Cloning And Polymorphism Analysis Of TaFLD And TaFPA Gene In Wheat Related To Flowering Time Of Autonomous Pathway

Plant flowing is the most important development stage from vegetative to produtive growth and the time of flowing occurred at an approppriate time plays a key role in the process of plant growth an development.Wheat(triticum aestivum L.)is one of the most important food crops in the world, its flowering is an important impact in the process of the wheat growth stages, the adaptability and yield have. In recent years, through the model plant Arabidopsis thaliana, there have been found four ways to achieve the ultimate plant flowering:long-day photoperiod pathway, vernalization pathway, gibberellin (GA) pathway and autonomous pathway. At present, long-day photoperiod pathway has been the focus of the study in rice and wheat, vernalization way has become a research focus in wheat, but for (GA) pathway and autonomous pathway of little , autonomous pathway is also an integral part of the process available flowering plants.This research is based on Arabidopsis autonomous pathway genes, to explore the types of wheat on the independent channels, according to the comparison Scafford, predicted gene sequences, using bioinformatics analysis of sequence features, design D genome specific primers, 68 were material PCR products were recovered, direct sequencing, development of molecular markers, and then Conduct the types of single-fold type, distribution and functional analysis of 196 breeding species,Cultivars and Wild species bred at home and abroad, the main results of this experiment are as follows:1.According to the mRNA sequence of autonomous pathway of Arabidopsis gene FLD and FPA, by sequence alignment, they were predicted homologous sequences in wheat, on the prediction of homologous sequences to sequence analysis, based on their functional domains initially found to wheat does contain two autonomous control flowering genes.2. Based on sequence analysis of the results,design D specific primers in the promoter region of the TaFLD and TaFPA gene, and make 68 copies of the material was recovered and PCR sequencing to identify the differences between different varieties.3. TaFLD gene detected four SNPs: S705, S815, S837, S1000, 1 deletion sites 831-862 (32bp). TaFPA gene detected three SNPs: S407, S429, S792, 2 deletion sites 857-1225bp (369bp) and 2018-2069 (52bp), 3 insertion sites 1748,1957 (11bp) and 2104 (6bp).4. Aimed at TaFLD gene's 32bp deletion and two SNPs to studied, the two SNPs have restriction Enzyme cutting site, you can use restriction endonuclease to detect whether there is polymorphism, for the insertion or deletion polymorphism sites used in the insertion or deletion sites at both ends of primers, if there have insertion or deletion, the amplification primers and the size of the amplified fragments of inconsistent goals, in order to determine whether there is polymorphism.5. According to the design of molecular markers, detected the polymorphism of 196 materials, using the second PCR, first use the D genome specific primers by TaFLD and TaFPA amplificate 196 copies of materials , then the development of molecular markers were determined.6. The result of molecular markers in 196 materials, the materials appear less polymorphism may be due to the two promoters have greater conservation and genetic background, making the material does not fully performance of the promoter region for expression analysis of gene function starts, but also the need for further study.