两近缘种茶尺蠖的抗菌肽基因在抵抗EoNPV感染过程中的表达差异

doi:10.16409/j.cnki.2095-039x.2017.04.006

中国生物防治学报 ›› 2017, Vol. 33 ›› Issue (4): 472-480.DOI: 10.16409/j.cnki.2095-039x.2017.04.006

两近缘种茶尺蠖的抗菌肽基因在抵抗EoNPV感染过程中的表达差异

毛腾飞, 付建玉, 孙亮, 周孝贵, 白家赫, 肖强

中国农业科学院茶叶研究所, 杭州 310008

收稿日期:2017-01-16 出版日期:2017-08-08 发布日期:2017-08-11
通讯作者: 肖强,研究员,E-mail:xqtea@vip.163.com
作者简介:毛腾飞,硕士研究生,E-mail:maotf2014@163.com
基金资助:
国家科技支撑计划（2011BAD01B02）；中国农业科学院科技创新工程（CAAS-ASTIP-2016-TRICAAS）

The Expression of the Antibacterial Peptide Genes from Two Sibling Species of Tea Geometrid was Different in Resistance to EoNPV Infection

MAO Tengfei, FU Jianyu, SUN Liang, ZHOU Xiaogui, BAI Jiahe, XIAO Qiang

Institute of Tea Research, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China

Received:2017-01-16 Online:2017-08-08 Published:2017-08-11

摘要/Abstract

摘要： 小茶尺蠖Ectropis obliqua Prout和灰茶尺蛾Ectropis grisescens Warren是危害茶园的两近缘种害虫。用小茶尺蠖核型多角体病毒E.obliqua nucleopolyhedrovirus（EoNPV）喂饲两近缘种茶尺蠖，10 d后小茶尺蠖的死亡率为100%，而灰茶尺蛾的死亡率仅为35.5%。以感染病毒的两近缘种3龄幼虫的cDNA为模板，采用RT-PCR技术克隆获得了gloverin、attacin和moricin 3个抗菌肽基因，通过邻位连接法构建系统发育树聚类分析显示小茶尺蠖和灰茶尺蛾的Gloverin、Attacin和Moricin蛋白均分别归为一个分支；进一步采用qPCR检测发现，病毒处理后gloverin、attacin和moricin 3个基因在两近缘种茶尺蠖体内均表达上调，gloverin基因在12 h达到最大值，moricin基因在48 h达到最大值，且gloverin和moricin基因在灰茶尺蛾中表达量显著高于小茶尺蠖，而attacin基因在病毒侵入后的不同时期在两近缘种茶尺蠖体内表达量是变化的，推测gloverin、moricin和attacin基因可能与病毒的侵入有关。这一研究结果为进一步揭示抗菌肽基因在昆虫与病毒互作中的作用打下了基础。

关键词: 灰茶尺蛾, 小茶尺蠖, EoNPV, 抗菌肽

Abstract: Ectropis obliqua and Ectropis grisescens are two sibling species, which are severe defoliator pests in tea plantations. The mortality rate of E. obliqua and E. grisescens were 100% and 35.5% after infection with the E. obliqua nucleopolyhedrovirus (EoNPV) for 10 days, respectively. Three antimicrobial peptides genes, including gloverin, attacin and moricin, were cloned from the third-instar larvae of the two infected sibling species by an overlapping PCR method. Phylogenetic analysis suggested that Gloverin, Attacin, Moricin proteins of E. obliqua and E. grisescens were classified into an independent branch using a Neighbor-joining method, respectively. Further determination by qRT-PCR showed that the transcript levels of gloverin, attacin and moricin genes were up-regulated in the two sibling species after infection with EoNPV. The highest expression level of gloverin was detected at 12 h group, while moricin was expressed highest at 48 h group. The transcript levels of gloverin and moricin were significantly higher in E. grisescens than in E. obliqua. However, the transcript levels of attacin were changed at the different time of EoNPV infection between two sibling species, which indicates that gloverin, moricin and attacin are potentially associated with active resistance to virus infection. These results are of significance for further exploring the role of antimicrobial peptide genes in the interaction between insects and viruses.

Key words: Ectropis grisescens, Ectropis obliqua, EoNPV, antimicrobial peptide

中图分类号:

S435.711

毛腾飞, 付建玉, 孙亮, 周孝贵, 白家赫, 肖强. 两近缘种茶尺蠖的抗菌肽基因在抵抗EoNPV感染过程中的表达差异[J]. 中国生物防治学报, 2017, 33(4): 472-480.

MAO Tengfei, FU Jianyu, SUN Liang, ZHOU Xiaogui, BAI Jiahe, XIAO Qiang. The Expression of the Antibacterial Peptide Genes from Two Sibling Species of Tea Geometrid was Different in Resistance to EoNPV Infection[J]. journal1, 2017, 33(4): 472-480.

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两近缘种茶尺蠖的抗菌肽基因在抵抗EoNPV感染过程中的表达差异

The Expression of the Antibacterial Peptide Genes from Two Sibling Species of Tea Geometrid was Different in Resistance to EoNPV Infection

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