Construction of an integrated map and location of a bruchid resistance gene in mung bean

Bruchid beetle(Callosobruchus chinensis) poses a serious threat to the production and storage of mung bean(Vigna radiata). Mapping bruchid resistance(Br) will provide an important basis for cloning the responsible gene(s) and elucidating its functional mechanism, and will also facilitate marker-assisted selection in mung bean breeding. Here, we report the construction of the genetic linkage groups of mung bean and mapping of the Br1 locus using an RIL population derived from a cross between Berken, a bruchid-susceptible line, and ACC41, a bruchid-resistant line. A total of 560 markers were mapped onto 11 linkage groups,with 38.0% of the markers showing distorted segregation. The lengths of the linkage groups ranged from 45.2 to 117.0 c M with a total coverage of 732.9 c M and an average interval of1.3 c M between loci. Br1 was located on LG9 between BM202(0.7 c M) and Vr2-627(1.7 c M).Based on 270 shared SSR markers, most of the linkage groups were assigned to specific chromosomes. These results should further accelerate the genetic study of this crop.

Genetic diversity assessment of a set of introduced mung bean accessions(Vigna radiata L.)

Genetic resources from other countries or regions play an important role in broadening the genetic background of local breeding varieties. Here we describe observations on the adaptability of mung bean germplasm obtained from the United States Department of Agriculture and their genetic diversity assessment using SSR markers. Several accessions were shown to be mixtures,based on their phenotypes for some characters. Most accessions were able to complete their lifecycles when grown in Beijing, China, making them ideal for crossbreeding without day length control. High diversity was revealed by the SSR markers, with an average of 4.2 alleles per locus and a PIC value of 0.650 per locus. STRUCTURE analysis divided the accessions into six groups.There was no obvious trend of accessions forming groups according to their geographical origin,owing mainly to germplasm exchange and an uneven distribution of accessions. The present results indicate that this germplasm would enrich the local gene pool, and provide information for the further use of germplasm in breeding programs.

Genetic diversity and a population structure analysis of accessions in the Chinese cowpea [Vigna unguiculata(L.)Walp.]germplasm collection

Cowpea(Vigna imguicuiata) is an important legume crop with diverse uses. The species is presently a minor crop, and evaluation of its genetic diversity has been very limited. In this study, a total of 200 genic and 100 genomic simple sequence repeat(SSR) markers were developed from cowpea unigene and genome sequences, respectively. Among them, 27 genic and 27 genomic SSR markers were polymorphic and were used for assessment of genetic diversity and population structure in 105 selected cowpea accessions. A total of 155 alleles and 2.9 alleles per marker were identified, and the average polymorphic information content(PIC) value was 0.3615. The average PIC of genomic SSRs(0.3996) was higher than that of genic SSRs(0.3235), and most of the polymorphic genomic SSRs were composed of di-and trinucleotide repeats(51.9% and 37.0% of all loci, respectively). The low level of detected genetic diversity may be attributed to a severe genetic bottleneck that occurred during the cowpea domestication process. The accessions were classified by structure and cluster analysis into four subgroups that correlated well with their geographic origins or collection sites. The classification results were also consistent with the results from principal coordinate analysis and can be used as a guide during future germplasm collection and selection of accessions as breeding materials for cultivar improvement. The newly developed genic and genomic SSR markers described in this study will be valuable genomic resources for the assessment of genetic diversity, population structure, evaluation of germplasm accessions, construction of genetic maps, identification of genes of interest,and application of marker-assisted selection in cowpea breeding programs.

Analysis of simple sequence repeats in rice bean(Vigna umbellata) using an SSR-enriched library

Rice bean(Vigna umbellata Thunb.), a warm-season annual legume, is grown in Asia mainly for dried grain or fodder and plays an important role in human and animal nutrition because the grains are rich in protein and some essential fatty acids and minerals. With the aim of expediting the genetic improvement of rice bean, we initiated a project to develop genomic resources and tools for molecular breeding in this little-known but important crop.Here we report the construction of an SSR-enriched genomic library from DNA extracted from pooled young leaf tissues of 22 rice bean genotypes and developing SSR markers.In 433,562 reads generated by a Roche 454 GS-FLX sequencer, we identified 261,458 SSRs, of which 48.8% were of compound form. Dinucleotide repeats were predominant with an absolute proportion of 81.6%, followed by trinucleotides(17.8%). Other types together accounted for 0.6%. The motif AC/GT accounted for 77.7% of the total, followed by AAG/CTT(14.3%), and all others accounted for 12.0%. Among the flanking sequences, 2928 matched putative genes or gene models in the protein database of Arabidopsis thaliana, corresponding with 608 non-redundant Gene Ontology terms. Of these sequences, 11.2% were involved in cellular components, 24.2% were involved molecular functions, and 64.6% were associated with biological processes. Based on homolog analysis, 1595 flanking sequences were similar to mung bean and 500 to common bean genomic sequences. Comparative mapping was conducted using 350 sequences homologous to both mung bean and common bean sequences. Finally, a set of primer pairs were designed, and a validation test showed that58 of 220 new primers can be used in rice bean and 53 can be transferred to mung bean.However, only 11 were polymorphic when tested on 32 rice bean varieties. We propose that this study lays the groundwork for developing novel SSR markers and will enhance the mapping of qualitative and quantitative traits and marker-assisted selection in rice bean and other Vigna species.