Analysis Of Genetic Diversity On Lablab By InDels Molecular Markers

Lablab purpureus(L.) Sweet is an ancient legume species and important economical corp. Which has outstanding qualities. The lablab have wide-rage in application, which can be edible and medicinal in many ways, In addition, its stems and leaves also serves as feed for livestock. However, it is small minority crop and has few large areas of cultivation in China. So the basic research in genetics and breeding, quality improvement and other aspects are relative lacking. Hence, how to breed new varieties of lentils and improve quality are big problems to the current agricultural science and technology workers. We analyzed the genetic diversity of lentils, in favor of lentils germplasm collection, conservation and utilization, and could prove the genetic relationship among lentils material, afford theoretical basis in genetic improvement or breeding in latter.This research according to transcriptome sequencing results about Yu Cuibian No-1 and dolichos lablab. Obtaining 12365 indels locus, We picking out the insertion/deletion mutation sites of more than 5bp, and designing primers on either side of the mutation sites. Utilizing agarose gel electrophoresis to filter In Dels primers; making use of PAGE gel electrophoresis to verify the insertion/deletion of Yu Cuibian No-1 and dolichos lablab genome and identify the suspected hybrid lablab; above all, using capillary electrophoresis to analyze the genetic diversity of 21 varieties lentils. The main research results are following.1. According to the result of the transcriptome sequencing, we checking 112 unigenes which have more than 5bp of insertion/deletion sites. Making use of dolichos lablab and Yu Cuibian No-1 DNA as template, by agarose gel electrophoresis picking up 82 In Dels primers. Including 11 primers which have insertion/deletion more than 15 bp. Selecting these primers, analyzing the insertion/deletion in Yu Cuibian No-1 and dolichos genomes. And the results showing, 11 primers amplifying bands have obvious differences, so the genomes have distinct characteristics of the insertion/deletion, and it corroborate with transcriptome sequencing result.2. Using 11 In Dels primers which amplified fragments of insertion/deletion more than 15 bp, and identifying the suspected hybrid lablab. Detecting two hybrid varieties which were dolichos lablab(W)×Yu Cuibian No-1(R) and white moon-5(WM-5) ×Yu Cuibian No-1(R).3. Utilizing 82 In Dels primers amplify genome DNA of 21 varieties of lablab, and using capillary electrophoresis analysis, 82 locus were detecting 539 variable alleles, and each locus has 4-12 loci alleles, so the average of 6.57 of loci alleles. The effective alleles of primers was 2.29-8.35, The average of effective alleles was 4.59. Above all, the primers c39113, c35717, c30881 have the most alleles. The In Dels primers of polymorphism information content(PIC) was 0.565-0.88, the average of PIC was 0.745; The Shannon index was 1.032-2.316, and an average of it was 1.631. The PIC of each primer was more than 0.5. So, The 82 pairs of In Dels primers of experiment were high polymorphism primers. The genetic similarity coefficient of lablab are 0.699-0.824, with an average of 0.763. and the genetic similarity coefficient is small. Indicated that the genetic diversity of 21 lentils type difference was small, and they have close relatives or narrow genetic background.4. UPGAM was implied that 21 samples could be divided into five groups. The first group can also be divided into two sub-class. A sub-class included: white moon-1, white moon-7, white moon-4; red moon-4, red moon-5 and precocious red moon. B sub-class included: white moon-2, white moon-5, white moon-6, white moon-8, white moon-9, red moon-1 and grey dolicho lablab. The second group included: red moon-2 and RM-3. The third group included: white moon-3 and Yu Cuibian No-1. The fourth group included: dolichos lablab, small dolichos lablab and green jade. In addition to, the fifth group is small black. The genetic similarity coefficient was 0.824 between white moon-8 and white moon-9. However, The genetic similarity coefficient between dolichos lablab and Yu Cuibian was 0.753.5. At present, the study of genetic diversity, few In Dels marker have been applied. In other words, This study has provided new way for genetic diversity analysis.