The Role Of BvM14-GO Gene In The Antioxidant Enzyme System Of Sugar Beet M14 Line Under Salt Stress

Sugar beet M14 strain is a sweet menu body addition line obtained by Professor Guo Dedong using Beta vulgaris L.and diploid Beta.corollifiora Zoss.Related studies have confirmed that sugar beet M14 strain has the characteristics of drought resistance,cold resistance,salt tolerance and apomixis in wild sugar beet.The previous use of iTRAQ LC-MS/MS technology of qualitative and quantitative research of sugar beet M14 response to salt stress（0,200,400 mmol/L NaCl）differentially expressed protein,found under salt stress in leaves of sugar beet M14 relates to the antioxidant system of stress and defense of the ROS clearance mechanism in the 7 main enzymes in the most significant,of which 6 proteins up-regulated and 1 proteins down regulated.In this study,one of the up-regulated protein spots,encoding Glycolate oxidase（GO）and BvM14-GO gene,was applied to investigate the function and function of antioxidant enzymes under salt stress.The full-length BvM14-GO gene cDNA using PCR technology and bioinformatics analysis of the full length cDNA of total 1151 bp,ORF to 1113 bp（including the stop codon）,encoding 370 aa;phylogenetic analysis showed that Gen Bank and sequenced in sugar beet（Beta vulgaris）recently in the GO relationship.Tissue specific expression analysis of BvM14-GO gene was carried out using Real-time technique,and the results showed that the highest expression in leaves of sugar beet M14 lines.The Escherichia coli expression system of BvM14-GO gene was constructed.Under the condition of 0.5 mM,IPTG and 37 3h induction,the expression of BvM14-GO protein was the highest,and the fusion protease could be expressed as 27.934U/ml.Construction of expression vector of BvM14-GO gene,carries on the observation and determination of 150 mM NaCl under the stress of BvM14-GO transgenic Arabidopsis plants root length,fresh weight,four kinds of antioxidants H₂O₂,MDA,ASA,GSH of GO content;plant gene transcription and enzyme activity determination,to explore the function of BvM14-GO gene under salt stress.The results showed that BvM14-GO gene could express stably in plants,and transferred into BvM14-GO gene could enhance the resistance of plants to salt stress,and reduce the inhibition of salt stress on plant growth and development.The transcriptional level and enzyme activity of seven key enzymes encoding genes in antioxidant enzymes were investigated to investigate the role of BvM14-GO gene in antioxidant enzymes under salt stress.Based on the above results,we can draw the conclusion that..:（1）The antioxidant ability of Arabidopsis thaliana transformed by BvM14-GO gene increased.Under salt stress of BvM14-GO transgenic Arabidopsis overexpressing plant to increase the content of H₂O₂,MDA,ASA,GSH content decreased,because the increase of antioxidant system in plant As A-GSH cycle in three key enzymes MDAR,APX,DHAR3 content,MDA,ASA,caused by excessive consumption of GSH.（2）BvM14-GO gene can increase the expression of seven kinds of main enzyme system and antioxidant enzyme activity,including GO,CAT,APX,MDAR,DHAR3,Trx and Prx R changes significantly increased but compared.H₂O₂ is produced by acid catalyzed ethanol,which regulate other key enzymes of antioxidant system in gene expression and enzyme activity,promote plant body can scavenge reactive oxygen species,and to improve plant antioxidant capacity,reduce the inhibitory effect of stress on plant growth of salt.