豌豆蚜组织蛋白酶K基因(ApCtsk1)的鉴定及功能研究  

Identification and functional analysis of ApCtsk1 encoding cathepsin K in Acyrthosiphon pisum

作　　者：马瑞 吴宜亭 李岩 于展姿 刘磊 王森山 MA Rui;WU Yi-ting;LI Yan;YU Zhan-zi;LIU Lei;WANG Sen-shan(College of Plant Protection,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学植物保护学院,甘肃兰州730070

出　　处：《草业学报》2025年第2期174-183,共10页Acta Prataculturae Sinica

基　　金：甘肃省种业攻关计划项目(GYGG-2024-7);甘肃省自然科学基金项目(20JR5RA028);甘肃省高等学校创新基金项目(2022A-060);甘肃农业大学公招博士科研启动项目(GAN-KYQD-2019-28)资助。

摘　　要：为探究豌豆蚜ApCtsk1基因的生理功能,评估其作为豌豆蚜防治靶标的潜力,从而为防控策略的制定提供理论依据。本研究以豌豆蚜为对象,利用NCBI、ExPASy Proteomics Server、MEGA 11等在线网站和软件分析ApCtsk1的氨基酸序列特征并构建系统发育树;采用RT-qPCR技术解析其在不同发育阶段、不同组织的表达模式;通过RNAi研究ApCtsk1在豌豆蚜个体发育和繁殖中的生物学功能。结果表明:豌豆蚜ApCtsk1与半翅目蚜科昆虫的半胱氨酸蛋白酶聚为一支,其在4龄若蚜期显著高表达,并在成虫腹部、卵巢、表皮表达量较高。RNAi干扰3龄若蚜的ApCtsk1后,与对照组相比,豌豆蚜死亡率升高至22.22%,体长体宽分别减小12.56%、13.22%,体色异常,行动缓慢。此外,经dsApCtsk1处理的蚜虫出现蜕皮失败的致死现象,且干扰后豌豆蚜繁殖力也受到影响,表现为产蚜时间的推迟和一段时间产蚜量的降低。由此可见,ApCtsk1在豌豆蚜个体发育、存活和繁殖过程中具有重要的作用。The aim of this research was to explore the function of ApCtsk1 in pea aphid(Acyrthosiphon pisum)and evaluate its potential as a target for the prevention and control of this pest.The amino acid sequence characteristics of the putative ApCtsk1 protein were analyzed using tools at various websites(NCBI,ExPASy Proteomics Server)and a phylogenetic tree was constructed using MEGA 11.The transcript levels of its encoding gene were determined in aphids at different stages of development and in different tissues by RT-qPCR analysis.The biological function of ApCtsk1 in the development and reproduction of A.pisum was investigated using an RNA interference gene knock-down strategy.In the phylogenetic analysis,ApCtsk1 was clustered with cysteine proteases from other members of the Aphididae in the Hemiptera.High transcript levels of ApCtsk1 were detected in fourth instar nymphs,and in the abdomen,ovary,and cuticle of adult aphids.Compared with A.pisum in the control group,those in the ApCtsk1-knockdown group showed an increased mortality rate(22.22%higher),decreased body length and body width(decreased by 12.56%and 13.22%,respectively),abnormal body color,and slower movement.In addition,the aphids with knocked-down ApCtsk1 showed the lethal phenomenon of molting failure,which affected their fecundity.These changes resulted in delayed reproduction and decreased reproduction over time.Our findings show that ApCtsk1 plays an important role in the development,survival,and reproduction of A.pisum.

关 键 词：豌豆蚜 组织蛋白酶K 繁殖 RNAI 

分 类 号：S64[农业科学—蔬菜学]