The Study Of Protein Expression Profiling Of Fat Body In Bombyx Mori Induced By Cypermethrin

Bombyx mori is not only an important economic insect, but also an important mode of Lepidoptera insects. The majority of food crops and forest pests belong to Lepidoptera. Through the Bombyx mori physiology, pathology, genetics and other researches, we can basically eliminate the threat of pests in food crops and forest pest, achieving new breakthroughs in pest control. Insect fat body is an important tissue. It has a variety of physiological functions, such as storage, excretion and detoxification, and so on.At present, the study of Lepidoptera resistance genes goes further. In China, there is more and more proteomics research on different Bombyx mori tissues. However, there is only several study of the resistance mechanism from the protein level in large-scale . This article, we researched the protein expression profile characteristics of Bombyx mori fat body inducted by cypermethrin, obtaining cypermethrin resistance-related proteins in Bombyx mori. And then using mass spectrometry techniques, we identified kinds of differential proteins. On top of it, the characteristics of protein in the level of gene transcription characteristic was studied by semi-quantitative RT-PCR method. The results obtained are showned as follows:1 The LD50 of Male and female Bombyx mori to the cypermethrin.To determine the LD50 of different sex of Bombyx mori .We treatment the male and female Bombyx mori with different concentrations of cypermethrin. The results showed that the LD50 of male Bombyx mori was 56.336 ng, the LD50 of female was 49.557 ng.The resistance of the male Bombyx mori slightly higher than females. These data lay the foundation for the male and female Bombyx mori individual pesticide resistance studies. 2 Protein expression profiling of fat body in male Bombyx mori induced by cypermethrin.2.1 The establishment of patterns of fat body protein in male Bombyx mori induced by cypermethrin by two dimensional polyacrylamide gel electrophoresis. We compared the different protein of fat body protein in male induced by cypermethrin using two-dimensional electrophoresis technology. The ImageMaster 6.0 analysis showed that 432 protein spots were detected in the control group, 369 protein spots in the experimental group, and the matches were 342. Comparative analysis showed that the matched value between the two groups were 76.68%. Protein spots were mainly distributed in the molecular mass of 20.1-66 kD and the pI between 4-8.2.2 The spectrum identification of different protein in male Bombyx mori fat body induced by cypermethrin.The interested protein spots were identified by MALDI-TOF MS. The identified proteins were enolase, mitochondrial aldehyde dehydrogenase, chaperonin containing t-complex polypeptide 1 beta subunit, and actin, respectively. These proteins were related to different metabolic pathways, such as protein folding, respiratory chain reaction, glycolysis channels, microtubule assembly , and so on.2.3The enolase gene transcription activity of 5 instar larvae Bombyx mori induced by cypermethrin.Inducement effect of cypermethrin treatment on transcription level of Enolase gene in the 5th instar larvae was determined in seven tissues: brain, midgut, silk gland, fat body, testis, malpighian tubules and hemo by semi-quantitative RT-PCR method, respectively. The result showed that enolase gene expression reduced in seven tissues after induced by Cypermethrin and the specific value is 13.79%, 10.71%, 22.48%, 7.40%, 27.00%, 11.50% and 14.52%, respectively. Both gene expression and protein expression of fat body enolase showed a downward trend. by cypermethrin.3.1 The establishment of patterns of fat body protein in female Bombyx mori induced by cypermethrin by two dimensional polyacrylamide gel electrophoresis.We compared the different protein of fat body protein in male induced by cypermethrin using two-dimensional electrophoresis technology. The ImageMaster 6.0 analysis showed that 758 protein spots were detected in the control group, 675 protein spots in the experimental group, and the matches were 554. Comparative analysis showed that the matched value between the two groups were 72.84%. Protein spots were mainly distributed in the molecular mass of 20.1-66 kD and the pI between 4-8.3.2 The spectrum identification of different protein in female Bombyx mori fat body induced by cypermethrin.The interested protein spots were identified by MALDI-TOF MS. The identified proteins were Catalase, actin, cytochrome p450 protein, unknown protein, retrotransposons, hot spotsprotein, and so on. These proteins were related to different metabolic pathways, such as protein folding, respiratory chain reaction, glycolysis channels, microtubule assembly, and so on. These proteins related to different metabolic pathways may be related to Bombyx mori resistance-related.We have analyzed the protein expression profiling from different gender Bombyx mori induced by cypermethrin, finding multiple differences proteins. These differentially expressed proteins were identificated by mass spectrometry. The resut including aldehyde dehydrogenase, enolase, catalase, cytochrome p450, and so on. These enriched the Bombyx mori proteome database. These enzymes involved in different metabolic mechanisms and may be associated with resistance to cypermethrin. This research not only significant in researching the resistance metabolites mechanisms in Bombyx mori,but aslo provides a reference for the further study on biological control of insect pests. 3 Protein expression profiling of fat body in female Bombyx mori induced...