参与棕榈蓟马氯氰菊酯抗性的CYP450解毒酶基因克隆及表达量分析  

Gene Expression Analysis of CYP450 Detoxification Enzyme Involved in Cypermethrin Resistance Mechanism in Thrips palmi Karny

作　　者：包文学 Bao Wenxue(Forestry College of InnerMongolia Agricultural University,Hohhot,010019)

机构地区：[1]内蒙古农业大学林学院

出　　处：《分子植物育种》2019年第19期6314-6322,共9页Molecular Plant Breeding

基　　金：内蒙古自治区留学人员科技活动项目;内蒙古自治区人才开发基金项目(108-111059);内蒙古自治区林学“双一流”建设项目(203-170006)共同资助

摘　　要：棕榈蓟马(Thrips palmi Karny)是世界性蔬菜重要害虫。该害虫在日本对氯氰菊酯等拟除虫菊酯类杀虫剂产生不同程度的抗性。本研究中的冈山和高知品系用解毒酶活性阻隔剂试验结果,酶活性阻隔剂PBO处理的冈山和高知品系对氯氰菊酯致死中浓度(lethal concentration 50,LC50)值比未处理的LC50值,分别降低约1/3和1/14。CYP450均参与两个品系对拟除虫菊酯杀虫剂抗性。酶活性阻隔剂DEM和DEF处理后两个品系对氯氰菊酯LC50值未显著变化。棕榈蓟马GST和CE未参与拟除虫菊酯类杀虫剂解毒分解而产生抗性。由高知品系互补脱氧核糖核酸(complementary DNA,cDNA)扩增的基因片段进行克隆,60菌落(Clone)进行碱基序列测序。氨基酸相同性检索结果,29菌落(Clone)是CYP4家族基因,31菌落(Clone)是CYP6家族基因。CYP4家族基因的29菌落(Clone)归属于6组,CYP6家族基因的31菌落(Clone)归属于4组。各菌落(Clone)中均存在CYP450 C-末端领域特有的血红素结合区(FXXGXXXCXG)。冈山和高知品系之间的表达量进行比较,参与两品系之间的拟除虫菊酯类杀虫剂抗性差异的基因尚未发现。参与抗性基因的表达是杀虫剂处理过程时其表达量增加的可能性。本研究为棕榈蓟马杀虫剂抗性机制研究提供帮助。Thrips palmi Karny is a globally considered pest for vegetable.This species has developed different levels of resistance for various types of insecticides in Japan,including cypermethrin which is a insecticide belong to Pyrethroid.In the present study,compared to the control group,the LC50 value decreased 1/3 and 1/14 for Okayama and Kouchi strain,respectively,after the PBO treatment.The results indicated that the CYP450 might play a role in the insecticide resistance of Okayama and Kouchi strain.There was no significant change in the LC50 value of Okayama and Kouchi strain for Cypermethrin after the treatment with enzyme activity blocking agents(DEM and DEF).It seems that the GST and CE do not play any role in the Pyrethroid resistance level of thrips.The 60 colonies from the cDNA of Kouchi strain were cloned and sequenced.According to the consensus of amino acids,the colony 29 and 31 are belong to CYP4(6 Group)and CYP6(4 Group)family,respectively.The CYP450 C-endodomain-specific heme-binding domain(FXXGXXXCXG)was found in each colony.According to the gene expression analysis,we did not find any difference between the Okayama and Kouchi strain in terms of the gene responsible for the Pyrethroid resistance.The expression of resistance gene might increased during the use of insecticide.This study provides help for the study of the anti-sexual mechanism of the palm-and-horse pesticide.

关 键 词：棕榈蓟马(Thrips palmi Karny) 氯氰菊酯 CYP450 基因表达量 

分 类 号：S436.3[农业科学—农业昆虫与害虫防治]