利用mtCOI PCR-RFLP技术鉴别草地贪夜蛾与其他3种形态相近昆虫  

Rapid differentiation of Spodoptera furgiperda from the other three morphologically similar species using mtCOI PCR-RFLP technique

作　　者：孔子艺 王鹏 朱军生 国栋 迟秀丽 孔宝玉 胡磊 褚栋 KONG Ziyi;WANG Peng;ZHU Junsheng;GUO Dong;CHI Xiuli;KONG Baoyu;HU Lei;CHU Dong(Key Laboratory of Integrated Control of Plant Diseases and Insect Pests of Shandong Province,College of Plant Health&Medicine of Qingdao Agricultural University,Qingdao,Shandong 266109,China;The Vegetational Protection Central Station of Shandong Province,Jinan,Shandong 250100,China;The Vegetational Protection Station of Jiaozhou Agricultural Bureau,Jiaozhou,Shandong 266300,China)

机构地区：[1]山东省植物病虫害综合防控重点实验室,青岛农业大学植物医学学院,山东青岛266109 [2]山东省植物保护总站,山东济南250100 [3]胶州市农业局植保站,山东胶州266300

出　　处：《生物安全学报》2020年第1期16-22,共7页Journal of biosafety

基　　金：山东省现代农业产业技术体系水稻创新团队项目(SDAIT-17-07);山东省重点研发计划项目(2017GNC11111)。

摘　　要：【目的】田间调查发现草地贪夜蛾与甜菜夜蛾、斜纹夜蛾、粘虫常混合发生,传统的形态学鉴定方法不能快速鉴别出该虫,当前亟需快速鉴别该虫的方法。【方法】本研究分析了草地贪夜蛾与甜菜夜蛾、斜纹夜蛾、粘虫mtCOI基因序列的酶切位点,根据目的片段设计上游引物并进行PCR-RFLP验证。【结果】草地贪夜蛾个体在mtCOI片段的556~561 bp处均存在Sbf I内切酶酶切位点,斜纹夜蛾、甜菜夜蛾、粘虫均无Sbf I酶切位点。草地贪夜蛾PCR产物经过Sbf I内切酶酶切,可出现420 bp左右的特征带,斜纹夜蛾、甜菜夜蛾、粘虫种群均不能被Sbf I内切酶酶切。【结论】基于新设计引物扩增的mtCOI片段的PCR-RFLP方法可有效鉴别草地贪夜蛾与其他3个形态相近昆虫,研究结果为草地贪夜蛾的快速鉴别提供了方法。【Aim】Recent research has found that Spodoptera frugiperda,Spodoptera exigua,Spodoptera litura,and Mythimna separata often occur simultaneously in fields.Because of similar morphological characteristics of adults and larvae,it is difficult to differentiate S.frugiperda from the other three species using traditional morphological identification methods.It is necessary to develop a rapid method to differentiate S.frugiperda from the other morphologically similar species.【Method】This study analyzed the restriction enzyme recognition sites in the mtCOI(mitochondrial cytochrome oxidase subunit I)of S.frugiperda,S.exigua,S.litura,and M.separata.A new forward primer of the target fragment of mtCOI gene was designed,which was used to amplify the targeted production.【Result】In the 556-561 bp of mtCOI segment,Sbf I enzyme recognition site was found in all individuals of S.frugiperda,while there was no Sbf I restriction enzyme recognition sites in S.liitura,S.exigua,and M.separata.The results showed that the PCR products of S.frugiperda could be digested by Sbf I,and a characteristic band of 420 bp was found.In contrast,the PCR products of S.litura,S.exigua,and M.separata could not be digested by Sbf I.【Conclusion】The mtCOI PCR-RFLP technique developed in this study can differentiate S.frugiperda from the other three species effectively,which provides a rapid method for the identification of S.frugiperda.

关 键 词：草地贪夜蛾 农业入侵害虫 MTCOI PCR-RFLP 快速鉴别 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]