The Study Of Pectin Acetylesterase Gene In Seedless Litchi Genetic Transformation Of Tomato

Hainan seedless litchi is a best mutant.The fruit is bigger,bright red in color,non-seed,pulp thick and juicy, taste great,edible rate can reach up to about90%,the market competitive and high economic value.During in the ovule development period,the function of the embryo has already degenerated.This for us to study litchi ovule and embryonic development provides a good experimental materials.Dealing with the litchi embryonic development of research,the past the main focus on morphorlogy,anatomy, and physiology,biochemistry,only a few reports concering to the gene expression and regulation of signal transmission way during the embryo development stage.By using the normal ovule and abortive embryo for materials from our research group used SSH and SMART RACE techniques had got three integrated cDNA sequences:L53,L69and L117.By Genebank retrieval showed that were the new genes in litchi.Gene L117sequence (Genebank registration number:EU717680) the length of cDNA1473bp, contains a open reading frame between22bp to1221bp.Analysis showed that L117might be through coding pectin acetylesterase to degradate the pectin in the cell wall and damaged the cells.But the specific function is not confirmed.Based on the research results,this paper was to clone pectin actylesterase gene(L117gene)and through the plant expression vector pCAMBIA1304(by using two kinds of promoter:CAMV35S promoter and the ovule specifically expressed promoter Inner no outer INO)transformed into the tomato plant for expression,for the next step in the research of pectin acetylesterase gene for seedless litchi to explore the function of forming seedless during ovule developing periode.The following results were achieved in this paper:(1)the pectin actylesterase gene had been cloned and a plant expression vector pCAMBIA1304-L117had been constructed.(2)The inner no outer(INO) promoter had been isolated and cloned from arabidopsis,a plant expression vector pCAMBIA1304-INO-L117had been constructed.(3)Tomato transformation had been made by the two plant exprssion vectors and the transgenic plant had been detected,sixteen tomato transgenic plants had been achieved,among fourteen plant have plant expression vector pCAMBIA1304-L117,two have plant expression vector pCAMBIA1304-INO-L117.