棉蚜ATP合成酶基因AgoATPb的克隆与表达  

Cloning and expression analysis of AgoATPb gene in cotton-melon aphid,Aphis gossypi

作　　者：杨卫军[1,2] 董艳蕾 吴秋芳 张美玲[1,2] 韩丽滨 张元臣 YANG Weijun;DONG Yanlei;WU Qiufang;ZHANG Meiling;HAN Libin;ZHANG Yuanchen(College of Biological and Food Engineering,Anyang Institude of Technology,Anyang 455000,Henan,China;Taihang Mountain,Forest Pests Observation and Research Station of Henan Province,Linzhou 456550,Henan,China)

机构地区：[1]安阳工学院生物与食品工程学院,河南安阳455000 [2]河南省太行山林业有害生物野外科学观测研究站,河南林州456550

出　　处：《浙江农业学报》2022年第2期329-336,共8页Acta Agriculturae Zhejiangensis

基　　金：安阳工学院博士启动基金(BSJ2019001,BSJ2019021);国家自然科学基金(31802007)。

摘　　要：为确定棉蚜(Aphis gossypii)ATP合成酶B亚基基因在棉蚜不同组织和日龄,以及取食不同植物的表达情况,以棉蚜为研究对象,采用RT-PCR和RACE技术获得AgoATPb的全长cDNA序列,通过Expasy、SignalP-4.0 Server等在线工具对其进行了生物信息学分析,同时利用实时荧光定量PCR技术研究了该基因在棉蚜不同组织和不同日龄,以及在不同植物上取食的表达水平。结果表明,棉蚜AgoATPb基因全长cDNA序列为1247 bp,开放阅读框(ORF)长度为822 bp,编码274个氨基酸,5′端非编码区长128 bp,3′端非编码区长297 bp,理论分子量为31.40 ku,等电点为8.95,无信号肽和跨膜区域。氨基酸序列比对结果表明,棉蚜AgoATPb与其他昆虫同源基因编码蛋白的氨基酸序列一致性为47%~99%。除了不在胚胎表达外,AgoATPb基因在棉蚜其他日龄和不同组织均有表达,且在不同日龄间与不同组织表达水平存在显著差异。取食不同植物后棉蚜AgoATPb基因表达水平存在显著差异,取食棉花表达水平最高,其次是黄瓜和西葫芦,取食甜瓜表达水平最低,推测该基因可能与棉蚜适应寄主植物有关。To evaluate the effects of tissue,age and host plant on expression of ATP synthase subunit B gene(AgoATPb)of Aphis gossypii,cotton aphid was taken as the research object,full-length cDNA sequence of AgoATPb gene was obtained by RT-PCR and RACE(rapid amplification cDNA ends)technology,and the bioinformatics analysis was carried out using online tools such as Expasy and SignalP-4.0 Server.Expression level of AgoATPb gene was also studied by real-time quantitative PCR technology in different tissues and different day ages of cotton aphid and when feeding on different plants.Full-length cDNA sequence of AgoATPb gene was 1247 bp,and its open reading frame(ORF)was 822 bp,encoding 273 amino acids.The 5′noncoding region and 3′noncoding region of AgoATPb gene were 128 bp and 297 bp,respectively.Theoretical molecular weight and isoelectric point of amino acid sequence coded by AgoATPb gene were 31.40 ku and 8.95,respectively.There were no signal peptide and no transmembrane region in AgoATPb.Amino acid sequence analysis showed that the sequence identity of AgoATPb with the homologous protein from other insect were between 47%and 99%.Apart from not expressed in the embryonic stage,AgoATPb gene was transcribed throughout all developmental stages and tissues of A.gossypii,but expression levels significantly differed among the developmental stages and among the different tissues.After feeding on different plants,the highest levels of AgoATPb expression were observed on cotton,followed by cucumber and zucchin,and the lowest expression levels was observed on melon.It was speculated that AgoATPb gene might be related to adaptation of cotton aphids to host plants.

关 键 词：棉蚜 ATP合成酶B亚基 生物信息学 基因克隆 寄主 

分 类 号：S435.6221[农业科学—农业昆虫与害虫防治]