Diamondback Moth In The Intestinal Cry1ac Toxin-binding Protein Separation And Identification

The microbial insecticide, which had the biggest production and was used very widely all of the world, is Bacillus thuringiensis (Bt).Diamondback moth (DBM), Plutella xylostella, is a main pest of cruciferous vegetables. It has been found to be the first insect pest to develop resistance to Bt in the open field, and the resistance is still increasing now.The amplified the proteomic analysis approach for P. xylostella midgut proteins was established successfully. Then we separated and identified Cry1Ac binding protein of P. xylostella midgut by using proteomic approach,so as to find the possible resistant mechanism of Bt toxin. The results were as follows.The P. xylostella rearing in different ways were compared. The results showed that the rearing with cabbage coupled with radish was better than then rearing with cabbage and rearing with radish. P. xylostella rearing with cabbage coupled with radish had higher population and its development was more orderly.Two P. xylostella strains, which was resistant to Cry1Ac, was achieved by selection with Cry1Ac toxin, and they were 998.21-fold and 35.07-fold resistance to Cry1Ac. A P. xylostella strains, which was resistant to Bt, was achieved by selection with Bt, and it was 91.32-fold resistance to Bt.In this study, we utilized a proteomic approach involving two dimensional differential gel electrophoresis, mass spectrometry, and function-based activity profiling to examine the CrylAc binding proteins of P. xylostella. The results indicate the proteins of P. xylostella BBMV molecular weight blow 120 KD, and pI between pH 4 and 7. We identified six other CrylAc binding proteins in P. xylostella BBMV for the first time:prohibitin protein WPH, NADH dehydrogenase iron-sulfur protein 3, NADH dehydrogenase (ubiquinone) Fe-S protein 3 and three novel proteins.