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Suitable electrophoretic separation methods for 34 isozymes from 17 enzymatic systems and a study of the genetic control of certain loci allowed us to investigate the genetic variability and the main factors contributing to its maintenance for wild Phaseolus lunatus L. populations originated in the central valley of Costa Rica. An analysis of the combination of seed and plant numbers to sample per population showed that ten to 80 plants and one to two seeds per plant are necessary to collect the highest genetic diversity when we consider polymorphism at 5 % level. Using the designed sampling scheme and 29 populations, we quantified the genetic diversity at 22 loci. The genotypic composition of the analysed populations showed a deviation from the expected Hardy-Weinberg proportions (Fit = 0.878). This disequilibrium is due to either genetic differentiation between populations (Fst = 0.444) and non random mating within populations (Fis = 0.777). The estimates of the total heterozygosi ty (Ht), the intrapopulation genetic diversity (Hs), and the interpopulation genetic diversity (Dst) were 0.193, 0.082, and 0.111 respectively. About 52 % of the total genetic diversity is due to interpopulation genetic variation (Gst = 0.519), 48 % being relative to intrapopulation genetic variability. No relationships has been highlighted between the intrapopulation genetic variability and the populations size for the studied material. The maintenance of the genetic variability in populations with small size has been attributed to the soil seed bank that reduce the genetic drift effect by increasing the population effective size. An estimation of the mating system parameters and a detailed analysis of factors that influence these parameters allowed us to confirm that wild P. lunatus can be classified as mixed mating with predominantly selfing breeding system (t = 0.072). Analysing the spatial structure, we demonstrated that wild Lima bean populations are subdivided into sub-populatio ns. A study of the micro-geographical pattern of the genetic variability for 95 populations highlighted a structuration in the spatial distribution of allele frequencies. From all the obtained results. we suggested recommendations for P. lunatus genetic resources in situ conservation.

Phaseolus lunatus --- plant population --- genetic variation --- Resource conservation --- Costa Rica --- Costa Rica

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Phaseolus lunatus --- plant population --- population genetics --- Costa Rica --- Costa Rica

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BR Staff Publications [c] (book chapter or book) --- Costa Rica --- Phaseolus lunatus --- in situ conservation

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BR Staff Publications [a] (paper in international, peer reviewed journal) --- Phaseolus lunatus --- genetic diversity --- sampling

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Phaseolus lunatus --- Phaseolus lunatus --- Enzymes --- Enzymes --- Enzyme activity --- Enzyme activity --- Genetic polymorphism --- Genetic polymorphism --- genetic maps --- genetic maps --- Electrophoresis --- Electrophoresis