Study On The Cloninq Expression And Functional Analysis Of CsGRX Gene Family In Cucumber

Glutaredoxins are small heat-stable oxidoreductase that regulate protein reversible oxidation-reduction reaction.It have a important role in resisting physiological and biochemical process in the plant.Currently,it has not been reported in cucumber.In this study,we isolated the first cucumber glutaredoxin gene CsGRX1(based on electronic clone sequence of Arabidopsis AtGRX480 gene).On this basis,the homologous cloned CsGRXs gene family members,and made bioinformatics analysis.Studied the subcellular localization of CsGRX1 Expression patterns analysis of CsGRXs gene family in the different organs of cucumber.Preliminary studied CsGRXs gene function by the Arabidopsis mesophyll protoplast transient expression system.We analyzed CsGRXs gene family members interacting with the Arabidopsis TGA transcription factors by yeast two-hybrid experiments.Laid the foundation for intensive study studying of cucumber CsGRX gene function.The main results are as follows.1.We isolated eight CsGRX gene from cucumber,and bioinformatics analysis indicates that seven are CC type GRX and another one is CPYC type GRX.CC type CsGRXs contain CCMC/S domain,CsGRX1 G105 glycine residue structure ?,L**LL domains ? and C-terminal domain IV is AL/IWL/V.CC type CsGRX1 and CsGRX6 have lower homology with AtGRXC9(GRX480).However,CsGRX4 and CsGRX7 gene have higher homology about 70%with Arabidopsis and rice ROXY1,ROXY2.2.The studied of CsGRX1 gene cellular localization in onion epidermal cells showed that it exists in the cytoplasm preliminary.3.The real-time PCR analysis of CsGRXs gene in different tissues and fruit development process of cucumber showed that the expression of CsGRXs gene in various organs,growth and development stage of cucumber was different.The expression of CC type CsGRXs gene in flowers was obviously higher than in other tissues and organs,while CsGRX5 relative expression in flower was the highest.In different tissues of cucumber,CC type CsGRX had the highest expression quantity in the root,of which CsGRX5 and CsGRX8 gene expression levels were higher than other genes in the different tissues.The CsGRXs gene expression was upward trend with the leaves mature and senescence,and old leaves had higher expression.CsGRX1 gene expression level was higher in cucumber young flower sepals,as the flowers mature CsGRX1 gene expression quantity increased in the stamens and petals.4.The transient expression system of cucumber CsGRXs gene in Arabidopsis protoplast showed that CPYC type CsGRX2 can not inhibit LUC activity,while CC-type CsGRXs with ALWL and AIWV domains can inhibit LUC activity.However,CC-type CsGRXs with ALWV domains no inhibition to LUC activity.5.The analysis of CsGRXs protein interactions with Arabidopsis TGA transcription factor by yeast two-hybrid experiments showed that CC-type CsGRXs have interactions with Arabidopsis TGA transcription factors,while CPYC type CsGRX2 no interactions with TGA.