Expression Of GPD1 And Promoter Of Key Enzyme For Glycerol Synthesis From Saccharomyces Cerevisia In Tobacco

The problems of drought, salinization and soil erosion are very serious in China , so breeding to fight the drought and salt tolerance of New Varieties of Plants is of great strategic significance.Biosystem can respond to the change of environmental conditions by accumulating osmoprotectants, such as glycerol and proline which contribute to the re-establishment of homeostasis when exposed to various adverse environmental stresses, such as drought, salinity, high and low temperatures.Glycerin is glycerol ester of the three components of the skeleton; It owns some merits such as small molecular weight, easily dissolved in the physiological pH range without a net charge, to be cell membrane maintain the biological cells without any side effects and so on.Therefore, it,s an extremely desirable to protect cells against stress agent under the conditions of stress in the face of adversity.Tobacco as an important model plant which playS a positive role in genetics, physiology and metabolism,has become a research hot by the domestic and foreign scientists.At the same time, 3 - glycerol phosphate dehydrogenase (GPDH) as the important body of glycerol metabolism plays an main role in the speed of its direct bearing on the decomposition of glucose metabolism in the direction of glycerin to the material flow and distribution of glycerol Synthesis level.As tobacco has the poor and limited capacity of glycerine synthesis and only a certain level of stress tolerance,so this study aimed at breeding a tobacco can increase the capacity of glycerol transgenic plants and enhance its anti-stress.Firstly, the experiment cloned promoter of key enzyme gene gpd1 of glycerol metabolism by the way of PCR from Saccharomyces cerevisiae, then constructed the shuttling vector pYX212-zeocin-PScgpd1-GUS which was transferred into Saccharomyces cerivisiae by electric shocking. Homologous expression found: Saccharomyces cerevisiae detected within the GUS gene expression had a significant difference in different osmotic stress,and to a certain extent with the improvement of coercive pressure on the more notable differences. The study showed that PScgpd1 is the kind of promoter of osmotic stress-induced adjustment.What,s more,we got the plant vector of p3300-zeocin- PScgpd1-GUS which was transferred into Agrobacterium tumefaciens by electric shocking.Then the transgene tobacco was been attained by the way of leaf-disc with the help of A. tumefaciens-mediated transformation. We found that: the promoter of tobacco in different parts of the organization can start GUS gene expression, and to a certain extent, tobacco GUS in the gene expression of the intensity with the increase of stress increase in different osmotic stress with GUS reporter gene expression system in the promoter of tobacco of the functional analysis.Finally, the key enzyme gene GPD1 of glycerol metabolism of S. cerevisiae was transferred into tobacco by the way of A. tumefaciens-mediated transformation . the glycerine content and GPDH activity of transgenic tobacco could increase under different NaCl concentration osmotic stress treatment, and In a certain range, it can resist the external environment of salt stress.