Identification Of SRAP Molecular Markers Linked To Three Pistils Loci(Pis1) In Wheat

In this study, SRAP technique was used to study the molecular markers related to three pistils gene(Pis1) in wheat. The F2 population produced by from the crossed of Chuanmai 28(CM28) and its near isogentic line for three pistils CM28 TP were used as materials. The molecular marker associated with Pis1 was screened by bulked sergeant analysis(BSA) method and SRAP technology. The results were summarized as follows:1. The wild-type commercial wheat Chuanmai 28(CM28) was crossed with its near isogentic line for three pistils CM28 TP for study the inheritance of three pistils trait. All the F1 hybrids showed three pistils phenotypes, and the F2 populations showed statistically typical Mendelian segregation of 3:1. These results indicated that three pistils trait in wheat was controlled by a single dominance gene.2. The leaves of wheat were used as materials for DNA extraction by the methods of CTAB, Plant Genomic DNA Kit and Plant DNA Isolation Kit(for rich in polysaccharides and phenol). The results suggested that the Plant DNA Isolation Kit(for rich in polysaccharides and phenol) is the best method for wheat DNA extraction both in purity and concertation.3. According to the character of SRAP prime, 1936 pairs of SRAP primer was combined using 44 forward and 44 reverse primers, and the parents(CM28 and CM28TP) was screened using those primer combination. And 224 pairs of primer showed polymorphism among 1936 pairs of primer, the polymorphic percentage was 11.57%. Moreover, 32 pairs of primer combinations appeared stable differences in the normal pilstil DNA pool and three pilsils DNA pool. 11 normal pistil plants and 11 three pistils plants of the two pools were amplified using the 32 pairs of primer combinations, and only seven pairs of primer combinations were related to the three pistils phenotype. By using the primer me2-em36, a specific band of about 100 bp was amplified in 8 three pistils plants, but no band in 6 normal pistils plants; We obtained a specific band of about 200 bp in 9 normal pistil plant by using primer me3-em23, but no band in the 7 three pistils plants; By using the primer me16-em21, a specific band of about 50 bp was obtained in 8 normal pistils plants, but no band found in the same site in 7 three pistils plants; The primer combinations of me16-em25 can amplified a 50 bp band in 8 three pistils plants, but no band in the same site in 7 normal pistils plants; By using primer me16-em26, a specific band of about 80 bp was obtained in 10 normal pistils plants, but no band found in the same site in 9 three pistils plants; By using primer me43-em27, a specific band of about 80 bp was amplified in 8 three pistils plants, but no band found in 7 normal pistils plants in the same site; By using the primer me44-em39, a specific band of about 80 bp was amplified in 8 three pistils plants, but no band found in 8 normal pistils plants. 218 single plants of F2, which including 159 three pistils plants and 59 normal pistil plants were tested using the 7 pairs of primer combinations. The result indicated that only primer combinations me2-em36 and me16-em26 linked with Pis1 gene. We tentatively named them m2e36 and m2e36, and the genetic distance to Pis1 was 18.22 cM and 22.63 cM respectively. The two markers were on both sides of Pis1.