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东方粘虫(Mythimna separata Walker)β-肾上腺素能样章鱼胺受体的特性和功能分析。

Characterization and functional analysis of a β-adrenergic-like octopamine receptor from the oriental armyworm (Mythimna separata Walker).

机构信息

National Engineering Research Center of Sugarcane, Fujian Agricultural and Forestry University, Fuzhou, China.

Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing, China.

出版信息

Arch Insect Biochem Physiol. 2021 Apr;106(4):e21772. doi: 10.1002/arch.21772. Epub 2021 Mar 15.

Abstract

The β-adrenergic-like octopamine receptor (OA2B2), which binds the biogenic amine octopamine, belongs to the class of G-protein coupled receptors and significantly regulates many physiological and behavioral processes in insects. In this study, the putative open reading frame sequence of the MsOA2B2 gene in Mythimna separata was cloned, the full-length complementary DNA was 1191 bp and it encoded a 396-amino acid protein (GenBank accession number MN822800). Orthologous sequence alignment, phylogenetic tree analysis, and protein sequence analysis all showed that the cloned receptor belongs to the OA2B2 protein family. Real-time quantitative polymerase chain reaction of spatial and temporal expression analysis revealed that the MsOAB2 gene was expressed in all developmental stages of M. separata and was most abundant in egg stages and second and fourth instars compared with other developmental stages, while the expression level during the pupal stage was much lower than that at the other stages. Further analysis with sixth instar M. separata larvae showed that the MsOA2B2 gene was expressed 1.81 times higher in the head than in integument and gut tissues. Dietary ingestion of dsMsOA2B2 significantly reduced the messenger RNA level of MsOA2B2 and decreased mortality following amitraz treatment. This study provides both a pharmacological characterization and the gene expression patterns of OA2B2 in M. separata, facilitating further research for insecticides using MsOA2B2 as a target.

摘要

β-肾上腺素样章鱼胺受体(OA2B2)结合生物胺章鱼胺,属于 G 蛋白偶联受体类,能显著调节昆虫的许多生理和行为过程。本研究克隆了粘虫 MsOA2B2 基因的开放阅读框序列,全长 cDNA 为 1191bp,编码 396 个氨基酸的蛋白(GenBank 登录号 MN822800)。同源序列比对、系统进化树分析和蛋白序列分析均表明克隆的受体属于 OA2B2 蛋白家族。时空表达分析的实时定量聚合酶链反应显示,MsOAB2 基因在粘虫的所有发育阶段均有表达,在卵期和 2 龄、4 龄期最为丰富,而在蛹期的表达水平明显低于其他发育阶段。进一步用 6 龄粘虫幼虫分析表明,MsOA2B2 基因在头部的表达量比在表皮和肠道组织中高 1.81 倍。dsMsOA2B2 的膳食摄入显著降低了 MsOA2B2 的信使 RNA 水平,并降低了拟除虫菊酯处理后的死亡率。本研究为 OA2B2 在粘虫中的药理学特征和基因表达模式提供了依据,为以 MsOA2B2 为靶标的杀虫剂的进一步研究奠定了基础。

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