Study On Improvement Of Pepper (Capsicum Annuum L.) Genetic Transformation System And Introduction Of Transcription Factor JERF-33# Into Pepper

Pepper is one of the most important vegetables. Abiotic and biotic stress can do harm to pepper, and decrease their yield and quality. For this reason, many studies have being carried on to improve its synthetic resistance.In order to enhance the resistance of pepper, anti-disease or anti- adversity gene may be transferred into pepper. But the qualities of anti-disease and anti-adversity are usually controlled by different genes. Only transfer one function gene into plants, the synthetic resistance cannot be improved. While cloning two kinds of genes at one time is difficult. The transcription factors can regulate the expression of many genes. Compared with introduction of one function gene into pepper, introduction of transcription factor gene into pepper can improve the synthetic resistance of pepper.The JERF-33# gene which can encode transcription factors was isolated from tomato cDNA express library. Analysis of the protein sequences suggests that there is a conserved DNA binding domain in each of them. These DNA binding proteins belong to the EREBP/AP2 transcription factor sub family, which can interact with GCC-box and thus regulate the expression of genes responsive to aboitic and biotic stresses. To improve the synthetic resistance of pepper, the JERF-33# gene was transferred into pepper.A stable genetic transformation system must be established before the JERF-33# gene was transfered into pepper. While the biggest problem in pepper transformation was the rate of transformation. To study the effect of different factors on agrobac-terium tumefacient-mediated exogenous transferring into pepper, GFP gene was transferred into three varieties of pepper. And this paper discussed systematically about the effect of the pH of agrobacterium tumefaciens suspended cultivated medium, the concentration of AS in cocultivated medium and the time and temperature at cocultivation on transformation. In addition, the system of pepper agrobacterium-mediated genetic transformation was established.The major results were as follows.1 When the pH of agrobacterium tumfaciens suspended cultivated medium was 5.2,it was favorable to the transformation.2 When the addictive amount of AS was 200i M in cocultivated medium, the fluorescent expression rate of callus is the highest3 the optimum temperature was 22 C for the agrobacterium-mediated transformation.4 the optimum time were 4 days in cocultivation.On this condition, the improved pepper genetic transformation system was established, and we transferred JERF-33# gene into pepper for the first time. 18 transgenic seedlings showed PCR positive.