Genetic Transformation Of Antisense Polygalacturonase Gene For Processing Tomato

The processing tomatoes production was the main induction in Xinjiang province. Choosing the best and suitable processing tomato for production was the target for breeding workers. With the development of the technique of Genetic engineering, people more and more trended towards improving the characters, breeding new varieties for storing and transporting easily, and breeding the higher quality for processing tomato, in this way, the problems of rot during the storing and transporting were well resolved. Cotyledons and hypocotyls of BO4 processing tomato were used as explants to study different combinations of cytokinin (ZT, 6-BA or KT) with auxin (IAA, IBA or NAA) and silver nitrate on bud induction; meanwhile, Agrobacterium was used for transformation, different factors of the transformation were studied(pre-cultural time,invading time,comcentration of bacteria, co-cultural time). And then transgenic plantlets with polygalacturonase gene of processing tomato B04 were obtained, and the activity of polygalacturonase was decreased. PG antisense genes were transformed into processing tomato B04.The main progress was as follows:1. The asexual systems of B04 processing tomato were established in the study. Effects of different three cytokinins on the regeneration of cotyledons and hypocotyls were compared. ZT was really better than 6-BA, KT was the most worst, the best inducting medium was MS+2.0 mg/L ZT+0.20 mg/L IAA, the induction ratio was 83.6 %;Effects of different three cytokinins on the regeneration of cotyledons and hypocotyls were compared. IAA was better than IBA, NAA was bad to the induction; The induction ratio of cotyledons explants was higher than hypocotyls explants, and the cotyledon explants was the receptor in the study of the construction of transformation system for B04 processing tomato; The regenerated buds could develop normally after roots were induced on MS+IAA0.1 mg/L, and finally could form the normal plants.2. Different concentration of silver nitrate in regeneration medium did not improve the budding induction for BO4 processing tomato.3. The inducting buds of explants based on the study of sensitive concentration of kanamycin for BO4 processing tomato was 30 mg/L; then studied the effect on the Carb to restrain the bacteria,and the concentration was 300 mg/L.4. Established the transformation system for processing tomato B04, the time of pre-culturing, the time and the bacteria concentration for invading, the time of the co-culture for explants and the bacteria were made. The suit pre-cultural time for processing tomato was 48 hours; OD600=0.6 was the fitful bacteria concentration for invading; and the time for invading was 10 minutes; the explants and the bacteria had been best co-cultured for 48 hours. Transgenic processing tomato plants were obtained.5. By PCR analysis, 9 plants were positive in 41 plants, and the rate of transformation was 22 %, the results proved that the foreign genes had been integrated into B04 processing tomato.