| 棉铃虫过氧化物还原酶基因Prx4的鉴定及功能分析 |
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| 引用本文：田志强,查萌,祝琳,申忠健,刘孝明,刘小侠.棉铃虫过氧化物还原酶基因Prx4的鉴定及功能分析.植物保护学报,2021,48(5):1008-1015 |
| DOI：10.13802/j.cnki.zwbhxb.2021.2021852 |
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| 中文摘要:为探究过氧化物还原酶（peroxiredoxin，Prx）在棉铃虫Helicoverpa armigera幼虫体内的功能，利用转录组数据鉴定得到了棉铃虫Prx4基因，并对其进行生物信息学分析，采用实时荧光定量PCR（real-time quantitative polymerase chain reaction，qRT-PCR）测定该基因在棉铃虫不同发育阶段和5龄幼虫不同组织中的相对表达，利用原核表达和RNA干扰方法探究该基因的功能。结果表明，Prx4基因开放阅读框长为744 bp，编码248个氨基酸，系统进化树和同源序列多重比对结果显示该基因属于典型的2-Cys过氧化物酶家族；Prx4基因广泛分布于棉铃虫幼虫各组织和发育阶段中；当感染核型多角体病毒（nucleopolyhedrovirus，NPV）后，棉铃虫Prx4基因相对表达量上调；重组蛋白的体外试验表明Prx4蛋白具有一定的抗氧化活性；与对照相比，小干扰RNA处理6、8和10 d后感病棉铃虫的死亡率分别较对照显著提高了4.46%、22.42%和38.68%。表明Prx4基因作为一种抗氧化酶保护棉铃虫免受氧化损伤，同时在抵御NPV感染过程中发挥着重要的免疫作用。 |
| 中文关键词:棉铃虫 过氧化物还原酶 抗氧化活性 RNA干扰 免疫 基因表达 核型多角体病毒 |
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| Identification and functional analysis of Prx4 gene in cotton bollworm Helicoverpa armigera |
| Author Name | Affiliation | E-mail | | Tian Zhiqiang | Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China | | | Zha Meng | Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China | | | Zhu Lin | Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China | | | Shen Zhongjian | Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China | | | Liu Xiaoming | Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China | | | Liu Xiaoxia | Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China | liuxiaoxia611@cau.edu.cn |
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| Abstract:In order to explore the function of peroxiredoxin in cotton bollworm Helicoverpa armigera larvae, the peroxiredoxin (Prx) 4 was identified and analyzed by using bioinformatics methods based on our transcriptome data; then the temporal and spatial expression level of Prx4 was measured using real-time quantitative polymerase chain reaction (qRT-PCR), and the function of Prx4 was verified using prokaryotic expression and RNA interference (RNAi). The results showed that Prx4 gene contained an open reading frame of 744 bp, which encoded 248 amino acids. The phylogenetic analyses and homology indicated that Prx4 gene belonged to the atypical 2-Cys peroxidase family; qRT-PCR assay demonstrated that Prx4 gene was widely distributed in different tissues and at all developmental stages of larvae, and was upregulated after infection with nucleopolyhedrovirus (NPV). The in vitro experiments demonstrated that the Prx4 protein displayed an antioxidant activity. Compared with the control group, RNAi results showed that the mortality rate of larvae infected by NPV was significantly increased by 4.46%, 22.42% and 38.68% after six, eight and ten days of treatment with small interfering RNA, respectively. These data indicated that Prx4 gene could be used as an antioxidant enzyme to protect the cotton bollworm from oxidative damage and played an important role in NPV infection. |
| keywords:Helicoverpa armigera peroxiredoxin antioxidant RNA interference immunity gene expression nucleopolyhedrovirus (NPV) |
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