Flavonoids isolated from Medicago littoralis Rhode (Fabaceae): their ...

CARYOLOGIA Vol. 63, no. 1: 106-114, 2010 

Flavonoids isolated from Medicago littoralis Rhode (Fabaceae): 

their ecological and chemosystematic significance 

Bertoli 1 Alessandra, Daniela Ciccarelli 2* , Fabio Garbari 2 and Luisa Pistelli 1 

1 

Alessandra Bertoli, Luisa Pistelli, Department of Bioorganic Chemistry and Biopharmaceutics, University of 

Pisa , via Bonanno 33, I-56126 Pisa, Italy 

2 

Daniela Ciccarelli, Fabio Garbari, Department of Biology, University of Pisa, via Luca Ghini 5, I-56126 Pisa, 

Italy 

Dedicated to the memory of Prof. Ivano Morelli. 

Abstract — Three flavonoids, laricitrin 3,5’-di-O-β-glucopyranoside (1), genistein 7-O-β-glucopyranoside (2), 

and biochanin A 7-O-β-glucopyranoside (= sissotrin) (3), were isolated for the first time from the aerial parts 

of Medicago littoralis Rhode. The structures of the isolated compounds were established by NMR and LC- 

DAD-ESI-MS analyses. The chemosystematic of flavones and isoflavones identified was useful for discriminating 

different taxa inside the section Spirocarpos. In addition, the phytochemical profile of M. littoralis showed a 

behaviour due to the ecological conditions of sandy dunes, where this plant normally grows. 

Key words: Chemosystematic, Fabaceae, Flavones, Flavonoids, Isoflavones, Medicago. 

INTRODUCTION 

The genus Medicago L. (Fabaceae) belongs 

to the subfamily Papilionoideae, tribe Trifolieae. 

It consists of 50 species of annual or perennial 

herbs, rarely shrubs, of which Medicago sativa L. 

(alfalfa) is well-known since it is used as a forage 

crop widely. 

Several taxonomical analyses have been carried 

out to establish which characters could be 

useful to delimit the genus, expecially regarding 

the generic circumscription of Medicago and 

Trigonella L., both belonging to the tribe Trifolieae 

(see SMALL et al. 1987). Investigations involving 

pollen morphology (SMALL 1981), floral 

characters (SMALL et al. 1981a), asymmetries in 

Trifolieae leaves (SMALL et al. 1981b), haemolytic 

*Corresponding author: phone: +39(0)502211327; fax: 

+39(0)502211309; e-mail: dciccarelli@biologia.unipi.it 

saponins in the seeds (JURZYSTA et al. 1992), phenolic 

variation (CLASSEN et al. 1982; SALEH et al. 

1982), stigma morphology (SMALL and BROOKES 

1983), pollen-ovule pattern (SMALL 1986) or seed 

characters (SMALL and BROOKES 1990; DJEMEL et 

al. 2005) were performed in order to solve these 

taxonomical problems. Recently, molecular phylogeny 

analyses (BENA et al. 1998; BENA 2001) 

supported the transfer of 23 Trigonella species 

previously known as “medicagoids” to the genus 

Medicago, as proposed by SMALL et al. (1987) on 

the base of morphological data. The infrageneric 

classification of Medicago is reported in the Table 

1 following mainly the classifications of LES- 

INS and LESINS (1979), SMALL and JOMPHE (1989), 

SMALL (1990a-b), GILLESPIE and MCCOMB (1991). 

In the same table is also reported the geographical 

distribution of each species obtained from 

the main Floras (TUTIN 1968; HEYN and DAVIS 

1970; PIGNATTI 1982; GREUTER et al. 1989; SALES 

and HEDGE 2000; GRIN Taxonomy for plants, 

2006) in order to compare these taxa from an 

ecological point of view.

RUNNING TITLE: FLAVONOIDS IN MEDICAGO LITTORALIS RHODE 107 

TABLE 1 — The infrageneric classification and the geographical distribution of Medicago L. from the Mediterranean 

region, Middle East and Asia. Evidenced in bold the species living on maritime sands or extending their area of occupancy 

up to the seashore. 

Section Subsection Species Geographical distribution 

Dendrotelis M. arborea L. Rocky places. Mediterranean region, Middle East and Asia. 

M. strasseri Greuter, Matthäs & Risse Rocky places. Europe. 

Medicago M. cancellata M. Bieb. Steppe. Russian Federation. 

M. daghestanica Rupr. Steppe. Russian Federation. 

M. hybrida Trautv. Woods and mountain slopes. Corbiéres and Pyrenees. 

M. marina L. Maritime sands. Shores of the Mediterranean, Black Sea and Atlantic. 

M. papillosa Boiss. Middle East and Asia. 

M. pironae Vis. Rocky places. Italy and Balcans. 

M. prostrata Jacq. Dry grassland. From Italy and Austria to the Black Sea. 

M. rhodopaea Velen. Rocky places. Bulgaria. 

M. rupestris M. Bieb. Rocky places. Krym. 

M. sativa L. Cosmopolitan. 

M. saxatilis M. Bieb. Rocky places. Bulgaria and Krym. 

M. suffruticosa Raymond Rocky places. Pyrenees and Morocco. 

Carstienses M. carstiensis Jacq. Bushy places. From Italy to Bulgaria. 

Spirocarpos Pachyspirae M. constricta Durieu Grasslands. Europe and Asia. 

M. doliata Carmign. Dry lands. Mediterranean region and Middle East. 

M. italica Mill. (= M. tornata (L.) Miller) Dry grasslands. Mediterranean region extending to Portugal. 

M. lesinsii E. Small Mediterranean region. 

M. littoralis Rohde Mediterranean region extending to Portugal and W. France. 

M. murex Willd Mediterranean region extending to Portugal. 

M. rigidula L. Dry grasslands. South Europe. 

M. rigiduloides E. Small Middle East, Caucasus and Asia. 

M. sinskiae Uljanova Asia. 

M. soleirolii Duby Dry grasslands. Krym, probably introduced in Italy and France. 

M. syriaca E. Small Syria. 

M. truncatula Gaertn. Mediterranean region. 

M. turbinata L. Mediterranean region. 

Rotatae M. blancheana Boiss. Mediterranean region. 

M. noeana Boiss. Iraq and Turkey. 

M. rotata Boiss. Eastern Mediterranean region. 

M. rugosa Desr. Mediterranean region living also on maritime sands. 

M. scutellata L. Grasslands. South Europe. 

M. shepardii Post Turkey. 

Intertextae M. ciliaris L. 

Mediterranean region. 

M. granadensis Willd. Mediterranean region. 

M. intertexta L. Grasslands, expecially near the sea. Mediterranean region and Portugal. 

M. muricoleptis Tineo Grasslands, expecially near the sea. Mediterranean region. 

Leptospirae M. arabica L. 

Cosmopolitan. Mediterranean region. 

M. coronata L. Stenomediterranean. 

M. disciformis DC. Stenomediterranean. 

M. laciniata L. Naturalized locally in the Mediterranean region. 

M. lanigera C. Winkl. Afghanistan, Tajikistan and Turkmenistan. 

M. laxispira Heyn Iraq. 

M. minima L. Living on sandy dunes. Most of Europe. 

M. polymorpha L. Subcosmopolitan. South Europe. 

M. praecox DC. Dry lands and garrigues. Mediterranean region. 

M. sauvagei Nègre Morocco. 

M. tenoreana Ser. Mediterranean region.

108 

BERTOLI, CICCARELLI, GARBARI and PISTELLI 

Table 1 Contd. 

Section Subsection Species Geographical distribution 

Geocarpa M. hypogaea E. Small Eastern Mediterranean region. 

Lupularia M. lupulina L. Throughout Europe. 

M. secundiflora Durieu Mediterranean region. 

Heynianae M. heyniana Greuter Europe and Middle East. 

Orbiculares M. orbicularis L. South Europe. 

Hymenocarpos M. radiata L. Afghanistan, Iran, Iraq, Israel, Jordan, Lebanon, Syria, Turkey and Caucasus. 

Platycarpae M. archiducis-nicolai Širj. China. 

M. cretacea M. Bieb. Ukraine, Krym and Asia. 

M. edgeworthii Širj. Asia. 

M. ovalis Širj. Spain, Algeria and Morocco. 

M. platycarpa L. Russian Federation and Asia. 

M. plicata Širj. Turkey. 

M. popovii Širj. Soviet Middle Asia. 

M. ruthenica (L.) Ledebour Russian Federation and Asia. 

Lunatae M. biflora E. Small Turkey and Caucasus. 

M. brachycarpa M. Bieb. Iraq, Lebanon, Turkey and Caucasus. 

M. huberi E. Small Turkey. 

M. rostrata E. Small Turkey. 

Buceras Erectae M. arenicola E. Small Turkey. 

M. astroites Trautv. Iran, Iraq, Israel, Jordan, Lebanon, Syria, Turkey, Caucasus and Soviet Middle Asia. 

M. carica E. Small East Aegean islands and Asiatic Turkey. 

M. crassipes E. Small Iran, Iraq, Lebanon, Syria and Turkey. 

M. fischeriana Trautv. Bulgaria. 

M. halophila E. Small Turkey. 

M. heldreichii E. Small Turkey. 

M. medicaginoides E. Small Europe, Middle East and Asia. 

M. monantha Trautv. Afghanistan, Iran, Iraq, Israel, Jordan, Lebanon, Syria, Turkey, Caucasus and Soviet 

Middle Asia. 

M. orthoceras Trautv. Iran, Iraq, Turkey, Caucasus and Soviet Middle Asia. 

M. pamphylica E. Small Turkey. 

M. persica E. Small Iran. 

M. phrygia E. Small Iran, Iraq, Syria and Turkey. 

M. polyceratia Trautv. France, Portugal, Spain, Algeria, Morocco and Tunisia. 

M. rigida E. Small Turkey. 

Deflexae M. retrorsa E. Small Afghanistan. 

Reflexae M. monspeliaca Trautv. Mediterranean region. 

Isthmocarpae M. isthmocarpa E. Small Turkey. 

M. rhytidiocarpa E. Small Turkey. 

The present report represents the first result 

of a phytochemical study of Italian populations 

of Medicago littoralis Rhode (strand medic). 

This species is a therophyte growing on sandy 

soils near the sea and distributed in the Mediterranean 

region extending to Portugal and 

West France. Whole plant is covered by hair; 

the leaves are ternate with obovate or cordate, 

cuneate, dentate towards the apex leaflets. The 

yellow flowers are grouped in a terminal raceme; 

flowering in March-May. The legumes are in a 

very close spiral of 3-6 turns, discoid to cilindrical, 

glabrous, spiny or not (TUTIN 1968). 

There are few photochemical studies on the


TABLE 2 — Distribution of flavonoids in the genus Medicago. The species that living on maritime sands or extending 

their area up to the seashore are evidenced in bold. 

Species Flavonols Isoflavones Flavones Isoflavans Pterocarpans Coumestans 

Sec. Dendrotelis 

M. arborea a,b x x x 

Sec. Medicago 

M. cancellata a,b x 

M. hybrida a,b x 

M. marina a,b x x 

M. sativa a,b,d,e,f x x x x x x 

Sec. Spirocarpos 

Subsec. Pachyspirae 

M. doliata a,b x x x 

M. littoralis a,b x x x x x 

M. murex a,b x 

M. rigidula a,b x x 

M. truncatula a,b,c x x x x 

M. turbinata a,b x 

Subsec. Rotatae 

M. blancheana a,b x 

M. rotata a,b x 

M. scutellata a,b x x x 

Subsec. Intertextae 

M. ciliaris a,b x x x x 

M. intertexta a,b x x x x 

Subsec. Leptospirae 

M. arabica a,b x x x x x 

M. coronata a,b x x 

M. laciniata a,b x x x 

M. minima a,b x x x 

M. polymorpha a,b x x x x 

Sec. Geocarpa 

M. hypogaea a,b x x 

Sec. Lupularia 

M. lupulina a,b x x 

Sec. Orbiculares 

M. orbicularis a,b x x x x x 

Sec. Hymenocarpos 

M. radiata a,b x x x x x 

Sec. Platycarpae 

M. cretacea a,b x 

M. platycarpa a,b x x 

Sec. Buceras 

Subsec. Erectae 

M. medicaginoides a,b x 

M. monantha a,b x 

M. polyceratia a,b x 

Subsec. Reflexae 

M. monspeliaca a,b x x x 

Phytochemical data source: a = Southon (1994); b = Hegnauer and Hegnauer (2001); c = Kowalska et al. (2007); d = Stochmal 

et al. (2001a); e = Stochmal et al. (2001b); f = Stochmal et al. (2001c).

110 


(1) Laricitrin 3,5-O-β-diglucopyranoside 

(2) Genistein 7-O-β-glucopyranoside R=H 

(3) Biochanin A7-O-β-glucopyranoside R=Me 

Figure 1 — Chemical structures of the flavonoids (1-3) 

isolated from the aerial parts of Medicago littoralis. 

aerial green parts of M. littoralis collected out 

of Italy (INGHAM 1979; SALEH et al. 1982), which 

show the presence of several typical compounds 

such as daidzein, luteolin, medicarpin, sativan, 

3’,4’,7-trihydroxyisoflavone and vestitol. Previous 

phytochemical studies on the genus Medicago 

showed the presence of flavones, flavonols, 

isoflavones, isoflavans, coumestans and pterocarpans 

(SOUTHON 1994; HEGNAUER and HEG- 

NAUER 2001; STOCHMAL et al. 2001a-c; KOWALSKA 

et al. 2007). The aim of this paper was the isolation 

and the characterization of some typical 

flavonoids of M. littoralis aerial parts collected 

in sandy dunes to suggest chemosystematic considerations. 

MATERIALS AND METHODS 

Plant material - The plant grows on the back of 

sandy dunes of Marina di Vecchiano (Pisa, Italy) 

within the Natural Park of Migliarino - San Rossore 

- Massacciuccoli. Aerial parts of Medicago 

littoralis were randomly collected from the same 

population during the flowering phase at the 

end of April 2004. A voucher specimen (3688/4) 

was deposited in the Herbarium Horti Botanici 

Pisani (PI), Italy. 

Chemicals and equipments - Acetonitrile, 

HCOOH and methanol were HPLC grade solvents 

by Baker (Netherlands). HPLC-Water was 

purified by a Milli-Q Plus system (Millipore Milford, 

MA, USA). Reference chemical material including 

luteolin, daidzein, and the isolated compounds 

(laricitrin 3,5’-di-O-β-glucopyranoside, 

genistein 7-O-β-glucopyranoside and biochanin 

A 7-O- β-glucopyranoside) were part of an 

home-made database where each compound 

was used as standard with an HPLC grade purity 

of 98-99%. The analysis were performed by 

a Surveyor Thermofinnigan liquid chromatograph 

pump equipped with a Thermofinnigan 

Photodiode Array Detector and a LCQ Advantage 

mass detector. The semipreparative HPLC 

system consisted of a Waters 600E (Millipore) 

pump and 990 photodiode array detector (PDA) 

and a column Merck LiChroCART 250RP-18 

column (30 x 1 cm, 10 µm) was used. 

Extraction and LC-DAD-MS screening - The aerial 

parts (184 g) of Medicago littoralis, air-dried at 

room temperature and powdered, were extracted 

in a Soxhlet apparatus with n-hexane, CHCl 3 

and 

MeOH in turn. After removal of solvent under 

vacuum, the following residues were obtained: 

R H 

(5.49 g), R C 

(6.53 g), and R M 

(25.81 g). 

R M 

(3.73 g) was chromatographed on a Merck 

Lichroprep RP-8 column (40-63 mm) eluted 

with MeOH-H 2 

O (7:3) to obtain 19 crude fractions 

(M 1 

-M 19 

). Fractions were checked by TLC 

[RP-18, MeOH-H 2 

O (7:3)], Merck Kieselgel 60 

F 254 

. TLC chromatograms were visualized under 

UV light at 254 and 366 nm and sprayed with 

Naturstoffreagents-PEG. The most interesting 

purified fractions were screened by LC-DAD- 

ESI-MS using an analytical Lichrosorb RP-18 

column (250 x 4.6 mm i.d., 5μm, Merck). The 

analyses (20 μl, 2 mg/ml methanolic extract solution) 

were carried out by a linear gradient using 

water with 0.1% HCOOH (solvent A), CH- 

CN (solvent B) and MeOH (solvent C) at flow 

3 

rate of 1.0 ml/min in the following conditions: 

from 20:80 v/v (B-A) to 40:60 (B-A) in 20 min, 

then to 40:60 v/v (B-A) (20 min), and then conditioning 

to the initial condition (20:80 v/v B-A) 

for 10 min. The total analytical run time was 50 

min for each sample. The spectral data from the 

PDA detector were collected during the whole 

run in the range 210-500 nm and the peaks were 

detected at 330 nm for all analysed compounds. 

LC-ESI-MS analyses were performed in the same 

chromatographic conditions using these specific


electrospray ionization values: sheath gas flowrate 

72 psi, auxilary gas flow 10 psi, capillary 

voltage -16 V and capillary temperature 280°C 

. Full scan spectra from m/z 200 to 500 amu in 

the negative ion mode were obtained. The identification 

of each constituents was carried out by 

the comparison of each peak in the extracts with 

the retention times, UV and MS spectra of an 

home-made database of flavonoid compounds. 

Isolation and purification of flavonoids - The filtered 

solution of fraction M 2 

(1 g) was subjected 

to column chromatography over a Merck Lichroprep 

RP-8 column (40-63 µm) eluted with 

MeOH-H 2 

O (3:7) to obtain 11 subfractions 

(E 1 

-E 11 

). Subfraction E 7 

(30 mg) was purified 

by semipreparative HPLC-PDA. The gradient 

profiles were based on two solvents, denoted 

A (CH 3 

CN) and B (H 2 

O), were employed. Initial 

conditions were 20% A, 80% B with a linear 

gradient reaching 40% A, 60% B; this was 

followed by isocratic elution, after which the 

programme returned to the initial solvent composition 

(t = 40 min). Column was mantained 

at room temperature, and the flow rate was 1.3 

ml/min. Injection volumes were 100 μl, and UV 

detection was at 330 nm. This semipreparative 

HPLC/UV gave 4 subfractions (E 7.1 

-E 7.4 

). Fraction 

M 4 

was subjected to column chromatography 

under the same conditions of fraction M 2 

to 

obtain 2 subfractions (M 4.1 

-M 4.2 

). 

The nuclear magnetic resonance (NMR) 

spectra of the isolated flavonoids were registered 

on a Bruker AC-200 spectrometer and a 

Bruker Avance 400 spectrometer using CD 3 

OD 

or DMSO-d 6 

as solvent. TMS was used as internal 

standard. All the 1D and 2D NMR experiments 

were carried on using the standard 

Bruker library of microprograms. MS spectra of 

the isolated compounds were registered by direct 

infusion in LCQ Advantage ion trap mass 

spectrometer using a methanolic solution (1 μg/ 

ml, 5 μl/min) with the following ESI parameters: 

sheath gas flow-rate 72 arbitrary units, auxilary 

gas flow 10 arbitrary units, capillary voltage -16 

V and capillary temperature 280°C. 

RESULTS AND DISCUSSION 

The preliminary screening by LC-DAD-ESI- 

MS analyses of Medicago littoralis (aerial parts) 

collected during the flowering phase on the 

sandy dunes of Marina di Vecchiano (Pisa, Italy) 

showed the presence of some characteristic flavonoids 

such as lutein and daidzein previously 

reported in the literature (SOUTHON 1994; HEG- 

NAUER and HEGNAUER 2001). 

However, our phytochemical investigation 

on this species showed also laricitrin 3,5’-di- 

O-β-glucopyranoside (1), genistein 7-O-βglucopyranoside 

(2), and biochanin A 7-O-β 

-glucopyranoside (= sissotrin) (3) as characteristic 

compounds (Fig. 1). These flavonoids were 

isolated for the first time in Medicago littoralis. 

After the extraction, the methanolic extract 

was chromatographed by semi-preparative 

HPLC to isolate the constituents. Subfractions 

E 7.2 

(2 mg) and E 7.4 

(2 mg) gave pure compounds 

(1) and (2), respectively. Compound (3) was 

identified as pure constituent of subfraction M 4.1 

(2.8 mg). The structural elucidation of the isolated 

compounds were performed on the basis 

of NMR and ESI-MS experiments. 

The signals in the 1 H and 13 C NMR spectra 

of the isolates were superimposable on those reported 

in the literature for laricitrin 3,5’-di-O-βglucopyranoside 

(1) (TORCK et al. 1983; KOWAL- 

SKA et al. 2007), genistein 7-O-β-glucopyranoside 

(2) (AGRAVAL 1989), and biochanin A 7-O-βglucopyranoside 

(= sissotrin) (3) (GUGGOLZ et 

al. 1961). 

Except for laricitrin 3,5’-di-O-β-glucopyranoside, 

isolated in the floral parts of M. arborea 

L. (TORCK et al. 1983), M. lupulina L. (BURDA 

and JURZYSTA 1988) and M. truncatula Gaertn. 

(KOWALSKA et al. 2007), the other flavonoids 

had never been recorded in the genus Medicago. 

Previous phytochemical studies showed the 

presence of luteolin, daidzein and 3’,4’,7-trihydroxyisoflavone 

in the hydrolized plant of M. littoralis 

(SALEH et al. 1982), while sativan, vestitol 

and medicarpin were isolated from the leaves of 

the same species (INGHAM 1979). 

Flavonoids are well accepted as chemical 

markers in plant taxonomy as a useful tool for 

the characterization and classification of higher 

plants, because of their widespread occurrence 

and chemical stability (HARBORNE and TURNER 

1984; HEGNAUER and HEGNAUER 2001). 

Regarding a significant connection between 

the distribution of secondary metabolites and 

the infrageneric classification of Medicago (Table 

2), it seems clear that flavones and isoflavones 

may be particularly useful for discriminating different 

taxa inside the section Spirocarpos Ser. Isoflavones, 

in fact, occur in the subsections Intertextae 

(Urb.) Heyn and Leptospirae (Urb.) Heyn, 

but they are absent in Pachyspirae (Urb.) Heyn 

(except for M. littoralis). In the same way as re-

112 


TABLE 3 — Flavonoids from M. littoralis and their occurrence in the genus Medicago. The species that living on maritime 

sands or extending their area up to the seashore are evidenced in bold. 

Species Flavonols Isoflavones Flavones Isoflavans Pterocarpans 

1 2 3 4 5 6 7 8 9 

Sec. Dendrotelis 

M. arborea x x x 

Sec. Medicago 

M. cancellata 

M. hybrida 

M. marina x x 

M. sativa x x x x x x 

Sec. Spirocarpos 

Subsec. Pachyspirae 

M. doliata x x x 

M. littoralis x x x x x x x x x 

M. murex 

M. rigidula x x 

M. truncatula x x x x 

M. turbinata 

Subsec. Rotatae 

M. blancheana 

M. rotata 

M. scutellata x 

Subsec. Intertextae 

M. ciliaris x x x 

M. intertexta x x x x 

Subsec. Leptospirae 

M. arabica x x x x 

M. coronata x x x 

M. laciniata x x 

M. minima x x x 

M. polymorpha x x x 

Sec. Geocarpa 

M. hypogaea 

Sec. Lupularia 

M. lupulina x x 

Sec. Orbiculares 

M. orbicularis x x x x x x 

Sec. Hymenocarpos 

M. radiata x x x 

Sec. Platycarpae 

M. cretacea 

M. platycarpa x 

Sec. Buceras 

Subsec. Erectae 

M. medicaginoides 

M. monantha 

M. polyceratia 

Subsec. Reflexae 

M. monspeliaca x x x 

1 = Laricitrin 3,5’-O-glc; 2 = Genistein 7-O-glc; 3 = Sissotrin; 4 = Daidzein; 5 = 3’,4’,7-trihydroxyisoflavone; 6 = Luteolin; 7 

= Sativan; 8 = Vestitol; 9 = Medicarpin.


gards flavones, it is interesting that these compounds 

occur in the subsection Leptospirae, but 

they are absent in Pachyspirae except again for 

M. littoralis. Our results are partially in accordance 

with molecular phylogenetic studies (BENA 

et al. 1998) and biochemical analyses of seed 

composition that show M. truncatula and M. littoralis 

as very closely related species (DJEMEL et 

al. 2005). M. truncatula, in fact, shows the same 

phytochemical profile of M. littoralis except for 

isoflavones (KOWALSKA et al. 2007). 

Moreover, this study was carried out in order 

to find relationships between the phytochemical 

profile of M. littoralis and the ecological conditions 

of sandy dunes, where the plants grows. 

M. littoralis is able to synthesise flavonols, isoflavones, 

flavones, isoflavans and pterocarpans 

(SOUTHON 1994; HEGNAUER and HEGNAUER 

2001). This behaviour is also found in M. sativa, 

M. arabica, M. orbicularis and M. radiata (Table 

2), which belong to different sections or subsections 

and live in different habitats. Anyway, if 

we consider the specific flavonoid and not the 

chemical class (see Table 3), the mentioned species 

produce different flavonoids. 

All the plants that live on maritime sands 

were able to synthesize flavonols (except for M. 

marina L., M. coronata L. and M. minima L.) and 

most of them produced isoflavones and flavones 

depending on the subsection considered. On the 

other hand, considering flavonoids isolated from 

M. littoralis samples and their occurrence in the 

genus Medicago (see Table 2-3), a so large variety 

in the flavonoidic composition has never been 

observed for plants living near the sea yet. 

In conclusion, this study showed laricitrin 

3,5’-di-O-β-glucopyranoside (1), genistein 

7-O-β-glu co pyra noside (2), and sissotrin (3) as 

characteristic flavonoids of the aerial parts of M. 

littoralis collected on the sandy dunes during the 

flowering period. Therefore, the list of chemotaxonomic 

markers of M. littoralis can be added 

by these flavonoids (1-3). To evaluate more 

deeply the flavonoidic production in this species, 

the authors plan to study its phytochemical 

profile also during seasonal and growth-phase 

variations. 

REFERENCES 

AGRAVAL P.K., 1989 — Carbon-13 NMR of Flavonoids. 

Elsevier, Amsterdam. 

BENA G., 2001 — Molecular Phylogeny supports the 

morphologically based taxonomic transfer of the 

“medicagoid” Trigonella species to the genus Medicago 

L. Plant Systematics and Evolution, 229: 

217-236. 

BENA G., LEJEUNE B., PROSPERI J.M. and OLIVIERI I., 

1998 — Molecular phylogenetic approach for studying 

life-history evolution: the ambiguous example 

of the genus Medicago. Proceedings of the Royal 

Society London B, 265: 1141-1151. 

BURDA S. and JURZYSTA M., 1988 — Isolation and 

identification of flavonoids from Medicago lupulina 

L. flowers. Acta Societatis Botanicorum Poloniae, 

57: 563-571. 

CLASSEN D., NOZZOLILLO C. and SMALL E., 1982 — A 

phenolic-taxometric study of Medicago (Leguminosae). 

Canadian Journal of Botany, 60: 2477- 

2495. 

DJEMEL N., GUEDON D., LECHEVALIER A., SALON C., 

MIQUEL M., PROSPERI J.-M., ROCHAT C. and BOU- 

TIN J.-P., 2005 — Development and composition of 

the seeds of nine genotypes of the Medicago truncatula 

species complex. Plant Physiology and Biochemistry, 

43: 557-566. 

GILLESPIE D.J. and MCCOMB J.A., 1991 — Morphology 

and distribution of species in the Medicago 

murex complex. Canadian Journal of Botany, 69: 

2655-2662. 

GREUTER W., BURDET H.M. and LONG G., 1989 — 

Med-Checklist, vol. 4, pp 136-147. Conservatoire 

et Jardin botaniques, Genéve. 

GRIN Taxonomy for Plants. 2006. URL: http://www. 

ars-grin.gov/ 

GUGGOLZ J., LIVINGSTON A.L. and BICKOFF E.M., 

1961 — Detection of Daidzein, Formononetin, 

Genistein and Biochanin A in forages. Journal of 

Agricultural and Food Chemistry, 9: 330-332. 

HARBORNE J.B. and TURNER B.L., 1984 — Plant chemiosystematics. 

Harcourt Brace Jovanovich Publishers, 

Academic Press, London. 

HEGNAUER R. and HEGNAUER M., 2001 — Chemotaxonomie 

der Pflanzen. Band XIb-2: Leguminosae. 

Teil 3: Papilionoideae. Birkhäuser Verlag, Basel. 

HEYN C.C. and DAVIS P.H., 1970 — Medicago L. In: 

P.H. Davis (Ed.) “Flora of Turkey and the East 

Aegean Islands”, vol. 3, pp 483-511. Edinburgh 

University Press, Edinburgh. 

INGHAM J.L., 1979 — Isoflavonoid phytoalexins of 

the genus Medicago. Biochemical Systematics and 

Ecology, 7: 29-34. 

JURZYSTA M., BURDA S., OLESZEK W., PLOSZYNSKI M., 

SMALL E. and NOZZOLILLO C., 1992 — Chemical 

composition of seed saponins as a guide to the classification 

of Medicago species. Canadian Journal of 

Botany, 70: 1384-1387. 

KOWALSKA I., STOCHMAL A., KAPUSTA I., JANDA B., 

PIZZA C., PIACENTE S. and OLESZEK W., 2007 — 

Flavonoids from barrel medic (Medicago truncatula) 

aerial parts. Journal of Agricultural and Food 

Chemistry, 55: 2645-2652. 

LESINS K.A. and LESINS I., 1979 — Genus Medicago 

(Leguminosae), a taxogenetic study. The Hague, 

Junk. 

PIGNATTI S., 1982 — Flora d’Italia, vol. 1, pp 711-720. 

Edagricole, Bologna.

114 


SALEH N.A.M., BOULOS L., EL-NEGOUMY S.I. and AB- 

DALLA M.F., 1982 — A comparative study of the 

flavonoids of Medicago radiata with other Medicago 

and related Trigonella species. Biochemical 

Systematics and Ecology, 10: 33-36. 

SALES F. and HEDGE I.C., 2000 — Medicago L. In: S. 

TALAVERA, C. AEDO, S. CASTROVIEJO, A. HERRERO, 

C. ROMERO ZARCO, F.J. SALGUEIRO and M. VELAYOS 

(Eds.) “Flora Iberica”, vol. 7(2), pp 741-774. Real 

Jardín Botánico, Madrid. 

SMALL E., 1981 — Numerical analysis of major groupings 

in Medicago employing traditionally used 

characters. Canadian Journal of Botany, 59: 1553- 

1577. 

SMALL E., 1986 — Pollen-ovule patterns in tribe Trifolieae 

(Leguminosae). Plant Systematics and Evolution, 

160: 195-205. 

SMALL E., 1990a — Medicago syriaca, a new species. 

Canadian Journal of Botany, 68: 1473-1478. 

SMALL E., 1990b — Medicago rigiduloides, a new species 

segregated from M. rigidula. Canadian Journal 

of Botany, 68: 2614-2617. 

SMALL E., BASSETT I.J. and CROMPTON C.W., 1981a 

— Pollen variation in tribe Trigonelleae (Leguminosae) 

with special reference to Medicago. Pollen 

et Spores, 23: 295-320. 

SMALL E. and BROOKES B.S., 1983 — The systematic 

value of stigma morphology in the legume-tribe Trifolieae 

with particular reference to Medicago. Canadian 

Journal of Botany, 61: 2388-2404. 

SMALL E. and BROOKES B.S., 1990 — Circumscriptium 

of the genus Medicago (Leguminosae) by seed characters. 

Canadian Journal of Botany, 67: 613-629. 

SMALL E., CROMPTON C.W. and BROOKES B.S., 1981a 

— The taxonomic value of floral characters in tribe 

Trigonelleae (Leguminosae) with special reference 

to Medicago. Canadian Journal of Botany, 59: 

1578-1598. 

SMALL E., LEFKOVITCH L.P. and BROOKES B.S., 1981b 

— Remarkable asymmetries in trifoliolate leaves 

with particular reference to Medicago. Canadian 

Journal of Botany, 59: 662-671. 

SMALL E. and JOMPHE M., 1989 — A synopsis of the 

genus Medicago. Canadian Journal of Botany, 67: 

3260-3294. 

SMALL E., LASSEN P. and BROOKES B.S., 1987 — An 

expanded circumscriptium of Medicago (Leguminosae, 

Trifoliae) based on explosive flower tripping. 

Willdenowia, 16: 415-437. 

SOUTHON I.W., 1994 — In: F.A. BISBY, J. BUCKINGHAM 

and J.B. HARBORNE (Eds.) “Phytochemical Dictionary 

of the Leguminosae”, vol. 1, pp 449-468. 

Chapman & Hall, London. 

STOCHMAL A., PIACENTE S., PIZZA C., DE RICCARDIS F., 

LEITZ R. and OLESZEK W., 2001 — Alfalfa (Medicago 

sativa L.) flavonoids. 1. Apigenin and Luteolin 

glycosides from aerial parts. Journal of Agricultural 

and Food Chemistry, 49: 753-758. 

STOCHMAL A., SIMONET A.M., MACIAS F.A. and 

OLESZEK W., 2001b — Alfalfa (Medicago sativa 

L.) flavonoids. 2. Tricin and Chrysoeriol glycosides 

from aerial parts. Journal of Agricultural and Food 

Chemistry, 49: 5310-5314. 

STOCHMAL A., SIMONET A.M., MACIAS F.A., OLIVEI- 

RA M.A., ABREU J.M., NASH R. and OLESZEK W., 

2001c — Acylated apigenin glycosides from alfalfa 

(Medicago sativa L.) var. Artal. Phytochemistry, 

57: 1223-1226. 

TORCK M., PINKAS M., JAY M. and FAVRE-BONVIN J., 

1983 — Heterosides flavoniques de Medicago arborea 

L. Pharmazie, 38:783. 

TUTIN T.G., 1968 — Medicago L. In: T.G. Tutin, 

V.H. Heywood, N.A. Burges, D.M. Moore, D.H. 

Valentine, S.M. Walters and D.A. Webb (Eds.) 

“Flora Europaea”, vol. 2, pp 153-157. Cambridge 

University Press, Cambridge. 

Received October 20 th 2009; accepted February 2 th 2010 

Published May 30 th 2010 

Editore: Università degli Studi di Firenze 

Registrazione Tribunale di Firenze n. 478 del 13/7/1951 

Redazione: Dipartimento di Biologia Vegetale 

Via La Pira, 4 - 50121 FIRENZE 

Direttore Responsabile: Dr. ALESSIO PAPINI 

Stampato a Firenze da Edizioni Tassinari - Firenze - May 2010

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