Cloning And Functional Analysis Of A Stress-responsive Gene OsTIFY10B In Rice

Rice (Oiyza 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.The TIFY family is a novel plant-specific gene family recently researched in plants. This family has a conserved TIFY domain. Now, it is known that this family of proteins plays an important role in plant growth, development and response to environment changes. Recently, there is some research on JAZ subfamily, and the results suggest that it is functional as a transcription repressor in JA signal transduction, in addition, it also involved in plant responses to abiotic stress.In this study, the OsTIFY10b, belong to JAZ subfamily with TIFY domain and Jas domain, was screened by Fluorescent Differential Display and Macroarry. At first, we analyze the expression level of OsTIFY10b by semi-quantitative PCR; then, perform its transgenic rice by Agrobacterium method, and preliminary analysis its resistance to salination; in order to understand it’s function in stress signal transduction, we use the OsTIFY10b as bait to screen the rice cDNA library of cold and drought treatments, besides we also prepared and purified its GST-fusion protein for the further research.The main results as following:1. Analysis the sequence of OsTIFY10b, we found numbers of cis-elements involved in stress response in its promoter, and it may localized in nucleus; By the result of semi-quantitative PCR, it shows the OsTIFY10b can be induced by various stress and belong to various stress induced gene;2. Performed its transgenic rice, and get the homozygous T2overexpression lines. We found the germination of overexpression line was inhibited on MS medium contain high concentration salt compared with normal Nipponbare, and it suggest that this gene may increase the sensitivity of rice to salt;3. By using the yeast two hybrid system, we use the OsTIFY10b as bait to screen its binding proteins, and one of them has been successfully cloned and sequenced. Prepared and purified its fusion protein by GST-fusion system for the further research.