华南农业大学学报2022，Vol.43Issue(1)：59-66,8.DOI:10.7671/j.issn.1001-411X.202101008
基于种特异性线粒体细胞色素氧化酶I引物快速鉴定茄二十八星瓢虫和马铃薯瓢虫
Rapid identification of Henosepilachna vigintioctopunctata and Henosepilachna vigintioctomaculata based on species-specific mitochondrial cytochrome oxidase I primers
摘要
Abstract
[Objective]Henosepilachna vigintioctopunctata (Fabricius) and Henosepilachna vigintiocto-maculata (Motschulsky), two kinds of phytophagous ladybeetles in China, are destructive pests causing great damage to solanaceous plants. They are difficult to distinguish based on external morphological characteristics, and it is therefore necessary to develop a rapid and accurate method to differentiate them.[Method]We established a molecular identification technique using species-specific (SS) PCR primers based on the species-specificity of mitochondrial cytochrome oxidase I (mtCOI) of two ladybeetles. Two pairs of SS-mtCOI primers, Hvp and Hvm, were designed based on sequence variations in the mtCOI gene between H. vigintioctopunctata and H. vigintioctomaculata.[Result]PCR amplifications were conducted using these two primers and both had species-specific amplifications. Sensitivity assays were conducted under different DNA concentrations, and the results showed that Hvp primers for H. vigintioctopunctata had a detectable amplification band at a DNA concentration of 3.13 mg/L, while Hvm primers for H. vigintioctomaculata had a detectable amplification band at a DNA concentration of 2.43 mg/L. The egg or 1st instar of H. vigintioctopunctata and H. vigintioctoma-culata could also be accurately differentiated by Hvp and Hvm primers. Furthermore, six field populations of H. vigintioctopunctata collected from six provinces could be authenticated by the Hvp primers.[Conclusion]These two pairs of SS-mtCOI primers can differentiate H. vigintioctopunctata and H. vigintioctomaculata rapidly, accurately and sensitively.关键词
茄二十八星瓢虫/马铃薯瓢虫/SS-mtCOI引物/分子鉴定/敏感性检测Key words
Henosepilachna vigintioctopunctata/Henosepilachna vigintioctomaculata/Species-specific mtCOI primer/Molecular identification/Sensitivity detection引用本文复制引用
郭木娟,ZHOU Xuguo,潘慧鹏,林妙金,潘广,庾家琪,吕晶,郭威,张亚锋,杨春晓,邱宝利..基于种特异性线粒体细胞色素氧化酶I引物快速鉴定茄二十八星瓢虫和马铃薯瓢虫[J].华南农业大学学报,2022,43(1):59-66,8.基金项目
国家重点研发计划(2017YFD0200900) （2017YFD0200900）
国家自然科学基金(31972269) （31972269）
