Pak. J. Bot., 38(4): 901-915, 2006. POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE ANJUM PERVEEN AND MUHAMMAD QAISER Department of Botany, University of Karachi, Karachi - 75270, Pakistan Abstract Pollen morphology of 74 species belonging to 23 genera of the family Caryophyllaceae from Pakistan has been examined by light and scanning electron microscope. Pollen grains are usually radially symmetrical, apolar rarely isopolar, pantoporate rarely 3-10 colpate, spheroidal to prolate-spheroidal to sub-prolate occasionally prolate. Sexine thicker or thinner than nexine. Tectum spinulose-punctuate or scabrate-punctate often reticulate or reticulate-scabrate. On the basis of apertural type three distinct pollen types viz., Silene indica-type, Spergula arvensis-type and Stellaria media-type are recognized. Introduction Caryophyllaceae is a family of about 87 genera and 2300 species (Mabberley, 1987) occurring mainly in north temperate or warm temperate regions. In Pakistan it is represented by 26 genera and 110 species (Ghazanfar & Nasir, 1986). Plants are herbs or rarely subshrubs, leaves are simple, nearly always opposite and decussate, estipulate or sometimes with scarious stipules, flowers are actinomorphic and mostly bisexual rarely absent, stamens are in one or two whorls, gynoecium consists of a single compound pistil of 2-5 carpels, usually with an equal number of styles and superior ovary, fruit mostly capsule. Family includes several ornamental plants like Arenaria (sandwort), Cerastium (mouse-ear), Stellaria (chick weed), Silene (catch fly), Dianthus (carnation) and Gypsophila (baby breath). Pollen morphology of the family Caryophyllaceae has been examined by various workers such as, Cranwell (1962), Nair (1962), Erdtman (1952), Dimbleby (1974) Kuprianova & Alyoshina (1978), Ghanzafar (1984) and Bittrich (1994). Chand (1963) studied pollen morphology of some Scandinavian Caryophyllaceae. Nowicke (1976) examined pollen morphology of the family Caryophyllaceae while conductivity of palynological studies of the order Centrospermae. Moore & Webb (1978) also examined pollen morphology of few species of the family Caryophyllaceae. Pollen of Arenaria L., and Stellari L., species from Argentine have been examined by Volponi (1987). However, the most comprehensive study on pollen morphology of the family Caryophyllaceae is that of Punt & Hoen (1995). There are no reports on pollen morphology of the family Caryophyllaceae from Pakistan. Present investigations are based on the pollen morphology of 74 species representing 23 genera of the family Caryophyllaceae by light and scanning electron microscope. Materials and Methods Pollen samples were obtained from Karachi University Herbarium (KUH) or collected from the field. The list of voucher specimens is deposited in KUH. The pollen grains were prepared for light (LM) and scanning microscopy (SEM) by the standard 902 ANJUM PERVEEN & MUHAMMAD QAISER methods described by Erdtman (1952). For light microscopy, the pollen grains were mounted in unstained glycerin jelly and observations were made with a Nikon Type-2 microscope, under (E40, 0.65) and oil immersion (E100, 1.25), using 10x eye piece. For SEM studies, pollen grains suspended in a drop of water were directly transferred with a fine pipette to a metallic stub using double sided cello tape and coated with gold in a sputtering chamber (Ion-sputter JFC-1100). Coating was restricted to 150 A. The S.E.M examination was carried out on a Jeol microscope JSM-2. The measurements are based on 15-20 readings from each specimen. Pollen diameter, polar axis (P) and equatorial diameter (E), aperture size, apocolpium, mesocolpium and exine thickness were measured (Tables 1-3). The terminology used is in accordance with Erdtman (1952), Kremp (1965), Faegri & Iversen (1964) and Walker & Doyle (1975). General pollen characters of the family Caryophyllaceae Pollen grains are usually radially symmetrical, apolar rarely isopolar, pantoporate rarely 3-10 colpate spheroidal or prolate-spheroidal-sub-prolate to rarely prolate, Sexine thicker or thinner than nexine. Tectum spinulose-punctate or scabrate-punctate often reticulate or reticulate-scabrate. On the basis of apertural type three distinct pollen types are recognized viz., Silene indica-type, Spergula arvensis-type and Stellaria media-type. Key to the pollen types + Pollen grains colpate ……………………………………….. Spergula arvensis-type - Pollen grains porate ………………………………………………………………... 2 + Tectum reticulate ……………………...………………………… Silene indica-type - Tectum spinulose–punctate rarely scabrate-punctate ................. Stellaria media-type Pollen type: Silene indica - type (Fig. 1A & D) Pollen class: Porate P/E ratio: 1.00 Shape: Spheroidal Apertures: Pore more or less circular, operculate, annulate Exine: Sexine thicker than nexine. Ornamentation: Reticulate or reticulate – scabrate. Measurements: Size: Length = (32.5-) 35.12 ± 0.78 (-37.5) µm and breadth (35.1) 37± 0.82 (40) µm, pore (7.5–) 10.11 ± 0.42 (12.5) µm in diameter. Exine 2.5 µm thick, sexine thicker than nexine. Tectum reticulate or reticulate-scabrate. Species included: Cucubalus baccifer L., Silene gonosperma (Rupr.) Bocquet, S. indica Roxb.ex Otth, S. brahuica Boiss. Key to the species + Tectum reticulate–scabrate .………. Group-I (Cucubalus baccifer, Silene brahuica) - Tectum simply reticulate …..…………….. Group-II (Silene gonosperma, S. indica) POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE 903 904 ANJUM PERVEEN & MUHAMMAD QAISER POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE 905 906 ANJUM PERVEEN & MUHAMMAD QAISER POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE 907 908 ANJUM PERVEEN & MUHAMMAD QAISER Fig. 1. Scanning micrographs: Silene indica: A, Pollen grain; B, Exine pattern. Cucubalus baccifer: C, Pollen grain; D, Exine pattern. Gypsophila cerastioides: E, Pollen grain, F, Exine pattern. POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE 909 Pollen type: Spergula arvensis-type - (Fig. 2 A-F) Pollen class: Tricolpate, 4-10 colpate. P/E ratio: 0.85-1.73 Shape: Prolate-spheroidal–sub-prolate or prolate rarely spheroidal Apertures: Colpus long sunken with acute ends. Exine: sexine thicker or thinner than or as thick as nexine. Ornamentation: Scabrate-punctate or spinulose-punctate. Measurements: Size: Polar axis P = 21 (32 ± 1.2) 44, and equatorial diameter E =19 (28 ± 2.1) 37 µm. Colpi 16.61 (34 ± 1.4) 25 µm long. Exine 0.9-2.5 µm thick, sexine as thick as nexine. Tectum scabrate-punctate. Species included: Spergula fallax (Lowe) E.H.L. Kraus., S. arvensis L., Spergularia media (L.) Presl., S. diandra (Guss) Heldr. Sart, Spergula marina (L.) Griseb., Polycarpaea carymbosa (L.) Lam. , P. spicata Wight & Arn., Telephium imperati L. Key to the species 1 + Pollen grains 14-18 µm in polar length ……………………………………………... ……………………………………… Group-I (Polycarpaea carymbosa, P. spicata) - Pollen grains more than 18 µm in polar length ………………………………….… 2 2 + Pollen grains prolate or sub-prolate ……………………………………………….. 3 - Pollen grains prolate-spheroidal or spheroidal ……………………………………. 4 3 + Pollen grains subprolate ...………………………….………….…… Spergula fallax - Pollen grains prolate …………………………………………… Telephium imperati 4 + Pollen grains spheroidal ……………………………………..…………… S. diandra - Pollen grains prolate-spheroidal …………………………………………...……… 5 5 + Pollen grains 3-colpate .…………………………………………. Spergularia media - Pollen grains 3-10 colpate...……Group-II (Spergula arvensis, Spergularia marina) Pollen type: Stellaria media- type (Fig. 1E & F; Figs. 3 and 4A-F) Pollen class: Pantoporate P/E ratio: 1.00 Shape: Spheroidal Apertures: Pore more or less circular, operculate, annulate, or non-operculate. Exine: Sexine thicker than nexine. Ornamentation: Spinulose punctuate or scabrate-punctate. Measurements: Size: Length = (32.5-) 37.12 ± 0.78 (-37.5) µm and Breadth (35.1) 37± 0.82 (40) µm, pore (7.5–) 10.11 ± 0.42 (12.5) µm in diameter. Exine 2.5 µm thick, sexine thicker than nexine. Tectum Spinulose-punctate Species included: Acanthophyllum laxiflorum Boiss, A. squarrosum Bioss., A. sordidum Bunge ex Boiss., Arenaria serpyllifolia L., A. leptoclados (Reichb.) Guss., Arenaria. neelgherrensis Wight & Arn., A. orbiculata Royle ex Edgew., A. giiffithii Boiss., Cerastium cerastioides (L.) Britton, C. dichotomum L., C. glomeratum Thuill, C. thomsoni Hook.f., C. dahuricum Fisch., C. pusillum Ser., Dianthus anatolicus Boiss, D. crinitus Sm, D.orientalis Adams., D. angulatus Royle ex Benth., D. cachmericus Edgew., 910 ANJUM PERVEEN & MUHAMMAD QAISER Fig. 2. Scanning micrographs: Spergula fallax: A, Polar view; B, Exine pattern. Spergularia diandra: C, Polar view; D, Exine pattern. S. media: E, Equatorial view; F, Polar view. Scale bar = A, B, D & F = 10; A, C = 1 µm. POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE 911 Fig. 3. Scanning micrographs: Arenaria neelgherrensis: A, Pollen grains; B, Exine pattern. Minuartia biflora: C, Pollen grain. Minuartia hybrida: D, Exine pattern. Lepyrodiclis tenera: E, Pollen grain. L. stellarioides: F, Exine pattern. Scale bar = A-F = 10 µm. 912 ANJUM PERVEEN & MUHAMMAD QAISER Fig. 4. Scanning micrographs: Stellaria uliginosa: A, Pollen grains; B, Exine pattern. Stellaria blatteri: C, Pollen grain. D, Exine pattern. Holosteum umbellatum: E, Pollen grain; F, Exine pattern. Scale bar = A-F = 10 µm. POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE 913 D. jacquemontii Edgew ex Hook.f., Gypsophila bellidifolia Boiss., G. floribunda (Kar. & Kir.) Turcz, G. muralis (Klok.) Schischkin, G. cerastioides D. Don, G. alsinoides Bunge, G. sedifolia Kurz., Holosteum umbellatum L., Lepyrodiclis holosteoides (C.A. Meyer) Fenzl ex. F. & M., Lychnis coronaria (L.) Desr., Lepyrodiclis stellarioides F. & M., L. tenera Boiss., Minuartia kashmirica (Edgew.) Mattf., M. hybrida (Vill.) Schischkin, M. biflora (L.) Schinz-Thell, Myosoton aquaticum (L.) Moench, Petrorhagia alpina (Habl.) Ball & Heywood, Pseudostellaria heterantha (Maxim.) Pax, Sagina saginoides (L.) Karst., Saponaria subrosularis Rech.f., Silene kunawarensis Benth., S. viscosa (L.) Pers., Silene tenuis Willd., S. falconeriana Benth., Silene citrina Boiss., S. vulgaris (Moench) Garcke. S. moorcroftiana Wall. ex Benth., S. longisepala, S. nana Kar. & Kir., S. arenosa C. Koch, S. conoidea L., S. coelirosea (L.) Godron, Stellaria media (L.) Vill., S. monosperma Buch.–Ham. ex Don, S. decumbens Edgew., S. persica Boiss., S. subumbellata Edgew. ex Edgew & Hook.f., S. uliginosa Mur., S. kotschyana Fenzl ex Boiss, S. blatteri Mattf., Vaccaria hispanica (Miller) Rauchert and Velezia rigida L. Key to the species and species group + Pollen grains spinulose-punctuate …………………………………….……. Group-I Acanthophyllum laxiflorum Boiss, A. squarrosum Bioss., A. sordidum Bunge ex Boiss., Arenaria serpyllifolia L., A. leptoclados (Reichb.) Guss., Arenaria. neelgherrensis Wight & Arn., A. orbiculataI Royle ex Edgew., A. giiffithii Boiss., Cerastiumcerastioides (L.) Britton, C. dichotomum L., C. glomeratum Thuill, C. dahuricum Fisch., C. pusillum Ser., Dianthus anatolicus Boiss, D. crinitus Sm, D. orientalis Adams., D.angulatus Royle ex Benth., D. cachmericus Edgew, D. jacquemontii Edgew ex Hook.f., Gypsophila bellidifolia Boiss., G. floribunda (Kar. & Kir.) Turcz, G. muralis (Klok.) Schischkin, G. cerastioides D. Don, G. alsinoides Bunge, G. sedifolia Kurz., Holosteum, umbellatum L., Lepyrodiclis holosteoidu (C.A. Meyer) Fenzl ex. F. & M., Lychnis coronaria (L.) Desr., Lepyrodiclis stellarioides F. & M., L. tenera Boiss., Minuartia kashmirica (Edgew.) Mattf. Minuartia hybrida (Vill.) Schischkin, M. biflora (L.) Schinz-Thell Myosoton aquaticum (L.) Moench, Petrorhagia alpina (Habl.) Ball & Heywood, Pseudostellaria heterantha (Maxim.) Pax Sagina saginoides (L.) Karst., Saponaria subrosularis Rech.f., S. viscosa (L.) Pers., Silene tenuis Willd., S. falconeriana Benth, Silene citrina Boiss. S. vulgaris (Moench) Garcke, S. arenosa C. Koch, S. conoidea L., S. coelirosea (L.) Godron, Stellaria media (L.) Vill., S. monosperma Buch.–Ham. ex Don, S. decumbens Edgew., S. persica Boiss., S. subumbellata Edgew. ex Edgew & Hook.f., S. uliginosa Mur., S. kotschyana Fenzl ex Boiss, Velezia rigida L. - Pollen grains scabrate- punctuate …………………………………………. Group-II Stellaria blatteri Matts, Cerastium thomsonii Hook.f., Silene kunawarensis Benth., S. moorcroftiana Wall.ex Benth.,S. longisepala, S. nana Kar. & Kir., S. conoidea L. Discussion Caryophyllaceae is a stenopalynous family. Pollen grains are usually radially symmetrical, apolar rarely isopolar, pantoporate rarely 3-10 colpate, spheroidal or prolate-spheroidal-sub-prolate to rarely prolate. Tectum spinulose-punctate or scabratepunctate often reticulate-reticulate to scabrate. However, considerable variation in 914 ANJUM PERVEEN & MUHAMMAD QAISER apertural types and exine pattern has been observed. On the basis these characters three distinct pollen types are recognized viz., pollen type-I: Silene indica is recognized by having porate pollen with reticulate or reticulate-scabrate tectum. It comprises of 4 species (see key to the species). Pollen type-II: Spergula arvensis is characterized by colpate pollen with spinulose-punctate or scabrate-punctate tectum 8 species are included in this pollen type. Pollen type-III: Stellaria media is easily delimited by having porate pollen with spinulose-punctate tectum, 61 species are present in this type. Although, the species belonging to this pollen type are fairly uniform in different pollen characters but on the basis of tectum species of this types can be divided into two subtypes (see Account of pollen types. In the family Caryophyllaceae majority of the genera viz., Acanthophyllum, Arenaria, Dianthus, Cerastium, Gypsophila, Holosteum, Lepyrodiclis, Lychnis, Minuartia, Myosoton, Petrorhagia, Pseudostellaria, Stellaria, Sagina, Silene, Saponaria, Vaccaria and Velezia are fairly uniform by having pantoporate grains, with spinulosescabrate/ punctate tectum. However, in 3 species of Silene i.e. S. brahuica, S. gonosperma (Rupr.) Bocquet and S. indica Roxb. ex Otth and the genus Cucubalus reticulate tectum is present. Nowicke (1976) and Nowicke & Skvarla (1979) also reported similar type of pollen from these species. In contrast to this in Spergularia, Spergula, Polycarpaea and Telephium 3-10colpate grains are found. However, in the genus Telephium along colpate grains 3colporate grains are also found. Erdtman (1952) reported only 3 - colpate pollen grains in the same species. In the genus Polycarpaea and in 2 species of Spergularia i.e. S. media (L.) Presl and S. diandra (Guss.) Heldr. & Sart., only tricolpate grains are found, while in the genus Spergula and a single species of Spergularia (S. marina (L.) Griseb.) pantocolpate grains occur. Shape of the grain also varies from spheroidal to prolate-spheroidal mostly spheroidal, Kubitzki (1990) divided the family into three subfamilies viz., Paronychioideae, Alsinoideae and Caryophylloideae on the basis of floral characters and presence and absence of stipules. With few exceptions, the three pollen types recognized on the basis of apertural types and exine ornamentation correspond with the subfamilial classification of the family Caryophyllaceae. For instance, in the pollen type Silene indica species of subfamily Paronychioideae are found. However, the Pollen type Sterllaria media occur in both the subfamilies i.e., Alsinoideae and Caryophylloideae. Palynologically family Caryopyllaceae is more or less similar to Illeceberaceae both have porate rarely colpate pollen spinulose-punctate tectum (Perveen & Qaiser, 2003). References Bittrich, V. 1994. Caryophyllaceae. In: The families and genera of vascular plants, Vol. 2. Magnoliid, hamamelid, and caryophyllid families. (Eds.): K. Kubitzki, J. Rohwer & V. Bittrich. Berlin: Springer-Verlag, 206-236. Chand, S. 1962. On the pollen morphology of some Scandinavian Caryophyllaceae. Grana Palynologica, 3: 67-89. Cranwell, L.M. 1962. Subantarctic pollen and spores. I. Lyallia of Kerguelen. Pollen et Spores, 3: 11-20. Dimbleby, G. 1974. Plants and Archaedogy. John Baker, London, 187 pp. Erdtman, G. 1952. Pollen Morphology and Plant Taxonomy. Angiosperms. Chronica Botanica Co., Waltham, Massachusettes. Faegri, K. and J. Iversen. 1964. Test book of Pollen Analysis. Munksgaard, Copenhagen. POLLEN FLORA OF PAKISTAN-LI -CARYOPHYLLACEAE 915 Ghanzafar, S. 1984. Pollen Morphology of the Genus Silene L. (Caryophyllaceae), SectionSiphonomorpha Otth. and Auriculatae (Boiss.). New Phytologist, 98: 683-690 Ghanzafar, S. and Y.J. Nasir. 1974. Caryophyllaceae. In: Flora of Pakistan. (Ed.): E. Nasir & S.I. Ali, 175: 1-25. Kremp, G.O.W. 1965. Encyclopaedia of Pollen Morphology, Univ. Arizona Press, Tuscon, U.S.A. Kubitzki, K. 1990. The families and genera of vascular plants. Kuprianova, L.A. and L.A. Alyoshina. 1978. Pollen dicotyledoneaerum Florae Partis Europareae. URSS. Lamiaceae-Zygophyllaceae.(In Russian). Nauka 184 p. Akad. Sci. USSR .L. Komarov. Inst. Bot. Mabberley, D.I. 1987. The Plant Book. Camb. Univ. Press, Cambridge, New York. Moore, P.D. and J.A. Webb. 1978. An illustrated guide to pollen analysis. Hodder and Stoughton, London. Nair, P.K.K. 1962. Pollen grains of Indian plants. II. Bull.Gard. Lucknow, No. 60: 6-9. Nowicke, J.W. 1976 Pollen morphology in the order Centrospermae. Grana Palynologica, 15: 5177. Nowicke, J.W. and J.J. Skvarla. 1979. Pollen morphology: The potential influence in higher order systematics. Ann. Mo. Bot. Gard., 66: 633-699. Perveen, A. and M. Qaiser. 2003. Pollen Flora of Pakistan-XVII. Illecebraceae. Pak. J. Bot., 35(2): 141-144. Punt, W. and P.P. Hoen. 1995. Caryophyllaceae. The Northwest European Pollen Flora Punt, W., (Eds.): S. Blackmore, P. Hoen & P. Stafford. Volponi, C.R. 1987. Palynological study of Argentine species of Arenaria L. & Stellaria L. (Caryophyllaceae). Candollea, 42: 545-551. Walker, J.W. and J.A. Doyle. 1975. The basis of Angiosperm phylogeny: Palynology. Ann. Mo. Bot. Gard., 62: 666-723. (Received for publication 11 February 2006)