全缘叶绿绒蒿谷胱甘肽S-转移酶基因(MiGST)克隆与功能分析OA

In the present study,the glutathione S-transferase gene(MiGST)from Meconopsis integrifolia was cloned,and the biological function through Arabidopsis thaliana transformation was characterized.This study could provide the theoretical basis for flavonoid development and utilization in different tissues of M.integrifolia.The MiGST gene cloned was analyzed by using bioinformatics methods.Then,the tissue expression patterns were examined by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Subsequently,A.thaliana was transformed using the floral dip method,and the total flavonoid content in T1 transgenic lines was determined by the Al(NO3)3 colorimetric assay.The results showed that the coding sequence(CDS)of MiGST was 702 bp in length,which encoded a 234-amino acid protein.The protein molecular weight was 25 861.91 Da with a theoretical isoelectric point of 5.84.Phylogenetic analysis showed that MiGST protein was the most closely related with GST protein from Papaver somniferum.Meanwhile,MiGST gene exhibited the tissue-specific expression,with the highest level in flowers and the lowest in stems.Besides,the total flavonoid content within A.thaliana overexpressing MiGST gene was a 1.27-fold of that of the wild-type one.In conclusion,this study confirmed that overexpression of MiGST gene significantly enhanced the total flavonoid content in transgenic A.thaliana,suggesting that MiGST gene played an important role in the transmembrane transport and accumulation of flavonoids in M.integrifolia.