Gene Expression Analysis Of CLC Gene Families And Functional Verification Of ZmCLCa In Maize

As the largest crop in China,maize also has the highest usage of nitrogen fertilizer in crop.Therefore,it has important significance in production to improve the absorptivity of nitrogen in maize.CLC chloride ion channel family genes have the function of transportation of nitrate,which can effectively improve the absorptivity of nitrogen.Nowadays,there are many researches for CLC chloride ion channel family genes which concentrated in arabidopsis and rice,but less for maize.This study systematically analyzes the CLC chloride channel family genes in maize,and explores the regulation of the response of nitrogen absorb in the family genes,and validates the function of a ZmCLCa gene in the family.The main research methods and conclusions as follows:1.By comprehensive analysis the change of NO₃^-,Cl and the quantity of CLC family genes expression in maize,confirmed the change rule of the amount of gene expression in CLC family by the treatment of KNO3,NaCl and PEG.We found that,BT066984.1?GQ412103.1,NM001176231.1 and NM001136935.2,these four genes response to the nitrate obviously and are likely to have the effect of transportation for nitrate.2.Cloning get over ZmCLCa gene coding regions in maize.This gene encoding region is composed of 2379 bases,contains two exons and three introns,promoter in148 bp upstream.3.Using bioinformatics methods for the gene analysis,define the molecular weight of the protein,the isoelectric point of the protein,the stability coefficient of the protein,the fat coefficient,the hydrophilic,across the membrane protein structure,function and structure domain was analyzed,and speculated that it is a transmembrane proteins.4.According protoplast transformation mediated of maize by PEG,determine the gene expression in cell membranes.5.Through test the phenotype of ZmCLCa transgenic arabidopsis and detection of nitrate in the plant,preliminary validation ZmCLCa genes can make plantsaccumulate more nitrate,and helps the plants adapt to low nitrogen environment.Conclusions show that corn CLC family gene expression changes with the nitrate content,especially BT066984.1,GQ412103.1,NM001176231.1 and NM001136935.2 these four genes response to nitrate,is likely to have the effect of nitrate transport.The family genes ZmCLCa can play a role on cell membrane,accumulate higher concentration of nitrate in plants and provide a large amount of nitrogen for plant growth.It provides useful genetic resources for our country for cultivating nitrogen efficient use of maize varieties and improve maize yield.