Transformation And Identification Of OsPHS1Gene Related To Rice Meiosis

Polyploidization is one of the important ways to speciation, is widespread in the plant kingdom polyploidization phenomena, many plants are produced by means of polyploid. More than70%of angiosperms are polyploid. Plant after polyploidization, whether in production or quality will show obvious advantages, attaches great importance to the cause of breeders and geneticists.Although, from an evolutionary standpoint rice after polyploidization its yield and resistance will obviously increase like wheat, cotton, rape, however, after long-term of polyploid rice seed setting rate is low in the study is the bottleneck problem of its development. The reason is that the polyploidy plant meiosis, chromosome pairing will occur disorder, easy to cause the formation of multivalent and lagging chromosome, which led to the pollen fertility and seed setting rate reduced, so as to offset the advantage of its shape and failed to achieve the goal of dramatically improve production. Therefore, increase the seed setting rate is the key. By employing the so-called Strategic guidance"by distant hybridization and polyploidization of dual advantages of breeding super rice", after many years of hard work, our lab has bred two polyploid meiosis stability (PMeS) gene lines, the meiosis of normal, high seed setting rate, known as class Ph material. While crossed with other polyploid rice can get higher seed setting rate of offspring, breaking through the long plagued bottleneck of polyploid rice breeding. Created the conditions to achieve the polyploid rice breeding objective. However, the mechanism of the meiotic stability is unclear. Therefore, in-depth study of molecular mechanisms is necessary.OsPHSl gene is maize(Zea may L.) PHS1(poor homologous synapsis1) homologous gene. Studies have shown that PHS1gene on chromosome pairing, synapsis and recombination plays a very important role. The previous experiments we have completed the cloning of rice meiosis related gene OsPHS, induced the expression of target gene in E.coli, constructing OsPHS1gene as a target of over expression vector and RNAi vector and get the gene for RNAi carrier target transformation high rice HN2026-4X resistant plantlet.This research mainly completed the following contents:1) We use PCR and enzyme digestion to verify the former experiments have built carriers, validation results completely correct.2) In order to further explore the rice meiosis related gene OsPHS1function, we are from the excessive expression of the gene and the silent two ways to research, trying to verify the function of the gene from two forward and reverse.Firstly, we use have constructed OsPHS1gene over-express vector to induce callus tissue of lines BLL-2X and4X by Agrobacterium, optimizing transgene system, got resistant plants by resistance gene screening, and analyzed by PCR, Southern blotting detection showed that the target gene has been successfully transferred. Secondly, we carried out the analysis of agronomic traits of TO generation of transgenic plants, the expression of OsPHSl gene over what kind of impact on rice, results show that the BLL-2X average seed setting rate has not improved obviously, while BLL-4X seed setting rate increased significantly, compared with the control plants, the average seed setting rate increased by49.27%. At the same time, the RNAi vector through Agrobacterium mediated transformation of high seed rice HN2026-2X, through selecting, detection of PCR, preliminary evidence the transgenic plants were obtained, but its agronomic traits, especially fecundity, is still under study.