Agrobacterium Rhizogenes Mediated Bt Of Cry8e Transformation Of Soybean Resistance To The Holotrichia

Soybean as the most important alimentary crop and economic crop is widely used in industrial processing for its high oil and protein content. However, mostly, the insects influenced the soybean bio-mass and quality. For this reason, insecticide is customarily used for insects control, even it gives a health risk to human and animals and may lead to enviroment pollution. Further more, long period insecticide using can easily cause the resistance formation of insects which subsequently have no significant effect to kill the insects.Holotrichia, the larva of chafer, is one of the underground insects that hardly to be controlled. It takes 80% of the total amount of the whole underground insects. Therefore, it a crucial economic affair for our country to exploit new Bt-like genes and put those genes into transgenetic breeding and bio-insect projects. The utilization of the insect-resistant gene modified crops to prevent itself from being destroyed becomes a favor.Many researches demostrates soybean is a reluctant plant during transfomation, and soybean genotype plays an important role in in. Our research based on Agrobacterium rhizogenes mediated transformation of soybean, through the disscussing of the genotype and optimizing of the transformation, trying to establish a transformation method for obtaining new GM soybean for Holotrichia controlling. We have already got the molecular check and bioactive data of the positive roots that induced by Agrobacterium rhizogenes, both of which give to good result:1 the construction of plant expression vector pCAMBIA1305.1-35S-8e contains Cry8e. and further Agrobacterium rhizogenes mediated transformation of soybean to obtain transformed positive hair roots.2 ten different genotypes of soybean have been used to induce hair root, all of these can get hair root and jilinxiaolin was the best.3 choose four most sensitive genotype for co-cultication with/without light. Result shows that there is no significant effect between this two treatments.4 choose four most sensitive genotype for co-cultication in 23℃, 25℃, 28℃. Result shows that there is no significant effect among the three treatments.5 choose two most sensitive genotype for hygromycin screening (10mg/L, 12mg/L, 14mg/L, 16mg/L , 18mg/L) comparation.6 choose three most tranform favorable genotype to induce hair roots. For the positive roots we propagated and indentified the roots via PCR and WesterWestern blot analysis. The results show that the gene can be expressed in soybean hair root.7 enlarge the cultivation amount of the positive hair roots and feed the Holotrichias for its bioactive analysis. High kill proportion, 80%,shows that the transgentic soybean root ca significantly control the insects.