Cloning Antifreeze Relevant Transcription Factor Of Chaling Wild Rice And Study On Antifreeze Mechanism

Wild rice that play a unique role in resistant breeding is rich in excellent gene such as anti-worm, anti-low temperature and drought resistance, poor resistance, and so on. Today some model plants as Arabidopsis of which DREB transcription factor has made good progress in molecular biology, DREB transcription factor expressed by induction of abiotic stress, such as drought, high salinity, low temperature, freezing and other Stress, which play a regulatory role in response to stress and hormone.Using Chaling wild rice as material in the study to research DREB transcription factor, separated total RNA of wild rice under low temperature stress and reverse transcript it into cDNA, Sequencing analysis showed the cDNA sequence be cloned is 958bp, which can be translated into a putative 218 amino acids protein. The cones-quence of Blast showed:the sequence of rice (Oryza sativa) DREB gene homology 98%, and wheat (Triticum monococcum) CRT/DREB4 protein homology of 93%; and barley (Hordeum vulgare) CBF2A and CBFIVc-14.1 protein were 93%,92%. This indicated that DREB transcription factor sequence has been successfully cloned.To further study the DREB expression model in tissue of Chaling wild rice under low temperature stress at different time, so as to Actin sequence of rice genome DNA sequence as an internal control, make DREB transcription factor expression semi-quantitative analysis with RT-PCR technique. The result showed DREB genes have tissue-specific expression at different time of cold stress conditions and expression in roots is more stable, the highest expression level is detected in root. DREB transcription factor is release from the T vector By PCR, and constructed to the plant expression vector pWM101 vector.Compared with cultivated rice, the CAT (Catalase), SOD (superoxide), POD (Peroxidase dismutase) of Chaling wild rice is detected under low temperature at different time. The results found that the enzyme activity of wild rice is higher than the cultivated rice, when the stress accumulated to 24 hours the enzymes activity of wild rice is still maintained at a high level, which is similar with DREB transcription factors expression model under cold stress at different times.