Functional Analysis Of A Transcription Factor Gene OsAP212 Of Rice And Cold Shock-inducible Protein Gene TIP1 Of Yeast

When the global economy were developing so rapidly, the air pollution, land desertification, water pollution, forest rapid decreasing, poisonous trash and so on,having resulted in the global environmental pollution and ecological crisis, badly affected in plants'growth and development. Drought, high salinity and extremely temperature are the most severe disasters of abiotic stresses harm to crops. Plants have evolved to perceive different stress signals from their surroundings, to integrate them and to respond to these different stresses by modulating the expression of related genes in environmental stress conditions. Transcription factors play vital roles in stress signal transduction, gene expression modulation and phase transition during plant development. In plants, based on the structure character and DNA-binding specificity, TFs can be divided into several families, such as bZIP, AP2/ERF, MYB, WRKY and so on. Transcription Factor OsAP212 gene of rice in our research belongs to AP2/ERF family, which is a special transcription factor family that divided into several subgroups like DREB, ERF, AP2, RAV and so on.Used rice as research materials, a transcription factor gene OsAP212 was isolated that encoded complete ORF from cDNA library of Oryza sativa L. by yeast one-hybrid method. The full-length cDNA of OsAP212 has an 864bp open reading frame (ORF) and encodes an AP2/ERF group transcription factor consisting in 287 amino acids. The secondary structure analysis revealed that OsAP212 strongly resembled other ERF genes and OsAP212 was more close to ERF subgroup in its homologous evolutional tree. All these showed that OsAP212 belongs to ERF subgroup of AP2/ERF familys.The result of RT-PCR indicated that the expression of OsAP212 in rice was more in stem than in leaf, and could be induced by ABA and NaCl.We transferred OsAP212 gene into Arabidopsis thanliana and discovered that the transplant could improve the salt resistance and had some relations to ABA.We guessed that OsAP212 could participate in ABA-dependent signaling transducation pathway.According to amino acid sequence of cold-shock protain gene TIP1 from yeast and preferred codons in plant, we designed primer and successively synthesized TIP1 gene by PCR-based two-step DNA synthesis (PTDS). To analyze the function of TIP1 gene in Arabidopsis thaliana plants, through constructing the Agrobacterium tumefaciens containing plant binary expression vector. The floral dip method was used to transfer TIP1 gene into A. thaliana. The results of study showed that the overexpression TIP1 plants exhibited more improved resistance to cold environment than wild type control plants. We didn't make a conclusion simply with lack of testimony from molecular leval.