豌豆蚜特异性扩张分支气味受体基因的克隆及表达分析

Molecular cloning and expression analysis of specific expansive odorant receptors clade in Acyrthosiphon pisum(Hemiptera:Aphididae)

豌豆蚜Acyrthosiphon pisum气味受体家族存在明显扩张现象,推测在豌豆蚜寄主适应过程中起着重要作用。本研究旨在克隆豌豆蚜特异性扩张分支上的气味受体基因,明确这些气味受体基因在不同组织中的表达水平。通过PCR技术克隆特异性扩张分支上气味受体基因的全长序列,采用实时荧光定量RT-qPCR技术测定这些基因在触角及足中的表达水平。克隆获得豌豆蚜特异性扩张分支中4个气味受体基因全长序列(GenBank登录号:OL739156-OL739159),分别编码374~376个氨基酸,序列相似性较高,为68.09%~83.11%,均具有7个跨膜结构域。荧光定量RT-qPCR结果显示ApisOR6、ApisOR8、ApisOR13和ApisOR14基因在豌豆蚜成虫触角及足中均有表达,其中ApisOR8、ApisOR13和ApisOR14基因在触角中偏好表达,ApisOR13基因在触角中的表达量最高。本研究为解析豌豆蚜特异性扩张气味受体的功能提供了分子基础。

Gene family expansions of odorant receptors(OR)were found in the pea aphid,Acyrthosiphon pisum,and might play a vital role in the host plant adaptation.This study aims to clone odorant receptor genes of a specific expansive OR clade,and to assay their expression levels in different adult tissues.PCR was used to clone the full-length cDNA sequences of four OR genes.Real-time quantitative PCR(RT-qPCR)was used to assess the expression patterns of specific expansive OR genes in antennae and legs.The full-length cDNA sequences of four OR genes(GenBank accession no.:OL739156-OL739159)of the specific expansive OR clade in A.pisum were cloned,encoding 374-376 amino acids,having the high sequence similarities from 68.09%to 83.11%and all containing seven transmembrane domains.The RT-qPCR results showed that the four OR genes were expressed in both antennae and legs of A.pisum adult,among which,ApisOR8,ApisOR13,and ApisOR14 bias to express in the antennae,and the ApisOR13 has the highest expression level in the antennae.This study provides a basis for revealing the molecule mechanism of specific expansion ORs in A.pisum.