Embryo development in reciprocal crosses of Phaseolus vulgaris L. and P. coccineus Lam.

Summary Embryo development was examined in reciprocal crosses of Phaseolus vulgaris cv. Great Northern and P. coccineus cv. Scarlet Runner. The formation of abnormal (shrunken and underdeveloped) embryos constituted the primary crossing barrier between the two species when P. coccineus was the female parent. Plants of P. coccineus X P. vulgaris were obtained by embryo culture. Although the P. vulgaris X P. coccineus cross resulted in normal seed development, the fertility of the resulting hybrids was much lower (27%) than that of the reciprocal hybrids (81%). Three classes of F₂ embryos, normal, shrunken, and underdeveloped were formed on reciprocal F₁s and the frequencies did not differ between reciprocal populations. Thus, the interactions between embryo and endosperm and/or maternal parent rather than cytoplasmic-nuclear effects seem to be important in the determination of the extent of embryo growth. The examination of pollen fertility of F₂ plants and the development of F₂ and F₃ embryos suggests that the formation of abnormal embryos and reduced male fertility are independent events. The P. vulgaris — P. coccineus crosses may be useful in studying the possible involvement of interspecific differences in hormonal metabolism in the development of hybrid embryos.     

DNA restriction fragment length polymorphisms correlate with isozyme diversity in Phaseolus vulgaris L.

Summary Genetic variation in Phaseolus vulgaris L. (P. vulgaris) was investigated at the isozyme and DNA levels. We constructed a library of size-selected Pst I clones of P. vulgaris nuclear DNA. Clones from this library were used to examine 14 P. vulgaris accessions for restriction fragment length polymorphisms (RFLPs). DNAs from each accession were analyzed with three restriction enzymes and 18 single copy probes. The same accessions were also examined for variability at 16 isozyme loci. Accessions included four representatives of the T phaseolin group and five representatives each of the C and S phaseolin groups. One member of the S group (the breeding line XR-235-1-1) was derived from a cross between P. vulgaris and P. coccineus. Isozymes and RFLPs revealed very similar patterns of genetic variation. Little variation was observed among accessions with C and T phaseolin types or among those with the S phaseolin type. However, both isozyme and RFLP data grouped accessions with S phaseolin separately from those accessions with C or T phaseolin. The highest degree of polymorphism was observed between XR-235-1-1 and members of the C/T group. RFLP markers will supplement isozymes, increasing the number of polymorphic loci that can be analyzed in breeding, genetic, and evolutionary studies of Phaseolus.     

Use of chloroplast DNA polymorphisms for the phylogenetic study of seven Phaseolus taxa including P. vulgaris and P. coccineus

The genetic variability of seven Phaseolus taxa has been evaluated on the basis of molecular data and the results have used to clarify the phyletic relationships between several taxa of the P. coccineus L. complex. Chloroplast DNA (cpDNA) from 33 populations was digested with six restriction endonucleases, revealing some polymorphisms that made it possible to divide most of the taxa into two main groups: the subspecies of P. coccineus on the one hand, and P. vulgaris L., P. polyanthus Greenman and P. costaricensis (Freytag and Debouck) on the other hand. P. polyanthus is closer to P. vulgaris than the other taxa of the second group and should be considered as a separate species. The position of the wild species P. costaricensis is intermediate between P. coccineus and P. polyanthus. P. glabellus shows sufficient polymorphisms at the cpDNA level to be recognized as a separate species, as previously suggested from total seed-protein electrophoretic studies. These results favour the hypothesis of a common phylogeny for P. vulgaris, P. polyanthus, P. costaricensis and P. coccineus from a single wild ancestor. Although cpDNA is generally known to be uniform at the intraspecific level, some additional polymorphisms were also detected within P. vulgaris, P. polyanthus and P. coccineus. Further studies are required to understand the significance of the latter.     

Investigations into the inter-relationships ofPhaseolus vulgaris L. andP. Coccineus LAM

The crossability ofP. coccineus (self-incompatible) andP. vulgaris (self-compatible) was much higher whenvulgaris was used as the female parent. Reciprocal difference in crossability was due to the retarded development of embryo and endosperm tissue during the immediate post-fertilization period in the crossP. coccineus ×P. vulgaris . Backcrossing the reciprocal F₁ hybrids to the maternal species was more successful than to the paternal species.Two varieties ofP. vulgaris differed significantly in their ability to hybridize with six genotypes ofP. coccineus; this difference was shown to be genetical.The F₁ hybrids were readily distinguishable from the parent species at the seedling stage and could be classified into two distinct morphological types; one group showed and abnormal wrinkling of the leaves. The frequency of the abnormal progeny in the F₁ generation could be satisfactorily explained on the basis of either allelic or genic interaction. The importance of this genetic system in the divergence of the two species is discussed.     

Changes in DNA-methylation during zygotic embryogenesis in interspecific hybrids of beans (<Emphasis Type="Italic">Phaseolus</Emphasis> ssp.)

Hybrid embryos resulting from crosses between Phaseolus species often fail to reach maturity and some combinations frequently abort at early developmental stages. The genetic or molecular basis for these consistent developmental defects is at present not clear. However, an extremely complex genetic system, thought to be caused by major epigenetic changes associated with gene expression changes, has been shown to be active in plant species. We have investigated DNA methylation in two interspecific hybrids, Phaseolus vulgaris × Phaseolus coccineus and its reciprocal crosses, using methylation sensitive amplification polymorphism (MSAP). The potential use of MSAP for detecting methylation variation during embryogenesis in interspecific hybrids is discussed. Significant differences in the DNA methylation patterns were observed in abortive (interspecific hybrids) and non abortive (parental) genotypes. Taken together, our results strongly suggest that generalized alterations in DNA methylation profiles could play a causative role in early interspecific embryo abortion in vivo. A considerable change in the methylation pattern during embryogenesis could be involved in the disruption of the regulation or maintenance of the embryogenesis process of Phaseolus interspecific hybrids. The results also support the earlier hypothesis that DNA methylation is critical for the regulation of plant embryogenesis and gene expression.     

DNA content of Phaseolus coccineus x P. vulgaris suspensors

Summary At the globular stage of embryo development, the level of DNA, as determined from microspectrophotometric analysis of Feulgen-stained squashes, was significantly higher in the interspecific hybrid suspensor than in suspensors from self-pollination of Phaseolus coccineus, the maternal parent. However, at the early-heart and early-cotyledonary stages of development, DNA content of interspecific hybrid suspensors was significantly lower than that of suspensors formed after self-pollination of either P. coccineus or P. vulgaris. The relationship between DNA content and suspensor cell length for P. coccineus and P. vulgaris at all developmental stages and between DNA content and cell area for P. coccineus at the early-cotyledonary stage was altered in hybrid suspensor cells. Nuclei in large cells of interspecific suspensors exhibited uneven distribution of polytene chromosomes and no clear nuclear outline.Paper number 18,470 of the Scientific Journal Series, University of Minnsota Agricultural Experiment Station, St. Paul, MN. This research was funded by the USDA CRGO under grant number USDA-85-CRCR-1-1676     

Restriction fragment length polymorphisms of the phytohemagglutinin genes inPhaseolus andVigna (Leguminosae)

Restriction fragment length polymorphisms at the phytohemagglutinin (PHA) locus were determined among 21 genotypes ofPhaseolus vulgaris, P. coccineus, P. acutifolius, P. lunatus, and threeVigna species, using five restriction enzymes and one double digestion, in order to provide molecular evidence for their genetic relatedness. The dissimilarity between genotypes was estimated from binary RFLP data. The dissimilarity was high among species (from 0.75 to 0.95), and of variable extent among genotypes of the same species (0.33–0.89). InP. vulgaris, two different DNA hybridization patterns were found, giving further evidence for two major gene pools in that species. The restriction patterns ofP. vulgaris var.aborigineus, the putative ancestral form ofP. vulgaris, exhibit clear homology toP. vulgaris genotypes. An undefined landrace from Taiwan could be identified as aP. vulgaris genotype. RFLP-based trees for the phytohemagglutinin genes of the species studied were computed with several distance matrix and parsimony methods.     

Interspecific hybridization of phaseolus vulgaris with P. lunatus and P. acutifolius

Summary The influence of genotypic combinations on the growth of hybrid embryos between Phaseolus vulgaris and P. lunatus, and between P. vulgaris and P. acutifolius was examined. All embryos obtained from P. vulgaris × P. lunatus crosses developed only to a stage which appears to be comparable to the pre-heart-shape stage of selfed embryos. Reciprocal crosses were attempted, but pods abscised at a very early stage. Embryos derived from P. vulgaris × P. acutifolius and reciprocal crosses attained the cotyledon stage although no mature seeds were formed. A distinct characteristic of these embryos was the uneven development of the two cotyledons. The rate of growth and final size of these hybrid embryos seemed to be influenced by the genotypes of both parents.Immature embryos were cultured on defined medium and the effects of glutamine and gibberellin (GA₃) were examined. Glutamine was effective in increasing the survival rate; gibberellin had no apparent effect. Plants derived from cultured embryos of P. vulgaris × P. lunatus, P. vulgaris × P. acutifolius and P. acutifolius × P. vulgaris were obtained.Technical paper No. 00 of the Oregon Agricultural Experiment Station. Research was supported by the Oregon Agricultural Experiment Station, the Research Council of Oregon State University (National Institute of Health Biomedical Research Support Grant RR07079) and the Processor Research Council of Oregon. A.R. is supported by an African Graduate Fellowship from the African-American Institute     

Nodulation and Nitrogen Fixation Efficacy of Rhizobium fredii with Phaseolus vulgaris Genotypes

Phaseolus plant introduction (PI) genotypes (consisting of 684 P. vulgaris, 26 P. acutifolius, 39 P. lunatus, and 5 P. coccineus accessions) were evaluated for their ability to form effective symbioses with strains of six slow-growing (Bradyrhizobium) and four fast-growing (Rhizobium fredii) soybean rhizobia. Of the 684 P. vulgaris genotypes examined, three PIs were found to form effective nitrogen-fixing symbioses with the R. fredii strains. While none of the Bradyrhizobium strains nodulated any of the genotypes tested, some produced large numbers of undifferentiated root proliferations (hypertrophies). A symbiotic plasmid-cured R. fredii strain failed to nodulate the P. vulgaris PIs and cultivars, suggesting that P. vulgaris host range genes are Sym plasmid borne in the fast-growing soybean rhizobia.     

Chloroplast DNA as an evolutionary marker in thePhaseolus vulgaris complex

We have analyzed the changes occurring in the chloroplast DNA (cpDNA) of taxa belonging to thePhaseolus vulgaris complex to help clarify relationships among species of this complex. Two restriction maps for 11 restriction enzymes comprising the whole chloroplast genome from a wildP. vulgaris and a wildP. coccineus accession were constructed. These maps allowed us to compare a total of 330 restriction sites between the two genomes in order to identify polymorphisms, assess the type of mutations detected, and identify regions of high variability. A region, located in the large single-copy region near the borders with the inverted repeats, accounted for a large portion of the variation. Most of the mutations detected were due to restriction sites gains or losses. Variable and conserved regions were then evaluated in 30 accessions belonging to taxa of theP. vulgaris complex. Phylogenetic analyses were made using parsimony methods. Conclusions obtained from such analyses were the following: (1) there was high cpDNA variability withinP. coccineus but not inP. vulgaris. (2)P. coccineus subsp.glabellus showed a very distinct cpDNA type that strongly suggests that it actually belongs to a different but as yet undetermined section of the genus. Our cpDNA observations are supported by distinctive morphological traits and reproductive biology of this taxon. (3) InP. coccineus subsp.darwinianus (also classified asP. polyanthus), the cpDNA lineage was in disagreement with data obtained from nuclear markers and suggested a reticulated origin by hybridization betweenP. coccineus as the male parent and an ancestralP. polyanthus type, closely allied toP. vulgaris, as the seed parent. This initial cross was presumably followed by repeated backcrossing toP. coccineus. Our cpDNA studies illustrate the importance of molecular markers in elucidating phylogenetic relationships. They also indicate that accurate phylogenies will require analyses of both nuclear and cytoplasmic genomes.     

Is a decreased water potential after withholding oxygen to roots the cause of the decline of leaf-elongation rates in Zea mays L. and Phaseolus vulgaris L.?

Leaf-elongation rates of Zea mays L. and Phaseolus vulgaris L. were measured in plants grown for 4 d in nutrient solution bubbled with N₂ and in soil-grown waterlogged Phaseolus plants. Leaf water potential in both species was lower 3–4h after replacing aeration by N₂-bubbling. In Zea, the water potential after 24 h or more was the same in control plants and plants with N₂ treatment. In Phaseolus, the water potential of inundated plants and plants with N₂ treatment was always lower than those of control plants. The leaf-elongation rate of both species was always lower in plants treated with N₂, especially during light periods. In Zea, the elongation rate was lowest in the first 24 h, whilst in Phaseolus it was lowest on the last (fourth) day of treatment. There was no difference between N₂ treatment and inundation experiments. It is concluded that during the first hours of treatment the leaf-elongation rate was reduced as a consequence of the lower water potential. Thereafter, however, elongation rates were lower than could be expected on the basis of the plant's water relations.Abbreviations LER leaf elongation rate - PEG-200 polyethylene-glycol 200 - RWC relative water content     

RELATIONS D‘ACANTHOSCELIDES OBTECTUS AVEC DIFFÉRENTES ESPÈCES DE PHASEOLUS: INFLUENCE SUR LA FÉCONDITÉ ET POSSIBILITÉS DE DÉVELOPPEMENT LARVAIRE

Résumé La présence des gousses et graines de six espèces de Phaseolus déclenche une ponte importante chez A. obtectus et stimule significativement la production ovarienne. Cependant, les meilleures performances ne sont pas observées avec P. vulgaris, hôte habituel, mais avec P. coccineus.Pour toutes ces espèces le tégument de la graine représente une barrière importante pour la pénétration des larves. Chez certaines espèces sauvages, et dans les conditions expérimentales, il permet une protection totale de la graine. Cette protection semble diminuer avec la domestication chez P. vulgaris.Lorsque les graines sont perforées artificiellement, les larves (une par graine) pénètrent sans difficulté, quelle que soit l'espèce. Les graines de P. coccineus et de P. vulgaris permettent le développement complet de la majorité des larves, tandis que dans celles de P. metcalfei et de P. acutifolius ne peut se développer qu'une faible partie de la population larvaire. Certains Phaseolinae peuvent donc stimuler la ponte sans permettre le développement des larves.Les femelles de la F₁ provenant de P. coccineus ont des performances reproductrices supérieures à celles des femelles provenant de P. vulgaris (variétés sauvage ou cultivée). P. coccineus pourrait être considéré comme une des plantes-hôtes d'origine.

---

Summary The presence of pods and seeds of six Phaseolus species induces significant egg-laying in A. obtectus and strongly stimulates ovarian production. However, the best performances are not observed with P. vulgaris, the common host, but with P. coccineus.For all six species, the seed testa is an important barrier against larval penetration. For some wild species, it can give total protection of the seed, but this protection is reduced with domestication in P. vulgaris.For all species, when the seeds are artificially perforated, larvae (one per seed) penetrate easily. Most of the larvae can complete their development in seeds of P. coccineus and P. vulgaris, but only a small proportion of the larval population can develop in seeds of P. metcalfei and P. acutifolius. Thus certain Phaseolinae seeds stimulate egg-laying without allowing larval growth.Females of the F₁ reared in P. coccineus have superior reproductive capacities than those reared in P. vulgaris. P. coccineus could be considered as one of the original host-plants of A. obtectus.

     

Fertile,intermediate hybrids between Phaseolus vulgaris and P. acutifolius from congruity backcrossing

Summary Recurrent or introgressive backcrossing of Phaseolus vulgaris — P. acutifolius hybrids with either P. vulgaris or P. acutifolius increased fertility, as measured by pollen stainability and seed per pod from non-manipulated flowers, while invariably resulting in loss of traits from the non-recurrent parent. When hybrids were backcrossed with each of the parent species in alternate generations (congruity backcrossing), fertility decreased in early generations but gradually increased in later generations. By the fourth or fifth generation, congruity-backcross hybrids produced numbers of seeds per pod from non-manipulated flowers comparable to those of parent species, although the percent of stainable pollen tended to be lower. Congruity-backcross hybrids were intermediate to parent species when pedigrees contained equal representation of parents but favored the majority parent when pedigrees were unbalanced. Individuals exhibiting symptoms of developmental incongruity, such as abnormal leaflet number, foliar variagation, or unusual growth patterns, occurred in each generation. However, completely new characteristics appeared after the second backcross generation, presumably due to recombination. Backcrossing to both parent species in alternate generation reversed incongruity, allowing selection for fertility without loss of traits from one of the parent species.Scientific Journal Series Paper Number 15, 671 of the Minnesota Agricultural Experimental Station     

Molecular evidence for an Andean origin and a secondary gene pool for the Lima bean (Phaseolus lunatus L.) using chloroplast DNA

 Chloroplast DNA (cpDNA) diversity has been examined using PCR-RFLP and RFLP strategies for phylogenetic studies in the genus Phaseolus. Twenty-two species, including 4 of the 5 cultivated species (P. lunatus L., the Lima bean; P. vulgaris L., the common bean; P. coccineus L., the runner bean and P. polyanthus Greenman, the year-bean), represented by 86 accessions were included in the study. Six PCR primers designed from cpDNA and a total cpDNA probe were used for generating markers. Phylogenetic reconstruction using both Wagner parsimony and the neighbor-joining method was applied to the restriction fragment data obtained from each of the molecular approaches. P. vulgaris L. was shown to separate with several species of largely Mesoamerican distribution, including P. coccineus L. and P. polyanthus Greenman, whereas P. lunatus L. forms a complex with 3 Andean species (P. pachyrrhizoides Harms, P. augusti Harms and P. bolivianus Piper) co-evolving with a set of companion species with a Mesoamerican distribution. Andean forms of the Lima bean are found to be more closely related to the 3 Andean wild species than its Mesoamerican forms. An Andean origin of the Lima bean and a double derivative process during the evolution of P. lunatus are suggested. The 3 Andean species are proposed to constitute the secondary gene pool of P. lunatus, while its companion allies of Mesoamerican distribution can be considered as members of its tertiary gene pool. On the basis of these data, an overview on the evolution of the genus Phaseolus is also discussed. Received: 1 May 1998 / Accepted: 13 July 1998     

A unique cytoplasmic male sterility (CMS) determinant is present in three Phaseolus species characterized by different mitochondrial genomes

Previous results have shown that cytoplasmic male sterility (CMS) in lines from Phaseolus coccineus and Phaseolus vulgaris contain the same CMS-specific sequence, raising the question of whether this sequence rearrangement arose before divergence of the two species or afterward with subsequent transfer by introgression. Hybridization patterns of total DNA from eight P. vulgaris lines with cytoplasm from P. coccineus and three P. vulgaris lines were examined in order to analyze the mitochondrial DNA (mtDNA) diversity within each species and to determine differences between CMS lines derived from the two species. Three restriction enzymes and 17 heterologous mtDNA sequences were used. The analysis of the different hybridization patterns revealed a considerable diversity in mtDNA organization particularly within P. coccineus. We obtained distinctive hybridization patterns for the five CMS lines tested. The resulting classification showed that mitochondrial genomes from P. coccineus CMS lines group with those of fertile P. coccineus but not with CMS lines from P. vulgaris. The groupings concur with the taxonomic classification of these lines. The results support the hypothesis of a single ancient origin of the CMS determinant and exclude the transfer of cytoplasm by introgression from P. vulgaris to P. coccineus and P. coccineus ssp polyanthus.     

Genetic structure and mating systems in wild and cultivated populations of Phaseolus coccineus and P. vulgaris (Fabaceae)

We present a study of the genetic variation, genetic structure, and outcrossing rate of two wild populations of Phaseolus coccineus ssp. formosus (both from Central Mexico), two cultivated populations of P. coccineus ssp. coccineus (one from Central Mexico, the other from Chiapas), and one cultivated population of P. vulgaris (from Central Mexico), using seven electrophorectic loci. All the populations of P. coccineus showed high and similar levels of genetic variation, without differences among wild and cultivated populations (H ranged between 0.187 to 0.271). In contrast, P. vulgaris has low levels of genetic variation (H = 0.041). The three P. coccineus populations from Central Mexico were genetically similar, while the cultivated P. coccineus from Chiapas and the cultivated P. vulgaris from Central Mexico were very different. In all the loci and all the populations for both species, the fixation indices (F) were positive and differed significantly from random mating expectation in 59% of the cases for P. coccineus. The highest F was the estimated for P. vulgaris. For P. coccineus, the multilocus outcrossing rate estimates ranged from 0.592 to 0.698, and these did not differ significantly among populations. For P. coccineus, we also estimated the outcrossing rate for each of 60 different families, showing great variation, with a majority of families with a t between 0.8 and 1, but a substantial fraction of the families with a t as low as 0.12–0.20. The domestication process has neither eroded the levels of genetic variation of the cultivated populations of P. coccineus nor changed its mating system.     

Mitochondrial genome size variation and restriction fragment length polymorphisms in threePhaseolus species

Summary Restriction patterns of mitochondrial DNA (mtDNA) from threePhaseolus species were examined to estimate their relative genome sizes and to determine the level of interspecific variability and relatedness. Three restriction endonucleases that produced relatively simple profiles were identified and used to determine the genome size of the three species. Taking into account fragment stoichiometries, the average estimates across enzymes were 456, 324, and 400 kb, respectively, forP. vulgaris, P. coccineus, andP. acutifolius. Restriction fragment length polymorphisms (RFLPs) differentiated the species when the mtDNAs were digested with seven endonucleases and hybridized with five cosmid clones covering ca. 200 kb of mtDNA sequences. Proportions of shared restriction fragments between every two species were computed as F-values and demonstrated thatP. vulgaris andP. coccineus are more related to each other than either is toP. acutifolius, and that the latter has a similar degree of relationship to the other two species.     

In situ hybridization to metaphase chromosomes in six species ofPhaseolus andVigna using ribosomal DNA as the probe

In situ hybridization with a biotin-labeled rice ribosomal DNA (rDNA) probe to the somatic metaphase chromosomes of six species ofPhaseolus andVigna (P. angularis, P. calcaratus, P. coccineus, P. vulgaris, V. sesquipedalis andV. sinensis) was done to determine the sites of rDNA. Hybridization signals were present in the terminal and subterminal chromosome regions of each of the six species. The number of rDNA sites was two inP. angularis andP. calcaratus, four inP. coccineus andP. vulgaris, and six inV. sesquipedalis andV. sinensis.     

Meiosis and fertility of interspecific hybrids betweenPhaseolus vulgaris L. andP. acutifolius A. Gray

Summary Meiosis and fertility of interspecific hybrids obtained from reciprocal crosses between Phaseolus vulgaris and P. acutifolius were examined. Bivalents as well as univalents were found at Metaphase I. The majority of the microsporocytes had four or more univalents and the average was 6.3 univalents per cell. The average number of lagging chromosomes at Anaphase I was 2.3 per cell and the most frequent chromosome distribution at late Anaphase I was 10–12. The lower than expected number of lagging chromosomes as compared with the number of univalents at Metaphase I suggests the possible occurrence of precocious separation of bivalents. The male fertility as measured by pollen stainability was 17%, however, the frequency of pollen germination in selfing was 3.5%. Upon selfing of the interspecific hybrids, no dividing embryos were found even though 7 and 26% of the ovules were fertilized at 12 hours and four days after pollination. In backcrosses to P. vulgaris (male), 6 and 20% of the ovules were fertilized and 0 and 4% of the ovules contained dividing embryos at the same sampling times. When P. acutifolius was the male parent, respective values were 8 and 31% for fertilization and 0 and 13% for ovules with dividing embryos. The frequencies of backcross embryos recovered at 14–26 days were in agreement with the frequencies of dividing embryos at four days. The ability to obtain backcross plantlets demonstrates the feasibility to further utilize interspecific hybrids for the improvement of P. vulgaris Technical paper No. 5311 of the Oregon Agricultural Experiment Station. Research was supported by the Oregon Agricultural Experiment Station, the Science and Education Administration of the U.S. Department of Agriculture under Grant 5901-0410-8-0028-0 from the Competitive Research Grants Office, the Research Council of Oregon State University (NIH Biomedical Research Support Grant RR 07079) and the Processor Research Council of Oregon, A.R. and C T.S. are respectively supported by an African Graduate Fellowship from the African-American Institute and a fellowship from the National Science Council of the Republic of China     

Description and analysis of genetic diversity between commercial bean lines (Phaseolus vulgaris L.)

The effectiveness of RFLP, DAMD-PCR, ISSR and RAPD markers in assessing polymorphism and relationships between 24 commercial lines of Phaseolus vulgaris L.was evaluated. We have used a Phaseolus-specific minisatellite sequence as a probe, which enabled 23 of the bean lines tested to be fingerprinted. Based on the sequence information obtained, primers corresponding to the bean-specific minisatellite core sequence were used in subsequent PCR amplifications. Our observations indicated that while the DAMD-PCR was sensitive in detecting genetic variation between bean species and between accessions of P. vulgaris, when used alone it may be limited in its ability to detect genetic variation among cultivated bean lines due to the low number of loci amplified. Only one out of the five ISSR primers tested was efficient in generating multiple band profiles, which was insufficient to distinguish all the different bean lines. Reproducible RAPD profiles were obtained, and these allowed us to differentiate all the genotypes tested with seven primers. We ultimately used only results from RFLP and RAPD markers to explore the genetic diversity among commercial bean lines. Both analyses led to the same clustering of the bean lines according to their geographical origins (United States or Europe). With respect to the European lines, the results obtained from RAPD data also enable the lines to be clustered according to their creators. Received: 15 January 2000 / Accepted: 21 March 2000