on the amount of light and raining 

by small plant and trees (Anderson et al.1969). This has 

influence on the produce of forest ecosystems.

SOHRABI et al. – Diversity of Carpinus betulus stand in Golestan, Iran 27 

CONCUSION 

The increasing of diameter and height classes, decrease 

species diversity. Regeneration layers diversity has 

significant difference with trees layers. Thus, the study of 

biodiversity changes in different diameter and height 

category cause ecologically precise perspective in 

management of forest stands. 


Therefore I express gratitude to any ones who is useful 

in my life. By the way, I thank Ezazi to give us translation 

of this paper. 

REFERENCES 

Ahani H, Pourbabaei H, Bonyad AE. 2006. Investigation of trees species 

diversity based on diameter at breast height (dbh) class on Norway 

Maple (Acer platanoides L.) in Shafarood Forest (Guilan Province). J 

Agric Sci 12 (3): 525-533. 

Anderson RC, Loucks OL, Swain AM. 1969. Herbaceous response to as 

soil indicators in Oregon’s western cascades old-growth forests. 

Northwest boreal coniferous forests. Ecology 50: 255-263 

Beasapour D. 2000. Reconnaissance The best Indices biodiversity and Use 

them in Ecosystem Estimate. Articeles Collection of chronicology 

and biodiversity 285-291 

Costanza R, Fisher B, Muler K, Liu S, Christopher T. 2007. Biodiversity 

and ecosystem services: A multi-scale empirical study of the 

relationship between species richness and net primary production. 

Ecol Econ 61: 478-491. 

Ejtehadi H, Sepehry A, Akkafi HR. 2009. Method of measuring 

biodiversity. Ferdowsi University of Mashhad Publication No. 530. 

Mashhad, IR Iran. 

Krebs CJ. 1999. Ecological Methodology. 2 nd ed. Addison-Welsey. Menlo 

Park, CA. 

Liang J, Buongiorno J, Monserud RA, Kruger EL, Zhou M. 2007. Effects 

of diversity of tree species and size on forest basal area growth, 

recruitment, and mortality. For Ecol Manag 243 (2007) 116-127. 

Markandya A, Nunes PALD, Bräuer I, ten Brink P, Kuik O, Rayment M. 

2008. The economics of ecosystems and biodiversity – Phase 1 

(scoping) economic analysis and synthesis. Final Report for the 

European Commission, Venice, Italy. 

Markandya T, Nishimurab N, Yamamotoa S. 2003. Population structure 

and spatial patterns of major trees in a sub alpine old-growth 

coniferous forest, central Japan. For Ecol Manag 182: 259-272. 

Mohajer MM. 2005. Silviculture. Tehran University Press, Tehran, Iran. 

Oheimb GO, Westphal Ch, Tempel H, Hardtle W. 2005. Structural pattern 

of a nearnatural beech forest (Fagus sylvatica) (Serrahn, North-east 

Germany). For Ecol Manag 212: 253–263 

Pourbabaei H, Dado KH. 2000. Species diversity of woody plants in the 

district No. 1 forests, Kelardasht, Mazandaran province. J Iran Biol 

18: 306-322. 

Sohrabi V. 2010. Comparing of species and stracture diversity in gradient 

of between Shastkolateh Beech Forest and Loveh Oak Forest [M.Sc 

thesis]. Faculty Forestry, Gorgan University of Agriculture Sciences 

and Natural Resources. Gorgan, IR Iran. 

Stohlgren TJ, Falkner MB, Schell LD. 1995. A modified-whittaker nested 

vegetation sampling method. Vegetatio 117 (2): 113-121. 

Varga P, Chen HYH, Klinka K. 2005. Tree-size diversity between 

singleand mixed-species stands in three forest types in western 

Canada. Can J Forest Res 35 593-601. 

Youngblooda A, Maxb T, and Coe K. 2004. Stand structure in eastside 

old-growth ponderosa pine forests of Oregon and northern California. 

For Ecol Manag 195: 238-256. 

Zenner EK, Hibbs DE. 2000. A new method for modeling the 

heterogeneity of forest structure. For Ecol Manag 129: 75-87.

Vol. 3, No. 1, Pp.: 28-35 

March 2011 



Microanatomy alteration of gills and kidneys in freshwater mussel 

(Anodonta woodiana) due to cadmium exposure 

FUAD FITRIAWAN 1,♥ , SUTARNO², SUNARTO² 

¹ Open University, UPBJJ Bandar Lampung. Jl. Soekarno-Hatta No. 108 B Rajabasa, Bandar Lampung 35144, Lampung, Indonesia. Tel.: +92-721- 

704772. Fak.: +92-721-709026. E-mail: ut-bandarlampung@upbjj.ut.ac.id, maz_afid@yahoo.co.id 

² Bioscience Program, School of Graduates, Sebelas Maret University, Surakarta 57126, Central Java, Indonesia 

Manuscript received: 15 December 2010. Revision accepted: 26 February 2011. 

Abstract. Fitriawan F, Sutarno, Sunarto. 2011. Microanatomy alteration of gills and kidneys in freshwater mussel (Anodonta 

woodiana) due to cadmium exposure. Nusantara Bioscience 3: 28-35. The purpose of this study were to determine the level of Cd 

accumulation in the gills and kidneys, to khow the changes in microanatomic structure of A. woodiana after the various treatments of 

heavy metals. Completely randomized design pattern of 5 x 3 as used in this laboratory experiment. The amount of exposure of heavy 

metals Cd were (0 ppm, 0.5 ppm, 1 ppm, 5 ppm, 10 ppm), while the variation of length of exprosure time to Cd were (7 days, 14 days, 

and 30 days). The parameters of Cd accumulation in the gills and kidney was analyzed by using AAS method, while abnormalities of 

gills and kidney were detected by microanatomy structure. Data collected were then analyzed using the analysis of variance (ANOVA) 

and continued with further test the DMRT. The results indicated that there is a significant effect in 475.3 > 0.000 and 60150.3 >0.000 

with 5% significance level (P 0,000 dan 60150,3 > 0,000 dengan taraf signifikansi rata-rata 5% (P

FITRIAWAN et al. – Effect of Cd on freshwater mussels A. woodiana 29 

tissue which became the meeting place of active transport 

between organisms and the environment (Soegianto et al. 

2004). Renal function begins in the glomerular ultrafilter 

that is formed from the plasma. Ultrafilter will enter the 

Bowman's capsule and into the lumen of the tubule. 

Filtering through the various segments of the tubules 

causes changes in the volume and composition of fluid 

filtration as a result of the process of reabsorption and 

secretion along the tubules (Tresnati et al. 2007). 

Glomerulus is composed of blood capillaries to function as 

a selective filter from the blood mainly in the normal blood 

screening (Takashima and Hibiya 1995). Following 

through on glomerular filtration and being re-absorbed in 

the tubular, it produces urine as a result of secretion in 

normal circumstances (Tresnati et al. 2007). 

The purpose of this study are: (i) to know the content of 

Cd accumulation, (ii) changes in the microanatomy 

structure, and (iii) in gill and kidney of A. woodiana after 

treatment. 


Time and place 

The research of the Cd treatment on A. woodiana 

conducted at the Laboratory of Pharmacy and Food 

Academy Analyst Sunan Giri, Roxburgh. Analysis of 

heavy metal content by AAS method was carried out in sub 

lab Chemistry Laboratory of Mathematics and Science 

Center UNS Surakarta, while the preparation for the 

analysis was conducted in the laboratory animal anatomy 

Faculty of Veterinary Medicine, Gadjah Mada University 

in Yogyakarta. The experiment was conducted in October- 

November 2009. 

Materials 

Freshwater mussels (A. woodiana) were obtained from 

farms in the fishing village of the tourist site of Janti, 

Polanharjo Subdistrict, Klaten District, Central Java. 

Procedures 

Freshwater bivalve A. woodiana was selected based 

onthe maximum growth and uniform size. The shellfish 

was acclimatized for 15 days, after which it was examined 

by using the compound of Cd for 30 days with repeated 3 

times at day 7, 14 and 30. Physical-chemical parameters 

measured mencakuip pH, DO and water temperature where 

the experiment. Cd content of the examination conducted 

on the gill and kidney A. woodiana with AAS method. 

Preparation of gill and kidney preparations performed with 

haematoxylin-eosin (HE) method with treatment stages, ie 

trimming, dehydration, embedding, cutting, stainning, 

mounting and reading the results. 

Data analysis 

Environmental chemistry parameters (pH, DO, 

temperature) were described with a descriptive method. 

Effects of Cd exposure on the gill and kidney A. woodiana 

were analyzed by ANOVA one-way significance level of 

5% (P> 0.05), followed by a further test of significant 

difference or Duncan's multiple range test (DMRT). 

Abnormality in the microanatomy structure of gills and 

kidneys of A. woodiana was directly observed and 

described with a descriptive method. 


Water environmental parameters 

Examination of physical and chemical parameters of 

water quality used in this study include the degree of 

acidity (pH), dissolved oxygen (DO), and water temperature. 

Degree of acidity (pH) 

The degree of acidity or pH is a value that shows the 

activity of hydrogen ions in water. The pH of a water 

reflects the balance between acid and base in these waters. 

The pH range 1-14, pH 7 is the boundary halfway between 

the acid and alkaline (neutral). The higher the pH of the 

water, the greater the base nature will be, and the lower the 

pH the more acidic the water. PH value is influenced by 

several parameters, including biological activity, 

temperature, oxygen content and the ions. From the 

biological activity, CO2 gas is generated as a result of 

respiration. This gas will form a buffer or buffer ions to 

maintain the pH range in the waters in order to remain 

stable (Erland 2007). 

In this study, the pH is very important as water quality 

parameters, for controlling the type and rate of speed of 

reaction some materials in the water. In addition, A. 

woodiana live at a certain pH interval, so that by knowing 

the value of pH, it can be known whether or not the water 

supports their lives. Based on Figure 1A, it is known that 

the higher Cd concentration, the higher the value of the 

range of pH waters. On day 7, pH values ranged from 7.34 

to 8.44, on day 14 ranged from 7.37 to 8.40, and the dayto-30 

range from 7.31 to 8.68. According to Erland (2007), 

pH to function as an index of environmental conditions and 

limiting factors, where each organism has a different 

tolerance to pH maximum, minimum and optimal. 

According to Erland (2007) pH value of water has a 

special characteristic, the hydrogen ion concentration be 

measured by the balance between acids and bases. Acidfree 

mineral acid and carbonic acid will lower pH value 

(acid), while the carbonate (CO3), hydroxide (OH-) and 

bicarbonate to raise pH (alkaline). Rochyatun et al. (2006) 

states, that at a relatively high metal content will be 

alkaline pH values (pH 7.40 to 8.59), where the metal is 

difficult to dissolve and settle to the bottom of the water. 

When in the treatment, pH values from 0.5 to 10 ppm, in 

the study it ranged from 7.92 to 8.68, indicating water has 

been polluted quite heavily, with the level of alkalinity in 

excess of tolerance. According to Connell and Miller 

(1995) increase in pH in the waters will be followed by 

decreasing the solubility of heavy metals that tend to settle. 

Deposition can occur in sediments and food; the food will 

enter and accumulate in the body of A. woodiana. Given 

the Cd is a non-essential metal that cannot be degraded so 

that it will cause interference with the organs, such as the 

gill and kidney.

30 

3 (1): 28-35, March 2011 

Water solubility of oxygen (DO) 

Oxygen is one of the gases dissolved in natural waters 

with varying levels are influenced by temperature, salinity, 

water turbulence, and atmospheric pressure. Besides 

necessary for the survival of aquatic organisms, oxygen is 

also needed in the process of decomposition of organic 

compounds. Sources of dissolved oxygen are mainly 

derived from the diffusion of oxygen from the atmosphere. 

This diffusion occurs directly on stagnant conditions 

(silent), or because of agitation (water mass unrest) caused 

by waves or wind. 

Figure 1B shows that the higher concentration of Cd 

treatment, then progressively decreasing levels of dissolved 

oxygen (DO) in water. Ardi (2002) classified water quality 

based on the DO into four types namely; not contaminated 

(> 6.5 mg/L), lightly polluted (4.5 to 6.5 mg/L), being 

contaminated (2.0 to 4, 4 mg/L) and heavily polluted (


increased in order to defend themselves, so that 

automatically the oxygen demand is very much while on 

the other hand, given the concentration of heavy metals 

higher, thus increasing the concentration of heavy metals 

that enter the more carbon dioxide (CO 2 ) are released that 

cause the oxygen content dwindling waters so that the 

rising water temperature. 

According to Connell and Miller (1995) the role of 

water temperature is very important to help the body's 

metabolism of aquatic animals. The increase in water 

temperature can cause the immune system of aquatic biota 

to decrease. So if a toxic Cd 2+ enters the body of A. 

woodiana the biota will be very difficult to retain yourself 

from the poison. 

Accumulation of Cd in the gills of A. woodiana 

The result of the content of Cd in the gill and kidney A. 

woodiana with AAS method are shown in Table 1. From 

the results it is known that the increasing Cd treatment, the 

increase of Cd accumulatd in the gill of A. woodiana. In the 

control (0 ppm), Cd accumulation in the gills of A. 

woodiana was 0.12 ppm, the accumulation in the control is 

still below the maximum tolerance limit Cd accumulation 

in organs, as specified by the FAO (1972) and MOH 

(1989), namely a maximum accumulation of Cd in organs 

of 1 ppm. It is also in accordance with preliminary studies 

that have been made to the content of Cd in water samples, 

with the result that the content of Cd in Janti aquaculture 

that was still in the normal state that was 0.0028 ppm (IGR 

No. 82/2001; EPA 1986). 

After the examination after 7 days the average values 

obtained Cd accumulation in gill A. woodiana in the 

treatment of 0.5 ppm was 0.58 ppm, treatment of 1 ppm 

was 0.87 ppm, 5 ppm was 1.00 ppm and 10 ppm was 2.15 

ppm. Meanwhile, after the examination on day 14, obtained 

an average value of accumulated Cd at 0.5 ppm treatment 

was 0.78, treatment of 1 ppm 0.93 ppm, 5 ppm treatment at 

1.24 ppm, and on treatment 10 ppm at 2.34 ppm. After the 

examination on day-30, it was obtained an average value of 

Cd accumulation in the treatment of 0.5 ppm 1.43 ppm, 1 

ppm treatment at 1.01, 5 ppm treatment at 2.58, and the 

treatment of 10 ppm 3.49 ppm. 

Darmono (1995) states that the relationship between the 

amount of metal absorption and metal content in water is 

usually in proportion, the increase in metal content in the 

network in accordance with the increase of metal content in 

water. According Sunarto (2007) Cd will also experience 

the process of biotransformation and bioaccumulation in 

aquatic biota. Cadmium enters the body along the water or 

food consumed, but water or food has been contaminated 

by Cd. The amount of metal that accumulates in the gills 

will continue to increase, even very likely continue to enter 

through the accumulation of Cd digestive tract to the 

kidney; in addition to increasing levels of pollutants in the 

presence of Cd may also biomagnification process in the 

water body. If the amount of Cd that enters the body and 

has exceeded the threshold value, it will experience death 

and even extinction. 

Cd treatment against gill A. woodiana with a 

concentration of 0 ppm, 0.5 ppm, 1 ppm, 5 ppm and 10 

ppm for 7 days, 14 days and 30 days to yield significant 

results (P

32 

3 (1): 28-35, March 2011 

Analysis of treatment outcome ofthe renal Cd of A. 

woodiana 

Test results on the Cd content of the kidney A. 

woodiana after treatment is shown in Table 1. The average 

value of the kidney of A. woodiana in the control group 

amounted to 0.019 ppm (0.018933 ppm), the value 

accumulated in the control was still below the maximum 

tolerance limit Cd accumulation in organs, as specified by 

the FAO (1972) and MOH (1989) that is equal to 1 ppm. It 

is also in accordance with preliminary studies that have 

been made to the content of Cd in the water, that the 

content of Cd in the water of Janti aquaculture is still in the 

normal state is 0.0028 ppm (IGR No. 82/2001; EPA 1986). 

Later in the treatment of 0.5 ppm Cd in the kidneys 

after examination, AAS average accumulation after 7 days 

was at 0.020 ppm, after 14 days was at 0.029 ppm, after 30 

days was at 0.086 ppm. Later in the treatment of 1 ppm 

after 7 days accumulation of 0.030 ppm, 0.031 ppm after 

14 days, and after 30 days at 0.066 ppm. Later in the 

treatment of 5 ppm Cd accumulation after 7 days at 0.057 

ppm, after 14 days at 0.085 ppm, and after 30 days at 0.107 

ppm. And in the treatment of 10 ppm Cd accumulation in 

the kidney after 7 days at 0.116 ppm, after 14 days at 0.150 

ppm and after 30 days showed the exposure of 1.717 ppm. 

From the above data, it is known that the higher the 

concentration of Cd treatment A. woodiana, the higher the 

value of exposure to cadmium in the kidneys of A. 

woodiana. This is similar to what has been mentioned by 

Sunarto (2007) that Cd will also experience the process of 

biotransformation and bioaccumulation in aquatic biota. 

Cadmium enters the body along the water or food 

consumed, where water or food has been contaminated by 

Cd. The amount of metal that accumulates in the gills will 

continue to increase, even very likely continue to enter 

through the accumulation of Cd digestive tract to the kidney. 

Based on ANOVA statistical test, it is known that Cd 

treatment on the kidney of A. woodiana with a 

concentration of 0 ppm, 0.5 ppm, 1 ppm, 5 ppm and 10 

ppm for 7 days, 14 days and 30 days, yield significant 

results (P


bottom waters, such as A. woodiana (bivalves) will have a 

great opportunity for exposure to heavy metals that have 

been bound and form sediment. 

Table 3. Changes in cellular structure mikroanatomi A. Gill 

woodiana after exposure to heavy metals cadmium with HE 

staining preparation. 

Concentration 

(ppm) 

Time of 

surgery 

(days) 

Edema Hyperplasia 

Fusion 

of 

lamella 

Necrosis Atrophy 

0 7 - - - - - 

14 - - - - - 

30 - - - - - 

0,5 7 + - - - - 

14 ++ + - - - 

30 +++ ++ - - - 

1 7 ++ + - - - 

14 ++ ++ ++ - - 

30 ++++ +++ +++ - - 

5 7 ++++ +++ + - - 

14 ++++ ++++ +++ - - 

(dead) 30 ++++ ++++ ++++ +++ - 

10 7 ++++ +++ + - - 

14 ++++ ++++ ++++ ++++ ++ 

(mati) 30 ++++ ++++ ++++ ++++ +++ 

Note: -: no change in the microanatomical structure (0%); +: there 

was a slight change in the microanatomical structure (1% -25%); 

+ +: there are changes in the microanatomical structure (26% - 

50%); + + +: occurred many changes in the microanatomical 

structure (51% -75%); + + + +: there are very many changes in 

the microanatomical structure (76% -100%). 

Gill cellular conditions experiencing edema (Figure 

2B), visible basement membrane began to stretch out, the 

field narrowing Lacuna cell deficiency causes gill function 

and difficulty in breathing process, so that the metabolism 

of the body began to fail. Edema is swelling of the cell or 

excessive accumulation of fluid in body tissues (Laksman 

2003). The presence of edema can cause fusion of 

secondary lamella of the lamella. In this study the 

occurrence of edema caused by the influx of Cd into the 

gills of A. woodiana resultied in the cell cell irritating so 

that the cell would swell. 

The process of entry of Cd into the gills by Palar 

(1994), together with other metal ions and the food that has 

been accumulated Cd, and will form ions that can dissolve 

in fat. Ions were able to penetrate the gill cell membrane, 

so it can get into the gills, and then there will be a process 

of loss of volume regulation in the cell. In this treatment 

were also seen pillar cells began to separate from the 

bottom of epithelial cells (middle lamella). When 

experiencing edema,l Cd accumulation in gills occurred at 

the accumulation of 0.5111 ppm. 

Gills in Figure 2C has undergone thorough hyperplasia 

and the fusion is taking place in two parts of the middle 

lamella, with a marked by the epithelial cells started to 

scarp, accompanied by the loss widened Lacuna red blood 

cells and pillar cells apart. Laksman (2003) says that 

hyperplasia is a process of formation of excessive tissue 

due to the increase in cell volume. Hyperplasia caused by 

excessive edema so that red blood cells out of kapilernya 

and separated from the backers. In the event this hyperplasia 

Cd accumulation began at 0.6829 ppm exposure level. 

Condition of cells and gill tissue had fused to lamella 

(Figure 2D), and began to show marked necrosis with 

epithelial cells in each lamella started together with 

epithelial cells on the other lamella, Lacuna also began to 

rupture causing respiratory function failure which affects 

the metabolism of A. woodiana. Secondary lamella fusion 

caused by the swelling in the cells of the gills (edema). The 

occurrence of secondary lamella fusion resulting in 

impaired function of the secondary lamella in the case of 

oxygen-making process and therefore contributes to the 

death of A. woodiana (Susilowati 2005). At a concentration 

of 5 ppm after 30 days A. woodiana experience death. In 

this incident the gills accumulate heavy metals at a 

concentration of 0.9280 ppm. 

At that last stage of a gill would experience the highest 

levels of damage, this damage can lead A. woodiana to 

experience the death of the level of necrosis and atrophy. 

Condition of cells and gill tissue necrosis and atrophy 

experienced (Figure 2E), characterized by the merging of 

each cell in lamella and lamella with bone loss starting 

institutions. Atrophy is a reduction (shrinking) the size of a 

cell, tissue, organ or body part (Harjono 1996). In this 

study occurred atrophy in primary lamella. Atrophy occurs 

due to experimental animals exposed to cadmium at high 

concentrations and in a long exposure time. Cells in 

primary lamella shrinkage (atrophy). 

Laksman (2003) states that the necrosis is cell’s death 

that occurrs due to hyperplasia and excessive fusion of 

secondary lamella, so that the gill tissue is no longer intact 

form or in other words necrosis occurs accompanied with 

the death of a biota. In the event necrosis and atrophy of 

the accumulated Cd in the gills of A. woodiana started at 

2.1279 ppm exposure and atrophy starting at the level of 

accumulation of 2.337 ppm. 

A B C D E 

Figure 2. Structural changes in gill cells A. woodiana. Note: A. Tues normal gills, B. Tues gill edema, C. Tues gill hyperplasia, D. Tues 

fusion gill lamella, E. Tues gill necrosis.

34 

3 (1): 28-35, March 2011 

Microanatomical changes in kidney of A. woodiana after 

exposure to Cd 

Changes in the microanatomical structure of the kidney 

of A. woodiana after administration of Cd are shown in 

Table 4. From this table, it is known that changes in 

cellular structure mikroanatomi kidneys began to occur at a 

concentration of 0.5 ppm for 7 days, edema of the tubules 

begin to appear and be perfect edema at 30 days and began 

to show more than 25% hyperplasia. Perfect hyperplasia is 

shown at a concentration of 1 ppm after 14 days of 

inspection, then the fusion epithelium of the kidney evenly 

shown at concentrations of 5 ppm after 30 days of inspection. 

Table 4. Changes in cellular structure of kidney mikroanatomi A. 

woodiana after exposure to heavy metals cadmium with 

haematoxylin-eosin staining preparation. 

Concentration 

(ppm) 

Time of 

surgery 

(Days) 

Edema Hyperplasia 

Fusion 

of 

lamella 

Necrosis 

0 7 - - - - 

14 - - - - 

30 - - - - 

0.5 7 + - - - 

14 +++ - - - 

30 ++++ ++ - - 

1 7 +++ + - - 

14 ++++ ++++ ++ - 

30 ++++ ++++ +++ - 

5 7 ++++ +++ ++ - 

14 ++++ ++++ +++ + 

(dead) 30 ++++ ++++ ++++ +++ 

10 7 ++++ +++ +++ - 

14 ++++ ++++ ++++ +++ 

(dead) 30 ++++ ++++ ++++ ++++ 

Note: same as Table 9. 

The kidney cells have shown complete necrosis at a 

concentration of 10 ppm after 30 days, while the 

concentration of 5 ppm to 10 ppm starting from day 14 and 

day of the 30th state of A. woodiana have been many who 

experienced the death (LC50), so the kidneys and gills 

partially preserved in a freezer with a temperature of -4 ° C 

for further examination. In Figure 2A, are shown in the 

picture kidney that is still in normal circumstances from the 

control A. woodiana. 

Situation normal kidney cells and tissues in the control 

A. woodiana or the treatment of 0 ppm (Figure 3A), visible 

layer between cells in glomeruli and tubules and blood 

cells are still visible above and below normal. The metal 

accumulation in the kidney ranged from 0.0095 to 0.0242 

ppm. Cd pollution levels, according to FAO (1972) still in 

the normal category below the threshold of fishery water 

quality (1 ppm), so it can be said that the content of Cd in 

the kidneys of A. woodiana the control is still normal. State 

accumulation is still at normal levels it may also occur due 

to A. woodiana, located diantrara aduktor posterior kidney, 

heart and pericardium (Suwignyo et al. 2005). Kidney position 

located on the inside and is relatively protected from the 

environment cause the accumulation of Cd is relatively 

small when compared to the accumulation of Cd in the gills. 

In Figure 3B, indicated changes in cell structure that 

has undergone kidney mikroanatomi edema in all parts of 

the tubules to glomeruli (indicated by black color), and 

seems to bleed blood cells due to accumulated Cd logan 

continuously. In clinical edema in kidney cells caused by 

erasifikasi proteins in the renal tubules in the network, so 

that the urine comes out containing excessive protein 

(Anonymous 2008). In these conditions, the accumulation 

of Cd to the kidney began to be exposed at a concentration 

of 0.0200 ppm. 

Then on further changes, where the higher Cd exposure 

then suffered kidney cell hyperplasia (Figure 3C), which is 

marked by the outbreak of the tubules, and the resulting 

mixing of intra cell with extra fluid cell, and then also in 

the glomerulus looks very black, because the glomerulus 

has accumulated more Cd long, which will result in 

epitelnya cells will rupture at any time. Then the blood 

cells were also seen indicating blackish blood has been 

contaminated with Cd. The range of Cd accumulation in 

kidney condition hyperplasia began to occur on exposure of 

0.0849 ppm. 

Highest level of damage to the kidney, necrosis of 

kidney cells have shown in Figure 3D, which has entered 

the stage of renal cell necrosis seen any broken tubules, 

glomeruli also broken so that mixed the cells with extra 

fluid cells, and whole blood cells were blackened due to 

Acute accumulation of Cd. 

The content of Cd in kidney like this occured at the 

exposure of 0.0786 ppm. 

According to Atdjas (2008) accumulated Cd at the highest 

level will cause some kidney disorder that is poisoning the 

nephrons of the kidney (nephrotoxicity), proteinuria or 

protein in the form contained in the urine, diabetes where 

there is the content of glucose in the urine (glikosuria), and 

aminoasidiuria or amino acid content in the urine 

accompanied by a decline in kidney filtration rate glumerolus. 

A B C D 

Figure 3. Structural changes in renal cell woodiana. Notes: A. Tues normal gills, B. Tues gill edema, C. Tues fusion gill lamella, D. 

Tues gill necrosis


CONCLUSION 

There is significant heavy metal accumulation of Cd in 

each treatment against the gill and kidney A. woodiana as 

evidenced by the Anova test data for 475.3> 60150.3 0.000 

and> 0.000 with an average significance level of 5% (P 

Vol. 3, No. 1, Pp. 36-43 

March 2011 



Site suitability to tourist use or management programs South Marsa 

Alam, Red Sea, Egypt 

MOHAMMED SHOKRY AHMED AMMAR 1,♥ , MOHAMMED HASSANEIN 2 , 

HASHEM ABBAS MADKOUR 1 , AMRO ABD-ELHAMID ABD-ELGAWAD 2 

1 National Institute of Oceanography and Fisheries (NIOF), Suez, P.O. Box 182, Egypt. Tel. (Inst.) 0020 62 3360015. Fax. (Inst.) 0020 62 3360016. ♥ 

Email: shokry_1@yahoo.com 

2 

Tourism Development Authority, Cairo, Egypt 

Manuscript received: 3 February 2011. Revision accepted: 3 March 2011. 

Abstract. Ammar MSA, Hassanein M, Madkour HA, Abd-Elgawad AE. 2011. Site suitability to tourist use or management programs 

South Marsa Alam, Red Sea, Egypt. Nusantara Bioscience 3: 36-43. Twenty sites in the southern Egyptian Red Sea (Marsa Alam-Ras 

Banas sector) were surveyed principally for sensitivity significance throughout the periode 2002-2003. Sensitivity of the study area was 

derived from internationally known criteria, the key words of each criterion and a brief description of its use was described. The present 

study assigned for the first time a numerical total environmental significance score that gives a full sensitivity significance evaluation for 

any site to decide to select either for tourist use or management purposes. However, the results of the study still have the availability to 

arrange sites with respect to one criterion or only two or many of the used criteria whichever needed. Sites selected for protection are 

categorized as belonging to the following protected area categories: sites 7, 10 (category vi), site 18 (category ib), site 5 (category iv), 

sites 16, 17 (category ii). Sites selected for tourist uses are suggested to be classified into 2 categories: first category sites (sites 1, 3, 8, 

11, 13, 15) which are recommended as tourist use sites with management of the sensitive resources beside non consumptive recreational 

activities like swimming, diving, boating, surfing, wind-surfing, jet skiing, bird watching, snorkelling, etc.; second category sites (sites 

2, 4, 6, 9, 12, 14, 19, 20) which are recommended as tourist use sites with both non consumptive and managed consumptive recreational 

activities like fishing. 

Key words: sensitivity significance, selection criteria, tourist use, management programs, Marsa Alam, Red Sea, Egypt 

Abstrak. Ammar MSA, Hassanein M, Abd-Elmegid AE. 2011. Kesesuaian untuk lokasi wisata atau program manajemen Marsa Alam 

Selatan, Laut Merah, Mesir. Nusantara Bioscience 3: 36-43. Dua puluh situs di Laut Merah bagian selatan Mesir (sektor Marsa Alam- 

Ras Banas) disurvei terutama untuk signifikansi sensitivitas sepanjang periode 2002-2003. Sensitivitas suatu daerah penelitian 

merupakan kriteria yang dikenal secara internasional, kata kunci setiap kriteria dan deskripsi singkat tentang penggunaannya dijelaskan. 

Penelitian ini dilakukan untuk pertama kalinya berupa skor nilai total signifikansi lingkungan yang memberikan arti evaluasi sensitivitas 

penuh untuk situs apapun untuk memutuskan memilih baik untuk tujuan wisata atau tujuan pengelolaan lainnya. Namun, hasil penelitian 

ini masih memiliki ketersediaan untuk mengatur situs-situs yang berkaitan dengan satu kriteria, hanya dua atau banyak dari kriteria yang 

digunakan mana yang diperlukan. Situs dipilih untuk perlindungan dikategorikan sebagai milik kategori kawasan lindung sebagai 

berikut: situs 7, 10 (kategori vi), situs 18 (kategori ib), situs 5 (kategori iv), situs 16, 17 (kategori ii). Situs yang dipilih untuk keperluan 

wisatawan disarankan harus diklasifikasikan menjadi dua kategori: situs kategori pertama (situs 1, 3, 8, 11, 13, 15) yang 

direkomendasikan sebagai situs menggunakan wisata dengan manajemen sumber daya sensitif di samping kegiatan rekreasi non 

konsumtif seperti berenang, menyelam, berperahu, berselancar, wind-surfing, jet ski, mengamati burung, snorkeling, dan lain-lain; situs 

kategori kedua (situs 2, 4, 6, 9, 12, 14, 19, 20) yang direkomendasikan sebagai tempat wisata baik kegiatan non konsumtif atau kegiatan 

rekreasi non konsumtif yang dikelola seperti memancing. 

Kata kunci: signifikansi sensitivitas, kriteria seleksi, kegunaan wisata, program manajemen, Marsa Alam Selatan, Laut Merah 

INTRODUCTION 

South Marsa Alam’s diverse coastal and marine 

environments are valuable community resource which may 

be good sites providing recreation and pleasure for visitors 

and tourists or scientific materials for scientists to do 

monitoring and conservation programs. There is no getting 

around the fact that tourism is huge, already categorized as 

the world’s largest industry and will continue to be the 

dominant developing force in the 21 st century (Hill 1998). 

As environmental conservation and protection is critically 

important in some sites, sustainable tourism is critically 

important as well since it may provide source of finance for 

parks and conservation, serve as an economic justification 

for park protection, offer local people economically sound 

and sustainable alternatives to natural resource depletion or 

destruction, promote conservation and build support with 

commercial constituencies (Hawkins 1998). 

Tourist uses includes a diversity of activities that take 

place in both coastal zone and coastal waters (Watson et al. 

2000), which involve the development of tourism 

capacities (hotels, resorts, second homes, restaurants, etc.)

AMMAR et al. – Tourist and management of South Marsa Alam, Egypt 37 

and support infrastructures (ports, marinas, fishing, diving 

shops and other facilities). Coastal recreation activities 

include two main types: consumptive and non-consumptive 

ones: Activities such as fishing, shell fishing and shell 

collection, etc. belong to the consumptive recreational uses 

while the non consumptive activities include swimming, 

diving, boating, surfing, wind-surfing, jet skiing, bird 

watching, snorkelling, etc (Porter and Bright 2003). Tourist 

uses is based on a unique resource combination at the 

interface of land and sea offering amenities such as water, 

beaches, scenic beauty, rich terrestrial and marine 

biodiversity, diversified cultural and historic heritage, 

healthy food and good infrastructure. 

Management programs are the programs that are used 

for preserving an area to provide lasting protection for part 

or all of the natural marine environments therein. IUCN 

(1994) defined the protected area as an area of land and/or 

sea especially dedicated to the protection and maintenance 

of biological diversity, and of natural and associated 

cultural resources, and managed through legal or other 

effective means. To help improve understanding and 

promote awareness of protected area purposes, IUCN has 

developed a six category system of protected areas 

identified by their primary management objective (IUCN 

1994) as follows: 

I. Strict Nature Reserve/Wilderness Area: Protected area 

managed mainly for science or wilderness protection. 

Ia. Strict Nature Reserve: Protected area managed mainly 

for science. 

Ib. Wilderness Area: Protected area managed mainly for 

wilderness protection. 

II. National Park: Protected area managed mainly for 

ecosystem protection and recreation. 

III. Natural Monument: Protected area managed mainly for 

conservation of specific natural features. 

IV. Habitat/Species Management Area: Protected area 

managed mainly for conservation through management 

intervention. 

V. Protected Landscape/Seascape: Protected area managed 

mainly for landscape/seascape conservation and 

recreation. 

VI. Managed Resource Protected Area: Protected area 

managed mainly for the sustainable use of natural 

ecosystem 

Carrying capacity is important to discuss on dealing 

with coastal sustainable tourism. The term "carrying 

capacity" is the number of organisms the resources of a 

given area can support over a given time period (MPA 

NEWS 2004). Adapted to tourism management, it has a 

similar meaning: the number of people who can use a given 

area without an unacceptable alteration in the physical 

environment. Carrying capacity can differ from site to site. 

Dixon et al. (1994), on analyzing coral cover, they 

estimated that the diver carrying capacity threshold for the 

Bonaire Marine Park is between 4000 and 6000 dives per 

site per year. Surveying the percent of damaged coral 

colonies in the Red Sea Ras Mohammed National Park, 

Hawkins and Roberts (1997) suggest 5000 to 6000 dives 

per site per year in the absence of a site specific data. 

Sampling a suite of invertebrates (hard corals, soft corals, 

sea fans, branching hydrocorals, and erect sponges), 

Chadwick-Furman (1996) found the threshold for diving 

sites in the US Virgin Islands to be only 500 dives per site 

per year and attributed this significantly lower estimate to 

the fragility of the various reef organisms in the study area. 

However, effective diver education programs can allow 

coral reef managers to increase carrying capacities (Medio 

et al. 1997). Mooring buoys and the management of the 

number of vessels using mooring buoys with respect to 

time and location are other effective tools coral reef 

managers use in reducing the anchor and diver damage to 

coral reefs. 

The purpose of this study is not to replace existing 

criteria with a new set, but to use existing frameworks for 

site selection to classify south Marsa Alam sites either for 

tourist use or management programs in order to assign sites 

either to EEAA (Egyptian Environmental Affairs Agency) 

for management purposes or to TDA (Tourism 

Development Authority) for tourist uses. It is also aimed to 

develop a total numerical value of sensitivity significance 

(by scoring and summing techniques) that can be used for 

site selection (tourist use or management programs), then 

using single criteria scoring for a particular management or 

tourist use. 


Twenty sites in the southern Egyptian Red Sea 

(between Marsa Alam and Ras Banas) were surveyed 

principally for sensitivity significance. The survey was 

conducted throughout the periode 2002-2003. The sites 

were determined by fixing more or less equal distances 

between them, however determining the position of sites 

was done during the preliminary survey. The area of study 

is shown in Figure 1. 

The ecological survey was performed using Scuba 

diving. For corals and other benthic fauna and flora, the 

transect line method applied by Rogers et al. (1983) was 

used by using a 30 m long tape for surveying the percent 

cover. The intercepted lengths of every individual coral and 

any other benthic organism or habitat were measured; these 

lengths are then used to calculate the percent cover using 

the formula: 

% cover = (intercepted length/transect length) * 100 

Three transects were used per depth zone and the 

average was calculated for all transects. 

For fishes, the stationary fish census applied by 

Bohnsack and Bannerot (1986) was used by using a 50 m 

long transect for the survey. Transects were laid parallel to 

the shore at 4 m depth in the deep reefs or just above the 

reef patch in case of the patchy reefs. The survey was 

basically done at 4m depth since it is the area of maximum 

fish abundance. 

Sensitivity significance of the study area is derived 

from internationally known criteria, however the key words 

of each criterion and a brief description of its use can be 

described as follow: 

Diversity (Ratcliffe 1977; IEEM 2006): large numbers 

of species, particularly when represented by large popu-

38 

3 (1): 36-43, March 2011 

lations are to be valued. A high species diversity is usually 

also reflected by a high diversity of different communities 

which show variation in environmental conditions. 

Rarity (Tubbs and Blackwood 1971; Wittig et al. 1983; 

Edwards-Jones et al. 2000): Applied to habitats or species 

where areas are limited, population numbers low or the 

habitat or species limited in distribution. 

Fragility (Ratcliffe 1977; IEEM 2006): Habitats or 

species vulnerable to disturbance and loss because of small 

area, low population or reliance on a single key resource. 

Ecological functions (IEEM 2006): Loss of ecological 

function of the physical conditions can be measured by 

calculating the area of vegetation that is removed or the 

area of nearshore habitat that is covered by the pier 

structure. 

Typicalness (Fandiño 1996; Edwards-Jones et al. 

2000): A measure of how well a site reflects all the habitats 

that are expected to occur in that geographical region.The 

more representative a site is of a region, the better. 

Naturalness (Ratcliffe 1977; IEEM 2006): Habitats 

largely unmodified by human activity (e.g. salt marsh, 

blanket bog). 

Scientific value (Wright 1977, Edwards-Jones et al. 

2000): The degree of interest of a natural area in terms of 

current or potential research. It may also be related to the 

extent to which a site has been used for past research. Sites 

with good histories (e.g., description of ecosystems’ 

dynamics in the past 50 years) are more valuable to science 

because they enhance our understanding of ecology 

Environmental significance (IEEM 2006): 

Significance of the site to the environment where that 

significance is global, natural or local 

Scenic value (Ratcliffe 1977): The combination of 

landforms and habitats is identified as having high scenic 

value in the context of surrounding landscape 

Size (Ratcliffe 1977; IEEM 2006): In general, nature 

conservation value increases with size. Large sites in 

general contain more species and larger populations of 

animals and plants than small ones. Chance extinction of 

species, either as a result of natural or man-made factors, is 

reduced if a species is present in large numbers. 

Estimating sensitivity significance (developed by the 

author) 

An optimal sensitivity score (the optimal score) was 

supposed for each criterion; this was the score at which the 

site could be optimal. In addition, an estimated score was 

assigned to each crierion depending on how much the site 

meets the conditions of the optimal score, then all 

sensitivity scores for each site were summed to get the total 

sensitivity significance. Methods of how values have been 

assigned to each site per each criterion is described as 

follows (developed by the author) (Table 1). 

Diversity: Diversity value of 1 (according to Shannon- 

Wiener 1948 formula) was assigned a sensitivity 

significance score of 5, so .diverrsity value of 1.2 = 

estimated score of 6 (1.2*5) and so on. 

Rarity: Each 1% of rare biota, relative to the total 

abundance, was assigned a sensitivity significance score of 

10, so 0.2% rare biota or habitats = an estimated score of 2 

(0.2*10) and so on. 

Fragility: Each 1% fragile habitats (nesting, feeding, 

breeding), relative to the total cover, was given an optimal 

score of 10, so each 0.3% fragile habitats = an estimated 

score of 3 (0.3*10) and so on. 

Ecological function: Each 6.66% vital ecological 

function (vegetation or habitats not removed by physical 

conditions) was assigned a score of 1 (6.66/6.66), thus a 

vital ecological function of 26.64% will have an estimated 

score of 26.64/6.66 = an estimated score of 4 and so on. 

Typicalness: A site representing 80% of the number of 

the characteristic ecosystems of a geographical area was 

assigned a score of 10% (80/8), thus a site having 24% 

characteristic ecosystems will have an estimated score of 

24/8=3% and so on. 

Naturalness: A 10% virgin area (with no human 

caused alteration) was assigned a score of 1 (=10/10), thus 

a 30% virgin area has an estimated score of 30/10=3 and a 

virgin area of 50% has an estimated score of 50/10=5 and 

so on. 

Scientific value: A site used for scientific research for 

the past 10 years was assigned a score of 1 (=10/10), thus a 

site used for the past 30 years will have an estimated score 

of 3 (=30/10), a site used for the past 50 years will have an 

estimated score of 5 (=50/10) and so on. 

Environmental significance: Global significance was 

assigned a score of 3, each of national and local 

significance was given a score of 1. 

Scenic value: Scenic value of the landscape depends on 

the value of the following dimensions: 1-visual dimension 

2-geology 3-topography 4-soils 5-ecology 6-landscape 

history 7-Anthropology 8-architecture 9-culture 

associations 10-public places. A site that fulfil the scenic 

value with respect to those 10 items was assigned a score 

of 5 (=10/2), thus a site that fulfil 4 items will have an 

estimated score of 4/2=2, a site that fulfil 2 items will have 

an estimated score of 2/2=1 and so on. 

Size: Each 5000m 2 habitats was assigned a score of 1 

(=5000/5000), so a size of 10000m 2 will have an estimated 

score of 2 (10000/5000) and so on. 

List of sites and their positions 

Site 1. Marsa Nakry: 24 o 55`35.476”N, 34 o 57`40.993”E 

Site 2. between Marsa Nakry and Gabal Dorry: 

24 o 54`36.428”N, 34 o 58`25.453”E 

Site 3. 1 km south of Gabal Dorry: 24 o 47`33.942”N, 34 o 59` 

14.139”E 

Site 4. South Host Mark: 24 o 47`33.942”N, 35 o 01`58.197”E 

Site 5. Northern Sharmel Fokairy: 

Transect 1: 24 o 45`16.192”N, 35 o 03`55.792”E 

Transect 2: 24 o 45`22.126”N, 35 o 03`50.218” E 

Site 6. Southern Sharmel Fokairy: 24 o 38`20”N, 35 o 04`51” E 

Site 7. Sha’b North Ras Baghdadi:24 o 40`25``N,35 o 05`38``E 

Site 8. Northern Ras Baghdadi: 24 o 40`05.900”N, 

35 o 05`52.625”E 

Site 9. Southern Ras Baghdadi: 24 o 39`16.800”N, 

35 o 05`54.200”E 

Site 10. North Sharmel Loly: 24 o 36`50.460”N, 

35 o 06`59.248”E


Site 11. Southern Sharmel Loly: 24 o 36`39.2666”N, 

35 o 07`08.795”E 

Site 12. North Hankourab: 24 o 34`49.624”N, 35 o 08`40.185”E 

Site13. South Hankourab: 24 o 33`23.20”N, 35 o 09`02.405”E 

Site 14. North Ummel Abas: 24 o 30`44.200”N, 

35 o 08`16.927”E 

Site 15. Middle Ummel Abas: 24 o 30`46.024”N, 

35 o 08`16.300”E 

Site 16. South Ummel Abas: 24 o 30`24.642”N, 

35 o 08`31.717”E 

Site 17. Wadi El-Mahara: 24 o 24`27.674”N, 35 o 13` 41.471” E 

Site 18. a mangroove area: 24 o 16`32.400”N, 35 o 3`15.815”E 

Site 19. South Hamata city: 24 o 16` 32.400” N, 35 o 23` 

15.815” E 

Site 20: Lahmy; South El-Gharabawy: 24 o 12`09.494”N, 

35 o 25`37.744”E 


Site priorities for management and protection 

Dealing with the total assigned value of sensitivity 

significance and considering sensitivity significance score 

≥ 50 to be suitable for management purposes, the following 

site priorities are suggested for management purposes: sites 

10, 7, 18, 17, 5 and 16 having significant scores of 86, 77, 

73, 61, 57 and 54 respectively. However, dealing with each 

criterion separately, site 10 has first priority for managing 

diversity and rarity; site 18 for fragility; sites 7, 10, 18 for 

ecological functions, scientific value, and environmental 

significance; sites 7, 10 for typicalness and size; site 10 for 

naturalness; site 18 for scenic value. Moreover, if we used 

many few of the used criteria, we'll have different site 

priorities according to the criteria selected for comparison. 

1 

2 3 

4 5 

67 8 9 

10 

11 

12 

13 

14 

15 

16 

17 

18 

19 

20 

Figure 1. Location of the study area South Marsa Alam (from Marsa Alam to Ras Banas) on the Egyptian Red Sea.

40 

3 (1): 36-43, March 2011 

Also, choosing a higher number of criteria used for 

comparison gives rise to shefting the priority into the site 

that is appropriate to most of the used criteria. 

Site priorities for tourist uses 

Acoording to the total assigned sensitivity 

significance and considering a sensitivity 

significance score < 50 to be suitable for 

touristic use, 16 sites were selected are 

suggested for touristc uses. As many of these 

sites have some sensitive resources, these sites 

are suggested to be divided into two categories: 

first category sites includes sites with some 

sensitive resources and with 30 ≤ sensitivity 

significance < 50, second category sites include 

sites without sensitive resources and with 

sensitivity significance < 30. First category sites 

are sites 1, 3, 8, 11, 13, 15. Second category 

sites are sites 2, 4, 6, 9, 12, 14, 19, 20. Sensitive 

habitats for first category sites are rarity for site 

1, diversity and rarity for site 3, fragility for site 

8, diversity for site 11 and typicalness for site 

13. 

Site description 

Site 1 (Marsa Nakry) is characterized by a 

95% degraded reef flat, 38% dead corals with 

increase in the hydrocoral Millepora dichotoma 

at 1-5m zone. However, one threatened manta 

ray (Taenura lymma) was observed. Site 2 is 

very poor and far from the shore with increased 

algae, sands and dead corals. Site 3 has 45% 

dead corals and one threatened stingray 

(Taenura lymma). Site 4 is dominated with 

algae while site 5 is characterized by two main 

ecosystems: a coral reef ecosystem and a 

seagrass ecosystem, 38.5% live corals, 66.5% 

dead corals and one threatened threatened 

Manta Ray (Himantura uarnak). Site 6 has the 

shoreline heavily condensed with quite a lot 

amount of plastic bags, glass and plastic bottles, 

wood pieces, steel pieces, robes, old shoes, 

small and big canes with a very poor marine 

life. Site 7 has 65% live corals, 14% dead corals 

and one endangered reptile, (the green turtle 

Chelonia mydas. Site 8 has 20% live corals and 

55% dead corals, site 9 has 46.5% live corals 

and 50% dead corals while site 10 has 91% live 

corals and 5% dead corals. Sites 11 and 12 have 

a poor marine life except some algae and spots 

of corals while site 13 is a clean sandy beach 

with few small coral patches having 47% dead 

corals and 35% live corals. Site 14 is mostly 

sand with few patches of algae, sea grasses and 

a well developed reef. Site 15 has a seagrass 

patch, a small reef patch suffering from old 

dynamite fishing and a lot of deep crab niches 

on the shoreline while site 16 has a fringing 

reef, a patch reef and a barrier reef with a 

seagrass bed in between. Site 17 has a a 

degraded reef flat, with 40% live corals, 30% dead corals 

with juveniles of the threatened organ pipe coral Tubipora 

musica attaching rocks, dead corals and rubble on the reef 

crest, fishes were mostly of large sizes. while site 18 has 

heavily condensed mangrove trees on both land and water 

Table 1. Environmental sensitivity of each of the studied sites 

Zone 

Div Rar Frag EcFu Typ Nat SciVa EnSi SceVa Size Tot 

(15) (15) (15) (15) (10) (10) (5) (5) (5) (5) (100) 

Site 1 1 1 0.3 26.64 24 30 30 4 

5 10 3 4 3 3 3 3 2 2 38 

Site 2 1.2 0.2 0.2 13.32 24 30 20 4 

6 2 2 2 3 3 2 2 2 4 28 

Site 3 1.6 1 0.3 19.98 24 30 20 4 

8 10 3 3 3 3 2 2 2 4 40 

Site 4 0.4 0 0 6.66 0 0 10 4 

2 0 0 1 0 0 1 2 2 1 9 

Site 5 1.2 0.9 1 59.94 64 50 30 4 

6 9 10 9 8 5 3 2 2 3 57 

Site 6 1 0.2 0.2 6.66 0 30 20 2 

5 2 2 1 0 3 2 1 1 2 19 

Site 7 2.4 1 0.9 86.58 80 70 40 6 

12 10 9 13 10 7 4 4 3 5 77 

Site 8 1.4 0 0.9 39.96 48 30 30 4 

7 0 9 6 6 3 3 3 2 3 42 

Site 9 1 0 0 33.3 32 20 10 2 

5 0 0 5 4 2 1 1 1 3 22 

Site 10 3 1.4 1 86.58 80 80 40 6 

15 14 10 13 10 8 4 4 3 5 86 

Site 11 1.4 0.4 0.4 26.64 24 30 30 4 

7 4 4 4 3 3 3 3 2 3 36 

Site 12 0.6 

3 

0 

0 

0 

0 

0 

0 

16 

2 

30 

3 

20 

2 2 

6 

3 3 18 

Site 13 1 0.6 0.5 39.96 40 30 20 4 

5 6 5 6 5 3 2 2 2 2 38 

Site 14 0.6 0.1 0.3 19.98 24 10 10 2 

3 1 3 3 3 1 1 1 1 1 18 

Site 15 1.2 0.3 0.9 59.94 24 30 20 4 

6 3 9 9 3 3 2 2 2 2 41 

Site 16 1.6 0.3 1 79.92 48 20 30 6 

8 3 10 12 6 2 3 3 3 4 54 

Site 17 1.4 

7 

1.3 

13 

0.9 

9 

66.6 

10 

48 

6 

40 

4 

30 

3 3 

4 

2 4 61 

Site 18 2.6 0.4 1.3 86.58 64 60 40 8 

13 4 13 13 8 6 4 4 4 4 73 

Site 19 1.6 

8 

0 

0 

0 

0 

0 

0 

24 

3 

40 

4 

40 

4 2 

4 

2 3 26 

Site 20 0.8 

4 

0 

0 

0 

0 

0 

0 

16 

2 

30 

3 

20 

2 2 

2 

1 2 16 

Note: Div = diversity, Rar = rarity, Frag = fragility, EcFu = Ecological function, Typ = 

typicalness, Nat = naturalness, SciVa = scientific value, EnSi = environmental 

significance, SceVa = scenic value, Tot = total. Site 2 = between Marsa Nakry and 

Dorry. Values in parenthesis are the optimum score for each criterion. Diversity: upper 

value in the table is the Shannon estimate of diversity, lower value is the estimated 

score. Rarity: upper value in the table is the percent rare biota or habitats, lower value 

is the estimated score. Fragility: upper value is the percent fragile habitats, lower value 

is the estimated score. Ecological function: upper value is the percent non removed 

vegetation or habitats, lower value is the estimated score. Typicalness: upper value is 

the percent of characteristic ecosystems of a geographical area, lower value is the 

estimated score. Naturalness: upper value is the percent of area of no human caused 

alteration, lower value is the estimated score. Scientific value: upper value is the 

number of past years the site has been used for scientific researches, lower value is the 

estimated score. Environmental significance: values in the table are the estimated 

scores. Scenic value: upper value is the number of items the site fulfil for scenic value, 

lower value is the estimated score. Size: values in the table are the estimated scores.


beside having seagrasses (10% of the bottom cover). Site 

19 is characterized by a dirty shoreline full of plastic bags, 

robs, bottles and canes with a very poor reef while site 20 

is a typical example of sandy beach having also a trace of 

an old reef completely degraded and buried with sand. 

Approaching a total mathematical sensitivity 

significance score 

Evaluation of sensitivity significance criteria in the 

previous studies dealt just phonetically with each criterion 

separately like for example Ratcliffe (1977), IEEM (2006) 

for evaluation of diversity as high, medium or low, fragility 

as reversible or irreversible, naturalness as virgin, semivirgin 

or altered, size as large, medium or small. Other 

criteria were phonetically evaluated like Tubbs and 

Blackwood for evaluation of rarity; IEEM (2006) for 

ecological functions; Fandiño (1996) and Edwards-Jones et 

al. (2000) for typicalness; Wright (1977) and Edwards- 

Jones et al. (2000) for scientific value; IEEM (2006) for 

environmental significance; Ratcliffe (1977) for scenic 

value. Such phonetic evaluation can only deal with each 

criterion separately making it difficult to compare several 

sites for a group of criteria together, in turn making it 

difficult to arrange those group of sites according to their 

importance with respect to several criteria. The present 

study solved that problem by assigning for the first time a 

numerical score for each criterion (explained in the 

material and methods section), then summing all 

mathematical scores to give a total sensitivity significance 

score. However, the study still has the availability to 

arrange the sites with respect to one criterion or only two or 

many of the used criteria whichever needed according to 

the management purpose. Although Croom and Crosby 

(1998) mentioned that scoring and summing techniques 

was used to minimize the personal bias, he used scoring 

and summing techniques with respect to only one separate 

criterion e.g. rarity. Approaching a total sensitivity 

significance score in the present study is important to select 

a site that is much appropriate with most of the used 

criteria. Salm and Clark (1984) and Ray and Legates 

(1998) expected that extremely complicated scoring and 

summing techniques may seem the most objective and 

defensible way to choose a priority site. They further 

related the reason of using a simple assessment system to 

the fact that it is easier to use, requires fewer resources and 

can be evaluated by a diverse group of individuals with 

varying levels of expertise. 

Site priorities for management purposes 

Since priorities for site selection with respect to a single 

criterion differ from those given on using another criterion 

and from those given on using the total sensitivity 

significance; it is important, after selecting sites for 

management purposes, to use the appropriate criterion for 

selecting the appropriate site for the appropriate 

management. Parkes (1990) favoured the rating of 

individual assets, but differed in how multiple values at a 

site should be reconciled. He suggested that, where a site 

has several assets of varying levels of biological 

significance, the site rating should be based on the value of 

the dominant asset at the site, or the majority of assets at 

the site. Selection criteria can be used to order candidate 

sites according to priority in the selection process (Nilsson 

1998). However, the present study has been directed 

mainly to solve the struggle between EEAA (Egyptian 

Environmental Affairs Agency) and TDA (Tourist 

Development Authority) for attaining as many sites as 

possible to EEAA for management purposes or to TDA for 

tourist uses. Therefore, it was important to think in 

developing a numerical total environmental significance 

score by which we can decide either to assign the site for 

EEAA or for TDA. Latimer (2009) stated that the use of 

precise numerical criteria, or indices for the evaluation of 

size, diversity or rarity could provide a guideline reference 

scale, he further mentioned that professional judgement is 

also important. According to the purpose of the study and 

considering a total sensitivity significance ≥50 to be 

significant and appropriate for assigning the site for 

management (protection) purposes, priorities of site 

selection assigned for management purposes are site 10, 

site 7, site 18, site 17, site 5 and site 16, other sites are 

assigned for touristic uses. 

Categorization, carrying capacity and management 

objectives of sites selected for management purposes 

Although sites 7 and 10 have high sensitivity 

significance with respect to all criteria, they are 

recommended as managed resource protected areas 

(category VI) since they contain fishing communities and 

fishing activities. It is important to sustain fishery resources 

by restricting fishing activities seasonally or temporarily to 

let the areas recover. Areas managed to sustain fisheries are 

very rarely promoted to MPAs, but there are exceptions 

like the fish habitat reserves in Australia. Site 18 having the 

highest sensitivity significance with respect to fragility and 

ecological functions, and being inhabited with mangrove 

trees, is recommended as wilderness area (category Ib) 

which is managed mainly for wilderness protection. Sites 7, 

10 and 18 having fragile habitats should have a diver 

carrying capacity threshold of 500 dives per site per year 

according to Chadwick-Furman (1996). However, site 5 

has considerable sensitivity significance with respect to 

fragility and ecological functions, being inhabited with the 

fragile seagrasses, it is recommended as habitat/species 

management area (protected area, category IV). Similar to 

sites 7, 10, 18; site 5 should have a diver carrying capacity 

of 500 dives per site per year. Sites 16 and 17 though 

having considerable sensitivity significance with respect to 

diversity, rarity, fragility, ecological functions and 

typicalness, they are recommended as national park 

(protected areas, category II) since they have a significant 

size which will increase their diver carrying capacity so as 

to tolerate recreation. According to Dixon et al. (1994) in 

Bonaire Marine Park and Hawkins and Roberts (1997) in 

Ras Mohammed National Park, sites 16 and 17 should have 

a diver carrying capacity of 4000-6000 dives per site per 

year. A matrix of management objectives in the sites 

assigned as protected areas are explained (Table 2) 

according to IUCN (1994).

42 

3 (1): 36-43, March 2011 

Table 2. Management objectives of sites selected for management purposes 

Management objective 

Sites 7, 10 Site 18 Site 5 Sites 16, 17 

Category VI Category Ib Category IV Category II 

Scientific research 3 3 2 2 

Wilderness protection 2 1 3 2 

Preservation of species and genetic diversity (biodiversity) 1 2 1 1 

Maintenance of environmental services 1 1 1 1 

Protection of specific natural / cultural features 3 – 3 2 

Tourism and recreation 3 2 3 1 

Education 3 – 2 2 

Sustainable use of resources from natural ecosystems 1 3 2 3 

Maintenance of cultural/traditional attributes 2 – – – 

Note: 1 = Primary objective; 2 = Secondary objective; 3 = Potentially applicable objective; – = not applicable. 

Site priorities for tourist uses 

Sites classified as first category sites (sites 1, 3, 8, 11, 

13 and 15) are recommended as tourist use sites with 

management of the sensitive resources and non 

consumptive recreational activities like swimming, diving, 

boating, surfing, wind -surfing, jet skiing, bird watching, 

snorkelling, etc. Locations of recreational activities could 

have a carrying capacity of up to 6000 dives per site per 

year (Roberts 1997) while in the sensitive locations, it 

should not exceed 500 dives per site per year (Chadwick- 

Furman 1996). However, effective diver education 

programs can allow coral reef managers to increase 

carrying capacities (Medio et al. 1997), also mooring buoys 

and the management of the number of vessels using 

mooring buoys with respect to time and location are other 

effective tools coral reef managers use in reducing the 

anchor and diver damage to coral reefs. Management of 

sensitive habitats in first category of tourist use sites 

includes protection of rarity for sites 1, diversity and rarity 

for site 3, fragility for site 8, diversity for site 11 and 

typicalness for site 13. Second category sites (sites 2, 4, 6, 

9, 12, 14, 19 and 20) are recommended as tourist use sites 

with non consumptive and managed consumptive 

recreational activities like fishing. Diver carrying capacity 

of these sites could approach 6000 dives per site per year. 

Site 4 having the lowest sensitivity significance and most 

minimum values with respect to every sensitivity criterion 

is suggested to allocate a part of it for building an artificial 

reef to restore the damaged ones (Ammar 2009a). 

Site description 

Damaged reef flat in site 1 is due to the absence of reef 

access points to deep water. Ammar (2009b) indicated the 

importance of reef access points in his assessment of some 

coral reef sites along the Gulf of Aqaba, Egypt. Increased 

algae and sands in site 2 with increased dead corals agree 

with Pearson (1981) and Nezali et al. (1998) that algae are 

among the most important factors which can influence 

coral recolonization. The high percentage cover of the 

hydrocoral Millepora dichotoma at 1-5m depth in Marsa 

Nakry as well as in other sites having that species, agrees 

with the finding of Ammar (2004) that, Millepora sp. (a 

hydrocoral) prefers high illumination and has a strong 

skeletal density to tolerate strong waves. The relatively low 

sensitivity significance in spite of the presence of the 

threatened species (the blue spotted stingray Taenura 

lymma) in sites 1 and 3, indicates the importance of using a 

particular criterion when dealing with a particular 

management purpose. The green turtle Chelonia mydas 

found in site 7 is categorized as a taxon having an 

observed, estimated, inferred or suspected reduction of at 

least 80% over the last 10 years or three generations, 

whichever is the longer (IUCN 2002). The lower recorded 

amount of dead corals in site 10 (Sharm El Loly) though it 

is highly used by fishing boats, is due to the fact that these 

boats anchor on the inlet terminal, away from the reef and 

go to open water through the middle of the inlet. Reporting 

juveniles of the vulnerable organ pipe coral Tubipora 

musica in site 17 (Wadi El-Mahara) is the reason of 

increased sensitivity significance with respect to rarity in 

that site. Ammar (2005) categorized the organ pipe coral 

Tubipora musica as vulnerable according to IUCN (2001), 

as there is an estimated population size reduction of ≥ 50% 

over the last 10 years, based on the index of abundance and 

the decline in area of occupancy. Site 18 having a 

mangrove ecosystem, a seagrass ecosystem and a coral reef 

ecosystem integrating together helped to increase most of 

the selection criteria, in turn increasing the overall 

sensitivity significance. Broody (1998) stated that selection 

criteria help to provide a rational basis for choosing among 

potential sites. 

CONCLUSIONS 

The present study approached for the first time a 

numerical total sensitivity significance score for each site 

to select a site that is much appropriate with most of the 

used criteria. This is important to classify a group of sites 

to be suitable either for tourist use or management 

purposes. Since priorities for site selection differ from one 

sensitivity criterion to the other and from the total 

sensitivity significance, it is important, after selecting a site 

for management (using the total sensitivity significance), to 

specify the appropriate criterion for deciding the 

appropriate management purpose per site. Sites selected for 

management (protection) purposes are categorized as 

belonging to the following protected area categories: sites 

7, 10 (category vi), site 18 (category ib), site 5 (category 

iv), sites 16, 17 (category ii). Sites selected for tourist uses 

are classified into 2 categories: 1- First category sites (sites


1, 3, 8, 11, 13, 15) which are recommended as tourist use 

sites with management of the sensitive resources and non 

consumptive recreational activities like swimming, diving, 

boating, surfing, wind -surfing, jet skiing, bird watching, 

snorkelling, etc. 2- Second category sites (sites 2, 4, 6, 9, 

12, 14, 19, 20) which are recommended as tourist use sites 

with non consumptive and managed consumptive 

recreational activities like fishing. 

REFERENCES 

Ammar MSA. 2004. Zonation of coral communities and environmental 

sensitivity offshore a resort site at Marsa Alam, Red Sea, Egypt. 

Egypt J Zool 42: 67-18. 

Ammar MSA. 2005. An alarming threat to the red organ pipe coral 

Tubipora musica and suggested solutions. Ecol Res 20: 529-535. 

Ammar MSA. 2009a. Coral reef restoration and artificial reef 

management, future and economic. Open Environ Eng J 2: 37-49. 

Ammar MSA. 2009b. Assessment of present status and future needs of 

four coral reef sites along the Gulf of Aqaba, Egypt. Open Environ 

Poll Toxicol J 1: 34-42. 

Bohnsack JA, Bannerot SP. 1986. A stationary visual census technique for 

quantitatively assessing structure of coral reef fishes. NOAA Tech 

Rep NMFS 41: 1-15. 

Broody SD. 1998. Evaluating the role of site selection criteria for marine 

protected areas in the Gulf of Maine. Report 2, Gulf of Maine Marine 

Protected Area Project. Maine, USA. 

Chadwick-Furman NE. 1996. Effects of scuba diving on coral reef 

invertebrates in the US Virgin Islands: implications for the 

management of diving tourism. Proc. 6th Intl Coelenterate Biol Conf. 

Amsterdam, The Netherland. 

Croom M, Crosby M. 1998. Description of dimensionless analyses and 

delphic priority ranking methodologies for selecting marine and 

coastal protected areas. In: Crosby MP, Laffoley D, Mandor C, 

O’Sullivan G, Geenen K (eds). Proceedings of the Second 

International Symposium and Workshop on Marine and Coastal 

Protected Areas: Integrating Science and Management. Silver Spring, 

MD: Office of Ocean and Coastal Resource Management, National 

Oceanic and Atmospheric Administration. 

Dixon JA, Scura LF, van't Hof T. 1994. Ecology and microeconomics as 

"joint products'': the Bonaire Marine Park in the Caribbean. In: 

Perrings CA,. Mäler KG, Folke C, Holling CS, Jansson BO(eds) 

(eds) Biodiversity conservation: problems and politics. Kluwer. 

Dordrecht. 

Edwards-Jones G, Davies B, Hussain S. 2000. Ecological economics; an 

introduction. Blackwell. Cornwall, UK. 

Fandiño M. 1996. Framework for ecological evaluation. [PhD 

Dissertation]. University of Amsterdam. ITC publication 45. 

Enschede. 

Hawkins DE. 1998. The relationship of tourism-related revenue 

generation to coral reef conservation. In: Hatziolos ME, Hooten AJ, 

Fodor M (eds). Coral reefs: challenges and opportunities for 

sustainable management The International Bank for Reconstruction 

and Development/The World Bank. New York. 

Hawkins JP, Roberts CM. 1997. Estimating the carrying capacity of coral 

reefs. Proc 8th Int Coral Reef Symp 2: 1923-1926. 

Hill C. 1998. Green globe: The tourism industry and sustainability. In: 

Hatziolos ME, Hooten AJ, Fodor M (eds). Coral reefs: challenges and 

opportunities for sustainable management The International Bank for 

Reconstruction and Development/The World Bank. New York. 

IEEM [Institute of Ecology and Environmental Management]. 2006. 

Guidelines for ecological impact assessment in the United Kingdom. 

Institute of Ecology and Environmental Management. Winchester 

IUCN. 1994. Guidelines for Protected Area Management Categories. 

IUCN. Gland, Switzerland. 

IUCN. 2001. The IUCN red list of threatened species 2001. Categories 

and criteria (v. 3.1). In: CITES identification manual. IUCN. Gland, 

Switzerland. 

IUCN. 2002. 2002 IUCN red list of threatened species. IUCN. Gland, 

Switzerland. 

Latimer W. 2009. Assessment of biodiversity at the local scale for 

environmental impact assessment and land-use planning. Planning 

Practice & Research 24(3): 389-408. 

Medio D, Pearson M, Ormond RFG. 1997. Effect of briefings on rates of 

damage to corals by scuba divers. Biol Conserv 79: 91-95. 

MPA NEWS. 2004. Assessing the carrying capacity of marine protected 

areas. MPA NEWS 6 (2): August 2004. 

Nezali LM, Johnstone RW, Mgaya YD. 1998. Factors affecting 

scleractinian coral recruitment on a nearshore reef In Tanzania. 

Ambio 27 (8): 717-722. 

Nilsson P. 1998. Criteria for the selection of marine protected areas - an 

analysis. Report 4834 Research and Development Department, 

Swedish Environmental Protection Agency. Stockholm. 

Oliver JK. 1986. Stony corals: classes hydrozoa and anthozoa. IUCN. 

Gland, Switzerland. 

Parkes D. 1990. Guidelines for the Assessment of Biological Significance 

in Victoria. Department of Conservation and Environment, Flora and 

Fauna Survey and Management Group, Lands and Forests Division. 

East Melbourne. 

Pearson RG. 1981. Recovery and colonization of coral reefs. Mar Ecol 

Prog Ser 4: 105-122 

Porter R, Bright RD. 2003. Non consumptives outdoor recreation, activity 

meaning, and environmental concern. Proceedings of the 2003 

Northeastern Recreation Research Symposium, GTR-NE-317. 

Ratcliffe DA (ed). 1977. A nature conservation review. Cambridge 

University Press. Cambridge. 

Ray C, Legates D. 1998. Range of selection approaches for marine and 

coastal protected areas. In: Crosby MP, Laffoley D, Mandor C, 

O’Sullivan G, Geenen K (eds). Proceedings of the Second 

International Symposium and Workshop on Marine and Coastal 

Protected Areas: Integrating Science and Management. Office of 

Ocean and Coastal Resource Management, National Oceanic and 

Atmospheric Administration. Silver Spring, MD. 

Rogers CS, Gilnack M, Fitz HC. 1983. Monitoring of coral reefs with 

linear transects: a study of storm damage. J Exp Mar Biol Ecol 66: 

285-300. 

Salm R, Clark J. 1984. Marine and coastal protected areas: a guide for 

planners and managers. IUCN. Gland, Switzerland. 

Tubbs CR, Blackwood JW. 1971. Ecological evaluation of land for 

planning purposes. Biol Conserv 3 (3): 169-172. 

Watson AE, Cole DN, Turner DL, Reynolds PS. 2000. Wilderness 

recreation use estimation; a handbook of methods and systems. 

general technical report RMRSGTR-56. U.S. Department of 

Agriculture-Forest Service, Rocky Mountain Research Station. 

Ogden, Utah. 

Wittig R, Armitage MT, Firse MT, Moss D. 1983. A quick method for 

assessing the importance of open spaces in towns for urban nature 

conservation. Biol Conserv 26: 57-64. 

Wright DF. 1977. A site evaluation scheme for use in the assessment of 

potential nature reserves. Biol Conserv 11: 293-305.

Vol. 3, No. 1, Pp.: 44-58 

March 2011 



Review: Natural products from Genus Selaginella (Selaginellaceae) 

AHMAD DWI SETYAWAN ♥ 

Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University, Surakarta 57126. Jl. Ir. Sutami 36A Surakarta 57126, 

Tel./fax. +62-271-663375, email: volatileoils@gmail.com 

Manuscript received: 28 Augustus 2010. Revision accepted: 4 October 2010. 

Abstract. Setyawan AD. 2011. Natural products from Genus Selaginella (Selaginellaceae). Nusantara Bioscience 3: 44-58. Selaginella 

is a potent medicinal-stuff, which contains diverse of natural products such as alkaloid, phenolic (flavonoid), and terpenoid. This species 

is traditionally used to cure several diseases especially for wound, after childbirth, and menstrual disorder. Biflavonoid, a dimeric form 

of flavonoids, is the most valuable natural products of Selaginella, which constituted at least 13 compounds, namely amentoflavone, 

2',8''-biapigenin, delicaflavone, ginkgetin, heveaflavone, hinokiflavone, isocryptomerin, kayaflavone, ochnaflavone, podocarpusflavone 

A, robustaflavone, sumaflavone, and taiwaniaflavone. Ecologically, plants use biflavonoid to response environmental condition such as 

defense against pests, diseases, herbivory, and competitions; while human medically use biflavonoid especially for antioxidant, antiinflammatory, 

and anti carcinogenic. Selaginella also contains valuable disaccharide, namely trehalose that has long been known for 

protecting from desiccation and allows surviving severe environmental stress. The compound has very prospects as molecular stabilizer 

in the industries based bioresources. 

Key words: natural products, biflavonoid, trehalose, Selaginella. 

Abstrak. Setyawan AD. 2011. Bahan alam dari Genus Selaginella (Selaginellaceae). Nusantara Bioscience 3: 44-58. Selaginella adalah 

bahan baku obat yang potensial, yang mengandung beragam metabolit sekunder seperti alkaloid, fenolik (flavonoid), dan terpenoid. 

Spesies ini secara tradisional digunakan untuk menyembuhkan beberapa penyakit terutama untuk luka, nifas, dan gangguan haid. 

Biflavonoid, suatu bentuk dimer dari flavonoid, adalah salah satu produk alam yang paling berharga dari Selaginella, yang meliputi 

sekurang-kurangnya 13 senyawa, yaitu amentoflavone, 2',8''-biapigenin, delicaflavone, ginkgetin, heveaflavone, hinokiflavone, 

isocryptomerin, kayaflavone, ochnaflavone, podocarpusflavone A, robustaflavone, sumaflavone, dan taiwaniaflavone. Secara ekologis, 

tumbuhan menggunakan biflavonoid untuk merespon kondisi lingkungan seperti pertahanan terhadap hama, penyakit, herbivora, dan 

kompetisi, sedangkan manusia menggunakan biflavonoid secara medis terutama untuk antioksidan, anti-inflamasi, dan anti 

karsinogenik. Selaginella juga mengandung trehalosa suatu disakarida yang telah lama dikenal untuk melindungi dari pengeringan dan 

memungkinkan bertahan terhadap tekanan lingkungan hidup yang keras. Senyawa ini sangat berpotensi sebagai stabilizer molekul 

dalam industri berbasis sumberdaya hayati. 

Kata kunci: produk alami, biflavonoid, trehalosa, Selaginella. 

INTRODUCTION 

Medicinal plant is plant containing substance which can 

be used for the medication or become precursor of drug 

synthesis (Sofowora 1982). Medicinal plant has been 

source of human health since ancient time, whereas about 

60-75% of world populations require plant for carrying 

health (Farnsworth 1994; Joy et al. 1998; Harvey 2000). 

Plants and microbes are the main source of natural products 

(Hayashi et al. 1997; Armaka et al. 1999; Lin et al. 

1999a,b; Basso et al. 2005), and consistently become main 

source of the newest drugs (Harvey 2000). The drug 

development from natural sources are based on the 

bioassay-guided isolation of natural products, due to the 

traditional uses of local plants (ethnobotanical and 

ethanopharmacological applications) (Atta-ur-Rahman and 

Choudhary 1999). 

Traditional medication system by using plant medicines 

has been developed during thousands of year especially by 

Chinese (Wu-Hsing) and India (Ayurveda, Unani and 

Siddha) (Peter 2004; Ahmad et al. 2006), while the most 

advanced, widespread and oldest traditional medication 

system in Nusantara or Malay Archipelago (Malesia) is 

jamu which developed by Javanese. Jamu contains several 

recipes that compiled by about 30 plant species. Relief at 

Borobudur temple about making jamu indicates that jamu 

has been widely recognized since early 9 th century (Jansen 

1993). This system has been documented for centuries in 

many serat and primbon, Javanese literary (Soedibjo 1989, 

1990; Sutarjadi 1990); and spreaded by trading, migration 

and expansion of several kingdoms such as Mataram Hindu 

(Sanjaya), Srivijaya (Saylendra) and Majapahit. 

Selaginella Pal. Beauv. (Selaginellaceae Reichb.) has 

been used as complementary and alternative medicines in 

several traditional medication. This matter is traditionally 

used to cure wound, after childbirth, menstrual disorder, 

skin disease, headache, fever, infection of exhalation 

channel, infection of urethra, cirrhosis, cancer, rheumatism, 

bone fracture, etc. Part to be used is entire plant, though 

only referred as leaves or herbs (Setyawan 2009; Setyawan

SETYAWAN – Natural products of Selaginella 45 

and Darusman 2008). The usage can be conducted single or 

combination, fresh or dried, direct eaten or boiled 

(Dalimartha 1999; Wijayakusuma 2004). This plant has 

sweet taste and gives warm effect on the body (Bensky et 

al. 2004). The use of Selaginella as medicinal matter is 

occurred in the entire world. The largest usage is conducted 

by Chinese, especially for S. tamariscina, S. doederleinii, 

S. moellendorffii, S. uncinata, and S. involvens (Lin et al. 

1991; Chang et al. 2000; Wang and Wang 2001). 

Unfortunately, Selaginella is rarely exploited in Nusantara. 

Traditional jamu of Java use more cultivated spices and 

rhizomes than wild herbs or grasses. 

Plant medicinal properties are contributed by natural 

products or secondary metabolites, such as phenolic 

(flavonoid), alkaloid, terpenoid, as well as non protein 

amino acid (Smith 1976). Natural products are chemical 

compounds or substances produced by a living organism 

and found in nature that usually has a biological activity for 

use in pharmaceutical drug discovery and drug design 

(Cutler and Cutler 2000). In this following discourse, the 

authors studied diversity of natural products from 

Selaginella, especially biflavonoid and trehalose 

compounds; and biological activity of Selaginella’s 

bifavonoid in modern medication. 

NATURAL PRODUCTS DIVERSITY 

Previous phytochemical studies on the constituents of 

genus Selaginella leds to the discovery of many 

compounds, including biflavonoids, the main secondary 

metabolite of Selaginella (Sun et al. 1997; Silva et al. 1995; 

Chen et al. 2005b; Lin et al. 1994; 2000). Biflavonoid has 

also distributed to Selaginellales, Psilotales, and 

Gymnosperms (Seigler 1998), several Bryophytes and 

about 15 families of Angiosperms (DNP 1992). The other 

compounds are including lignin (White and Towers 1967); 

lignan (Lin et al. 1994), lignanoside (Lin et al. 1990; Zheng 

et al. 2004, 2008b), alkaloid (Zheng et al. 2004; Lin et al. 

1997), selaginellin (Zhang et al. 2007; Cheng et al. 2008), 

glycosides (Man and Takahashi 2002; Zhu et al. 2008), 

glucosides (Dai et al. 2006; Yuan et al. 2008), C- 

glycosylflavones (Richardson et al. 1989), etc. Selaginella 

species of Java contains alkaloid, phenolic (flavonoid, 

tannin, saponin), and terpenoid (triterpene, steroid) 

(Chikmawati and Miftahudin. 2008; Chikmawati et al. 

2008). Some species of Japan consist of a steroid type 

namely ekdisteroid (Takemoto et al. 1967, Hikino et al. 

1973; Yen et al. 1974). The diversity and content of other 

compound is relatively lower than biflavonoid, 

nevertheless they have also certain bioactivities. 

Water extracts of S. tamariscina also has several natural 

products such as ferulic acid, caffeic acid, vanillic acid, 

syringic acid, umbelliferone (Bi et al. 2004b); 

tamariscinoside A, tamariscinoside B, adenosine, 

guanosine, arbutin (Bi el al. 2004a); tamariscinoside C, 

tyrosine, D-mannitol, and shikimic acid (Zheng et al. 

2004). The EtOH extract of the whole herbs of S. 

tamariscina that fractionated by chloroform and ethyl 

acetate contains selaginellin A and selaginellin B (Cheng et 

al. 2008). The main constituen of S. tamariscina 

subsequently is amentoflavone, robustaflavone, bilobetin, 

hinokiflavone, isocryptomerin and an apigenin-diglucoside 

(Yuan et al. 2008). S. tamariscina has also many sterols 

that inhibit the growth of human leukemia HL-60 cells 

indicating anti cancer property (Gao et al. 2007). The aerial 

parts of S. pulvinata has steroid constituent (Zheng et al. 

2007), and several Selaginella has also sterol (Chiu et al. 

1988). Steroid compound namely ekdisteroid has been 

found in Japanese species of S. deliculata, S. doederleinii, 

S. moellendorffii, S. nipponica, S. involvens (= S. 

pachystachys), S. stauntoniana (= S. pseudo-involvens), S. 

remotifolia var. japonica, S. tamariscina, and S. uncinata 

(Takemoto et al. 1967; Hikino et al. 1973; Yen et al. 1974). 

Methanolic extract of S. lepidophylla contains 3- 

methylenhydroxy-5-methoxy-2,4-dihydroxy tetrahydrofurane, 

which can a slight inhibitory effect on the uterus 

contraction (Perez et al. 1994). S. lepidophylla is also 

reported contain volatile oils (Andrade-Cetto and Heinrich 

2005). The acetone extract of S. sinensis contains 

selaginellin A, an unusual flavonoid pigment (Zhang et al. 

2007). S. sinensis has a glucoside, namely selaginoside 

(Dai et al. 2006), a sesquilignan, namely sinensiol A (Wang 

et al. 2007), secolignans, namely styraxlignolide D and 

neolloydosin (Feng et al. 2009), and (+)-pinoresinol 

(Umezawa 2003a,b). S. uncinata also has chromone 

glycosides, namely uncinoside A and uncinoside B (Man 

and Takahashi 2002), which shows antiviral activities 

against RSV and PIV-3 (Ma et al. 2003). Ethanol extract 

of S. uncinata also contains flavonoids that possessing a 

benzoic acid substituent (Zheng et al. 2008a). 

S. doederleinii contains several phenolic compounds 

such as (+)-matairesinol, (-)-lirioresinol A, (-)-lirioresinol 

B, (-)-nortracheloside (Lin et al. 1994), and (-)- 

matairesinol, (+)-syringaresinol, (+)-wikstromol, (+)- 

nortrachelogenin (Umezawa 2003a,b). The (-)-matairesinol 

has inhibitory activity against cAMP and acts as an 

insecticide synergist, while (+)-syringaresinol has cytotoxic 

effect (Harborne et al. 1999). S. doederleinii also contains a 

glycosidic hordenine (Markham et al. 1992), which 

increases hypertension (Lin et al. 1991). 

S. caulescens, S. involvens, and S. uncinata contain 

about 0.2% silicon, higher than the most of other club 

mosses and true ferns (Ma and Takahashi 2002), which 

may improve plant tolerant to disease, drought, and metal 

toxicities (Epstein 1999; Richmond and Sussman 2003; Ma 

2004). S. labordei contains 4'-methylether robustaflavone, 

robustaflavone, eriodictyol and amentoflavone (Tan et al 

2009). S. apoda yields substantial amounts of 3-O-methyl- 

D-galactose (Popper et al. 2001). S. moellendorfii contains 

several pyrrolidinoindoline alkaloids (Wang et al. 2009). 

Other natural products, beside biflavonoid and trehalose, 

also have several molecular properties that can increase 

human health and have economical values; and need for 

further observation. 

Natural products of Selaginella can vary depend on 

climate, location, and soil factors; as well as harvesting and 

extraction procedure (Nahrstedt and Butterweck 1997); and 

also plant species or variety, parts to be extracted and age. 

The different species of Selaginella shows different HPLC 

fingerprint characteristic. The samples of the similar

46 

3 (1): 44-58, March 2011 

species, collected in different period, different environment 

or different locations shows certain difference in 

fingerprints. However, it also generate main fingerprint 

peaks, which can be used to evaluate and distinguish the 

different species or infra species (Fan et al. 2007). 

BIFLAVONOID 

Selaginella species have a large number of bioactive 

compounds, the most important being biflavonoids (Silva 

et al. 1995; Lin et al. 1999). Biflavonoids are naturally 

occurring compounds that are ubiquitous in all vascular 

plants and have many favorable biological and 

pharmacological effects (Lee et al. 1996; Baureithel et al. 

1997; Lobstein-Guth et al. 1998). One of flavonoid 

structure that has high medicinal valuable is biflavonoid; a 

dimeric form of flavonoid which formed by binding of two 

flavone units or mixture between flavone and flavanon or 

aurone (Geiger and Quinn 1976; DNP 1992; Ferreira et al. 

2006). 

Flavonoid (or flavanoid) is widespread plant natural 

products (5-10%); its chemical structure and biological role 

are very diverse (Macheix et al. 1990). This compound is 

formed by shikimate and phenylpropanoid pathways 

(Harborne 1989), with a few alternative biosynthesis 

(Robards and Antolovich 1997). Flavonoid is derived from 

phenols having basic structure of phenylbenzopiron 

(tocopherol) (Middleton et al. 2000); distinguished by 15 

carbon skeletons (C6-C3-C6) consisted of one oxygenated 

ring and two aromatic rings (Figure 1). Substitution of 

chemical group at flavonoid is generally hydroxylation, 

methoxylation, methylation and glycosilation (Harborne 

1980). Flavonoid is classified diversely; among them are 

flavone, flavonone, isoflavone, flavanol, flavanon, 

anthocyanin, and chalchone (Porter 1994; Ferreira and 

Bekker 1996; Ferreira et al. 1999a,b). More than 6467 

flavonoid compounds have been identified and amount of 

new discovery is consistently increasing (Harborne and 

Baxter 1999). This compound is playing important role in 

determining color, favor, aroma, and quality of nutritional 

food (Macheix et al. 1990). Flavonoid is mostly monomeric 

form, but there is also dimer (biflavonoid), trimer, tetramer, 

and polymer (Perruchon 2004). 

Biflavonoid (or biflavonil, flavandiol) is a dimeric form 

of flavonoid which formed by bonding of two flavone units 

or mixture between flavone and flavanon or aurone (Geiger 

and Quinn 1976; DNP 1992; Ferreira et al. 2006). Basic 

structure of biflavonoid is 2,3-dihydroapigeninil-(I-3’,II- 

3’)-apigenin (Figure 1.). This compound has interflavanil 

C-C bond between carbon C-3’ at each flavone group. 

There is also some biflavonoid with interflavanil C-O-C 

bonding (Bennie et al. 2000, 2001, 2002; Ferreira et al. 

2006). Locksley (1973) suggest generic term ‘biflavanoid’ 

to replace ‘biflavonil’ which is early used. Term 

‘biflavanoid’ is assumed more accurate than ‘biflavonoid’ 

because indicating saturated in nature. Suffix ‘oid’ 

indicates homogeneous dimeric type, including biflavanon, 

biflavon, biflavan, etc. However, term ‘biflavonoid’ is 

more regularly used because articulated easier. 

Phenol 

C 

Flavonoid 

B 

Biflavonoid 

Figure 1. Basic structure of phenol, flavanoid and biflavanoid. 

Bicyclic ring system is named A and C rings, while unicyclic ring 

is named B ring. The two unit of monomeric biflavonoid is 

marked by Roman number I and II. Position number at each 

monomer is started from containing oxygen atom ring, position of 

C-9 and C-10 indicate unification of them (Rahman et al. 2007; ). 

Biflavonoid is found at fruit, vegetable, and other part 

of plant. This compound is originally found by Furukawa 

in 1929 (Lin et al. 1997) from leaf extract of G. biloba in 

form of yellow colored compound, later named ginkgetin 

(I-4’, I-7-dimetoxy, II-4’, I-5, II-5, II-7-tetrahydroxy I-3’, 

II-8 biflavone) (Baker and Simmonds 1940). Nowdays, 

amount of biflavonoid which isolated and characterized 

from nature continually increase (Oliveira et al. 2002; 

Ariyasena et al. 2004; Chen et al. 2005a), but learning to 

bioactivity is still limited. The most observed biflavonoid is 

ginkgetin, isoginkgetin, amentoflavone, morelloflavone, 

robustaflavone, hinokiflavone, and ochnaflavone. Those 

compounds have similar basic structure, i.e. 5,7,4’- 

trihydroxy flavonoid, but differing at nature and position of 

flavonoid bond (Rahman et al. 2007). 

Biflavonoid has several namenclaturing systems, such 

as Loksley, IUPAC, and vernacular name. The first of two 

systems is the most systematic, but the most used is 

vernacular name. Locksley (1973) standardize 

nomenclature and position number of biflavonil ring 

skeleton. Every monomer unit is marked by Roman 

numerals I and II that indicate bonding between monomer, 

followed by Arabic numerals indicate that bonding 

position. The two numeral from two monomer unit 

compiled dimeric, than paired with hyphen to show 

bonding position of two monomer. Number of substitution 

group at monomer unit follow IUPAC system for flavone. 

In Locksley system, amentoflavone named I-4’, II-4’, I-5, 

II-5, I-7, II-7-hexahydroxy I-3’, II-8 biflavone, while 

hinokiflavone which its flavone unit bonded with an 

oxygen is named by II-4’, I-5, II-5, I-7, II-7-pentahydroxy 

I-4’-O-II-6 biflavone. This system is intuitive, logical, and 

depicts the chemical structure. In IUPAC, amentoflavone is 

named by 8-5-(5,7-dihydroxy-4-oxo-4H-chromen-2-il)-2- 

hydroxyphenyl-5,7-dihydroxy- 2-(4-hydroxy-phenyl)- 

chromen-4-on, while hinokiflavone is 6-4-(5,7-dihydroxy- 

4-oxo-4H-chromen-2-il)-phenoxy- 5,7-dihydroxy-2-(4-


hydroxyphenyl)- chromen-4-on. Basic difference between 

two systems is reference of structural skeleton. Locksley 

use flavanoid structure, while IUPAC use chromen 

structure that more complex (Rahman et al. 2007). The 

above two nomenclature is rarely used because its 

complication. Vernacular name that given by each inventor 

is often used because simpler and easier, though it is not 

systematic and does not depict chemical structure, such as 

amentoflavone, hinokiflavone, ginkgetin, etc. 

In vivo biosynthesis of flavonoid in nature is relatively 

mysterious, but there are some approaches by in vitro to 

explain biosynthesis. According to Rahman et al. (2007) 

there are nine pathways of biflavonoid synthesis, namely: 

(i) Ullmann coupling halogenated flavones; (ii) synthesis of 

biflavones via 1,1’-biphenyls; (iii) metal catalyzed cross 

coupling of flavones; (iv) Wessely-Moser rearrangements; 

(v) phenol oxidative coupling of flavones; (vi) Ullmann 

condensation with flavone salts; (vii) nucleophilic 

substitution; (viii) dehydrogenation of biflavanones into 

biflavones; and (ix) dehydrogenation of biflavone into 

biflavanone. 

In East Asia, biflavonoid is usually produced from leaf 

of Ginkgo biloba which main constituent is ginkgetin 

(Krauze-Baranowska and Wiart 2002; Dubber 2005). In 

sub Sahara-Africa, it is especially produced from seed of 

Garcinia cola which main constituent is kolaviron (Iwu 

and Igboko 1982; Iwu 1985, 1999; Iwu et al. 1987, 1990; 

Braide 1989, 1993; Han et al. 2006; Farombi et al. 2005; 

Adaramoye and Medeiros 2009). The biflavanones are the 

most dominant in the most Garcinia species (Waterman 

and Hussain 1983), pericarp of Javanese mangosteen (G. 

mangoestana) contains amentoflavone and other flavonoids 

(ADS 2008, data not be shown). In Europe, biflavonoid is 

commonly produced from herbs of Hypericum perforatum 

which main constituent is amentoflavone (Berghofer and 

Holzl 1987, 1989; Nahrstedt and Butterweck 1997; Borlis 

et al. 1998; Tolonen 2003; Kraus 2005). Selaginella has 

potent as source of biflavonoid, which can yield various 

biflavonoid compounds depending on species. It has 

cosmopolitanly distributed and able to cultivate almost all 

the words depending on species. 

DIVERSITY OF BIFLAVONOID 

Selaginella is one of the potential medicinal plants as a 

source biflavonoid in Nusantara, where 200 of the 700-750 

species from the entire world are found (Setyawan 2008). 

A total of 13 biflavonoid compounds have been isolated 

from Selaginella, including amentoflavone (3',8”- 

biapigenin), 2',8''-biapigenin, delicaflavone, ginkgetin, 

heveaflavone, hinokiflavone, isocryptomerin, kayaflavone, 

ochnaflavone, podocarpusflavone A, robustaflavone, 

sumaflavone, and taiwaniaflavone (Figure 2). In Setyawan 

and Darusman (2008) mentioned that the number is only 12 

biflavonoid compounds. Some biflavonoid are easily found 

at various species of Selaginella, but the other is only 

found at certain species. Amentoflavone and ginkgetin is 

biflavonoid compound of the most Selaginella, while 

sumaflavone is only reported from S. tamariscina (Yang et 

al. 2006; Lee et al. 2008) and delicaflavone is only reported 

from S. delcatula (Andersen and Markham 2006). At least 

11 species of Selaginella have been tested by 

amentoflavone content (Sun et al. 2006). There are also 

biflavonoid which is rarely found at Selaginella but it is 

commonly found at other species. Preliminary study shows 

that amentoflavone is found in high content (> 20%) at two 

of about 35 species of Malesian Selaginella, namely S. 

subalpina and S. involvens (ADS 2008, data not be shown). 

In Selaginella, taiwaniaflavone is only reported from S. 

tamariscina (Pokharel et al. 2006), while this is also found 

at other plant such as Taiwania cryptomerioides (Kamil et 

al. 1981). 

Selaginella is generally extracted from whole plant, 

though it is only conceived as herbs or leaves. Extraction 

can be conducted by various solvent, i.e. polar, semi-polar 

and non polar. For example: boiling in water, extraction by 

using methanol, ethanol, buthanol, ethyl acetate, 

chloroform, or extraction by using solvent mixture such as 

alcohol-water, ethanol-ethyl acetate, and ethanolchloroform. 

Methanol and ethanol are the most solvent 

used for biflavonoid extraction. Solvent types and 

extraction procedure can influence obtaining chemical 

structure and bioactivity of extract. Disease which is most 

treated by Selaginella extract is cancer. Besides, 

Selaginella extract also has many other usefulness, namely 

antioxidant, anti-inflammatory, antimicrobial (virus, 

bacterium, fungi, and protozoa), anti UV irradiation, anti 

allergy, vasorelaxation, anti diabetes, blood pressure 

stability, anti hemorrhagic, and antinociceptive. 

Biflavonoid needs evaluation for its medical and nutritional 

value (Harborne and Williams 2000). Selaginella contains 

various biflavonoid with difference medical properties 

(Table 2). 

Amentoflavone. Amentoflavone, the most common 

biflavonoid of Selaginella, has various biological and 

pharmacological effects, including antioxidant (Mora et al. 

1990; Cholbi et al. 1991; Shi et al. 2008), anti cancer (Silva 

et al. 1995; Lee et al. 1996; Lin et al. 2000; 

Guruvayoorappan and Kuttan 2007), anti-inflammatory 

(Gambhir et al. 1978; Baureithel et al. 1997; Gil et al. 

1997; Kim et al. 1998; Lin et al.. 2000; Woo et al.. 2005), 

antimicrobial (Woo et al. 2005; Jung et al. 2007), antivirus 

such as influenza (A, B), hepatitis (B), human 

immunodeficiency virus (HIV-1), herpes (HSV-1, HSV-2), 

herpes zoster (VZV), measles (Lin et al. 1998, 1999a,b, 

2002; Flavin et al. 2001, 2002), and respiratory syncytial 

virus (RSV) (Lin et al. 1999a,b; Ma et al. 2001), 

vasorelaxation (Kang et al. 2004), anti-urcerogenic 

(Gambhir et al. 1987), anti stomachic-ache (Kim et al. 

1998), anti depressant (Baureithel et al. 1997), anxiolytic 

(Cassels et al. 1998, 1999), analgesic (Silva et al. 2001), 

and anti-angiogenesis agent (Lee et al. 2009c). 

2',8''-biapigenin. 2',8''-biapigenin is an anticancer, 

which inhibit transactivation of iNOS gene and 

cyclooxigenase-2 (COX-2) through inactivate nuclear 

factor-κB (NF-κB) and prevent translocation of p65 (Chen 

et al. 2005b; Woo et al. 2006); and anti-inflammatory 

(Grijalva et al. 2004; Woo et al. 2005 2006; Pokharel et al. 

2006).

48 

3 (1): 44-58, March 2011 

OH 

OH 

HO 

O 

2’ 

HO 

8” 

O 

OH 

HO 

OH 

O 

HO 

OH 

O 

2’,8”-biapigenin 

Delicaflavone 

Amentoflavone 

(3',8”-biapigenin) 

OH 

OCH 3 

CH 3 O 

HO 

OCH 3 

OH 

CH 3O 

CH 3O 

Hinokiflavon 

OH 

OH 

O 

Hinokiflavone 

Ginkgetin 

Heveaflavone 

OCH 3 

OCH 3 

CH 3O 

HO 

CH 3O 

Ochnaflavone 

Isocryptomerin 

Kayaflavone 

OCH 3 

HO 

HO 

Podocarpusflavone A 

Sumaflavone 

OH 

OH 

HO 

OH 

OH 

HO 

Robustaflavone 

Taiwaniaflavone 

Figure 2. Structure of biflavonoid from Selaginella, namely: amentoflavone, 2',8''-biapigenin, delicaflavone, ginkgetin, heveaflavone, 

hinokiflavone, Isocryptomerin, kayaflavone, ochnaflavone, podocarpusflavone A, robustaflavone, sumaflavone, and taiwaniaflavone.


Delicaflavone. Its bioactivity is not observed yet from 

Selaginella. 

Ginkgetin. This compound is the second most studied 

biflavonoid of Selaginella beside amentoflavone. It has 

several properties including antioxidant (Su et al. 2000; 

Sah et al. 2005; Shi et al. 2008), anti-inflammatory 

(Grijalva et al. 2004; Woo et al. 2005, 2006; Pokharel et al. 

2006), anti viral such as herpes and cytomegalovirus 

(Hayashi et al. 1992); anti protozoan such as Trypanosoma 

cruzi (Weniger et al. 2006); anti cancer (Sun et al. 1997; 

Kim and Park 2002; Yang et al. 2007), such as such as 

ovarian adenocarcinoma (OVCAR-3), cervical carcinoma 

(HeLa) and foreskin fibroblast (FS-5) (Su et al. 2000). 

Ginkgetin is the strongest biflavonoid that inhibit cancer 

(Kim and Park 2002). Besides, this matter increase activity 

of neuroprotective against cytotoxic stress, and has potent 

for curing neurodegenerative disease such as stroke and 

Alzheimer (Kang et al. 2004; Han et al. 2006). Ginkgetin 

can also replace caffeine in food-stuff and medicines 

without generating addiction (Zhou 2002). 

Heveaflavone. Heveaflavone has cytotoxic activity 

against cancer cell of murine L 929 (Lin et al. 1994). 

Hinokiflavone. Hinokiflavone has antioxidant, antiviral 

and anti protozoan effect. This matter assists cell growth 

and protect from free radical cased by hydrogen peroxide 

(H 2 O 2 ) (Sah et al. 2005). It also inhibit sialidase influenza 

virus (Yamada et al. 2007; Miki et al. 2008); has high 

resistance to HIV-1 by in vivo and to polymerase HIV-1 

RTASE by in vitro (Lin et al. 1997). Lin et al. (1998, 

1999a,b, 2002) and Flavin et al. (2001, 2002) is patenting 

antiviral effect of hinokiflavone and others to influenza 

virus (A, B), hepatitis (B), human immunodeficiency virus 

(HIV-1), herpes (HSV-1, HSV-2), herpes zoster (VZV), 

and measles. It has antiprotozoan activity by in 

vitro against Plasmodium falciparum, Leishmania 

donovani and Trypanosoma sp. (Kunert et al. 2008). 

Isocryptomerin. Isocryptomerin has anti cancer 

property as well as anti-inflammatory, immunosuppressant 

and analgesic (Kang et al. 1998, 2001). It has cytotoxic 

activity against various cancer cells (Silva et al. 1995), 

including P-388 and HT-29 (Chen et al. 2005b). It has 

antibacterial activity against Gram-positive and Gramnegative 

bacteria (Lee et al. 2009b); and also has antifungal 

properties, which can depolarize fungal plasma membrane 

of Candida albicans (Lee et al. 2009a). 

Kayaflavone. Kayaflavone has moderately anti cancer 

property (Sun et al. 1997; Yang et al. 2007) and 

antioxidant, such as depleting H 2 O 2 (Su et al. 2000). 

Ochnaflavone. Ochnaflavone derivatives may have 

antioxidant activity that inhibits expression of gene COX-2 

at colon cancer cell (Chen et al. 2005b). 

Podocarpusflavone A. It has moderately anti cancer 

(Sun et al. 1997; Yang et al. 2007) and antioxidant 

properties (Su et al. 2000; Shi et al. 2008). 

Robustaflavone. Robustaflavone has anti cancer and 

anti virus properties. This matter significantly cytotoxic to 

various cancer cells (Silva et al. 1995) and significantly 

inhibits tumor cell of Raji and Calu-1 (Lin et al. 2000), 

cancer cell of P-388 and HT-29 (Chen et al. 2005b). It has 

also antiviral properties, which indicates high resistance to 

polymerase HIV-1 RTASE by in vitro (Lin et al. 1997) and 

also influenza virus (A, B), hepatitis (B), human 

immunodeficiency virus (HIV-1), herpes (HSV-1, HSV-2), 

herpes zoster (VZV), and measles (Lin et al. 1998, 1999a,b 

2002; Flavin et al. 2001, 2002). 

Sumaflavone. Sumaflavone has anti-inflammatory 

property that able to inhibit production of NO, by mean 

blocking lipopolysaccharide formation that induces iNOS 

gene expression (Yang et al. 2006). It can also significantly 

inhibit ability of UV irradiation to induce matrix 

metalloprotease-1 and -2 (MMP-1 and -2) activities at 

fibroblast of primary human skin (Lee et al. 2008). 

Taiwaniaflavone. It has anti-inflammatory, such as 

induce iNOS and COX-2 at macrophage of RAW 264.7 

(Pokharel, et al. 2006). 

MOLECULAR BIOACTIVITIES 

Selaginella is traditionally treated to cure several 

disease depending on species, such as cancer or tumor 

(uterus, nasopharyngeal, lung, etc), wound, after childbirth, 

menstrual disorder, female reproduction disease, expulsion 

of the placenta, tonic (for after childbirth, increase body 

endurance, anti ageing, etc), pneumonia, respiratory 

infection, exhalation channel infection, inflamed lung, 

cough, tonsil inflammation, asthma, urethra infection, 

bladder infection, kidney stone, cirrhosis, hepatitis, cystisis, 

bone fracture, rheumatism, headache, fever, skin diseases, 

eczema, depurative, vertigo, toothache, backache, blood 

purify, blood coagulation, amenorrhea, hemorrhage 

(resulting menstrual/obstetrical hemorrhage, stomachic, 

pile or prolepses of the rectum), diarrhea, stomach-ache, 

sedative, gastric ulcers, gastro-intestinal disorder, rectocele, 

itches, ringworm, bacterial disease, bellyache, neutralize 

poison caused by snakebite or sprained, bruise, paralysis, 

fatigue, dyspepsia, spleen disease (diabetic mellitus), 

emmenagogue, diuretic, and to refuse black magic 

(Martinez 1961; Bouquet et al. 1971; Dixit and Bhatt 1974; 

Ahmad and Raji 1992; Bourdy and Lee et al. 1992; Bourdy 

and Walter 1992; Nasution 1993; Lin et al. 1994; Kambuou 

1996; Caniago and Siebert 1998; Sequiera 1998; 

Dalimartha 1999; Mathew et al. 1999; Abu-Shamah et al. 

2000; van Andel 2000; Uluk et al. 2001; Harada et al. 

2002; Man and Takahashi 2002; Warintek 2002; Winter 

and Jansen 2003; ARCBC 2004; Batugal et al. 2004; de 

Almeida-Agra and Dantas 2004; DeFilipps et al. 2004; 

Wijayakusuma 2004; Mamedov 2005; Khare 2007; Pam. 

2008; Setyawan and Darusman 2008) (Table 1). This plant 

has sweet taste, and gives warm effect on the body (Bensky 

et al. 2004). 

Plants ecologically use biflavonoid to response 

environmental condition such as defense against pests, 

diseases, herbivory, and competitions; while human 

medically use as antioxidant, anti-inflammatory, anti 

cancer, anti allergy, antimicrobial, antifungal, antibacterial, 

antivirus, antiprotozoan, protection to UV irradiation, 

vasorelaxation (vasorelaxant), heart strengthener, anti 

hypertension, anti blood coagulation, and influence enzyme

50 

3 (1): 44-58, March 2011 

metabolism (Havsteen 1983, 2002; Kandaswami and 

Middleton 1993, 1994; Lale et al. 1996; Bisnack et al. 

2001; Duarte et al. 2001; Kromhout 2001; Kang et al. 

2004; Moltke et al. 2004; Arts and Hollman 2005; Martens 

and Mithofer 2005; Yamaguchi et al. 2005). The 

antioxidant, anti cancer and anti-inflammatory are the most 

important bioactivities of this secondary metabolite. 

Selaginella is known possess various molecular 

bioactivities depending on species, but only a few species 

has been detailed observe in the advanced research. Several 

species that also distributed in Nusantara are observed, 

such as S. tamariscina, S. doederleinii, S. involvens, S. 

moellendorffii, S. uncinata, and S. willdenowii; while the 

most distributed Selaginella in Nusantara namely S. plana 

has not been investigated yet (Table 2). 

S. tamariscina is the most powerful and the most useful 

plant Selaginella in the world. This herb is widely used as 

anti cancer, antioxidant and anti-inflammatory; and also 

used as anti UV irradiation, anti allergy, vasorelaxation, 

anti diabetic, immunosuppressant, analgesic, neuro 

protectant, antibacteria, antifungal, and possess estrogenic 

activity. As anti cancer, S. tamariscina can decrease 

expression of MMP-2 and -9, urokinase plasminogen 

activator, and inhibits growth of metastasis A549 cell and 

Lewis lung carcinoma (LLC) (Yang et al. 2007); inhibits 

proliferation of mesangial cell which activated by IL-1β 

and IL-6 (Kuo et al. 1998); inhibits leukemia cancer cell of 

HL-60 cell (Lee et al. 1999); induces expression of tumor 

suppressor gene of p53 (Lee et al. 1996); degrades 

leukemia cancer cell of U937 (Lee et al. 1996; Yang et al. 

2007); reduces proliferation nucleus antigen cell from 

stomach epithelium (Lee et al. 1999); chemopreventive for 

gastric cancer (Lee et al. 1999); induces apoptosis of cancer 

cell trough DNA fragmentation and nucleus clotting (Ahn 

et al. 2006); and induces breast cancer apoptosis through 

blockade of fatty acid synthesis (Lee et al. 2009c). This 

property is mostly given by amentoflavone and 

isocryptomerin (Kang et al. 1998, 2001; Lee et al. 2009c), 

while ginkgetin is also acted as anti cancer to OVCAR-3 

(Sun et al. 1997). As antioxidant, amentoflavone from S. 

tamariscina inhibits production of NO, which inactivates 

NF-κB, while sumaflavone blocks lipopolysaccharide 

formation that induces iNOS gene expression (Yang et al. 

2006). As anti-inflammatory, amentoflavone, 

taiwaniaflavone and ginkgetin from S. tamariscina inhibit 

inflammation that induce iNOS and COX-2 at macrophage 

RAW 264.7 which stimulated by lipopolysaccharide 

(Grijalva et al. 2004; Woo et al. 2005; Pokharel et al. 

2006). Amentoflavone inhibits activity of phospholipase 

Cγ1 (Lee et al. 1996); phospholipase A-2 (PLA-2) and 

COX-2 (Kim et al. 1998), while 2',8''-biapigenin inhibits 

transactivation of iNOS gene and COX-2 through 

inactivate NF-κB and prevent translocation of p65 (Woo et 

al. 2006). 

Amentoflavone from S. tamariscina inhibits fungi (Junk 

et al. 2006), anti influenza and resist to HSV-1 and -2 

(Rayne and Mazza 2007); hinokiflavone inhibits sialidase 

influenza virus (Yamada et al. 2007; Miki et al. 2008) and 

resists to HIV-1 (Lin et al. 1997); robustaflavone and 

hinokiflavone resist to polymerase HIV-1 RTASE (Lin et 

al. 1997); ginkgetin inhibits herpes and cytomegalovirus 

(Hayashi et al. 1992), by degrading protein synthesis of 

virus and depress gene transcription (Middleton et al. 

2000). Isocryptomerin from S. tamariscina shows potent 

antibacterial activity against Gram-positive and Gramnegative 

(Lee et al. 2009b). Amentoflavone from S. 

tamariscina inhibits several pathogenic fungi (Woo et al. 

2005; Jung et al. 2007). Isocryptomerin from S. 

tamariscina can depolarize fungal plasma membrane of C. 

albicans (Lee et al. 2009a). 

S. tamariscina is effective ingredient to prevent and 

cure acute brain degenerative disease, such as stroke and 

dementia (Han et al. 2006). Capability to prevent brain 

damage is especially given by amentoflavone (Kang et al. 

1998). S. tamariscina can elastic vascular smooth muscle 

through endothelium related to nitric oxide (NO) activity 

(Yin et al. 2005). Amentoflavone from S. tamariscina 

induces relaxation of phenylephrin which responsible to 

aorta contraction (Kang et al. 2004; Yin et al. 2005). S. 

tamariscina containing sumaflavone and amentoflavone 

inhibit ability of UV irradiation to induce MMP-1 and -2 at 

fibroblast (Lee et al. 2008). S. tamariscina reduces 

histamine from peritoneal mast cell causing allergic 

reaction (Dai et al. 2005). S. tamariscina decreases sugar 

blood and lipid peroxide, and also increases insulin 

concentration (Miao et al. 1996). Amentoflavone from S. 

tamariscina inhibits activity of tyrosine phosphatase 1B to 

maintain type-2 diabetic and obesity (Na et al. 2007). 

S. articulate is treated as anti hemorrhagic. Water 

extract of this matter can moderately neutralize 

hemorrhagic effect and inhibits proteolysis of casein by 

venom (Otero et al. 2000; Winter and Jansen 2003). 

S. bryopteris acts as antioxidant, anti-inflammatory, 

antiprotozoan, anti UV-irradiation and anti spasmodic. 

Water extract of S. bryopteris increases endurance to 

oxidative stress; and assists cell growth and protects from 

free radical stress caused by H 2 O 2 (Sah et al. 2005). S. 

bryopteris is treated as anti-inflammatory and cures veneral 

disease (Agarwal and Singh 1999). Amentoflavone and 

hinokiflavone from S. bryopteris have antiprotozoan 

activity against P. falciparum, L. donovani and 

Trypanosoma sp (Kunert et al. 2008). Water extract of S. 

bryopteris also significantly reduces potent cell dying 

caused by UV irradiation (Sah et al. 2005), while ethanolic 

extract can cure stomachic (Pandey et al. 1993). 

S. delicatula acts as anti cancer and antioxidant. Water 

extract of S. delicatula has antioxidant characteristic and 

degrades blood cholesterol (Gayathri et al. 2005). Extract 

of S. delicatula that contained by robustaflavone and 

amentoflavone or its derivatives is cytotoxic against cancer 

cell of P-388, HT-29 (Chen et al. 2005b), Raji, Calu-1, 

lymphoma and leukemia (Lin et al. 2000) 

S. doederleinii is usually treated as anti cancer, but also 

acts as antiviral and anti-inflammatory. Water extract of S. 

doederleinii has antimutagenic against both picrolonic 

acid- and benzo[α]pyrene-induced mutation to cancer cell 

(Lee and Lin 1988). Ethanolic extract of S. doederleinii 

that is amentoflavone and heveaflavone has cytotoxic 

activity against cancer cell of murine L 929 (Lin et al. 

1994). Extract of S. doederleinii also has cytotoxic against


the three human cancer cell lines, HCT, NCI-H358, and 

K562 (Lee et al. 2008), and has anti mutagenic effect 

against cholangiocarcinoma cancer, but my cause bone 

marrow depression (Pan et al. 2001). Amentoflavone from 

S. doederleinii has potent as antiviral and antiinflammatory 

agents (Lin et al. 2000). However, hordenine 

that isolated from S. doederleinii increases hypertension 

(Lin et al. 1991). 

S. involvens has characteristics as antioxidant, antiinflammatory 

and anti bacteria. Extract of S. involvens can 

inhibit production and effect of free radicals of NO and 

expression of iNOS/IL-1β (Joo et al. 2007). Water extract 

of S. involvens has significantly antioxidant effect to lipid 

peroxides (EC50 = 2 ug/mL). This extract is non toxic and 

degrades blood cholesterol (Gayathri et al. 2005). Water 

extract of S. involvens kills the various Leptospira strains, 

which causes infectious of leptospirosis diseases (Wang et 

al. 1963). Extract of S. involvens depresses activity of 

Propionibacterium acnes (> 100 ug/mL), which responses 

to acne inflammation; although has no antibiotic property 

(Joo et al. 2007). Beside, water extract of S. involvens may 

have analgesic activity (ECMM 1997; Ko et al. 2007). 

S. labordei indicates antioxidant, anti cancer, and anti 

virus characteristics. S. labordei can inhibit activity of 

xanthine oxidase (XOD) and lipoxygenase (LOX), and 

absorb free radical (Chen et al. 2005b; Tan et al. 2009). It 

also down-regulate COX-2 gene expression in human 

colon adenocarcinoma CaCo-2 cells (Chen et al. 2005b). 

Robustaflavone of S. labordei can inhibit hepatitis B virus 

(Tan et al. 2009) 

S. lepidophylla has hypoglycemic property (Andrade- 

Cetto and Heinrich 2005); while non-biflavonoid 

compound from methanolic extract of S. lepidophylla, 3- 

methylenhydroxy-5-methoxy-2,4-dihydroxy 

tetrahydrofuran, has moderate resistance to uterus 

contraction (Perez et al. 1994). 

S. moellendorffii contains antioxidant and anti cancer 

properties. Ethyl acetate extract of S. moellendorffii 

contains amentoflavone, hinokiflavone, podocarpusflavone 

A, and ginkgetin that has antioxidant properties (Shi et al. 

2008). Ginkgetin that extracted by ethanol or ethyl acetate 

from S. moellendorffii can inhibit cancer cell growth of 

OVCAR-3, HeLa, and FS-5 (Sun et al. 1997; Su et al. 

2000). It also act as anti-metastasis at lung cancer cell of 

A549 and LLC (Yang et al. 2007); and apoptosis resulting 

caspase activation by H 2 O 2 (Su et al. 2000); while 

amentoflavone and its derivatives, kayaflavone, and 

podocarpusflavone A, have no this bioactivity (Sun et al. 

1997). 

S. pallescens has moderately antimicrobials and anti 

spasmodic activities. S. pallescens contains an endophytic 

Fusarium sp. that produce pentaketide anti fungal agent, 

CR377 (Brady and Clardy 2000). Chloroform-methanolic 

extract of S. pallescens can inhibit spontaneously 

contraction of ileum muscle (Rojas et al. 1999). 

S. rupestris contains amentoflavone which has 

antispasmodic effect to ileum; and strengthening heart in 

case of normodinamic and hypodinamic (Chakravarthy et 

al. 1981) 

S. sinensis contains amentoflavone which has antiviral 

actifity against RSV (Ma et al. 2001) 

S. uncinata has activity as anti virus but generated by 

non biflavonoid compounds. S. uncinata has chromone 

glycosides, namely uncinoside A and B (Man and 

Takahashi 2002), which showed antiviral activities against 

RSV and PIV-3 (Ma et al. 2003). 

S. willdenowii contains isocryptomerin and derivatives 

of amentoflavone and robustaflavone which significantly 

cytotoxic against various cancer cell (Silva et al. 1995). 

TREHALOSE 

Trehalose is formed by α,α-1,1-glycosidic linkage of 

two low energy hexose moieties (Paiva and Panek 1996; 

Elbein et al 2003; Grennan 2007). This matter is a unique 

simple sugar which non reactive, very stable, colorless, 

odor-free, non-reducing disaccharide, and capable to 

protect biomolecules against environmental stress 

(Schiraldi et al. 2002). Therefore, this compound is a 

natural product, although not as commonly secondary 

metabolites of natural products. It works as osmoprotectant 

during desiccation stress (Adams et al. 1990); such as 

compatible solute in the stabilization of biological 

structures under abiotic stress (Garg et al. 2002); serves as 

a source of energy and carbon (Elbein et al 2003; 

Schluepmann et al. 2003); serves as signaling molecule to 

control certain metabolic pathways (Muller et al. 2001; 

Elbein et al 2003; Avonce et al 2005); protects proteins and 

cellular membranes from inactivation or denaturation 

caused by harsh environmental stress, such as desiccation, 

dehydration (drought), thermal heat, cold freezing, 

oxidation, nutrient starvation, and salt (Avigad 1982; 

Elbein et al. 2003; Wu et al. 2006). Trehalose acts as a 

global protectant against abiotic stress (Jang et al. 2003). 

This matter is proved to be an active stabilizer of enzymes, 

proteins, biomasses, pharmaceutical preparations and even 

organs for transplantation (Schiraldi et al. 2002), and very 

prospects as molecular bio stabilizer in cosmetic, pharmacy 

and food (Roser 1991; Kidd and Devorak 1994). These 

multiple effects of trehalose on protein stability and folding 

suggest promising applications (Singer and Lindquist 

1998). 

Trehalose has long been known for protecting certain 

organisms from desiccation. The accumulation of the 

disaccharide trehalose in anhydrobiotic organisms allows 

them to survive severe environmental stress (Zentella et al. 

1999). Trehalose also promotes survival under extreme 

heat conditions, by enabling proteins to retain their native 

conformation at elevated temperatures and suppressing the 

aggregation of denatured proteins (Singer and Lindquist 

1998). Desiccation can reduce the lipid component in 

thylakoid membranes (Guschina et al. 2002). However, in 

desiccation-tolerant plants, membrane integrity appears not 

to be affected during drought-stress. S. lepidophylla retain 

their structural organization as intact bilayers (Platt et al. 

1994) and often referred as resurrection plant because able 

to live on long drought and recovery through rehydration

52 

3 (1): 44-58, March 2011 

process (Crowe et al. 1992), even when the most water 

body (99%) is evaporated (Schiraldi et al. 2002; van Dijck 

et al. 2002). Another species, S. tamariscina, can also 

remain alive in a desiccated state and resurrect when water 

becomes available (Liu et al. 2008). The drought can 

change fluorescence and pigmentation, but can not cause 

dying (Casper et al. 1993). 

Trehalose exists in a wide variety of organisms, 

including bacteria, yeast, fungi, insects, invertebrates, and 

lower and higher plants (Elbein 1974; Crowe et al. 1984; 

Elbein et al. 2003), but rarely find in Angiosperms (Muller 

et al. 1995) and does not find in mammals (Teramoto et al 

2008), and it is not accumulated to detectable levels in the 

most plants (Garg et al. 2002). This sugar plays important 

roles in cryptobiosis of Selaginella and other organisms, 

which revive with water from a state of suspended 

animation induced by desiccation (Teramoto et al 2008). 

Trehalose is the major sugar formed in photosynthesis of 

Selaginella (White and Towers 1967). Some Selaginella 

contains high concentration of trehalose, such as S. 

lepidophylla (Adams et al. 1990; Mueller et al. 1995; 

Zentella et al. 1995), S. sartorii (Iturriaga et al. 2000), S. 

martensii (Roberts and Tovey 1969), S. densa, and S. 

wallacei (White and Towers 1967). Trehalose can reach 

10-15% of cell dry weight (Grba et al. 1975). 

Trehalose is not merely chemical compounds that 

responsible to resurrection ability of Selaginella. The 

protective effect of trehalose is correlated with a trapping 

of the protein in a harmonic potential, even at relatively 

high temperature (Cordone et al. 1999). Deeba et al (2009) 

suggest that S. bryopteris, one kind of resurrection plants, 

has about 250 proteins that expressed in response to 

dehydration and rehydration, and involved in transport, 

targeting and degradation in the desiccated fronds. Harten 

and Eickmeier (1986) suggest that several conservationed 

enzymes are beneficial for rapid resumption of metabolic 

activity of S. lepidophyla. Furthermore, Eickmeier (1979; 

1982) suggests that both organelle- and cytoplasm-directed 

protein syntheses are necessary for full photosynthetic 

recovery during rehydration of S. lepidophyla. 

FUTURE RESEARCH 

Research on Selaginella is still widely challenging. In 

the most elementary study of plant taxonomy, the high 

morphological variation of Selaginella causes several 

misidentification of this taxon. In ecology, global warming, 

habitat fragmentation and degradation that affected on 

sustainability of this resource need to be observed. In 

physiology, changes of fluorescens and pigmentation 

caused by environmental factor and age need to be 

explained. In biochemistry, several natural products are not 

exploited yet. One of non-biflavonoid compound from 

Selaginella that needs to be further investigated is 

trehalose. Molecular study is also required clarifying 

certain identity and phylogenetics relationship. 

In Indonesia, several authors often misidentify 

Selaginella species, especially on popular article. This 

matter is often identified as S. doederleinii, including 

Javanese wild species. The most authors agree that S. 

doederleinii is recognized as non native plant of Indonesia, 

which natural distribution is India, Burma, Thailand, Laos, 

Cambodia, Vietnam, Malaya, Chinese, Hong Kong, 

Taiwan, and Japan (Huang 2006; USDA 2008). Java has no 

species of S. doederleinii according to Alston (1935a) and 

observation on Selaginella collection of Herbarium 

Bogoriense, through several Kalimantan collection is 

suspected and has morphological similarity to this species 

(ADS 2007, data is not shown). This matter is possibility 

caused by referring to Dalimartha (1999), which include S. 

doederleinii in Indonesian plant medicines. Harada et al. 

(2002) conduct similar misidentification, which cite S. 

plana as one of plant medicine in Mount Halimun NP 

(nowadays Mount Halimun-Salak NP), but the main picture 

presented is S. willdenowii. Field survey indicate that S. 

willdenowii is easily found in road side to Cikaniki 

Research Station of Mount Halimun-Salak NP, at rice field, 

shrubs land, primary and secondary forest, while S. plana is 

easier to be found in countrifield at lower height (ADS 

2008, personal observation). 

Species misidentification impacts on drug properties, 

because each species differ chemical constituent. Natural 

products content of Selaginella highly vary depending on 

species, although does not always congruent with 

traditional medical recipes. Sundanese people of Mount 

Halimun-Salak NP complementarily or substitutionally 

uses several Selaginella for treatment of after childbirth 

including S. ornata, S. willdenowii, S. involvens, and S. 

intermedia, but for similar recipe Sundanese around Bogor 

only uses S. plana (ADS 2008, personal observation). 

Morphological diversity at infra specific level, and changes 

on pigmentation caused by age, drought and other 

environmental factors able to entangle identify base on 

morphological characteristics. It needs identification base 

on molecular characteristic, such as Korall et al. (1999) and 

Korall and Kenrick (2002, 2004). Beside, taxonomy of 

Malesian Selaginella needs to revise, because still based on 

old literature namely Alderwereld van Rosenburgh 

(1915a,b; 1916, 1917, 1918, 1920, 1922) and Alston (1934, 

1935a,b; 1937, 1940). In a research brief about the 

traditional utilization of Selaginella in Indonesia, Setyawan 

(2009) collect at least 40 species of which half are 

estimated to new species or new records. 

Completely research on variability of biflavonoid 

compounds of various Selaginella species with various 

solvent have not conducted yet. This matter is only 

conducted to certain species, compounds, and solvents. 

Natural products of certain plant determine economical 

value that required in industrial scale of modern pharmacy. 

Species with various low content of natural products less 

value than species with restricted high content, because 

modern pharmacy exploits natural products at molecular 

level. However, this matter is not always become 

consideration in traditional medication, because it generally 

uses simplicia that can be easily substituted by each others. 

In phytochemistry and chemotaxonomy, high variety of 

natural products can assist identification, though each has 

no high content. However, a very low compound is not


significantly important for identification, because often 

influenced by environmental factors, not merely to genetic 

factor. Bioactivity of each biflavonoid also requires to be 

observed because nowadays only bioactivity of 

amentoflavone and ginkgetin has been completely studied. 

HPLC is potent method for analyzing of natural products of 

Selaginella (Fan et al. 2007). 

Besides, trehalose observation on Selaginella is still 

restricted on a few species, and need to be conducted to 

amount of other species caused by potent economic value 

that can be generated. It can preliminary indicated by 

species that incurling leaves in hot weather or drough 

condition. 

Biflavonoid study of Selaginella is still require attention 

such as: (i) the importance of assuring species identity 

caused by height morphological variety including by using 

molecular method; (ii) the importance of extending 

research coverage most of biflavonoid type, species, and 

extraction method; and also (iii) the importance of 

extending investigation on bioactivity, including nonbiflavonoid 

compound, which also have high economic 

potent such as trehalose. 

CONCLUSION 

Selaginella is a potent medicinal matter, which mostly 

contains phenolic (flavonoid), alkaloid, and terpenoid. This 

matter is traditionally used to cure several diseases 

especially for wound, after childbirth, and menstrual 

disorder. Biflavonoid, a dimeric form of flavonoids, is one 

of the most valuable natural products of Selaginella, which 

constituted at least 13 compounds, namely amentoflavone, 

2',8''-biapigenin, delicaflavone, ginkgetin, heveaflavone, 

hinokiflavone, isocryptomerin, kayaflavone, ochnaflavone, 

podocarpusflavone A, robustaflavone, sumaflavone, and 

taiwaniaflavone. Human medically use biflavonoid 

especially for antioxidant, anti-inflammatory, and anti 

cancer. Selaginella also contains several natural products, 

such as trehalose which valuable for bioindustry. 

Selaginella research exhaustively needs to be conducted to 

explore all natural products constituents and their 

bioactivities. 


I would like to thank Prof. Dr. Keon Wook Kang from 

Chosun University, Gwangju, South Korea which delivered 

a number of valuable articles on chemistry of biflavonoid. I 

also thank Prof. Dr. Umesh R. Desai from Commonwealth 

University, Virginia, USA for permitting to cite a very 

interesting article on biosynthesis of biflavonoid. Grateful 

thank to Prof. Dr. Raphael Ghogomu-Tih from University 

of Yaoundé, Cameron for suggesting to write this article in 

international language. Sincere thanks are expressed to the 

late Dr. Muhammad Ahkam Soebroto from Research 

Center for Biotechnology, Indonesian Institute of Science, 

Cibinong-Bogor and anonymous peer reviewer of this 

article for their criticism. I also would like to thank Prof. 

Dr. Wasmen Manalu from SEAMEO-BIOTROP Bogor 

which had invited me for training in writing scientific 

articles. 

REFERENCES 

Abu-Shamah Z, Faridz ZF, Rizal MK, Fadhilah ZW, Abdullah I. 2000. 

Ethnobotanical study in Kuala Nerang, Kedah. In: Chang YS, Mohtar 

M, Bramaniam V, Abu-Samah Z (eds) Towards bridging science and 

herbal industry; Proceedings of the Seminar on Medicinal and 

Aromatic Plants. Forest Research Institute Malaysia, Kepong, Kuala 

Lumpur, 12-13 September 2000 

Adams RP, Kendall E, Kartha KK. 1990. Comparison of free sugars in 

growing and desiccated plants of Selaginella lepidophylla. Biochem 

Sys Ecol 18: 107-110 

Adaramoye OA, IA Medeiros. 2009. Endothelium-independent 

vasodilation induced by kolaviron, a biflavonoid complex from 

Garcinia kola seeds, in rat superior mesenteric arteries. J Smooth 

Muscle Res 45 (1): 39-53 

Agarwal SS, Singh VK. 1999. Immunomodulators: a review of studies on 

Indian medicinal plants and synthetic peptides; part 1. medicinal 

plants. Pinsa 65 (3&4): 179-204 

Ahmad F bin, Raji H. 1992. Medicinal plants of the Murut community in 

Sabah. In: Ghazzaly I, Siraj O, Murtedza M (eds) Forest biology and 

conservation in Borneo. Centre for Borneo Studies, Kota Kinabalu 

Ahmad I, Aqil F, Ahmad F, Owais M (ed.). 2006. Modern phytomedicine, 

turning medicinal plants into drugs. Wiley-Vch Verlag GmbH & Co. 

KGaA, Weinheim 

Ahn, SH, Mun YJ, Lee SW, Kwak S, Choi MK, Baik SK, Kim YM, Woo 

WH. 2006. Selaginella tamariscina induces apoptosis via a caspase-3- 

mediated mechanism in human promyelocytic leukemia cells. J Med 

Food 9 (2): 138-144 

Alderwereld van Rosenburgh CRWK van. 1915a. Malayan Fern Allies. 

Department of Agriculture, Industry and Commerce, Batavia 

Alderwereld van Rosenburgh CRWK van. 1915b. New or interesting 

Malay ferns 7. Bull Jard Bot Buitenzorg 2 (20): 1-28 

Alderwereld van Rosenburgh CRWK van. 1916. New or interesting 










Alston AHG. 1934. The genus Selaginella in the Malay Peninsula. Gard 

Bull Strait Settl 8: 41-62 

Alston AHG. 1935a. The Selaginella of the Malay Islands: I. Java and the 

Lesser Sunda Islands. Bull Jard Bot Buitenzorg 3 (13): 432-442 

Alston AHG. 1935b. The Philippines species of Selaginella. Philip J Sci 

58: 359-383 

Alston AHG. 1937. The Selaginella of the Malay Islands: II. Sumatra. 

Bull Jard Bot Buitenzorg 3 (14): 175-186 

Alston AHG. 1940. The Selaginella of the Malay Islands: III. Celebes and 

the Moluccas. Bull Jard Bot Buitenzorg 3 (16): 343-350 

Andersen ØM, Markham KR. 2006. Flavonoids: chemistry, biochemistry, 

and applications. CRC Press, Taylor & Francis Group, Boca Raton, 

FL 

Andrade-Cetto A, Heinrich M. 2005. Mexican plants with hypoglycaemic 

effect used in the treatment of diabetes. J Ethnopharmacol 99: 325- 

348 

ARCBC [ASEAN Regional Centre for Biodiversity Conservation]. 2004. 

Checklist of medicinal plants in Southeast Asia. ASEAN Regional 

Centre for Biodiversity Conservation, Manila 

www.aseanbiodiversity.org/medicinal_plants/page7.htm 

Ariyasena J, Baek S-H, Perry NB, Weavers RT. 2004. Ether-linked 

biflavonoids from Quintinia acutifolia. J Nat Prod 67: 693-696 

Arts IC, Hollman PC. 2005. Polyphenols and disease risk in 

epidemiologic studies. Amer J Clin Nutr 81: 317-325

54 

3 (1): 44-58, March 2011 

Atta-ur-Rahman, Choudhary MI. 1999. Recent studies on bioactive 

natural products. Pure Appl Chem 71 (6): 1079-1081 

Avigad G. 1982. Sucrose and other disaccharides. In: Loewus FA, Tanner 

W (eds) Encyclopedia of plant physiology. Springer, New York 

Avonce N, Leyman B, Thevelein J, Iturriaga G.. 2005. Trehalose 

metabolism and glucose sensing in plants. Biochem Soc Trans 33 (1): 

276-279 

Baker W, Simmonds WHC. 1940. Derivatives of 5,6,4’- and 5,8,4’- 

trihydroxyflavones, and a note on the structure of ginkgetin. J Chem 

Soc 1370-1374 

Basso, LA, da Silva LH, Fett-Neto AG, de Azevedo Jr WF, de Moreira IS, 

Palma MS, Calixto JB, Astolfi-Filho S, dos Santos RR, Soares MB, 

Santos DS. 2005. The use of biodiversity as source of new chemical 

entities against defined molecular targets for treatment of malaria, 

tuberculosis, and T-cell mediated diseases; A Review. Mem Inst 

Oswaldo Cruz 100: 475-506 

Batugal, PA, Kanniah J, Young LS, Oliver JT (eds). 2004. Medicinal 

plants research in Asia, Vol 1: The framework and project workplans. 

International Plant Genetic Resources Institute-Regional Office for 

Asia, the Pacific and Oceania (IPGRI-APO). Serdang, Selangor DE, 

Malaysia 

Baureithel KH, Buter KB, Engesser A, Burkard W, Schaffner W. 1997. 

Inhibition of benzodiazepine binding in vitro by amentoflavone, a 

constituent of various species of Hypericum. Pharm Acta Helv 72 (3): 

153-157 

Bennie L, Coetzee J, Malan E, Ferreira D. 2001. Structure and 

stereochemistry of triflavanoids containing both ether and carboncarbon 

interflavanyl bonds. Phytochem 57: 1023-1034 

Bennie L, Coetzee J, Malan E, Ferreira D. 2002. (4-6)-Coupled 

proteracacinidins and promelacacinidins from Acacia galpinii and 

Acacia caffra. Phytochem 60: 521-532 

Bennie L, Malan E, Coetzee J, Ferreira D. 2000. Structure and synthesis 

of ether-linked proteracacinidin and promelacacinidin 

proanthocyanidins from Acacia caffra. Phytochem 53: 785-793 

Bensky D, Clavey S, Stöger E. 2004. Chinese herbal medicine-materia 

medica, 3rd ed. Eastland Press, Seattle, W.A 

Berghofer R, Holzl J. 1987. Biflavonoids in Hypericum perforatum. Part 

1. Isolation of I3,II8-biapigenin. Planta Med 216-217 

Berghofer R, Holzl J. 1989. Isolation of I3',II8-biapigenin 

(amentoflavone) from Hypericum perforatum. Planta Med 55: 91 

Bi YF, Zheng XK, Feng WS, Shi SP, Wang JF, Niu JZ. 2004b. Study on 

the chemical constituents of Selaginella tamariscina (Beauv.) Spring. 

Yao Xue Xue Bao 39 (4): 266-268 

Bi YF, Zheng XK, Feng WS, Shi SP. 2004a. Isolation and structural 

identification of chemical constituents from Selaginella tamariscina 

(Beauv.) Spring. Yao Xue Xue Bao. 39 (1): 41-45 

Bisnack R, Boersma BJ, Patel RP, Kirk M, White CR, Darley-Usmar V, 

Barnes S, Zhou F, Parks DA. 2001. Enhanced antioxidant activity 

after chlorination of quercetin by hipochlorous acid. Alcohol Clin Exp 

Res 25 (6): 434-443 

Borris RP. 1996. Natural products research: perspectives from a major 

pharmaceutical company. J Ethnopharmacol 51: 29-38 

Bouquet A, Cave A, Paris R. 1971. Plantes medicinales du Congo- 

Brazzaville (III) plantes medicinales et phytotherapie. Tome 5 (2): 

154-158 

Bourdy G, Walter A. 1992. Maternity and medicinal plants in Vanuatu I. 

The cycle of reproduction. J Ethnopharmacol 37: 179-196 

Brady SF, Clardy J. 2000. CR377, a new pentaketide antifungal agent 

isolated from an endophytic fungus. J. Nat. Prod. 63:1447-1448 

Braide VB. 1989. Antispasmodic extracts from seeds of Garcinia kola. 

Fitoterapia 9: 123 

Braide VB. 1993. Antiinflammatory effect of kolaviron, a biflavonoid 

extract of Garcinia kola. Fitoterapia 64 (5): 433 

Caniago I, Siebert S. 1998. Medicinal plant ecology, knowledge and 

conservation in Kalimantan, Indonesia. Economic Botany 52 (3): 229- 

250. 

Casper C, Eickmeier WG, Osmond CB. 1993. Changes of fluorescence 

and xanthophyll pigments during dehydration in the resurrection plant 

Selaginella lepidophylla in low and medium light intensities. Oecol 

94: 528-533 

Cassels BK, Dajas FJ, Medina JH, Paladini AC, Silveira RH. 1998. 

Flavonoid and biflavonoid derivatives, their pharmaceutical 

compositions, their anxiolytic activity. United States Patent 5756538 

(May 26, 1998) 

Cassels BK, Dajas FJ, Medina JH, Paladini AC, Silveira RH. 1999. 

Flavonoid and biflavonoid derivatives, their pharmaceutical 

compositions, their anxiolytic activity. United States Patent 6004998 

(December 21, 1999) 

Chakravarthy BK, Rao YV, Gambhir SS, Gode KD. 1981. Isolation of 

amentoflavone from Selaginella rupestris and its pharmacological 

activity on central nervous system, smooth muscles and isolated frog 

heart preparations. Planta Med 43 (9): 64-70 

Chang CY, Chen XD, Xiao XY, Lin RC. 2000. Studies on 

micromorphology and its significance in anatomy and identification 

of Selaginella. J Med Anal 20 (2): 75-78 

Chen JJ, Duh CY, Chen JF. 2005a. New cytotoxic biflavonoids from 

Selaginella delicatula. Planta Med 71 (7): 659-665 

Cheng XL, Ma SC, Yu JD, Yang SY, Xiao XY, Hu JH, Lu Y, Shaw PC, 

But PPH, Lin RC. 2008. Selaginellin A and B, two novel natural 

pigments isolated from Selaginella tamariscina. Chem Pharm Bull 56 

(7): 982-984 

Chikmawati T, Miftahudin. 2008. Biodiversity and potent of genus 

Selaginella as anti oxidant and anti cancer. LPPM IPB, Bogor 

Chikmawati T, Setyawan AD, Miftahudin. 2008. Phytochemical 

constituent of plant extract of Selaginella in Java. 8th Seminary and 

Congress of Indonesian Plant Taxonomy Association (“PTTI”), 

Cibinong Science Center, Bogor-Indonesia, 21-23 October 2008 

Chiu PL, Patterson GW, Salt TA. 1988. Sterol composition of 

pteridophytes. Phytochem 27 (3): 819-822 

Cholbi MR, Paya M, Alcaraz MJ. 1991. Inhibitory effect of phenolic 

compounds on CCl 4 induced microsomal lipid peroxidation. 

Experientia 47: 195-199 

Cordone L, Ferrand M, Vitrano E, Zaccai G. 1999. Harmonic behavior of 

trehalose-coated carbon-monoxy-myoglobin at high temperature. 

Biophysic J 76: 1043-1047 

Crowe JF, Hoekstra FA, Crowe LM. 1992. Anhydrobiosis. Ann Rev 

Physiol 54: 579-599 

Crowe JH, Crowe LM, Chapman D. 1984. Preservation of membranes in 

anhydrobiotic organisms: the role of trehalose. Science 223: 701-703 

Cutler SJ, Cutler HG. 2000. Biologically active natural products: 

pharmaceuticals. CRC Press. Boca Raton, CA. 

Dai Y, But PH, Chu LM, Chan YP. 2005. Inhibitory effects of Selaginella 

tamariscina on immediate allergic reactions. Amer J Chin Med 33 

(6): 957-966 

Dai Z, Ma SC, Wang GL, Wang F, Lin RC. 2006. A new glucoside from 

Selaginella sinensis. J Asian Nat Prod Res 8 (6): 529-533 

Dalimartha S. 1999. Atlas tumbuhan obat Indonesia. Trubus Agriwidya, 

Yogyakarta 

de Almeida-Agra C, Dantas IC. 2004. Identificacao dos fitoterapicos 

indicados pelos raizeiros e utilizados pelas mulheres no combate a 

enfermidades do aparelho geniturinario na cidade de Campina Grande 

- PB. Universidade Estadual da Paraiba, Paraiba 

Deeba F, Pandey V, Pathre U, Kanojiya S. 2009. Proteome analysis of 

detached fronds from a resurrection plant Selaginella bryopteris - 

response to dehydration and rehydration. J Proteomics Bioinform 2 

(2): 108-116 

DeFilipps RA, Maina SL, Crepin J. 2004. Medicinal Plants Index of the 

Guianas (Guyana, Surinam, French Guiana). Smithsonian Institution, 

Washington, D.C. 

Dixit RD, Bhatt GK. 1974. Ferns: A much neglected group of medicinal 

plants II. J Res Indian Med 9: 59-68 

DNP [Dictionary of Natural Products]. 1992. Dictionary of natural 

products. Chapman and Hall, New York 

Dreschner EE, Ruperto J, Wong G, Newmark HL. 1991. Quercetin and 

rutin as inhibitors of azoxymethanol-induced colonic neoplasia. 

Carcinogenesis 7: 1193-1196 

Duarte J, Perez-Palencia R, Vargas F, Ocete MA, Perez-Viscaio F, 

Zarzuelo A, Tamargo J. 2001. Antihypertensive effects of the 

flavonoids quercetin in spontaneously hypertensive rats. Br J 

Pharmacol 133 (1): 117-124 

Dubber MJ. 2005. Application of CE, HPLC and LC-MS-MS for the 

analysis and quality control of Ginkgo biloba dosage forms. 

[Dissertation]. Rhodes University, Rhodes 

ECMM [Encyclopedia of Chinese Materia Medica]. 1977. Encyclopedia 

of Chinese materia medica (Zhongyao dacidian). Shanghai Science 

and Technology Press, Shanghai 

Eickmeier WG. 1979. Photosynthetic recovery in the resurrection plant 

Selaginella lepidophylla after wetting. Oecol 39: 93-106 

Eickmeier WG. 1982. Protein synthesis and photosynthetic recovery in the 

resurrection plant, Selaginella lepidophylla. Plant Physiol 69: 135- 

138


Elbein AD, Pan YT, Pastuszak I, Carroll D. 2003. Review: New insights 

on trehalose: a multifunctional molecule. Glycobiol 13 (4). DOI: 

10.1093/glycob/cwg047 

Elbein AD. 1974. The metabolism of alpha-trehalose. Adv Carbohydr 

Chem Biochem 30: 227-256 

Epstein E. 1999. Silicon. Ann Rev Pl Physiol Pl Mol Biol 50: 641-664 

Fan XL, Wan DR, Ye CJ, Chen KL. 2007. Study on HPLC fingerprint 

characteristics of Selaginella plants. Zhongguo Zhong Yao Za Zhi 32 

(20):2102-2106 

Farnsworth NR. 1994. Ethnobotany and the search for new drugs. John 

Wiley and Sons, New York 

Farombi EO, Adepoju BF, Ola-Davies OE, Emerole GO. 2005. 

Chemoprevention of aflatoxin B1-induced genotoxicity and hepatic 

oxidative damage in rats by kolaviron, a natural biflavonoid of 

Garcinia kola seeds. Eur J Cancer Prev 14 (3): 207-214 

Feng, WS, Chen H, Zheng XK, Wang YZ, Gao L, Li HW. 2009. Two new 

secolignans from Selaginella sinensis (Desv.) Spring. J Asian Nat 

Prod Res 11 (7): 658-662 

Ferreira D, Bekker R. 1996. Oligomeric proanthocyanidins: naturally 

occurring O-heterocycles. Nat Prod Rep 13: 411-433 

Ferreira D, Nel RJ, Bekker R. 1999a. In: Barton DHR, Nakanishi K, 

Meth-Cohn O, Pinto BM (eds) Comprehensive natural products 

chemistry. Elsevier, New York 

Ferreira D, Slade D, Marais JPJ. 2006. Flavans and proanthocyanidins. In: 

Anderson OM, Markham KR (eds) Flavonoids: chemistry, 

biochemistry and applications. CRC Press, Boca Raton, FL 

Ferreira DE, Brandt V, Coetzee J, Malan E. 1999b. Condensed tannins. In: 

Zechmeister L, Herz W, Falk H, Kirby GW, Moore RE (eds) The 

chemistry of organic natural products. Springer, Wien 

Flavin MT, Lin YM, Zembower DE, Schure R, Zhao GX. 2002. 

Biflavanoids and derivatives thereof as antiviral agents. United States 

Patent 6399654 (June 4, 2002) 

Flavin MT, Lin YM, Zembower DE, Zhang H. 2001. Robustaflavone, 

intermediates and analogues and method for preparation there of and 

a method for synthesizing the same are provided. United States Patent 

6225481 (May 1, 2001) 

Freeman BA, Crapo JD. 1982. Biology of diseases, free radicals and tissue 

injury. Lab Invest 47: 412-426 

Gambhir SS, Goel RK, Dasgupta G. 1987. Anti-inflammatory and 

antiulcerogenic activity of amentoflavone. Indian J Med Res 85: 689- 

693 

Gao LL, Yin SL, Li ZL, Sha Y, Pei YH, Shi G, Jing YK, Hua HM. 2007. 

Three novel sterols isolated from Selaginella tamariscina with 

antiproliferative activity in leukemia cells. Planta Med 73: 1112-1115 

Garg AK, Kim JK, Owens TG, Ranwala AP, Choi YD, Kochian LV, Wu 

RJ. 2002. Trehalose accumulation in rice plants confers high 

tolerance levels to different abiotic stresses. Proc Natl Acad Sci 

U.S.A 99 (25): 15898-15903 

Gayathri V, Asha V, Subromaniam A. 2005. Preliminary studies on the 

immunomodulatory and antioxidant properties of Selaginella species. 

Indian J Pharmacol 37 (6): 381-385 

Gil B, Sanz MJ, Terencio MC, Gunasegaran R, Paya M, Alcaraz MJ. 

1997. Morelloflavone, a novel biflavonoid inhibitor of human 

secretory phospholipase A2 with antiinflammatory activity. Biochem 

Pharmacol 53: 733-740 

Gordana SC, Sonja MD, Jasna MC, Vesna TT. 2004). Antioxidant 

properties of marigold extracts. Food Res Int 37: 643-650 

Grba S, Oura E, Suomalainen H. 1975. On the formation of glycogen and 

trehalose in baker’s yeast. Eur J Appl Microbiol 2: 29-31 

Grennan AK. 2007. The role of trehalose biosynthesis in plants. Pl Physiol 

144: 3-5 

Grijalva V, Navarro S, Rhule A, Shepherd DM. 2004. The 

immunomodulatory effects of amentoflavone on cultured 

macrophages (RAW264.7) and dendritic cells (DC2.4). In: Abstracts 

of 21 st Annual Meeting. Pacific Northwest Association of 

Toxicologists. Oregon, September 17-19, 2004 

Guruvayoorappan C, Kuttan G. 2007. Effect of amentoflavone on the 

inhibition of pulmonary metastasis induced by B16F-10 melanoma 

cells in C57BL/6 mice. Integr Cancer Ther 6 (2): 185-197 

Guschina IA, Harwood JL, Smith M, Beckett RP. 2002. Abscisic acid 

modifies the changes in lipids brought about by water stress in the 

moss Atrichum androgynum. New Phytol 156: 255-264 

Han BH, Kang SS, Son KH. 2006. Composition for preventing or treating 

acute or chronic degenerative brain diseases including extract of 

Selaginella tamariscina Spring. World Intellectual Property 

Organization Publication Number: WO/2006/001664 (January 5, 

2006) 

Harada K, Rahayu M, Muzakkir A. 2002. Medicinal plants of Gunung 

Halimun National Park, West Java, Indonesia. Biodiversity 

Conservation Project JICA, PHPA & LIPI, Bogor 

Harborne JB, Baxter H. 1999. Handbook of natural flavonoids. John 

Wiley and Sons, Chichester 

Harborne JB, Williams CA. 2000. Advances in flavonoid research since 

1992. Phytochem 55: 481-504 

Harborne JB. 1980. Plant phenolics. In: Bella EA, Charlwood BV (eds) 

Encyclopedia of plant physiology. Springer, Berlin 

Harten JB, Eickmeier WG. 1986. Enzyme dynamics of the resurrection 

plant Selaginella lepidophylla (Hook. & Grev.) Spring during 

rehydration. Plant Physiol 82: 61-64. 

Harvey A. 2000. Strategies for discovering drugs from previously 

unexplored natural products. Drug Discov Today 5 (7): 294-300 

Havsteen B. 1983. Flavonoids, a class of natural products of high 

pharmacological potency. Biochem Pharmacol 32 (7): 1141-1148 

Havsteen BH. 2002. The biochemistry and medical significance of the 

flavonoids. Pharmacol Ther 96: 67-202 

Hayashi K, Hayashi T, Morita N. 1992. Mechanism of action of the 

antiherpes virus biflavone ginkgetin. Antimicrob Agents Chemother 

36: 1890-1893 

Hayashi K, Hayashi T, Otsuka H (et al.) 1997. Antiviral activity of 5,6,7- 

trimethoxyflavone and its potential of the antiherpes activity of 

acyclovir. J Antimicrob Chemother 39: 821-824 

Hikino H, Okuyama T, Jin H, Takemoto T. 1973. Screening of Japanese 

ferns for phytoecdysones. I. Chem Pharm Bull 21: 2292-2302 

Huang T-C. 2006. Flora of Taiwan. Vol. 6. 2nd ed. National University of 

Taiwan, Taipei 

Iturriaga G, Gaff DF, Zentella R. 2000. New desiccation-tolerant plants, 

including a grass, in the central highlands of Mexico, accumulate 

trehalose. Aust J Bot 48: 153-158 

Iwu MM, Igboko OA, Okunji CO, Tempesta MS. 1990. Antidiabetic and 

aldose reductase activities of biflavones of Garcinia kola. J Pharm 

Pharmacol 42: 290-292 

Iwu MM, Igboko OA, Onwuchekwa UA, Olaniyi CO. 1987. Evaluation of 

the hepatotoxic activity of the biflavonoids of G. kola seed. J 

Ethnopharmacol 21 (2): 127-138 

Iwu MM, Igboko OA. 1982. Flavonoids of Garcinia kola seeds. J Nat 

Prod 45: 650-651 

Iwu MM. 1985. Antihepatotoxic constituents of Garcinia kola seeds. 

Experientia 41: 699-670 

Iwu MM. 1999. Garcinia kola: a new adaptogen with remarkable 

immunostimulant, anti-infective and antinflammatory properties. 

Abstract of the International Conference on Ethnomedicine and Drug 

Discovery; Silver Spring, Maryland, USA, November 3-5, 1999 

Jang IC, Oh SJ, Seo JS, Choi WB, Song SI, Kim CH, Kim YS, Seo HS, 

Choi YD, Nahm BH, Kim JK. 2003. Expression of a bifunctional 

fusion of the Escherichia coli genes for trehalose-6-phosphate 

synthase and trehalose-6-phosphate phosphatase in transgenic rice 

plants increases trehalose accumulation and abiotic stress tolerance 

without stunting growth. Pl Physiol 131: 516-524 

Jansen L. 1993. Indigenous knowledge systems in health care: A 

traditional medical system in Indonesia. Thesis. University of 

Amsterdam, The Netherland 

Joo SS, Jang SK, Kim SG, Choi JS, Hwang KW, Lee DI. 2007. Anti-acne 

activity of Selaginella involvens extract and its non-antibiotic 

antimicrobial potential on Propionibacterium acnes. Phytother Res. 

www3.interscience.wiley.com/cgi-bin/abstract/116328562/ 

ABSTRACT 

Joy PP, Thomas J, Mathew S, Skaria BP. 1998. Medicinal plants. Kerala 

Agricultural University, Kerala 

Jung HJ, Park K, Lee IS, Kim HS, Yeo SH, Woo ER, Lee DG. 2007. S- 

Phase accumulation of Candida albicans by anticandidal effect of 

amentoflavone isolated from Selaginella tamariscina. Biol Pharm 

Bull 30 (10): 1969-1971 

Jung HJ, Sung WS, Yeo SH, Kim HS, Lee IS, Woo ER, Lee DG. 2006. 

Antifungal effect of amentoflavone derived from Selaginella 

tamariscina. Arch Pharmacal Res 29 (9): 746-751 

Kambuou RN. 1996. Papua New Guinea: country report. FAO 

International Technical Conference on Plant Genetic Resources, 

Leipzig, Germany, June 17-23, 1996 

Kamil M, Ilyas M, Rahman W, Hasaka N, Okigawa M, Kawano N. 1981. 

Taiwaniaflavone and its derivatives: a new series of biflavones from

56 

3 (1): 44-58, March 2011 

Taiwania cryptomerioides Hayata. J Chem Soc Perkin Transact 1: 

553-559 

Kandaswami C, Middleton E. 1994. Free radical scavenging and 

antioxidant activity of plant flavonoids. In: Armstrong D (ed). Free 

radicals in diagnostic medicine. Plenum Press. New York 

Kang DG, Yin MH, Oh H, Lee DH, Lee HS. 2004. Vasorelaxation by 

amentoflavone isolated from Selaginella tamariscina. Planta Med 70 

(8): 718-722 

Kang SS (et al) 1998. Korean Patent No. 137179 (April 25, 1998) 

Kang SS (et al) 2001. Korean Patent No. 267060 (March 2, 2001) 

Khare C. 2007. Indian medicinal plants - an illustrated dictionary. 

Springer. New York. 

Kidd G, Devorak J. 1994. Trehalose is a sweet target for agrobiotech. 

Biotechnol 12: 1328-1329 

Kim HK, Son KH, Chang HW, Kang SS, Kim HP. 1998. Amentoflavone, 

a plant biflavone: a new potential anti-inflammatory agent. Arch 

Pharmacol Res 21 (4): 406-410 

Kim J, Park EJ. 2002. Cytotoxic anticancer candidates from natural 

resources. Curr Med Chem Anti-Cancer Agents 2: 485-537 

Ko YJ, Park SH, Lee Y-H, Park BC, Hur JH, Min YD, Kim JK, Kim J-A . 

2007. Inhibitory effects of water extract of Selaginella involvens on 

the tube formation and invasion of human umbilical vein endothelial 

cells. Yakhak Hoeji 51 (1): 51-55 

Korall P, Kenrick P, Therrien JP. 1999. Phylogeny of Selaginellaceae: 

evaluation of generic/subgeneric relationships based on rbcL gene 

sequences. Intl J Plant Sci 160: 585-594 

Korall P, Kenrick P. 2002. Phylogenetic relationships in Selaginellaceae 

based on rbcL sequences. Amer J Bot 89: 506-517 

Korall P, Kenrick P. 2004. The phylogenetic history of Selaginellaceae 

based on DNA sequences from the plastid and nucleus: extreme 

substitution rates and rate heterogeneity. Mol Phylogenet Evol 31: 

852-864 

Kraus B. 2005. Der Einfluss von Johanniskraut (Hypericum perforatum 

L.) - Extrakt auf immunologische Prozesse in Mikroglia-Zellkulturen 

[Disertation]. Technischen Universität München, Munchen 

Krauze-Baranowska M, Wiwart M. 2002, Antifungal activity of 

biflavones from Taxus baccata and Ginkgo biloba. Z Naturforschung 

58: 65-69 

Kromhout D. 2001. Diet and cardiovascular diseases. J Nutr Health Aging 

5: 144-149 

Kunert O, Swamy RC, Kaiser M, Presser A, Buzzi S, Appa-Rao AVN, 

Schühly W. 2008. Antiplasmodial and leishmanicidal activity of 

biflavonoids from Indian Selaginella bryopteris. Phytochem Lett 1 (4) 

171-174 

Lale A, Herbert JM, Augereau JM, Billon M, Leconte M, Gleye JJ. 1996. 

J Nat Prod 59: 273-276 

Lee CW, Choi HJ, Kim HS, Kim DH, Chang IS, Moon HT, Lee SY, Oh 

WK, Woo ER. 2008. Biflavonoids isolated from Selaginella 

tamariscina regulate the expression of matrix metalloproteinase in 

human skin fibroblasts. Bioorg Med Chem 16 (2): 732-738 

Lee H, Lin JY. 1988. Antimutagenic activity of extracts from anticancer 

drugs in Chinese medicine. Mutation Res 204 (2): 229-234 

Lee HS, Oh WK, Kim BY, Ahn SC, Kang DO, Shin DI, Kim J, Mheen TI, 

Ahn JS. 1996. Inhibition of phospholipase Cγ1 activity by 

amentoflavone isolated from Selaginella tamariscina. Planta Med 62 

(4): 293-296 

Lee IS, Nishikawa A, Furukawa F, Kasahara K, Kim SU. 1999. Effects of 

Selaginella tamariscina on in vitro tumor cell growth, p53 expression, 

G1 arrest and in vivo gastric cell proliferation. Cancer Lett 144 (1): 

93-99 

Lee IS, Park SH, Rhee IJ. 1996. Molecular-based sensitivity of human 

leukemia cell line U937 to antineoplastic activity in a traditional 

medicinal plants (Selaginella tamariscina). J Fd Hyg Safety 11 (1): 

71-75 

Lee J, Choi Y, Woo ER, Lee DG. 2009a. Isocryptomerin, a novel 

membrane-active antifungal compound from Selaginella tamariscina. 

Biochem Biophys Res Comm 379 (3): 676-680 

Lee J, Choi Y, Woo ER, Lee DG. 2009b. Antibacterial and synergistic 

activity of isocryptomerin isolated from Selaginella tamariscina. J 

Microbiol Biotechnol 19 (2):204-207 

Lee JS, Lee MS, Oh WK, Sul JY. 2009c. Fatty acid synthase inhibition by 

amentoflavone induces apoptosis and antiproliferation in human 

breast cancer cells. Biol Pharm Bull 32 (8): 1427-1432 

Lin LC, Kuo YC, Chou CJ. 2000. Cytotoxic biflavonoids from Selaginella 

delicatula. J Nat Prod 63 (5): 627-630 

Lin R, Skaltsounis AL, Seguin E, Tillequin F, Koch M. 1994. Phenolic 

constituents of Selaginella doederleinii. Planta Med 60 (2): 168-170 

Lin RC, Peyroux J, Seguin E, Koch M. 1991. Hypertensive effect of 

glycosidic derivatives of hordenine isolated from Selaginella 

doederleinii Hieron and structural analogues in rats. Phytotherapy Res 

5 (4): 188-190 

Lin RC, Skaltsounis AL, Seguin E, Tillequin F, Koch M. 1990. The 

isolation and identification of a new lignanoside from Selaginella 

tamariscina (Beauv.) assay for anticancer-drug screening. J Natl 

Cancer Inst 82: 

Lin YM, Anderson H, Flavin MT, Pai YH, Mata-Greenwood E, 

Pengsuparp T, Pezzuto JM, Schinazi RF, Hughes SH, Chen FC. 1997. 

In vitro anti-HIV activity of biflavonoids isolated from Rhus 

succedanea and Garcinia multiflora. J Nat Prod 60: 884-888 

Lin YM, Flavin MT, Schure R, Chen FC, Sidwell R, Barnard DL, 

Huffman JH, Kern ER. 1999a. Antiviral activities of bioflavonoids. 

Planta Med 65: 120-125 

Lin YM, Flavin MT, Schure R, Zembower DE, Zhao GX. 1998. 

Biflavanoids and derivatives thereof as antiviral agents. United States 

Patent 5773462 (June 30, 1998) 

Lin YM, Flavin MT, Schure R, Zembower DE, Zhao GX. 1999b. 

Biflavanoids and derivatives there of as antiviral agents. United States 

Patent 5948918 (September 7, 1999) 

Lin YM, Zembower DE, Flavin MT, Schure R, Zhao GX. 2002. 

Biflavanoids and derivatives there of as antiviral agents. United States 

Patent 6339654 B1 (June 4, 2002) 

Liu MS, Chien CT, Lin TP. 2008. Constitutive components and induced 

gene expression are involved in the desiccation tolerance of 

Selaginella tamariscina. Plant Cell Physiol 49 (4): 653-663 

Lobstein-Guth A, Briancon-Scheid F, Victoire C, Haag-Berrurier M, 

Anton R. 1998. Isolation of amentoflavone from Ginkgo biloba. 

Planta Med 54: 555-556 

Locksley HD. 1973. Fortschritte der Chemie Organischer Naturstoffe 30: 

207-312 

Ma JF, Takahashi E. 2002. Soil, fertilizer, and silicon research in Japan. 

Elsevier, Amsterdam. 

Ma JF. 2004. Role of silicon in enhancing the resistence of plant to biotic 

and abiotic stresses. Soil Sci Pl Nutr 50: 11-18 

Ma LY, Wei F, Ma SC, Lin RC. 2002. Two new chromone glycosides 

from Selaginella uncinata. Chin Chem Lett 13 (8): 748-751 

Ma SC, But PP, Ooi VE, He YH, Lee SH, Lee SF, Lin RC. 2001. 

Antiviral amentoflavone from Selaginella sinensis. Biol Pharm Bull 

24 (3): 311-312 

Macheix JJ, Fleuriet A, Billot J. 1990. Fruit Phenolics. CRC Press, Boca 

Raton, FL 

Mamedov N. 2005. Adaptogenic, geriatric, stimulant and antidepressant 

plants of Russian Far East. J Cell Mol Biol 4: 71-75 

Markham KR, Gerger H, Jaggy H. 1992. Phytochemrstry 31: 009 

Martens S, Mithofer A. 2005. Flavones and flavone synthases. Phytochem 

66: 2399-2407 

Mathew PJ, Mathew D, Unnithan CM, Pushpangadan P. 1999. 

Ethnomedical information of some Pterydophytes of Kerala sector of 

Western Ghats. In: Sasikumar B, Krishnamurti B, Rema J, Ravindren 

PN, Peter KV (eds) Biodiversity conservation and utilization of 

spices, medicinal and aromatic plants. Indian Institute of Spices 

Research, Calicut 

Maxwell SR, Lip GY. 1997. Free radicals and antioxidants in 

cardiovascular disease. Br J Clin Pharmacol 44: 307-317 

Miao N, Tao H, Tong C, Xuan H, Zhang G. 1996. The Selaginella 

tamariscina (Beauv.) Spring complex in the treatment of 

experimental diabetes and its effect on blood rheology. Zhongguo 

Zhong Yao Za Zhi 21 (8): 493-495 

Middleton E, Kandaswami C, Theoharides TC. 2000. The effects of plant 

flavonoids on mammalian cells: implications for inflammation, heart 

disease, and cancer. Pharmacol Rev 52: 673-751 

Miki K, Nagai T, Nakamura T, Tuji M, Koyama K, Kinoshita K, Furuhata 

K, Yamada H, Takahashi K. 2008. Synthesis and evaluation of 

influenza virus sialidase inhibitory activity of hinokiflavone-sialic 

acid conjugates. Heterocycles 75 (4): (published online January 11, 

2008) 

Moltke LL von, Weemhoff JL, Bedir E, Khan IA, Harmatz JS, Goldman 

P, Greenblatt DJ. 2004. Inhibition of human cytochromes P450 by 

components of Ginkgo biloba. J Pharm Pharmacol 56: 1039-1044 

Mora A, Paya M, Rios JL, Alcaraz MJ. 1990. Structure-activity 

relationships of polymethoxyflavones and other flavonoids as


inhibitors of non-enzymic lipid peroxidation. Biochem Pharmacol 40: 

793-797 

Mueller J, Boller T, Wiemken A. 1995. Trehalose and trehalase in plants: 

recent developments. Plant Sci 112: 1-9 

Muller D, Fleury JP. 1991. A new strategy for the synthesis of 

biflavonoids via arylboronic acids. Tetrahedron Lett 32: 2229-2232 

Muller J, Aeschbacher RA, Wingler A, Boller T, Wiemken A. 2001. 

Trehalose and trehalase in Arabidopsis. Pl Physiol 125: 1086-1093, 

Na M, Kim KA, Oh H, Kim BY, Oh WK, Ahn JS. 2007. Protein tyrosine 

phosphatase 1B inhibitory activity of amentoflavone and its cellular 

effect on tyrosine phosphorylation of insulin receptors. Biol Pharm 

Bull 30 (2): 379-381 

Nahrstedt A, Butterweck V. 1997. Biologically active and other chemical 

constituents of the herb of Hypericum perforatum L. 

Pharmacopsychiatry 30: 129-134 

Nasution RE. 1993. Proceeding of the 2 nd National Seminary and 

Workshop on Ethnobotany. Book 1: Medicinal Plant. Yogyakarta, 24- 

25 January 1993. Puslitbang Biologi LIPI, Fakultas Biologi UGM & 

Ikatan Pustakawan Indonesia (IPI), Jakarta 

Oliveira MCC de, Carvalho MG de, da Silva CJ, Werle AA. 2002. New 

biflavonoid and other constituents from Luxemburgia nobilis 

(EICHL). J Brazilian Chem Soc 13 (1): 119-123 

Otero R, Núñez V Barona, J, Fonnegra R, Jiménez SL, Osorio RG, 

Saldarriaga M, Díaz A. 2000. Snakebites and ethnobotany in the 

northwest region of Colombia. Part III: neutralization of the 

haemorrhagic effect of Bothrops atrox venom. J Ethnopharmacol 73 

(1-2): 233-241 

Paiva CLA, Panek AD. 1996. Biotechnological applications of the 

disaccharide trehalose. Biotechnol Ann Rev 2: 293-314 

Pan KY, Lin JL, Chen JS. 2001. Severe reversible bone marrow 

suppression induced by Selaginella doederleinii. J Clin Toxicol 39 

(6): 637-639 

Pandey S, Khan AA, Shankar K, Singh N. 1993. An experimental study 

on the anti-stress and antioxidant activity of Selaginnella bryopteris. J 

Biol Chem Res 12: 128-129 

Perez S, Perez RM, Perez C, Zavala MA, Vargas R. 1994. Inhibitory 

activity of 3-methylenhydroxy-5-methoxy-2,4-dihydroxy 

tetrahydrofurane isolated from Selaginella lepidophylla on smooth 

muscle of Wistar rat. Pharm Acta Helv 69 (3): 149-152 

Perruchon S. 2004. Synthese und struktur-aktivitäts-beziehungen von 

flavonoiden. [Dissertation]. Technischen Universität Darmstadt, 

Darmstadt 

Peter KV (ed) 2004. Handbook of herbs and spices. Vol 2. CRC Press, 

Boca Raton 

Platt KA, Oliver MJ and Thomson WW. 1994). Membranes and 

organelles of dehydrated Selaginella and Tortula retain their normal 

configuration and structural integrity; Freeze facture evidence. 

Protoplasma 178: 57-65 

Pokharel YR, JW Yang, JY Kim, HW Ohb, HG Jeong, ER Woo, KW 

Kang. 2006. Potent inhibition of the inductions of inducible nitric 

oxide synthase and cyclooxygenase-2 by taiwaniaflavone. Nitric 

Oxide 15 (3): 217-225 

Popper ZA, Sadler IH, Fry SC. 2001. 3-O-Methyl-D-galactose residues in 

lycophyte primary cell walls. Phytochem 57 (5): 711-719 

Porter LJ. 1994. Flavans and proanthocyanidins. In: Harborne, JB (ed). 

The flavonoids, advances in research since 1986. Chapman and Hall, 

New York 

Rahman M, Riaz M, Desai UR. 2007. Synthesis of biologically relevant 

biflavanoids-A review. Chem Biodiv 4: 2495-2527 

Rayne S, Mazza G. 2007. Biological activities of extracts from sumac 

(Rhus spp.): a review. National Bioproducts and Bioprocesses 

Program, Pacific Agri-Food Research Centre, Agriculture and Agri- 

Food Canada, Summerland BC 

Richardson PM. 1989. Flavonoids of the ‘Fern Allies’. Biochem Sys Ecol 

17 (2): 155-160 

Richmond KE, Sussman M. 2003. Got silicon? The non-essential 

beneficial plant nutrient. Curr Op Pl Biol 6: 268-272 

Robards K, Antolovich M. 1997. Analytical chemistry of fruit 

bioflavonoids. Analyst 122: 11R-34R 

Roberts WM, Tovey KC. 1969. Trehalase activity in Selaginella martensii. 

Arch Biochem Biophys 133: 408-412 

Rojas A, Bah M, Rojas JI, Serrano V, Pacheco S. 1999. Spasmolytic 

activity of some plants used by the Otomi Indians of Quéretaro 

(México) for the treatment of gastrointestinal disorders. Phytomed 6 

(5): 367-371 

Rose WM, Creighton MO, Stewart DHPJ, Sanwal M, Trevithick GR. 

1982. In vivo effects of vitamin E on cataractogenesis in diabetic rats. 

Can J Ophtalmol 17: 61-66 

Roser B. 1991. Trehalose a new approach to premium dry food. Trends 

Food Sci Technol 7: 166-169 

Sah NK, Singh SNP, Sahdev S, Banerji S, Jha V, Khan Z, Hasnain SE. 

2005. Indian herb sanjeevani (Selaginella bryopteris) can promote 

growth and protect against heat shock and apoptotic activities of ultra 

violet and oxidative stress. J Biosci 30 (4): 499-505 

Sah NK, Singh SNP, Sahdev S, Banerji S, Jha V, Khan Z, Hasnain SE. 

2005. Indian herb 'Sanjeevani' (Selaginella bryopteris) can promote 

growth and protect against heat shock and apoptotic activities of ultra 

violet and oxidative stress. J Biosci 30 (4): 499-505. 

Schiraldi C, Di Lernia I, De Rosa M. 2002. Trehalose production: 

exploiting novel approaches. Trends Biotechnol 20 (10): 420-425 

Schluepmann H, Pellny T, van Dijken A, Smeeekens S, Paul M. 2003. 

Trehalose 6-phosphate is indispensable for carbohydrate utilization 

and growth in Arabidopsis thaliana. Proc Natl Acad Sci USA 100: 

6849-6854 

Seigler DS. 1998. Plant secondary metabolism. Kluwer, Dodrecht 

Sequiera KM. 1998. Diversity, systematics, distribution, and taxonomy of 

epiphytic pterydophytes of Kerala part of Western Ghats, South India. 

Indian Fern J 15: 106-130 

Setyawan AD, Darusman LK. 2008. Review: Biflavonoid compounds of 

Selaginella Pal. Beauv. and its benefit. Biodiversitas 9 (1): 64-81 

Setyawan AD. 2008. Species richness and geographical distribution of 

Malesia Selaginella. 8th Seminary and Congress of Indonesian Plant 

Taxonomy Association (“PTTI”), Cibinong Science Center, Bogor- 

Indonesia, 21-23 October 2008 

Setyawan AD. 2009. Traditionally utilization of Selaginella; field research 

and literature review. Nus Biosci 1: 146-155. 

Shi S, Zhou H, Zhang Y, Huang K. 2008). Hyphenated HSCCC-DPPH for 

rapid preparative isolation and screening of antioxidants from 

Selaginella moellendorffii. Chromatographia 68: 173-178 

Silva GL, Chai H, Gupta MP, Farnsworth NR, Cordell GA, Pezzuto JM, 

Beecher CW, Kinghorn AD. 1995. Cytotoxic biflavonoids from 

Selaginella willdenowii. Phytochem 40 (1): 129-134 

Silva KL da, dos Santos AR, Mattos PE, Yunes RA, Delle-Monache F, 

Cechinel-Filho V. 2001. Chemical composition and analgesic activity 

of Calophyllum brasiliense leaves. Therapie 56: 431-434 

Singer MA, Lindquist S. 1998. Thermotolerance in Saccharomyces 

cerevisiae: the Yin and Yang of trehalose. Trends Biotechnol 16 (11): 

460-468 

Smith PM. 1976. The chemotaxonomy of plants. Edward Arnold, London 

Soedibyo M. 1989. Philosophy of herbal medicine as the point of 

development of traditional medicine. Symposium on Cosmetics and 

Traditional Medicine II. Department of Health,, Jakarta [Indonesia] 

Soedibyo M. 1990. Javanese traditional medicine. Congress on Traditional 

Medicine and Medicinal Plants. Denpasar, Bali, Indonesia, October 

15-17, 1990 [Indonesia] 

Sofowora A. 1982. Medicinal plants and traditional medicine in Africa. 

Spectrum Books Ltd. Ibadan, Nigeria 

Su Y, Sun CM, Chuang HH, Chang PT. 2000. Studies on the cytotoxic 

mechanisms of ginkgetin in a human ovarian adenocarcinoma cell 

line. Naunyn Schmiedebergs Arch Pharmacol 362 (1): 82-90 

Sun CM, Syu WJ, Huang YT, Chen CC, Ou JC. 1997. Selective 

cytotoxicity of ginkgetin from Selaginella moellendorffii. J Nat Prod 

60 (4): 382-384 

Sun DM, Luo WH, Li ZY. 2006. Determination of amentoflavone in 11 

species of Selaginella medicinal material by HPLC. Zhong-Yao-Cai 

29 (1): 26-28 

Sutarjadi. 1990. Research and development Indonesian jamu science. 

Symposium to Increasing Image and Benefit of Jamu and Traditional 

Drugs, and the Security to Human Body Healt. Universitas Airlangga, 

Surabaya 

Takemoto T, Ogawa S, Nishimoto N, Arihara S, Bue K. 1967. Insect 

moulting activity of crude drugs and plants (1). Yakugaku Zasshi 87: 

1414-1418. 

Tan WJ, Xu JC, Li L, Chen KL. 2009. Bioactive compounds of inhibiting 

xanthine oxidase from Selaginella labordei. Nat Prod Res 23 (4): 

393-398. 

Teramoto N, Sachinvala ND, Shibata M. 2008. Trehalose and trehalosebased 

polymers for environmentally benign, biocompatible and 

bioactive materials. Molecules 13 (8): 1773-816

58 

3 (1): 44-58, March 2011 

Tolonen A. 2003. Analisys of secondary metabolites in plant and cell 

culture tissue of Hypericum perforatum L. and Rhodiola rosea L. 

[Dissertation]. Faculty of Science, University of Oulu, Oulu 

Uluk A, Sudana M, Wollenberg E. 2001. Dependence of Dayaks society 

to forest around Kayan Mentarang National Park. Cifor, Bogor 

Umezawa T. 2003a. Diversity in lignan biosynthesis. Phytochem Rev 

2: 371-390. 

Umezawa T. 2003b. Phylogenetic distribution of lignan producing 

plants. Wood Res 90: 27-110. 

van Andel TR. 2000. Non-timber forest products of the north-west district 

of Guyana. part I & part II (a field guide). Tropenbos-Guyana 

Programme, Georgetown, Guyana 

van Dijck P, Mascorro-Gallardo JO, de Bus M, Royackers K, Iturriaga G, 

Thevelein JM. 2002. Truncation of Arabidopsis thaliana and 

Selaginella lepidophylla trehalose-6-phosphate synthase unlocks high 

catalytic activity and supports high trehalose levels on expression in 

yeast. Biochem J 366: 63-71 

Wang CC, Lu SH, Chen J, Wei H. 1963. The effect of certain wild plants 

and fertilizer in the killing of the genus Leptospira of orde 

Spirochetes. Chung-hua I-hsueh Tsa-chih 49 (6): 393-396 

Wang YH, Long CL, Yang FM, Wang X, Sun QY, Wang HS, Shi 

YN, Tang GH. 2009. Pyrrolidinoindoline alkaloids from Selaginella 

moellendorfii. J Nat rod. 2009 May 7. 

Wang YZ, Chen H, Zheng XK, Feng WS. 2007. A new sesquilignan from 

Selaginella sinensis (Desv.) Spring. Chin Chem Lett 18 (10): 1224- 

1226 

Warintek. 2002. CD-ROM mencerdaskan bangsa seri kedua. 

http://ftp.ui.edu/bebas/v12/data/top.htm 

Waterman PG, Hussain RA. 1983). Systematic significance of xanthones, 

benzophenones and biflavonoids in Garcinia. Biochem Sys Ecol 11: 

21-30. 

Weniger B, Vonthron-Senecheau C, Kaiser M, Brun R, Anton R. 2006). 

Comparative antiplasmodial, leishmanicidal and antitrypanosomal 

activities of several biflavonoids. Phytomed 13: 176-180 

White E, Towers GHN. 1967. Comparative biochemistry of the Lycopods. 

Phytochem 6 (5): 663-667 

Wiart C. 2007. Anti-inflammatory plants. In: Ethnopharmacology of 

medicinal plants Asia and the Pacific. Humana Press, Berlin 

Wijayakusuma HM. 2004. To contend and to prevent lung cancer as 

naturally. http://cybermed.cbn.net.id 

Winter WP de, Jansen PCM. 2003. Selaginella Pal. Beauv. In: Winter WP 

de, Amoroso VB (eds) Plant resources of south-east Asia No. 15 (2) 

Cryptogams: fern and fern allies. Bakhuys Publisher, Leiden 

Woo ER, Lee JY, Cho IJ, Kim SG, Kang KW. 2005. Amentoflavone 

inhibits the induction of nitric oxide synthase by inhibiting NF-κB 

activation in macrophages. Pharmacol Res 51 (6): 539-546 

Woo ER, Pokharel YR, Yang JW, Lee SY, Kang KW. 2006. Inhibition of 

nuclear factor-κB activation by 2',8''-biapigenin. Biol Pharm Bull 29 

(5): 976-980 

Wu W, Pang Y, Shen GA, Lu J, Lin J, Wang J, Sun X, Tang K. 2006. 

Molecular cloning, characterization and expression of a novel 

trehalose-6-phosphate synthase homologue from Ginkgo biloba. J 

Biochem Mol Biol 39 (2): 158-166 

Yamada H, Nagai T, Takahashi K. 2007. Anti-influenza virus compound 

comprising biflavonoid-sialic acid glycoside. United States Patent & 

Trademark Office (USPTO) No. 20070004649 (January 4. 2007) 

Yamaguchi LF, Vassao DG, Kato MJ, Mascio P. 2005. Biflavonoids from 

Brazilian pine Araucaria angustifolia as potentials protective agents 

against DNA damage and lipoperoxidation. Phytochem 66: 2238- 

2247 

Yang JW, Pokharel YR, Kim MR, Woo ER, Choi HK, Kang KW. 2006. 

Inhibition of inducible nitric oxide synthase by sumaflavone isolated 

from Selaginella tamariscina. J Ethnopharmacol 105 (1-2): 107-113 

Yang SF, Chu SC, Liu SJ, Chen YC, Chang YZ, Hsieh YS. 2007. 

Antimetastatic activities of Selaginella tamariscina (Beauv.) on lung 

cancer cells in vitro and in vivo. J Ethnopharmacol 110 (3): 483-489 

Yen KY, Yang LL, Okuyama T, Hikino H, Takemoto T. 1974. Screening 

of Formosan ferns for phytoecdysones. Chem Pharm Bull 22: 805- 

808 

Yin MH, Kang DG, Choi DH, Kwon TO, Lee HS. 2005. Screening of 

vasorelaxant activity of some medicinal plants used in Oriental 

medicines. J Ethnopharmacol 99 (1): 113-117 

Yuan Y, Wang B, Chen L, Luo H, Fisher D, Sutherland IA, Wei Y. 2008. 

How to realize the linear scale-up process for rapid purification using 

high-performance counter-current chromatography. J Chromatogr A 

1194 (2): 192-198 

Zentella R, Mascorro-Gallardo JO, van Dijck P, Folch-Mallol J, Bonini B, 

van Vaeck C, Gaxiola R, Covarrubias AA, Nieto-Sotelo J, Thevelein 

JM, Iturriaga G. 1999. A Selaginella lepidophylla trehalose-6- 

phosphate synthase complements growth and stress-tolerance defects 

in a yeast tps1 mutant. Pl Physiol 119: 1473-1482 

Zhang LP, Liang YM, Wei XC, Cheng DL. 2007. A new unusual natural 

pigment from Selaginella sinensis and its noticeable physicochemical 

properties. J Org Chem 72: 3921-3924 

Zheng JX, Wang NL, Gao H, Liu HW, Chen HF, Fan M, Yao XS. 2008a) 

A new flavonoid with a benzoic acid substituent from Selaginella 

uncinata. Chin Chem Lett 19 (9): 1093-1095 

Zheng JX, Zhou TC, Xu K, Li LN. 2006. Effect of Selaginella combined 

with radiotherapy on nasopharyngeal carcinoma. Nan Fang Yi Ke Da 

Xue Xue Bao 26 (2): 247-248 

Zheng X, Du J, Xu Y, Zhu B, Liao D. 2007. A new steroid from 

Selaginella pulvinata. Fitoterapia 78 (7-8): 598-599 

Zheng XK, Li KK, Wang YZ, Feng WS. 2008b. A new 

dihydrobenzofuran lignanoside fro om Selaginella moellendorffii 

Hieron. Chin Chem Lett 19 (1): 79-81 

Zheng XK, Shi SP, Bi YF, Feng WF, Wang JF, Niu JZ. 2004. The 

isolation and identification of a new lignanoside from Selaginella 

tamariscina (Beauv.) Spring. Yao Xue Xue Bao 39 (9): 719-721 

Zhou JH. 2002. Herbal caffeine replacement composition and food 

products incorporating same. United States Patent 6416806 (July 9, 

2002) 

Zhu TM, Chen KL, Zhou WB. 2008. A new flavone glycoside from 

Selaginella moellendorffii Hieron. Chin Chem Lett 19 (12): 1456- 

1458

THIS PAGE INTENTIONALLY LEFT BLANK


ISSN 2087‐3940 (PRINT) | ISSN 2087‐3956 (ELECTRONIC) 

I S E A Journal o f B i o l o g i c a l S c i e n c e s 

Optimization of DNA extraction of physic nut (Jatropha curcas) by selecting the 

appropriate leaf 

EDI PRAYITNO, EINSTIVINA NURYANDANI 

Characterisation of taro (Colocasia esculenta) based on morphological and isozymic 

patterns markers 

TRIMANTO, SAJIDAN, SUGIYARTO 

Study on floristic and plant species diversity of the Lebanon oak site (Quercus libani) in 

Chenareh, Marivan, Kordestan Province, western Iran 

HASSAN POURBABAEI, SHIVA ZANDI NAVGRAN 

Evaluation structural diversity of Carpinus betulus stand in Golestan Province, North of 

Iran 

VAHAB SOHRABI, RAMIN RAHMANI, SHAHROKH JABBARI, HADI MOAYERI 

Microanatomy alteration of gills and kidneys in freshwater mussel (Anodonta woodiana) 

due to cadmium exposure 

FUAD FITRIAWAN, SUTARNO, SUNARTO 

Site suitability to tourist use or management programs South Marsa Alam, Red Sea, Egypt 

MOHAMMED SHOKRY AHMED AMMAR, MOHAMMED HASSANEIN, HASHEM ABBAS 

MADKOUR, AMRO ABD‐ELHAMID ABD‐ELGAWAD 

Review: Natural products from Genus Selaginella (Selaginellaceae) 

AHMAD DWI SETYAWAN 

1‐6 

7‐14 

15‐22 

23‐27 

28‐35 

36‐43 

44‐58 

Published three times in one year 

PRINTED IN INDONESIA 

ISSN 2087‐3940 (print) 

ISSN 2087‐3956 (electronic)

ISSN 2087-3940 (PRINT) | ISSN 2087-3956 ... - Biodiversitas

Create successful ePaper yourself

Delete template?

Save as template?