Genetic Analysis Of T-type CMS Restore Gene And Cloning And Functional Identity Of Fertility-Relative Genes In Wheat

At present, heterosis application can' t make great progress in wheat. Firstly, wheat is allopoly hexaploid and self-pollination plant. Furthermore, wheat genome is very huge. As the result of artificial selection long ages the heterosis isn' t obvious between varieties. Secondly, T-type (T. timopheevii) CMS line is maintained easily but is restored difficultly. So it is important to culture the new restore line and strengthen screening the hybrid combination in wheat. In the paper the genetic rule of restore gene was analysed and the fertil i ty relative gene were cloned. This would establish the theory and experiment base for wheat heterosis applying in agriculture production.The experiment materials are CMS line 75-3369A, restore line 7269-10 and F2 population and F3 family of 75-3369A/7269-10 combination. The F2 fertile isolations data of continuous three years were analysed. The analysis result showed there were two pairs of dominant major genes and there was additive effect between non-allele genes. The fertility restoring exits quantity character and quality character. The results indicated that the fertility restoring of T-type CMS was controlled by the two pairs of major genes together with some minor genes. The imitate near-isogenic lines (INIL) and near-isogenic lines (NIL)of Rf gene were developed utilizing crossing, backcrossing and self-pollinating.The fertility relative genes were cloned from the single-nuclear pollen of F2 fertility plants using subtractive hybridization. Bulked segregant analysis (BSA) method was used to weaken the inherit background. Two bulks representing extremely fertile (100%) and sterile (0%) plants were constructed. The fertile bulk is the Tester, the sterile bulk is the Driver. Hybridizations were taken three times in order to hybrid adequately. The final hybrid production amplified by LD-PCR was inserted into T-vector to construct the subtractive library. Screening the false positive recombination by reverse northern. The positive clones were used to sequence, BLAST analysis, PCR identify and function analysis.