白背飞虱SfABCD1对杀虫剂胁迫的响应表达  

Expression of SfABCD1 in the white-backed planthopper,Sogatella furcifera(Hemiptera:Delphacidae)in response to insecticide stress

作　　者：曾庆会 周操 杨熙彬 杨洪[1,3] 金道超 龙贵云[1] ZENG Qing-Hui;ZHOU Cao;YANG Xi-Bin;YANG Hong;JIN Dao-Chao;LONG Gui-Yun(Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region, Scientific Observing and Experimental Station of Crop Pests in Guiyang, Ministry of Agriculture and Rural Affairs, Institute of Entomology, Guizhou University, Guiyang 550025, China;College of Life Sciences, Chongqing Normal University, Chongqing 401331, China;College of Tobacco Science, Guizhou University, Guiyang 550025, China)

机构地区：[1]贵州大学昆虫研究所,贵州山地农业病虫害重点实验室,农业农村部贵阳作物有害生物科学观测试验站,贵阳550025 [2]重庆师范大学生命科学学院,重庆401331 [3]贵州大学烟草学院,贵阳550025

出　　处：《昆虫学报》2022年第1期21-30,共10页Acta Entomologica Sinica

基　　金：国家自然科学地区基金项目(31960537,31560522);重庆师范大学基金项目(21XLB010);贵州省研究生科研基金立项课题(黔教合YJSCXJH(2020)075)。

摘　　要：【目的】ABC转运蛋白(ATP-binding cassette transporter,ABC transporter)是一个庞大的膜转运蛋白超家族,在昆虫生长发育和抗药性方面具有重要作用。本研究旨在通过克隆白背飞虱Sogatella furcifera ABCD1基因,并测定其在杀虫剂胁迫下的表达模式,为后续的功能研究提供理论依据。【方法】采用RT-PCR技术克隆白背飞虱ABCD1基因cDNA序列并进行生物信息学分析;通过RT-qPCR技术测定其在白背飞虱不同发育时期(1-5龄若虫和羽化后第1-4天雌雄成虫)、5龄若虫和成虫的不同组织(头部、体壁、脂肪体、肠道、足、翅、精巢和卵巢)中及不同浓度(LC_(10),LC_(25),LC_(50)和LC_(90))噻虫嗪、噻嗪酮和阿维菌素胁迫下3龄若虫中的表达量。【结果】获得一个白背飞虱ABC转运蛋白基因,命名为SfABCD1(GenBank登录号:MW701394),其开放阅读框长2220 bp,编码739个氨基酸,预测分子量为82.08 kD,理论等电点为9.32;SfABCD1与灰飞虱Laodelphax striatellus和褐飞虱Nilaparvata lugens的ABCD1亲缘关系较近。RT-qPCR结果表明,SfABCD1在5龄若虫中表达量最高,且在5龄若虫和成虫各组织中均有表达,在5龄若虫肠道和头部中表达量最高。3龄若虫受杀虫剂胁迫48 h后,与清水处理的对照组相比,噻嗪酮(LC_(10),LC_(50)和LC_(90)),噻虫嗪(LC_(10),LC_(25)和LC_(50))和阿维菌素(LC_(50)和LC_(90))处理组中的SfABCD1表达量均显著上调;LC_(10),LC_(25),LC_(50)和LC_(90)4个浓度的3种杀虫剂处理不同时间后,SfABCD1的表达量在杀虫剂胁迫48 h时与对照差异最大,胁迫96 h时表达差异下降,处理120 h后该基因的表达水平接近对照组。【结论】白背飞虱可通过上调SfABCD1表达量来应对噻嗪酮和噻虫嗪的胁迫,SfABCD1表达量随噻虫嗪浓度的升高而降低;另外,白背飞虱也通过调节该基因的转录水平来响应高浓度阿维菌素的毒害。【Aim】ATP-binding cassette transporter(ABC transporter),a huge superfamily of membrane transporters,plays an important role in insect growth,development and insecticide resistance.This study aims to provide theoretical basis for subsequent functional studies by cloning ABCD1 of the white-backed planthopper,Sogatella furcifera and determining its expression pattern under insecticide stress.【Methods】The cDNA sequence of ABCD1 of S.furcifera was cloned by RT-PCR,and subjected to bioinformatics analysis.The expression levels of ABCD1 in S.furcifera at different developmental stages(1st-5th instar nymphs and 1-4 day-old male and female adults),in different tissues(head,integument,fat body,gut,leg,wing,testis and ovary)of the 5th instar nymphs and adults,and in the 3rd instar nymphs exposed to different concentrations(LC_(10),LC_(25),LC_(50) and LC_(90))of thiamethoxam,buprofezin,and abamectin were detected by RT-qPCR.【Results】An ABC transporter gene of S.furcifera was obtained and named SfABCD1(GenBank accession no.:MW701394),with an open reading frame of 2220 bp in length.It encodes a putative protein of 739 amino acid residues with the predicted molecular weight of 82.08 kD and isoelectric point of 9.32.SfABCD1 is closely related to ABCD1 of Laodelphax striatellus and Nilaparvata lugens.RT-qPCR results showed that SfABCD1 had the highest expression level in the 5th instar nymphs and was expressed in various tissues of the 5th instar nymphs and adults with the highest expression level in the gut and head of the 5th instar nymphs.After the 3rd instar nymphs were exposed to insecticides for 48 h,the expression levels of SfABCD1 in the treatment groups of buprofezin at the concentrations of LC_(10),LC_(50) and LC_(90),thiamethoxam at the concentrations of LC_(10),LC_(25) and LC_(50) and abamectin at the concentrations of LC_(50) and LC_(90) were significantly up-regulated as compared to that in the control group treated with water.When the 3rd instar nymphs were exposed to the three insecticides at the four c

关 键 词：白背飞虱 ABCD1基因 杀虫剂胁迫 表达分析 抗药性 

分 类 号：Q966[生物学—昆虫学]