桃蛀螟细胞色素P450基因CYP4G113时空表达及功能研究

doi:10.16409/j.cnki.2095-039x.2021.03.023

摘要/Abstract

摘要： CYP家族基因是昆虫体内重要的解毒酶基因，本研究拟明确桃蛀螟体内解毒酶基因CYP4G113的基本序列特征、表达模式及其在桃蛀螟适应性进化中的功能。基于桃蛀螟转录组数据信息，克隆获得桃蛀螟CYP4G113的开放阅读框；利用实时荧光定量PCR（Quantitative real-time PCR，qRT-PCR）技术，测定CYP4G113在桃蛀螟各发育阶段和幼虫不同组织内的表达水平；利用RNAi（RNA interference）技术研究该基因被干扰后对桃蛀螟生长发育的影响。获得了桃蛀螟CYP4G113全长cDNA序列（GenBank登录号：MT721971），其开放阅读框为1707 bp，编码568个氨基酸，预测蛋白分子量约为64.59 kDa，理论等电点为8.54；qRT-PCR结果显示，该基因在桃蛀螟各发育阶段均有表达，其中1龄幼虫期表达量最高，卵和5龄幼虫中表达量最低；4龄幼虫不同组织中，唾液腺中的相对表达量显著高于其他组织；该基因被干扰后，桃蛀螟幼虫在其适宜寄主植物-玉米上的存活率及化蛹率均显著下降。桃蛀螟CYP4G113在低龄幼虫中表达量相对较高且主要在唾液腺中表达，该基因被干扰后的幼虫生长发育受到显著影响，显示该基因在桃蛀螟寄主适应中发挥重要作用。本研究结果可以为基于RNAi的害虫控制提供新的靶标基因。

关键词: 桃蛀螟, 细胞色素P450, 荧光定量PCR, RNAi, 基因功能

Abstract: CYP family genes encode important detoxifying enzymes in insects. This study was designed to clarify the basic sequence characteristics, expression patterns and function of the CYP4G113 in the adaptive evolution of Conogethes punctiferalis. The open reading frame of C. punctiferalis CYP4G113 was cloned based on C. punctiferalis transcriptome data, and CYP4G113 expression levels were determined in various developmental stages and larval tissues using quantitative real-time PCR (qRT-PCR). CYP4G113 expression was silenced using RNA interference (RNAi) technology and the RNAi effect on C. punctiferalis growth and development was examined. The full-length C. punctiferalis CYP4G113 cDNA sequence, with an open reading frame of 1707 bp, was obtained (GenBank accession number:MT721971). This open reading frame encodes 568 amino acids, with a predicted molecular weight of approximate 64.59 kDa, and a theoretical isoelectric point of 8.54. qRT-PCR results showed that CYP4G113 was expressed in all developmental stages of C. punctiferalis, with the highest expression in 1^st instar larvae, and the lowest expression in the eggs and 5^th instar larvae. Meanwhile, the expression of CYP4G113 was the highest in the salivary gland among the tissues of 4^th instar larvae. The survival and pupation rates of C. punctiferalis larvae on a suitable host plant, maize, were significantly reduced after CYP4G113 silencing. In C. punctiferalis, CYP4G113 is expressed at relatively high levels in young larval insects and is mainly expressed in the salivary gland. In addition, the growth and development of C. punctiferalis larvae are significantly affected by CYP4G113 silencing, indicating that this gene plays an important role in C. punctiferalis host adaptation. The results of this study will provide a new potential target gene for controlling insects using RNAi technology.

Key words: Conogethes punctiferalis, cytochrome P450, quantitative real-time PCR, RNAi, gene function

中图分类号:

S476

袁星星, 董少奇, 王鑫辉, 郭线茹, 王高平, 李为争, 张利娟, 赵曼. 桃蛀螟细胞色素P450基因CYP4G113时空表达及功能研究[J]. 中国生物防治学报, 2022, 38(1): 196-204.

YUAN Xingxing, DONG Shaoqi, WANG Xinhui, GUO Xianru, WANG Gaoping, LI Weizheng, ZHANG Lijuan, ZHAO Man. Spatio-temporal Expression and Function of Cytochrome P450 Gene CYP4G113 in Conogethes punctiferalis[J]. Chinese Journal of Biological Control, 2022, 38(1): 196-204.

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