Characterization Of OsCaMBP Interaction Protein And Function Analysis Of OsSCL30b In Rice

Abiotic stress such as heat shock, cold, drought, salination can influence the plant's growth and development, reduce its yield and even cause their death. In recent years, the global enviroment is becoming worse and worse, drought, flood, cold and other atrocious weather seriously threat food security. Rice is one of main grain crops and wildly growed in our country, its planting area and yield ranks first among other grains. But worse and worse enviroment restrict the output of rice seriously, therefore, the study on stress tolerance become a focus of rice research. Using method of molecular biology to study stress tolerance of plant, finding the process of signal transduction and gene of regulating signal under abiotic stress has a very important significance.Calcium(Ca2+) is a important second messenger in animal/plant cell and joining many physiological reaction. In the plant cell,the concentration of Ca2+ will specific changes at different time and space, which is so-called induction Ca2+singal.In cells, calmodulin(CaM) is one of most important signal transduction molecules in Ca2+signal transduction pathway, which is extensively exist and highly conserved. CaM can regulate the physiological function by interact with CaM binding proteins (CaMBP). The researching CaM is important content of revealing mechanism of CaM, key of proving system of CaM signal, key of further understanding the function of CaM in stress singal pathway, significance of clarifying network of intracellar signaling pathways.In this study, the OsCaMBP whose N-terminal had a IQ CaM binding constraction was sreened by Fluorescent Differential Display (FDD) and Macroarry, IQ motif was the first be discovered Ca2+independent CaMBD, so we verified the interacton between OsCaMBP and OsCaM by yeast two hybrid system (Y2H); and took OsCaM as bait protein to screen its interact protein from the low temperature and drought cDNA library of rice by PEG/LiAc method to transform yeast cell, and then analyze the function of new gene. 1 The position 140-150 amino acid of OsCaMBP form a typical IQ CaM binding motif,which had a feature of Ca2+independent binding protein. Whether OsCaMBP can interact with OsCaM was verified by Y2H. The result show this OsCaM can binding specific OsCaM, and the interaction has some specificity.2 According to Y2H, took OsCaMBP as bait protein to screen low temperature and drought cDNA library of rice by PEG/LiAc method to transform yeast cell. We screen 8 protein interact with OsCaMBP.3 The alternative splicing factor OsSCL30b is one of screened protien by taking OsCaMBP as bait. After RT-PCR and sequencing, a 1250bp cDNA was obtained, its ORF is from 87bp to 878bp and encodes a 263 amino acids protein which is predict 30KD, the 38 to 110 amino acid residues encode a RNA recognition motif(RRM). RRM is highly enrichment in the process of post-transcriptional expression in eukarya; 40,80, 82 amino acid residues is RNA recognition locus.4 In order to study the physiological function of this gene in stress response in rice, we construct the overexpression and anti-sense vectors of OsSCL30b'cDNA fragment, and transform them to callus of rice by agrobacterium.