Optimizing The Techno-system Of Genetic Transformation Mediated By Agrobacterium Tumefaciens And LeJA2 Gene Function Research Of Tomato

Currently, the high-dose application of pesticides and herbicide had caused great destruction to environmentand also caused a bad influence on the human body, and the quality of life.Hiher plant has established the compli-cated and elaborate defense mechanism against insert attack during the long-term evolution.Having deeper understanding of the molecular mechanism of plant resistance to herbivores will provide theoretical evidence to establish the environment-friendly agricultural herbivores management by employing the resistance ability of plant itself.The signal molecules that have been ap-praisaled including peptide signal molecules system and plant hormone jas-monic acid coming from the unsaturated fatty acid,making genetics analysis resistance in molecular level mediated by system and JA., identifying one of the important components(functional gene) and doing further research for its function.. This research optimized agrobacterium mediated tomato genetic transformation system. Using reverse genetics technology, constructed the ex-cessive expression LeJA2 vector and RNAi expression vector. By transgenic technology mediated by agrobacterium , after seedling two types of transgenic , identified the two types of transgenic seedlings and the determination of PPO activity, the function of LeJA2 was predicted preliminary. Main research re-sults are as follows:1.Established agrobacterium mediated tomato genetic transformation systemOptimized agrobacterium mediated tomato transformed LeJA2 system. Study the influence of three factors, Agrobacterium tumfaciens infection time , in-fection temperature and infection concentration ,to tomato callus formation. Seeking the main factors affecting gene transformation. Obtained the optimal combination of tomato agrobacterium-mediated infection system .2 Obtained transgenic tomato plantsTake cotyledons as conversion, using Efficient agrobacterium mediated ge-netic transformation system transfer LeJA2 genes and LeJA2 - RANi gene to tomatoes, acquired 11 LeJA2 gene excessively expresses resistant tomato plants and 35 LeJA2 gene silencing resistant plants. PCR detection results show that kanamycin resistant plants can be amplified the 1050bp and 517bp purpose Gershgorim band.This Prove that genes has been integrated into the tomato genome.3 LeJA2 gene function validation.Tested PPO activity of transgenic tomato plants preliminary. By PPO ac-tivity determination results obtain that:Under normal conditions,excessive expression LeJA2 gene transgenic plants present brown,while control and LeJA2 gene silencing expression transgenic plants still appear normal juice green after PPO dyeing.;Do mechanical damage with scissors,excessive expression LeJA2 gene transgenic plants,control and LeJA2 gene silencing expression transgenic plants appear brown after PPO dyeing. Experiment re-sults show that LeJA2 gene plays a role of regulation upstream in signal transmission channelmediated by jasmonic acid of tomato, regulating the ex-pression of resistance related gene.