Insect The Gene Cry1c ~ * Early Japonica Transformation

Stemborers and leaffolders were two groups of Lepidopteran pests that caused severe damage to rice in many areas of the world. Kongyu131, being the most popular rice variety of Heilongjiang province, was highly susceptible to Lepidopteran insects, which greatly reduced the expected yield of Kongyu131. In this study, cry1C^* gene encoding Bacillus thuringiensis(Bt) d-endotoxin was synthesized based on codon optimization as one step toward gene stacking in the resistance management strategy of transgenic rice. The gene was transformed into Kongyu131 (Oryza sativa L.) via Agrobacterium-mediated method. The integration of the cry1C^* gene in rice genome was detected with PCR analyses. Meanwhile, the activity of EHA105B was compared to EHA105N and the optimized hardening method of regenerated plantlets in winter was improved in Heilongjiang province. The main results were as follows:1. The vector pBar-cry1C^* was confirmed in E.coli DH5αwith restriction enzyme digestion and PCR detection.2. The activity of the stain EHA105B of Agrobacterium tumefaciens was more powerful than the strain EHA105N and the EHA105N was suitable for the Agrobacterium-mediated transformation.3. The optimized hardening method of regenerated plantlets elevated the living rate of transgenic plants in winter in Heilongjiang province.4. 123 plants of the rice Kongyu131 transformed with cry1C^* gene from 20 independent resistant calli were regenerated and 115 living transgenic plants were obtained after optimized hardening.5. PCR detection indicated that the cry1C^* gene was integrated into the genome of 120 transgenic plants of T₀ generation of the rice variety Kongyu131.