Cloning And Characterization Of Glutathione Synthetase Gene From Potato

Potato is one of the staple crops in China.Glutathione(GSH)regulates the plant growth and development,and plays important roles in plant response to biotic and abiotic stresses.Whether potato has genes for GSH synthesis,could these genes regulate in potato growth and development,what is the related molecular mechanism,are still unclear.Glutathione synthetase(GS)is one of the key enzymes for GSH synthesis in plants.In this paper,the potato glutathione synthetase gene St GS was cloned,the potato genetic transformation system was established,the transgenic St GS function was characterized,and the St GS tissue localization was analyzed.In this experiment,the potato glutathione synthetase gene St GS was successfully cloned by homologous cloning method,and the vector of overexpression,interference and tissue location of St GS gene was constructed.The potato genetic transformation research system was improved,and the screening,identification and functional verification of the transgenic plants with overexpression of St GS were carried out.At the same time,the tissue location of St GS in potato was analyzed.The results showed that the length of c DNA of the gene was 1452 bp,encoding 484 amino acids;phylogenetic tree analysis showed that St GS protein had the highest homology with GS protein of wild panari tomato and tomato;four transgenic plants with over expression were successfully screened out,and phenotypic analysis showed that the fresh weight of root,stem and leaf of the transgenic lines OX-21 and OX-26 were higher than that of wild type,especially that of OX-26 The results showed that the content of GSH in the leaves of OX-21,OX-26 and wild type was significantly higher than that of wild type,in which the content of GSH in the leaves of OX-21 and OX-26 was 3.12 times higher than that of wild type,and the content of GSH in the leaves of OX-21 and OX-26 was 2.88 times higher than that of wild type.Gus staining showed that St GS was mainly located in leaves,roots,meristem and old stem.RNAi research on St GS is still in progress.The results lay a foundation for further understanding the function of GSH synthesis gene and its molecular mechanism of regulating GSH metabolism in potato.