摘要
【目的】明确烟粉虱Bemisia tabaci MED隐种抑制蛋白基因Btarrestin是否参与吡虫啉耐药性。【方法】根据已有烟粉虱转录组数据,利用RT-PCR克隆Btarrestin的全长序列,进行生物信息学分析。通过qPCR分析Btarrestin在烟粉虱MED隐种各个龄期(卵,1-2龄、3龄、4龄若虫,雌、雄成虫)以及吡虫啉处理(100 mg/L)后成虫中的表达量变化,明确其时空表达模式。利用RNAi技术沉默Btarrestin,观察沉默前后Btarrestin表达量和烟粉虱MED隐种成虫死亡率变化。【结果】成功克隆烟粉虱MED隐种Btarrestin的cDNA序列(GenBank登录号: MK204377),编码区全长1 227 bp,编码409个氨基酸,预测所编码的蛋白分子量约为45.33 kD,理论等电点(pI)为8.38。保守结构域分析表明,Btarrestin具有Arrestin_N和Arrestin_C两个超家族保守结构域,符合抑制蛋白家族特征。分子系统树分析表明,Btarrestin与褐飞虱Nilaparvata lugens的Arrestin亲缘关系最近。Btarrestin的表达量随烟粉虱MED隐种的生长发育逐渐升高,成虫期表达量最高。100 mg/L吡虫啉处理成虫24 h后Btarrestin的表达量较对照增加了2.39倍。RNAi干扰Btarrestin后进行生物测定,烟粉虱MED隐种成虫的死亡率上升了31.27%。【结论】Btarrestin可能与烟粉虱MED隐种对吡虫啉的耐药性有关。
【Aim】 To clarify the relationship between arrestin gene Btarrestin and imidacloprid resistance in the whitefly Bemisia tabaci MED. 【Methods】 Based on the previous transcriptome data of B. tabaci, the full-length cDNA sequence of Btarrestin was cloned by RT-PCR in B. tabaci MED and then subjected to bioinformatics analysis. The expression levels of Btarrestin at different developmental stages (egg, 1st-2nd instar, 3rd instar and 4th instar nymph, and female and male adult) and in adults exposed to imidacloprid (100 mg/L) were detected by qPCR. After silencing Btarrestin by RNAi, the expression level of Btarrestin and the mortality of B. tabaci MED adults were detected. 【Results】 The cDNA sequence of Btarrestin (GenBank accession no.: MK204377) was cloned from B. tabaci MED. The complete cDNA is 1 127 bp in length encoding 409 amino acids, with the molecular weight of 45.33 kD and the pI value of 8.38. Conserved domain analysis indicated that Btarrestin has conserved domains of the two superfamilies Arrestin_N and Arrestin_C, consistent with the family characteristics of arrestins. Phylogenetic tree analysis showed that Btarrestin shares high homology with the arrestin of Nilaparvata lugens. Developmental stage-specific expression results revealed that the expression level of Btarrestin was increased gradually with its development and reached the peak at the adult stage. The expression level of Btarrestin in adults exposed to 100 mg/L imidacloprid for 24 h increased by 2.39-fold as compared with that in the control. The RNAi results showed that knockdown of Btarrestin in B. tabaci MED increased the mortality of imidacloprid-treated adults by 31.27%. 【Conclusion】 Btarrestin might be involved in imidacloprid resistance in B. tabaci MED.
作者
梁金金
何超
刘少楠
谢文
张友军
LIANG Jin-Jin;HE Chao;LIU Shao-Nan;XIE Wen;ZHANG You-Jun(College of Plant Protection of Hunan Agricultural University, Changsha 410128, China;Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China)
出处
《昆虫学报》
CAS
CSCD
北大核心
2019年第10期1129-1139,共11页
Acta Entomologica Sinica
基金
国家重点研发计划项目(2018YFD0201201)
现代农业产业技术体系(CARS-24-C-02)
蔬菜有害生物控制与优质栽培北京市重点实验室
中国农业科学院科技创新工程(CAAS-ASTIP-IVFCAAS)
