Comparison of Seed Proteins in Some Representatives of the Genusvigna
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BIOLOGIA PLANTARUM (PRAHA) 27 (1) : 70--73, 1985 BRIEF COMMUNICATION Comparison of Seed Proteins in Some Representatives of the Genus Vigna V~.RA TURKOV.~ and EvA KLOZOV~ Institute of Experimental Botany, Czechoslovak Academy of Sciences, Praha* Abstract. Seed protein patterns were compared in 10 species of the genus Vigna, and in the genera Macroptilium, Strophostyles, Macrotyloma, Psophocarpus and Phaseolus. Three Vigna groups could be clearly separated using immunochemical methods: Vigna hosei and Vigna luteola; Vigna mungo, Vigna angularis, Vigna umbellata, Vigna trilobata and Vigna radiata ; Vigna sinensis, Viqna unquiculata and Vigna aconitifolia. Among the other investigated genera Psophocarpus appears to be very distant, the others showing partial similarity in seed protein patterns. With regard to the present lack of nutritionally valuable protein sources, the genus Vigna is of great nutritional importance for the population of Asia. The protein content of the seeds of cultivated species is 22 to 30% and, what is especially important, the lysine content is high. Thus Vigna crop is at present one of the research priorities. Its taxonomic classification, the relationship t)haseolus -- Vigna and the assignment of particular species to these genera have been continuously studied (DANIMIHARDJA and LESTER 1974). Separation of these two genera was not simple because theb wild forms are related, although flower morphology on which the separation has been based is distinct. All the Asiatic species of the genus Phaseolus, section Ceratotropis, have been successively classified to the genus Vigna (JAIN and MEm~A 1978, MA~ECHAL et al. 1978). In addition to the Asiatic group, there is a group of African species which is very heterogeneous (MASC~ERPA 1976). The definition of the genus Vigna, originally based only on flower morpho- logy, appeared as insufficient (V~RDCOVl~T 1970) and for this reason also chemical, cytological and palynological criteria were included in the taxo- nomic evaluation. Received April 15, 1984; accepted August 3, 1984 *Address: Na Pernik~ce 15, 160 00 Praha 6, Czechoslovakia. 70 SEED PROTEINS IN GENUS VIGNA 71 The genus Phaseolus and the genus Vigna are clearly distinct from the other genera of the family Leguminosae (C~ISTOrOLrm and P~RI 1983). The original genus Phaseolus, very heterogeneous, with Asiatic and American species, was successively divided and segregated species were assigned to the genera Azukia, Strophostyles, Disolobium, Macroptilium and Vigna. The genus PhaseoIus has been limited exclusively to the American species and has become a natural group of species (KI~zovX et al. 1983), whereas the genus Vigna is very wide, heterogeneous, and includes approximately 100 to 150 species of the Asiatic and African regions (SMA~TT 1978). Species of the orig- inal genus Phaseolus from the Old World appear to be related to the genus Vigna rather than to the g. Phaseolus from the New World. Thus, for example, Phaseolus radiatus and Phaseolus campestris similarly as Phaseolu8 mungo have finally been classified to the genus Vigna (Vv,~DCOVl~T 1970). Also studies carried out by Kloz and his coworkers supported the hypothesis on the hetero- geneity of the g. Phaseolus, and on the basis of similarity of protein patterns showed the existence of 3 groups corresponding to the new division together with the classification of the Asiatic species of the g. Phaseolus to the g. Vigna (KLoz and TvRxovX 1963, KLozovA. 1965, KLoz et al. 1966). The aim of this paper was to appreciate a set of several species of the genus Vigna on the basis of comparison of their seed protein patterns by means of immunochemical methods and to evaluate their relationship to some other genera of the tribe Phaseoleae as e.g. Macroptilium, Strophostyles, Pha- aeolus, Macrotyloma and Psotahocarpus. Plant material: Vigna sinensis (L.) SAvI -- origin GDR Vigna aconitifolia (JAcQ.) -- India Vigna angularis (WIL~.) -- Japan Vigna luteola (JAcQ.) -- Mexico Vigna mungo (L.) -- Zaire Vigna hosei var. hosei -- Indonesia Vigna radiata var. radiata -- Ghana Vigna trilobata (L.) -- India Vigna unquiculata ssp. unquiculata -- Zaire Vigna umbellata (Tr[u~.) -- Costarica Macrotyloma axilari (L.) Psophocarpus tetragonolobus (L.) Dc. Macroptilium lathyroides L. vat. semierectum Strophostyles umbellata (WILD.) Phaseolus vulgaris cv. Veltrusks Saxa Seed proteins were extracted with buffered physiological solution pH 7.2, immunoelectrophoresis was carried out under standard conditions in agarose- starch gel (KLOz and KLOZOVX 1974). The seed protein mixtures were evaluated immunoelectrophoretically using antdsera against the albumin and the globulin complexes of Vigna sinensis and Vigna radiata (= Phaseolus aureus) seeds. The albumin antisera showed weak reactivity, the globulin antisera gave more clean-cut reactions. Vigna hosei and Vigna luteola are clearly distinct from the other investigated species. The other species can be arranged with regard to protein similarity as follows: Vigna mungo, Vigna angularis, Vigna umbellata, Vigna trilobata 72 V. TURKOVA, E. KLOZOVA and Vigna radiata, and finally Vigna sinensis, Vigna aconitifolia, and Vigna unquiculata. These three groups differ in both the number of precipitation lines in cathodic and anodic regions and eloetrophoretic mobility; some particular species could also be distinguished. The species Vigna aconitifolia, V. mungo, V. radiata, V. angularis and V. umbellata are widely distributed over the entire Asiatic continent. JAIx and M~m~A (1978) have pointed out the great similarity of V. angularis, V. umbellata and V. radiata. Those species form a taxonomic complex. Their protein patterns are also similar but not identical. Similar conclusions were drawn by IsmKv~A etal. (1981) from studies on flavonoids in some lines of Vigna mungo, Vigna radiata vat. radiata and var. sublobata. In contrast, Vigna aconitifolia distinctly differs from V. radiata. This has also boon verified by the results of our studies in which V. trilobata was found to differ from the former representatives of the genus Phaseolus. Cytological and hybridization studies have confirmed the hypothesis that V. radiata vat. sublobata (= Ph. sublobatus) is an ancestor of the species V. radiata (Ph. aureus) and V. mungo (Ph. mungo) (S~AI~TT 1978). Their seed proteins are also very similar. Justi- fication of the classification of the former Ph. aureus Rox~., now Vigna radiata L. (WILcz~x) to the genus Vigna and their phylogenetic proximity is supported by the findings of Cm~ISr~LS and BAIIMGAI~TN~I~ (1978), who characterized a trypsin inhibitor in Vigna radiata, and immunochemically positive reaction between Ph. aureus and Vigna mungo found by means of double diffusion. When investigating the similarity or dissimilarity of seed proteins of the other studied genera, we found the most marked dissimilarity in the g. Psopho- carpus and Phaseolus vulgaris. The genera Macrotyloma, Macroptilium, and Strophostyles showed a partial similarity of seed protein patterns. This work touched only broadly some taxonomic problems of the set of species classified to the genera Vigna and Phaseolus and confirmed the necessity to employ not only morphological but also biochemical criteria in taxonomic studies. REFERENCES CHliIS1)EELS, •[. J., BAUMGAI~T.'CEIr B. : Trypsin inhibitor in mung bean cotyledons. Purification, characteristics, subeellular localization and metabolism. -- Plant Physiol. 61 : 617--623, 1978. CRIS~OFOLIXI, G., PER1, P.: Immunochemistry and phylogeny in selected Leguminosae tribes. -- In: JENSEN, V., FAIRBROTHERS, D. E. (ed.): Protein and Nucleic Acids in Plant Systematies. Pp. 324-- 340. Springer Verlag, Berlin-- Heidelberg-- New York 1983. ]DANIMIHAI~I)JA, S., LESTER, ]~. N.: Comparative studies of some Asiatic species of Phaseolus using eleetrophoretic analyses of seed proteins. -- Ann. bogoriensis 5 (4) : 159-- 171, 1974. ISHIKURA, ~., IWATA, M., NIYAZAKI,S.: Flavonoids of some Vigna-plants in Leguminosae. -- Bet. 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