The Optimization Of Tissue Culture And Microtuber Induction Of Potato

The potato is the forth-largest crop in the world, and it is the major source of the food, vegetable, forage and the process and production of raw materials in our country. It has a very distribution for its high yield, rich nutrition and strong adaptability. However, during the current annual planting, viral disease has impeded the growth of potatoes. It not only causes the degradation of potato varieties and production, but also affects the quality of potatoes. At present, traditional breeding methods can no longer meet the requirement of the production of potatoes. Therefore, it is really urgent to develop new varieties of potato cultivation using genetic engineering technique combined with traditional breeding technique.The results offered the materials,test basic and science reference for the microtuber formation in the future research work.The experimental results indicated that:1. Adding a certain amount of exogenous Hormone was in favor of the proliferation of potato plantlet. The sequence that single hormone improved the proliferation on potato plantlets was ZT > 6-BA > 2,4-D > IAA > KT > GA. The optimal ZT concentration was 1.0 mg.L^-1 . The best multiple hormone combination was 1.0 mg.L^-12,4-D + 2.0 mg.L^-1IAA + 4.0 mg.L^-1 6-BA + 2.0 mg.L^-1ZT + 2.0 mg.L^-1KT.2. Adding a certain amount of exogenous Cytokinins was in favor of the formation of micro-tuber and the advance of the yield; The best medium for inducement and growth of micro-tuber of Solanum tuberosum L. is MS+10% sucrose(PH 5.8) + 5 mg.L^-1 KT, with which the micro-tuber inducement rate could reach to 100% and micro-tuber grew vigorously. Different position of materials gave different effects on microtuber induction,basal part is better.No microtuber formed in the segments that containing the apicial site because of the apicial dominance.3. A simple,rapid protocol was established for Agrobacterium-mediated transformation with microtuber of Chuan potato 27.Adventitious buds differentiation occurred after 2-3 weeks on selective medium containing 75 mg/L kanamycin, and transgenic plants could be obtained in 4-5 weeks.By combining the best treatments ,especially using 2 mg.L^-1 zeatin riboside in the shoot induction medium,this protocol yieded transformation efficiacy of 42.8%.4. The sequence that single hormone improved root induction and sound seedling of potato plantlets was NAA > IAA > IBA,The optimal NAA concentration was 1.5 mg.L^-1 .The best multiple hormone combination was 1.0mg.L^-12,4-D + 1.5mg.L^-1NAA + 2.0mg.L^-1IAA + 0.5mg.L^-1IBA + 1.5mg.L^-1Met and 1.5mg.L^-12,4-D + 1.5mg.L^-1NAA + 0.5mg.L^-1IAA + 1.0mg.L^-1IBA + 2.0mg.L^-1Met.