Cloning And Functional Analysis Of Gene BI-1 In Rice

Rice [Oryza sativa L.] is one of most important crops in this world. Abiotic stress such as heat shock, cold, drought and salination not only influence the growth and development of rice but also reduce its yield. In recent years, with the worsening of the global environment, various extreme weather become a serious threat to the human food production safety, and this make the breeding of the rice with resistance become a key research area today. At present, the research of stress genes and signal transduction by molecular biology can provide some important information to rice breeding, so this is very significance, and The main results of this thesis are as follows:1. According to the approaches of yeast two-hybrid system of the laboratory, In the rice cDNA library was screened with a similar Bax Inhibitor-I gene OsBI-1, and on this basis, Using Agrobacterium mediated rice callus transformation method to obtain obtain transgenicsense and antisense OsBI-1 transgenic rice, to provide materials for further research the expression change of OsBI-lgene in biotic and abiotic stress and stress mechanism.2. The use of online database analysis of OsBI-1 and find the gene open reading frame (ORF) 795bp and encoded 264 amino acid residues. The predicted molecular weight of about28.76KD.Suchasisoelectricpoint7.14.The protein functionprediction that regulator se quence has anti apoptotic protein GAAPcell apoptosis in a 21-261 amino acid. Promoter analysis revealed that the gene contains seven stress induced cis acting element, suggesting that the gene could be a variety of stress induced.3. By semi quantitative RT-PCR analysis OsBI-1, found in the low temperature, high temperature, high salt, drought and abscisic acid of different adversity stress is expressed. And in different tissues of rice in different physiological periods have different degrees of expression, the gene showed differences in spatial and temporal and tissue expression. Suggests that OsBI-1 may belong to as many stress inducible genes.4. constructed a green fluorescent protein vector OsBI-1, PEG mediated transient tran sformation to protoplasts in rice, The subcellular localization of the gene; The Escheric hia coli (BL21) gene expression system, construct the fusion protein expression vector, Induced by expression of fusion protein about 28KD, lay the foundation for further st udies of the gene.5. to obtain a homozygous OsBI-1 transgenic rice plants, through a variety of differentstress treatments on the, Such as:low temperature, high salt, ABA, results show that the over expression lines of the gene enhanced resistance to high salt stress, Inhibition of the expression of plants more sensitive to low temperature stress, furtherconfirmed that the gene is a stress inducible gene.