The Distribution Of Bacillus Thuringiensis In The Brown Rice Planthopper And The Impact Of Related Enzymes In The Body

This study aimed at providing evidences for security evaluation of non-target organisms which were affecting by Bt transgenic rice. A Series of related endoenzymes activity of Nilaparvata lugens were measured, such as superoxide dismutase (SOD), peroxidase (POD), Glutathione peroxide enzyme (GSH), catalase (CAT), acetylcholin esterase (AchE), FE esterase and carboxylesterase (CES), including their activity alteration in different body parts. The pests Nilaparvata lugens were feed by CrylAb transgenic rice (shanyou 63) planted in greenhouse, the samples of different body parts at different periods were took to detected the enzymes total activity by euzymelinked immunosorbent assay (ELISA). The various cellular structures were obtained by differential centrifugation. Then the enzymes activity of the pest intracorporal organs, cell debris, mitochondria and subcellular organelles were detected by using spectrophotometer or microplate reader. The results are showed as follows:1. The concentration of Cry1Ab protein in Cry1Ab transgenic rice (shanyou 63) was 12.8385ng/L, detected by the CrylAb protein assay kit.2. The CrylAb protein was distributed in cell debris, mitochondria and subcellular organelle in mature pest body. The content of Bt protein in different cellular parts was ubcellular organelle>mitochondria>cell debris. In addition, Bt proteins could be detected in those three parts within 1-15days.3. Under the stress of CrylAb, the total CAT enzyme activity reduced at beginning 10 days and reached the minimum value, while 15 days later, it can rise rapidly until recovery. The total GST enzyme activity reached the minimum value at beginning 5 days and then it increased. However, the POD and AchE enzyme activity went up at early stage and declined in later period, it can recover to a level in which was non-Bt rice breeding stage. The SOD enzyme activity rose slightly over the feeding time, on the contrary, the CES enzyme activity lowered gradually. While the FE enzyme activity descended all the time.4. Analyzed by synthesis, since various biochemistry reactions were responded to the stimulation of exogenous proteins (or active materials), different functional enzymes (POD, GST, CAT, SOD, CES, AchE, FE) activity in Nilaparvata lugens after being feed by Bt protein rice would change corresponding. Even the experimental group had significant different from the control group. However, with the feeding time extending, those enzyme activity changes can recover gradually to the initial level. One possible reason is that the creature immune system can make self-protect physiological reactions to allogenic material. The other possible reason is that after the Bt protein enter the organism through the food chain, it has harmful effects on some functions of creatures which cause some irritable physiological reactions that can neutralize or alleviate harmful effects to the organism.