| Targeting Induced Local Lession in Genomes (Tilling) is a high-throughput reverse genetics method, which can detect associated gene alleles of the biological communities and can identify different gene loci quickly and effectively. In this study, an EMS mutagenesed library of oat was constructed using oat cultivar"Baiyan 2"as the material. The biological characters and agronomic traits of the M2 population were investigated to identify the mutants in field condition, as the genetic variation among some of the M2 population by PCR with intron spliced junction primers and long random primers as well. At the same time, SNPs in the beta glucan synthetic genetic gene AsCsLF6 among the oat mutant materials were detected by Tilling technology based on the agarose gel electrophoresis. The main results were summarized as follows:(1) The phenotype and molecular marker analysis of mutant population showed that there were rich genetic variations among the M2 mutants in biological characters and agronomic traits with total variation of 7.17% in agronomic traits. A total of 99 fragments were amplified from the genomic DNA of 195 individuals of the M2 mutation population with 15 intron spliced junction primers and long random primers. Of which, 65 fragments were polymorphic with a frequency of 65.7%. The genetic similarity coefficients among those M2 mutation individuals ranged in 0.667 to 0.973, which indicated extensive genetic variations among the individuals.(2) SNPs in the beta glucan synthetic genetic gene AsCsLF6 among the oat mutant materials were detected by agarose gel Tilling. The result showed each DNA mixed pool have the same digestion fragments with wide type, showed that there is not mutant in this gene loci among the mutants detected.(3) Cloning analysis for the oat VRN gene, obtained genome sequences 676bp, which has the highest similarity 92% with wheat VRN-A1 genes. Conservative structure domain is MADS and K- box contained VRN - A1 genes. |
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