Analysis Of Genes Expression In The Parathion Inducible Drosophila Cells And The Relation Of Differentially Expressed Genes To Resistance

Organophosphorus pesticide is one of the most production and widely use of pesticide in the world. Parathion is widely used for its strong toxicity and quickly action. It makes the resistance for a large amount of unreasonable use for a long time. But the mechanisms of stress for parathion are unclear. The stress mechanism and function targets of parathion were uncovered at the genome level in this paper.The cytotoxicity and survival rate of parathion stressed cell was detected by use of cell counting kit-8 (CCK-8) and cell count plate in Drosophila S2 model cell. The difference of genes expression between parathion stressed cells and normal cells were compared, and the genes related to parathion stress were screened through gene expression spectrum and real-time fluorescence quantitative PCR (qRT-PCR). The results shown that the death of cell processed at 35μg/mL(ppm) parathion is minima by many experiments of CCK-8 kit and cell count plate detection. The expression of 534 genes obviously changed between 35ppm parathion treated cells and control cells (only added methanol solvent processing cells and added none normal cells) by digital gene expression profiles detection. Of them, the cytochrome P450 gene, such as Cyp6a8 and Cyp4el gene expression were increased 3.34 times and 2.04 times, Cyp4adl, Cyp-4d8, Cyp313a4 have specific expression. GST genes GSTE6 expression increase 2 times and GSTE7 has specific expression. An odorant binding protein gene Obp99a, transporters gene MFS3 and CG5321 gene expression increased about 2.5 times. But Five kinds of NADH gene:ND3, ND4, ND5, ND6, ND4L expression was decreased about 2-3.11 times. Many redox enzymes dehydrogenase genes also appeared different expression.We chose eight different expressed genes to check the reliability of digital gene expression profile and selection of resistant concentration by qRT-PCR. The results shown the consistence of fluorescence quantitative with gene expression profiles, and the resistant concentration selected by CCK-8 kit with detected by fluorescence quantitative was also consistence. In order to verify the correlation of gene expression with the detoxification reaction, we respectively transfected Obp99a and GSTE6 gene into S2 cells to detect the cell toxicity under parathion procession after the gene over-expressing. The results showed that the resistant to parathion obviously increased for Obp99a and GSTE6 gene over-expression.Of all, the resistance of insect to parathion is controlled by genes. The level of high expression, low expression and specific expression of some genes may affect the cell resistance. These results provided the experimental basis for the further study the resistant mechanism of insect to parathion and screen the new target to parathion.