Develop Of High Starch Content And Anti-diseases Potato (Solanum Tuberosum L.) Lines By Gene Transformation

Potato (Solanum tuberosum L.) is the important crop that can be used as staple food and vegetables in the world. It plays very important role in daily life and national economy. But serious disease has greatly hampered the development of the potato industry. The enhancement of consumer's demand for processing potato varieties bring out researchers new objectives. In conventional sexual hybridization of potato, the segregation of homology tetraploid of potato cultivars made it very time consuming and low efficiency in improving characters of potato. The application of genetic transformation is a promising strategy for potato genetic improvement, with which the desire gene could be efficiently introduced into the genome without altering the good characters and identity of the cultivars.In this research, Agrobacterium tumefaciens mediated transformation method was performed to develop two aspect studies: Firstly, the key enzyme gene in the starch-biosynthesis named glgC was introduced into potato cultivars; Secondly, the hrp gene which induced hypersensitive reaction of nonhost plant was introduced into potato cultivars, and then to validate the feasibility of using wound-induced promoter and Me13'enhancer as control element in broad-spectrum resistance breeding of potato.Analyzing the transformation effects of different recipient material, the leaf discs and stem segments were used as the experimental materials. The high effective regeneration systems were obtained by the comparison of different regeneration system. The system used for leaf discs and stem segments culture: MS base medium complemented with 2.5mg/L 6-BA +0.6mg/L 2,4-D (for callus induction), the percentage of explants reducing callus can reaching 100%; MS base medium complements with 2.5mg/L 6-BA +0.25mg/L IAA +0.25mg/L 2,4-D (for shoots generation), the shoots inducing rate reaching 65.33%. Effects of the explant growth state, Kan sensibility, the period of preculture, bacterial infection and co-cultivation on the transgenic efficiency were investigated. The result showed that the best explant was stem segments which growing on 1/2 MS liquid medium, the concentration of kanamycin was 75mg/L for the stage of callus and shoot formation and 50mg/L for root formation, the optimized period for preculture was 2 days (resistant callus inducing rate reaching 48.24%), for bacterial infection was OD600 0.5 and 8 minutes...