Cloning And Genetic Transformation Of Pod-shattering Gene In Soybean

Pod shattering is a natural attribute of the soybean. It is one of the important factors which influence the production of soybean. The purpose of this study is to find the soybean cracking control in large pod genes, and through transgenic technology put the gene into soybean yellow 10, observe the phenotypic changes. The early stage of the experiment through to the related literature search, were found two genesFULLand PpMADS6 in arabidopsis and peach. Research shows that the two genes and floral organ development in arabidopsis thaliana and pod cracking properties related, so compare the genes with the soybean genome, thus found an soybean AGL8 gene which is high similarity Experiment will let AGL8 gene and expression vector pCAMBIA 1300::EYFP connection and by the method of agrobacterium infect soybean cotyledon section yellow 10 for genetic transformation of soybean, and bioinformatics analysis of soybean AGL8 gene. The experimental results show that through DNAMAN compare AGL8 with the two genes FRUITFULL and PpMADS6. The result is they are similarity 78% and 80% respectively. By PCR amplification of soybean yellow 10, put the AGL8 genes sequence in the NCBI and alignment gengbank, The genes sequence is similarity of 99.84% of gengbank. Using agrobacterium mediated soybean cotyledon section of genetic variations that will put AGL8 gene into soybean. Experimental test of hygromycin selection agent concentration, ultimately determine the growing medium concentration of hygromycin for 8 to 10 mg/1, elongation medium concentration of hyg romycin is 5 mg/1. By PCR detection and laser confocal microscopy proved gene AGL8 have been successfully transferred to soybean yellow 10. Through the To generation of transgenic plants, agronomic characters observation found that the To generation transformant flowering is later than Wt, the number of flowering is increased, and we also found the situation of Flower abortive in To.In terms of pod cracking character, mature To generation pod has not yet appeared cracking character. Through bioinformatics analysis, we found that the total length of gene AGL8 is 855 bp, CDS area is 735 bp, containing MEF2 structure, DNA binding sites, assume that the phosphorylation sites and peptide binding sites. Evolutionary tree analysis found that soybean and bean tian qing similarity is higher, close relationship in the evolutionary tree.