The Preliminary Research On The Rice LHR1 Homologous Gene Function

Polyamine,as a polycation,plays an essential roles in regulating genes expression and proteins activity at multi-level of cells.Polyamine transporters could specifically mediate the polyamines transmembrane transport and function in adjusting concentration of polyamine,regulating expression of resistance gene,maintaining level of mRNA,enhancing resistance of plant.Now,a little functions of LHR1 has been known that are encoding polyamine transporter protein in Arabidopsis thaliana,e.g.AtPUT3 controls absorption of the polyamine and takes part in the molecular regulation process of plant heat-stress early responses.However,the role is not clear in rice,especially in early responses of heat-stress of rice.By sequence alignment,we found that Os02g47210 and the Os03g37984 are function homologous gene of the AtPUT3 in rice.In order to explore the function of the function of rice polyamine transport proteins LHR1 in improving the ability of the plant heat residence,bioinformatics,biochemistry and molecular biology,genetics test were carried out.The main results are as follows:1?The full-length CDS of OsLHR1?Os03g37984.1?2?3and Os02g47210?gene was cloned,and the result of bioinformation analysis effectively of the genes encoding protein showed,Os03g37984 has five different cutting mode,it can encoding OsPUT3.Os03g37984.1 has 1653 bp,Os03g37984.2has 1479 bp,Os03g37984.3 has 1428 bp.Compare Os03g37984.4andOs03g37984 to Os03g37984.3,which have higer overlap-rate.Os02g47210 has 1695 bp,it can coding OsPUT1.OsLHR1?Os03g37984.1?2?3and Os02g47210?has 4 structures take part in the transporter and transmembrane of polyamine.AA_permease₂ is a carrier protein,in the APC superfamily,AA_permease₂ is transporters specific for amino acids,polyamines.PotE is a amino acid carrier protein,it can regulate transport and metabolism of amino acid.2A0306,it can transport and band the proteins,amino acids,peptides and amine derivatives.PRK10644 is a spermine reverse vector,it can transport and regulate of spermine.2?The GFP expression vector p35s::OsLHR1-GFP?p35s::OsOs03g37984.1?2?3-GFP and p35s::Os02g47210-GFP?was constructed successfully,and the OsLHR1 were transferred into onion epidermal cell by Agrobacterium-mediated transformation.Using fluorescence microscope,we found OsLHR1 were positioned on cytomembrane and karyotheca of onion epidermal cell.3?The over-expression vector p35s::OsLHR1?p35s::Os03g37984.1 ? 2 ? 3 and p35s::Os02g47210?were constructed successfully,and the OsLHR1 were transferred into Arabidopsis thaliana mutant lhr1 by Agrobacterium-mediated transformation.We got transgenic seeding by the selection of vector plant resistance.4?Functional complementation of Arabidopsis thaliana mutant lhr1.Cloning OsLHR1,and transfer it into Arabidopsis thaliana,screening of homozygous.Then,the transgenic plants returned to paraquat sensitive phenotype,in 1 ? M paraquat resistance medium.