昆虫学报 ›› 2017, Vol. 60 ›› Issue (3): 254-263.doi: 10.16380/j.kcxb.2017.03.003

• 研究论文 • 上一篇    下一篇

豌豆蚜体内参与抵御细菌感染引起的氧化胁迫的过氧化物氧化还原酶2的鉴定及分子特性分析(英文)

张永栋1, 李君兰1, 陈金红1, 马力1, 陈康康1, 吕志强1,2,*   

  1. (1. 西北农林科技大学植物保护学院昆虫学系, 陕西杨凌 712100; 2. 西北农林科技大学植保资源与病虫害治理教育部重点实验室, 陕西杨凌 712100)
  • 出版日期:2017-03-20 发布日期:2017-03-20

Identification and characterization of peroxiredoxin 2 involved in the antioxidative response induced by bacterial infection in the pea aphid, Acyrthosiphon pisum (Hemiptera: Aphididae) (In English

ZHANG Yong-Dong1, LI JunLan1, CHEN Jin-Hong1, MA Li1, CHEN Kang-Kang1, LÜ Zhi-Qiang1,2,*   

  1. (1. Department of Entomology, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China; 2. Key Laboratory of Plant Protection Resources and Pest Management, Ministry of Education, Northwest A&F University, Yangling, Shaanxi 712100, China)
  • Online:2017-03-20 Published:2017-03-20

摘要: 【目的】本研究旨在揭示豌豆蚜Acyrthosiphon pisum体内过氧化物氧化还原酶2 (ApPrx2)在豌豆蚜应对细菌感染中的作用。【方法】克隆并在大肠杆菌Escherichia coli中异源表达ApPrx2的开放阅读框;对重组蛋白的抗氧化活性进行鉴定;测定绿脓杆菌Pseudomonas aeruginosa和金黄色葡萄球菌Staphylococcus aureus感染豌豆蚜后豌豆蚜体内H2O2浓度和ApPrx2的转录水平;通过RNA干扰降低ApPrx2的表达;降低ApPrx2的表达后检测豌豆蚜体内H2O2浓度、细菌数目和豌豆蚜的存活率。【结果】 序列比对结果表明,ApPrx2与其他物种2-Cys Prxs具有较高的相似性。重组表达的ApPrx2蛋白能够有效降解H2O2,表达ApPrx2蛋白的大肠杆菌E. coli对H2O2的抗性更高。绿脓杆菌P. aeruginosa和金黄色葡萄球菌S. aureus感染引起豌豆蚜体内H2O2水平及ApPrx2转录水平的升高。通过RNA干扰降低ApPrx2表达之后,金黄色葡萄球菌感染的蚜虫体内的H2O2水平显著上升,其体内的细菌数显著低于对照,蚜虫的存活率显著降低。【结论】ApPrx2作为抗氧化蛋白能够帮助豌豆蚜抵御因细菌特别是金黄色葡萄球菌感染引起的氧化胁迫。

关键词: 豌豆蚜, 过氧化物氧化还原酶, 活性氧, 抗氧化反应, 细菌感染

Abstract: 【Aim】 The aim of this study is to reveal the function of peroxiredoxin 2 (ApPrx2) in the pea aphid, Acyrthosiphon pisum during bacterial infection. 【Methods】 The open reading frame of ApPrx2 was cloned and heterologously expressed in Escherichia coli. The antioxidative function of the purified recombinant ApPrx2 was characterized. The H2O2 concentration and ApPrx2 transcription level were detected in pea aphids after bacterial infections of Pseudomonas aeruginosa and Staphylococcus aureus. RNA interference was used to knockdown the expression of ApPrx2. H2O2 concentration, bacterial load and the survival rate of aphids were evaluated after the knockdown of ApPrx2. 【Results】 Sequence alignment showed that ApPrx2 is highly similar to 2-Cys Prxs from other insect species. In vitro assays demonstrated that the recombinant ApPrx2 protein degraded H2O2, and ApPrx2 expression rendered E. coli cells resistant to H2O2. Infections of the bacteria P. aeruginosa and S. aureus increased the H2O2 concentration and ApPrx2 expression in the pea aphids. The knockdown of ApPrx2 expression increased the H2O2 concentration, reduced the bacteria load and increased the mortality rate of aphids infected with S. aureus. 【Conclusion】 ApPrx2 functions as an antioxidative protein protecting pea aphids from stress induced by infection with bacteria, particularly S. aureus.

Key words: Acyrthosiphon pisum, peroxiredoxin, reactive oxygen species (ROS), antioxidative response, bacterial infection