马铃薯瓢虫与茄二十八星瓢虫SCAR标记的建立及应用  

Establishment and application of SCAR marker for Henosepilachna vigintioctomaculata(Motschulsky)and Henosepilachna vigintioctopunctata(Fabricius)

作　　者：郭木娟 陈诗敏 庾家琪 叶翠怡 邱宝利 李文香[2] 潘慧鹏 GUO Mu-Juan;CHEN Shi-Min;YU Jia-Qi;LE Cui-Yi;QIU Bao-Li;LI Wen-Xiang;PAN Hui-Peng(Agricultural College of South China Agricultural University/Key Laboratory of Biopesticide Creation and Application of Guangdong Province/Guangdong Agricultural Pest Biocontrol Engineering Research Center,Guangzhou 510642,China;College of Agriculture and Forestry Science and Technology,Hebei North University,Zhangjiakou 075000,Hebei Province,China)

机构地区：[1]华南农业大学农学院/广东省生物农药创制与应用重点实验室/广东省农业害虫生物防治工程技术研究中心,广州510642 [2]河北北方学院农林科技学院,河北张家口075000

出　　处：《环境昆虫学报》2020年第2期434-440,共7页Journal of Environmental Entomology

基　　金：国家重点研发计划(2017YFD0200900);广州市科技计划项目(201804020070)。

摘　　要：为了探索快速鉴定马铃薯瓢虫Henosepilachna vigintioctomaculata(Motschulsky)和茄二十八星瓢虫Henosepilachna vigintioctopunctata(Fabricius)的分子生物学方法,本研究在随机扩增多态性DNA(random amplified polymorphic DNA,RAPD)的基础上,分别设计了可以鉴别两个物种的序列特征扩增区域(sequence characterized amplified regions,SCAR)标记。从随机合成的60条引物中筛选出来2条特异性引物(分别为OPI-6和OPJ-15),引物OPI-6在马铃薯瓢虫中扩增出约750 bp的特异性条带,引物OPJ-15在茄二十八星中扩增出约750 bp的特异性条带,根据测序结果设计了两对SCAR引物对筛选结果进行验证,发现根据OPI-6的测序结果所设计的SCAR引物(OPI-6 test)仅能在马铃薯瓢虫中扩增出645 bp的条带,而根据OPJ-15的测序结果所设计的SCAR引物(OPJ-15 test)仅能在茄二十八星瓢虫中扩增出436 bp的条带。这两对SCAR引物能够准确、稳定且快速地区分马铃薯瓢虫与茄二十八星瓢虫,对这两种害虫的精准防控具有重要意义。To explore a molecular method for the rapid identification of Henosepilachna vigintioctomaculata(Motschulsky)and Henosepilachna vigintioctopunctata(Fabricius),sequence characteristic amplified regions(SCAR)makers were designed respectively to identify two ladybird beetles based on random amplified polymorphic DNA(RAPD).Two specific primers(OPI-6 and OPJ-15)were screened out from 60 randomly synthesized RAPD primers.The primer OPI-6 only amplified approximately 750 bp amplicon in H.vigintioctomaculata,whereas the primer OPJ-15 only amplified approximately 750 bp amplicon in H.vigintioctopunctata.Two pairs of SCAR primers were designed based on the sequencing results to validate the screening results.It was found that the SCAR primers(OPI-6 test)based on OPI-6 could amplify 645 bp amplicon in H.vigintioctomaculata,whereas SCAR primers(OPJ-15 test)based on OPJ-15 could amplify 436 bp amplicon in H.vigintioctopunctata.These two pairs of SCAR primers can distinguish H.vigintioctomaculata and H.vigintioctopunctata accurately,steadily,and rapidly,which is of great significance for the accurate control of these two pests in the field.

关 键 词：马铃薯瓢虫 茄二十八星瓢虫 RAPD标记 SCAR标记 分子鉴定 

分 类 号：Q968.1[生物学—昆虫学] S433[农业科学—农业昆虫与害虫防治]