Analyse The Expression Of Gene Osh69,SGR And OsDos In Rice And Construct Its RNA Interference Vectors

The research used beta-actin as reference by fluorescence quantitative PCR technology,identified the expression of related regulation gene to senescence of rice from experimental results,and verified these genes’ specific role through experiment.The two breeds’ leaves of rice as the sample,the premature aging(Pei ai 64S,An nong S-1 and Y58S)and the stay green(R287,HD9802s and Liang you 287),to analyze expression level of gene Osh69,OsDos and SGR in heading stage,milk stage and the lately period,and construct the vector of RNA interference,the results show that:(1)Analysis by T-test,the expression of gene Osh69 has not a highly significant increase in the stay green rice,a max increase of 41 percent in Liang you 287;The relative expression amount were increased by 3.7 times and 1.7 times in An nong S-1 and Y58S,having a significant difference.Through the analysis we suggest this gene may contribute to the senescence of leaves in rice.(2)Analysis by T-test,the expression of gene SGR has an increase in the stay green rice,a min increase of 3 percent in R287,an extremely significant difference in different period in HD9802s;The expression amount have a significant difference in An nong S-1 and Y58S.Through the analysis we don’t suggest this gene may contribute to the senescence of leaves in rice.(3)Analysis by T-test,the expression of gene OsDos has a decline in the two breeds of rice,a max decline amount of 27 percent in R287,having a significant difference.Meanwhile,HD9802s and Liang you 287 respectively decreased the percent of 11 and 13;The respectively degressive amount of 28 percent and 80 percent in Pei ai 64S and Y58S.(4)On the basis of the expression vector of pTCK303,successfully built the RNA interference vector of the three purpose gene by using the means of enzyme digestion and enzyme linked,pTCK303-Osh69、pTCK303-SGR and pTCK303-OsDos,and successful genetic transformation in rice cv.Orzysa and R287.