灰飞虱雌激素相关受体基因的克隆、序列分析及在氟啶虫胺腈胁迫下的表达模式

doi:10.16380/j.kcxb.2017.03.004

昆虫学报 ›› 2017, Vol. 60 ›› Issue (3): 264-273.doi: 10.16380/j.kcxb.2017.03.004

灰飞虱雌激素相关受体基因的克隆、序列分析及在氟啶虫胺腈胁迫下的表达模式

徐鹿^1，2, 赵春青², 徐德进¹, 徐广春¹, 许小龙¹, 张亚楠³, 韩召军², 顾中言^1，*

（1. 江苏省农业科学院植物保护研究所, 南京210014; 2. 南京农业大学植物保护学院, 农作物生物灾害综合治理教育部重点实验室, 南京 210095; 3. 淮北师范大学生命科学学院, 安徽淮北235000）

出版日期:2017-03-20 发布日期:2017-03-20

Molecular cloning and sequence analysis of the estrogen-related receptor (ERR) gene in the small brown planthopper, Laodelphax striatellus (Delphacidae: Hemiptera) and its expression profiles under the stress of sulfoxaflor

XU Lu^1，2, ZHAO Chun-Qing², XU De-Jin¹, XU Guang-Chun¹, XU Xiao-Long¹, ZHANG Ya-Nan³, HAN Zhao-Jun², GU Zhong-Yan^1，*

(1. Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 2. Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China; 3. College of Life Sciences, Huaibei Normal University, Huaibei, Anhui 235000, China)

Online:2017-03-20 Published:2017-03-20

摘要/Abstract

摘要： 【目的】雌激素相关受体（estrogen-related receptor， ERR）是一类依赖配体激活的转录因子，能感受外源物质胁迫调控靶基因的转录，参与外源物质的代谢过程。本研究旨在通过克隆灰飞虱Laodelphax striatellus ERR基因，分析其序列和在氟啶虫胺腈胁迫下表达谱，探析其生理功能及在杀虫剂代谢中的作用。【方法】根据灰飞虱转录组数据信息，利用RT-PCR克隆灰飞虱ERR基因，并进行生物信息学分析；通过实时荧光定量PCR，分析暴露于氟啶虫胺腈（0.76 ng/头）后，灰飞虱4龄若虫ERR基因在不同时间点和不同组织中的表达量。【结果】从灰飞虱中克隆到ERR基因，命名为LsERR（GenBank登录号： KY210878），其cDNA序列全长1 854 bp，开放阅读框长1 260 bp，编码419个氨基酸，预测编码蛋白的分子量为47.70 kD。序列分析显示，LsERR具有核受体（nuclear receptors， NRs）家族成员的共同特征性结构：DNA结合区（DNA-binding domain， DBD）(第75-168位氨基酸）和配体结合区（ligand-binding domain， LBD）(第194-416位氨基酸）。采用APSSP2法预测LsERR蛋白二级结构，其中α螺旋占43.53%，β折叠占5.49%，无规则卷曲占50.98%。其中，在DBD区，有6个β折叠和3个α螺旋；在LBD区，有3个β折叠和11个α螺旋。系统发育分析表明，LsERR与褐飞虱Nilaparvata lugens ERR亲缘关系最近。灰飞虱4龄若虫暴露于氟啶虫胺腈后，12 h时其体内LsERR基因上调表达，24 h时达到表达高峰，48 h时表达量下调；LsERR在头部微弱表达，在腹部特异性高表达。【结论】LsERR是参与灰飞虱代谢氟啶虫胺腈的候选基因。本研究为解毒酶基因的调控和新分子靶标农药的研制提供分子基础。

Abstract: 【Aim】 Estrogen-related receptors (ERRs) are a class of transcription factors that are dependent on ligand activation, which can respond to the stress of xenobiotic by regulating transcription levels of target genes and being involved in xenobiotic metabolism. In order to define the physiological functions of ERR in Laodelphax striatellus and its roles in insecticide metabolism, we cloned and characterized its ERR gene and analyzed its expression profiles after exposure to sulfoxaflor. 【Methods】 The ERR gene of L. striatellus was cloned using RT-PCR according to transcriptome data of L. striatellus and analyzed by bioinformatic tools, and its mRNA levels at different time and in different tissues of the 4th instar nymphs of L. striatellus after exposure to sulfoxaflor (0.76 ng/individual) were detected via real-time PCR. 【Results】 The ERR gene was cloned from L. striatellus， and named LsERR (GenBank accession no.: KY210878), its complete cDNA is 1 854 bp in length with a 1 260 bp of open reading frame， encoding a 47.70 kD protein with 419 amino acids. LsERR has the common conserved domains of NRs family, including the DNA-binding domain (DBD) (75-168 aa) and the ligand-binding domain (LBD) (194-416 aa). By using APSSP2 method, the predicted secondary structure of LsERR showed that the alpha helix accounts for 43.53%， the beta fold accounts for 5.49%, and the random coil accounts for 50.98%. There are six beta folds and three alpha helices in the DBD region while there are three beta folds and 11 alpha helices in the LBD region. Phylogenetic tree analysis indicated that LsERR is most closely related to Nilaparvata lugens ERR. In the 4th instar nymphs of L. striatellus exposed to sulfoxaflor, the expression level of LsERR increased at 12 h, reached the peak at 24 h, and decreased at 48 h. LsERR had weak expression in the head and high expression in the abdomen. 【Conclusion】 LsERR is a candidate gene involved in sulfoxaflor metabolism of L. striatellus. This study provides a molecular basis for the study of regulation of detoxification enzyme genes and the development of a novel molecular target of insecticides.

Key words: Laodelphax striatellus, estrogen-related receptor, gene cloning, sequence analysis, expression profile, sulfoxaflor

徐鹿, 赵春青, 徐德进, 徐广春, 许小龙, 张亚楠, 韩召军, 顾中言. 灰飞虱雌激素相关受体基因的克隆、序列分析及在氟啶虫胺腈胁迫下的表达模式[J]. 昆虫学报, 2017, 60(3): 264-273.

XU Lu, Zhao Chun-Qing, Xu De-Jin, Xu Guang-Chun, Xu Xiao-Long, Zhang Ya-Nan, Han Zhao-Jun, Gu Zhong-Yan. Molecular cloning and sequence analysis of the estrogen-related receptor (ERR) gene in the small brown planthopper, Laodelphax striatellus (Delphacidae: Hemiptera) and its expression profiles under the stress of sulfoxaflor[J]. Acta Entomologica Sinica, 2017, 60(3): 264-273.

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链接本文: http://www.insect.org.cn/CN/10.16380/j.kcxb.2017.03.004

http://www.insect.org.cn/CN/Y2017/V60/I3/264

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灰飞虱雌激素相关受体基因的克隆、序列分析及在氟啶虫胺腈胁迫下的表达模式

Molecular cloning and sequence analysis of the estrogen-related receptor (ERR) gene in the small brown planthopper, Laodelphax striatellus (Delphacidae: Hemiptera) and its expression profiles under the stress of sulfoxaflor

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