吡唑硫磷对映体对东亚飞蝗的对映选择毒性及其机制研究  

Study on the mechanism of the enantioselective toxicity of pyraclofos to Locusta migratoria manilensis(Meyen)

作　　者：钮利喜[1] 朱欣凯[1] 王萍[1] 赵美蓉[2] 

机构地区：[1]山西大学生物技术研究所,山西太原030006 [2]浙江工业大学生物及环境工程学院环境科学研究中心,浙江杭州310032

出　　处：《山西农业大学学报（自然科学版）》2016年第5期357-363,共7页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：国家自然科学基金重点项目(21337005)

摘　　要：通过比较吡唑硫磷(pyraclofos)两种对映体对东亚飞蝗1型乙酰胆碱酯酶基因Lm-ace1转录水平、乙酰胆碱酯酶(AChE)活性影响以及其对AChE活性抑制的差异,探究吡唑硫磷对映体在东亚飞蝗体内毒性差异产生的机理。利用HPLC法拆分吡唑硫磷。分别对三龄若虫点滴得到的两种手性对映体,24h后计算LD50值,并检测实验虫体各组AChE的活性,采用RT-PCR检测Lm-ace1mRNA的表达水平。提取对照组AChE粗酶液进行体外抑制实验检测。结果表明:(+)/(-)-吡唑硫磷的半致死量LD50分别为0.398mg·kg-1虫重和36.535mg·kg-1虫重。在AChE体外活性抑制实验中,随着(+)/(-)-pyraclofos浓度的增加,农药对东亚飞蝗AChE的抑制程度也在逐渐增强。吡唑硫磷对映体对东亚飞蝗具有显著的毒力差异,两种对映体对飞蝗Lm-ace1不同的诱导表达活性以及对AChE不同的抑制活性是造成这种毒性差异的部分原因。In order to reveal the generation mechanism of the enantioselective toxicity of pyraclofos to L.migratoria manilensis,the transcription of acetylcholinesterase 1gene（Lm-ace1）,the activity acetylcholinesterase（AChE）in vivo and the inhibition of AChE in vitro were investigated.Chiral separation of pyraclofos racemates was achieved by HPLC.The individual enantiomers with different concentration were topically applied to 3th-instar nymphs and the median lethal dose（LD50）of each pyraclofos enantiomer was calculated by SPSS 19.0software 24 hlater.The expression level of Lm-ace1 gene was examined by RT-qPCR and AChE activity was assayed in control group and experimental groups.AChE inhibition test was measured using crude enzyme extracted from the control group.LD50of（＋）/（-）-enantiomer of pyraclofos toward L.migratoria manilensis were 0.398mg·kg-1 and 36.535mg·kg-1 respectively.In vitro,the inhibition activity of（＋）/（-）-pyraclofos to AChE increased with the increase of the individual enantiomer concentration.The different inducing activity to the expression level of Lm-ace1 mRNA and the different inhabitation activity to AChE in Locust have some contribution to the significant toxicity difference between the two enantiomers of pyraclofos.

关 键 词：东亚飞蝗 吡唑硫磷 对映选择毒性 乙酰胆碱酯酶 

分 类 号：S482.3[农业科学—农药学]