Genetics Analysis and Cloning of Associated Gene Fragments for Large Grain Size Mutant in Wheat
|School||Shandong Agricultural University|
|Keywords||Wheat EMS Mutant EST-SSR SSR SSH|
Traditional chemical mutagenesis method can be induced to produce a high density of point mutations, mutant similar genetic background. Can not only solve the problem of the lack of wheat breeding germplasm resources, but also for the fine mapping of the gene cloning and gene function analysis provides the basis for materials. In this study, EMS mutagenesis wheat Yannong 15 large grain, high pole mutant 8008 study material, genetic analysis of the mutated grain length, grain width and weight traits, try to initial positioning of the mutant gene and gene fragments Cloning, to reveal the Grain Weight genetic mechanism at the molecular level to lay the foundation. (1) large grain, tall mutants in 8008 with the receptor farmers 15 hybridization to build the genetic analysis groups. Analysis found that high grain weight and grain weight low, wide grain with a narrow grain, tall and dwarf the three pairs of traits in the F2 population showed a segregation ratio of 3:1, and the phenotype of the F2 and F3 generation hybrid groups by chi-square test of difference was very significant, are a single dominant gene control; Correlation analysis showed that grain width and grain weight, grain length and grain weight, high pole and grain weight were significantly correlated. (2) mutant 8008 and farmers 15 to 860 pairs of SSR' dissertation">EST-SSR primer material and 960 pairs of SSR primers material carried screening, the resulting differences primer secondary screening in F2 populations selected strains, 59 pairs of primers in a group can amplified the total 99 differential bands. 59 pairs of primers on chromosome 16, to the B genome distribution bit point up to a total of 27; chromosome 16 1B, 2B, the bit point up to 7B three chromosome distribution, respectively, there are seven , 6, 6. (3) 8008 and receptor farmers mutant 15 as the test side and driven by suppression subtractive hybridization (SSH) to build two-way subtractive cDNA library. Positive clones were picked randomly in the two-way library sequencing and bioinformatics analysis in the GenBank database. Received a total of 12 ESTs were highly homologous with the known functional genes, including: photosynthesis-related genes, such as RuBP carboxylase, phytochrome, chlorophyll; role of metabolism-related genes, such as cysteine ??hydrolase, cysteine ??protease, fructosyltransferase, sucrose synthase I; RNA function-related genes, such as RNA-binding proteins, ribosomal protein body. 5 EST sequences with unknown function the cereal crops cDNA fragment highly homologous, the 7 EST failed to find homology match.