Analysis Of OsSCL30b And Cloning And Stress Resistance Analysis Of OsVHA16 Gene In Rice

Through the previous work of this laboratory,the gene OsSCL30b was transferred into rice to obtain T2 transgenic plants.This protein domain is critical for the binding of RNA molecules.RNA recognition motifs are generally relatively conserved in sequence and are protein domains widely found in eukaryotes.In the process of transcriptional regulation,multiple RRM proteins are involved in every step of regulation including pre-mRNA processing,substitutional splicing and editing.The OsSCL30b transgenic plants showed a delayed growth phase in the field.To study the delayed growth mechanism of OsSCL30b transgenic plants,OsVHA16 gene was obtained by yeast two-hybrid screening,and the OsVHA16 gene studied in this study was named as VHA16 because it encodes rice V-type H~+-ATPase.H~+-ATPase exists in the plasma membrane and various endometrial systems of almost all animal and plant cells,and plays a role in establishing a transmembrane proton gradient,regulating ion balance in plant cells,promoting vacuolar Na~+regionalization,and improving plant salt tolerance.Playing an important role.Through bioinformatics analysis,in the promoter region of this gene,there are many stress-related elements,suggesting that it may play a role in abiotic stress.The main findings are as follows:1?The acquired T2 generation of OsSCL30b transgenic plants was subjected to stress treatment,and the growth of transgenic plants expressing OsSCL30b protein under adverse conditions was investigated.The results showed that the expression of this gene can enhance the tolerance of rice to high salt,low temperature and high temperature.It is not obvious to the drought.Under the treatment of plant growth regulators,the expression level of OsSCL30b was greatly affected by ABA..2?The OsVHA16 gene was cloned from the rice cDNA library.Bioinformatics analysis revealed that the gene has an isoelectric point of 5,a predicted molecular weight of 16.57 KD,an open reading frame of 501 bp,encoding 167 amino acid residues,and the upstream promoter region includes multiple stress-related cis-acting elements such as ABRE,TATA,ATCT,CGTCA,TGACG.3?The expression quantity of OsVHA16 in leaves under various abiotic stresses was analyzed by quantitative PCR.It was found that OsVHA16 gene had different degrees of response under different stresses,and the expression level of OsVHA16 was induced to the highest at high temperature.Drought also has a higher degree of impact.4?We constructed the prokaryotic expression vector PET-32a:OsVHA16 and induced the expression of the OsVHA16 fusion protein in vitro.Based on this,we explored the response of OsVHA16 protein to adversity conditions in vitro,it was found that the expression of OsVHA16 protein in E.coli can significantly enhance the tolerance to high temperature and low temperature.5?In order to study the expression mechanism in rice,we constructed an overexpression vector and an inhibitory expression vector of OsVHA16 gene,and transferred it to Nipponbare callus by Agrobacterium infection to obtain transgenic plants.The hygromycin test and 35S primer detection are correct transgenic plants,providing materials for further study of the role of OsVHA16 protein in eukaryotic rice.