Cloning And Preliminary Research Of A New Rice Receptor-Like Protein Kinase OsRLK197

Rice is a kind of important grain crops,which is the stable food of about half of the population in the world,and to reveal its gene function is of great significance to maintain the global food security.Plants receptor protein kinase(receptor-like protein kinase,RLKs)is one of the gene families that belongs to the protein kinase family and was composed of the extracellular domain structure,the structure of the transmembrane domain and intracellular kinase domain.Plant cells can sense the stimulation of the biological and non-biological factors of the outside and make proper response by adjusting the signal transduction pathways and finally regulating the transcriptome and proteome.The intracellular kinase area of RLK response to the external stimulation by opening or closing the downstream target protein by phosphorylation or phosphorylation and switching the extracellular signals into cytoplasm,leading to the starting of the corresponding physiological and biochemical reaction.The study of protein kinase family is of great significance to reveal the mechanism of plant disease resistance and signal transduction;to analyze the mechanism of stimulating of plant response to the outside;to find the receptors during the signal transduction and has become a research hotspot in recent years.OsRLK197 belongs to the sub-family of LRR-RLKs,and whether OsRLK197 responses to the hormone signal transduction and biological and abiotic stresses was studied and the results were as follows:(1)Analysing the basic physical and chemical characters,the orientation in the cells and the expression in different tissues of OsRLK197 using bioinformatics.OsRLK197 was confirmed to belong to the sub-family of LRR-RLKs and have a single transmembrane domain and leucine-rich repeats domain.OsRLK197 is a kind of hydrophilic protein,have no signal peptide,and its kinase domain may have 29 phosphorylation sites which may send signals by changing the conformation.OsRLK197 expresses in different tissues and organs and have the elements that response to ABA and adversity srresses.(2)Five over-expressed OsRLK197 transgenetic lines were screened out via genomic DNA PCR,building over-expressed recombinant vector,conversion of agrobacterium and infection of rice callus.The agronomy traits including plant height,effective ear,thousand seed weight and yield of a single plant was statistics.It was showed that the OsRLK197 overexpression lines had higher plant height,more production of a single ear,less effective ear and no obvious difference of thousand seed weight compared with the wild type.(3)OsRLK197 was confirmed to locate in the cell membrane via transient expression of conversion of arabidopsis protoplast and this is consistent with the prediction results of bioinformatics analysis.The expression of OsRLK197 in different tissues and organs was analyzed through real-time fluorescence quantitative PCR and consistent with the prediction of bioinformatics analysis,OsRLK197 was expressed in every tissue and especially higher in young leaves and seed and lower in root.(4)It is known that OsRLK197 genes has intracellular kinase domain.We built the kinase domain prokaryotic expression vector and inducted the expression of prokaryotic recombinant and purified the protein,which prepared for the subsequent experiments.(5)The over-expressed OsRLK197 transgenetic lines were dealt with 150 mM NaCl,10 uM ABA and drought stress to test the expression of OsRLK197.The results showed that the expression of OsRLK197 increased with the increase of processing time by NaCl and ABA and reduced with the increase of processing time by drought stress.Speculating that the expression of OsRLK197 was induced by NaCl,ABA and drought treatment and OsRLK197 might be involved in adversity slope response.In conclusion,we get the overexpression transgenic plants of OsRLK197,which is a new gene and belongs to leucine rich repeat receptor protein kinase gene family.Analysing the related domain,protein structure and the expression in different tissues of OsRLK197 using bioinformatics.The expression of OsRLK197 in different tissues and organs was analyzed and OsRLK197 was expressed in every tissue and organs.The over-expressed OsRLK197 transgenetic lines were dealt with NaCl,ABA and drought stress and the expression of OsRLK197 was induced by NaCl,ABA and drought treatment.The primary study of OsRLK19 lay a foundation for the research of the mechanism of OsRLK19 in response to ABA signaling pathways and the functions of adversity stress,and provide some experimental basis to reveal the functions of rice receptor protein kinase in plant growth and development,hormone signal transduction and art stress,and helps to reveal the function and the mechanism of more LRR-RLKs in plants.