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BIOCHEMICAL EVIDENCE FOR A FIFTH CULTIGEN WITHIN THE GENUS PHASEOLUS. D.G. Debouck, V. Schmit, D. Libreros & H. Ramirez * In the Phaseolus coccineus complex, two biological materials are still currently grown by the American Indians in the humid highlands of Mesoamerica and the Northern Andes: P. coccineus L. and P. polyanthus Greenman (1,2,3). Although evident genetic affinities exist between these two taxa (4,5), morphological differences were judged significant enough to use a separate taxonomical treatment (3,6). Significantly enough, in almost every place where the two taxa are grown together, they are designated by different vernacular names by each ethnic group (3,7). That situation raises the question of the identity and origin of P. polyanthus. Recent electrophoretic evidence (8) showed that cultivated P. polyanthus is not a between P. vulgaris L. and P. coccineus as it was thought previously (7), but a variant of P. coccineus. Recent findings of a wild bean morphologically close to P. polyanthus in central Guatemala (9) suggest that this cultigen would have arisen ft-om a wild ancestor present in that country through a process different from the ones of P. vulgaris and P. coccineus. According to that scheme, the P. polyanthus growing feral in Mesoamerica (3) and in the Northern Andes (10; where it has been named P. flavescens) would be an escape from early cultivation. We will report here about testing these hypotheses on a collection of 163 populations (with a total of 356 individuals) covering the range of distribution of P. polyanthus from Veracruz, Mexico to Cajamarca, Peru (see Table). protein extracts were run in one- dimensional SDS/PAGE for phaseolin (13.5% acrylamide, for 12 hours). By running first 12 individual of 13 populations from 10 different regions, it was possible to appreciate better the internal variation due to outcrossing. Results and Discussion Four major proteinic fractions can be observed on the gels, with molecular weights of 45, 33, 28 and 24 kd. They were tentatively classified as phaseolin, lectin I, lectin II and albumin respectively according to what has been recently defined in P. vulgaris (11,12). Our results show first the great similarity between all populations of the range independently from their biological status. Some polymorphisms have been detected however in the lectin I, lectin II and albumin fractions. The Mesoamerican materials commonly have two groups of polypeptides of about 24 kd, while the North Andean types have three. Moreover the frequent pattern found in wild forms of Guatemala (b) is dominant in the Mesoamerican cultivated forms (in 115 individuals/127 or 91%, from Mexico, Guatemala and Costa Rica). Second, the highest variability of patterns is found in the wild populations of Guatemala, and much less in the cultivated types; despite a lower number for the former, this fact would be indicative of a reduction of variability upon domestication. Third, nearly all South American forms irrespective of their biological status, display the 'k' pattern and are similar to P. flavescens (from the type locality in Colombia; included in the escapes in the Table). Fourth, although P. flavescens at the type locality usually shows three albuminic bands of about 24 kd (the 'k' pattern), 3 display a typical 'b' pattern with two bands. These results indicate that we are dealing with a single species and a single gene pool. The identity of pattern found in the wild and cultivated materials fort most of the plants from Mesoamerica demonstrates that this wild species is the ancestor of P. polyanthus. Of particular significance is the higher polymorphism in the wild types of Guatemala, and on the other hand the lack of any noteworthy difference at both biochemical and morphological levels between the cultivated and the weedy types in the Northern Andes. The later m.aterials would thus be a kind of early escapes growing in habitats mimicking the original montane rain forests (9). The high frequence of the 'b' pattern in Mesoamerica (151 out of 202 individuals or 75 % of plants examined) and of the 'k' 107 pattern (145 out of 154 individuals or 94% of plants examined) in the Nonhem Andes suggest that a genetical drift is progressively taking place between the two zones. Table. Geographic frequency distribution of seed protein patterns among Phaseolus polyanthus Greenman materials.

Patterns Ctry Status Accès, a b c d e f g h k ][ Individu. % MEX C 53 83 1 1 86 24.2 GTA C 21 30 30 8.5 E 4 4 4 1.1 W 7 13 31 1 1 1 52 14.6 CRA C 11 2 9 11 3.1 E 2 1 18 19 5.3 VNZ C 2 11 11 3.1 CLB C 27 5 61 66 18.5 E 7 3 21 24 6.7 ECD C 3 14 14 3.9 E 2 2 2 0.6 PER C 14 27 27 7.6 E 10 9 ] 1 10 2.8 Total 163 13 159 5 1 2 1 1 1 172 1 356 MEX= Mexico; GTA= Guatemala; CRA= Costa Rica; VNZ= Venezuela; CLB= Colombia; ECD= Ecuador; PER= Peru. C= cultivated; E= escape; W= wild. Literature Cited 1. Baudet JC. 1977. Bull. Soc. Roy. Bot. Belg. 110: 65-76. 2. Schmit V & Baudoin JP. 1987. Bull. Rech. Agron. Gembloux 22(3): 235-253. 3. Delgado S., A. 1988. In: "Genetic Resources of Phaseolus beans", P. Gepts (ed.), Kluwer Academic Publishers, Dordrecht, Holland, p. 441-463. 4. Maréchal R. 1971. Bull. Rech. Agron. Gembloux 6(3-4): 461-489. 5. Smartt J. 1973. Euphytica 22: 424-426. 6. Maréchal R, Mascherpa JM & Stainier F. 1978. Boissiera 28: 273p. 7. Hernandez X., E, Miranda C, S & Prywer C. 1959. Rev. Soc. Mex. Hist. Nat. 20(1-4): 99-121. 8. Pinero D & Eguiarte L. 1988. Euphytica 37: 199-203. 9. Debouck DG & Soto JJ. 1988. Tikaiia 6(1): 17-34. 10. Berglund-Brucher O & Brucher H. 1974. Angew. Bot. 48(3-4): 209-220. 11. Brown JWS, Osborn TC, Bliss FA & Hall TC. 1981. Theor. Appl. Genet. 60: 245- 250. 12. Osborn TC. 1988. CRC Critical Rev. Sei. 7(2): 93-116.

* CIAT/IBPGR Research Programme, CIAT-GRU/University of Gembloux Project, Universidad Nacional de Colombia, CIAT Biotechnology Research Unit, respectively, CIAT, A.A. 6713, Cah, COLOMBIA.