Polymorphism of resistance gene analogs, peroxidase and wrky transcription factors gene families in the genus lens miller

Resistance gene analogs (RGAs), peroxidases and WRKY transcription factors are multigene families that play roles in resistance to various biotic and abiotic stresses during evolution. In this study, the polymorphism of these gene families are determined in the genus Lens Miller and Vicia montbretii (Fish. & Mey) Davis ve Plitmann. As genetic material, total of 32 genotypes of 7 lentil taxa including four accessions of L. lamottei Czefr., four accessions of L. nigricans (M.Bieb.) Godr, four accessions of L. odemensis Ladiz., eleven accessions of L. orientalis (Boiss.) Ponert, three L. tomentosus Ladiz. and two accessions of V. montbretii were used. Specific degenerate primers were designed from conserved motifs of resistance gene analogs (RGAs), peroxidases and WRKY gene families were used with PCR to amplify targeted regions from genomic DNA of 32 genotypes and obtained monomorphic and polymorphic bands. Genetic similarity between species was determined using UPGMA dendograms. 123 RGA, 22 peroxidases and 10 WRKY primer combinations were tested and 103 RGA, 66 peroxidases and 52 WRKY bands screened. Polymorphism rate of RGAs was 83% and genetic similarity was between 0.62 and 0.90. On the other hand the polymorphism rate of peroxidise genes was 89%. The genetic similarity was between 0.13 and 0.62. In WRKY genes, 47 of the evaluated 52 bands were polymorphic and the polymorphism rate was 90%. The genetic similarity was between 0.07 and 0.63. Results indicated that the gene families have evolved continuously in response to biotic/abiotic stresses. In the comparative analysis with the polymorphism of whole genome, it is apparent that the gene families have evolved at a faster rate than genome average. The results justify targeted gene families approach in studying evolution as compared to the use of random or anonymous loci. The PCR-based RGA, peroxidase, and WRKY markers described in this study may also have potential for linkage mapping and differential gene expression studies in lentils.