福建新记录入侵害虫木瓜秀粉蚧的分子检测鉴定  

A molecular detection and identification method for papaya mealybug (Paracoccus marginatus), the first recorded invasive pest in Fujian province of China

作　　者：林凌鸿 郑丽祯[2] 史梦竹[2] 李建宇[2] 王秋月 李銮 傅建炜[2,3] 吴梅香 LIN Linghong;ZHENG Lizhen;SHI Mengzhu;LI Jianyu;WANG Qiuyue;LI Luan;FU Jianwei;WU Meixiang(College of Plant Protection,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Institute of Plant Protection,Fujian Academy of Agricultural Science,Fuzhou 350013,Fujian,China;Institute of Quality Standards&Testing Technology for Agro-Products,Fujian Academy of Agricultural Science,Fuzhou 350013,Fujian,China;State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学植物保护学院,福州350002 [2]福建省农业科学院植物保护研究所,福州350013 [3]福建省农业科学院农业质量标准与检测技术研究所,福州350013 [4]福建农林大学省部共建闽台作物有害生物防控国家重点实验室,福州350002

出　　处：《果树学报》2019年第9期1130-1139,共10页Journal of Fruit Science

基　　金：福建省科技重大专项(2017NZ0003-1-1);国家重点研发计划(2016YFC1202101-4);福建省农业科学院科技创新团队建设项目(STIT2017-1-12);福建省星火计划项目(2017S0029)

摘　　要：【目的】基于核糖体28S rDNA基因序列的种特异性PCR方法,快速鉴定福建新纪录入侵性害虫——木瓜秀粉蚧(Paracoccus marginatus Williams and Granara de Willink)。【方法】通过1对扩增粉蚧28S rDNA的通用引物(S3660/A335)扩增以木瓜秀粉蚧为靶标,以野外采集的另外10种粉蚧为对照的11种粉蚧的基因片段序列。根据所得基因序列结果并参照GeneBank数据库中已知粉蚧的28S rDNA基因序列,设计出木瓜秀粉蚧28S rDNA的种特异性引物(28S-ParF/28S-MarR),对该种特异性引物的效果进行检验。【结果】该种特异性引物对木瓜秀粉蚧的28S rDNA基因具有扩增能力,得到产物大小为446 bp的扩增片段,对其他10种粉蚧均无扩增效果。该引物不仅对木瓜秀粉蚧雌成虫具有稳定的扩增效果,对不同虫态、不同寄主来源以及不同地区来源的木瓜秀粉蚧均有稳定的扩增效果。【结论】对福建发现的木瓜秀粉蚧进行分子鉴定并建立了木瓜秀粉蚧快速分子检测技术,可用于识别和防控木瓜秀粉蚧,有助于遏制木瓜秀粉蚧的入侵危害。【Objective】Paracoccus marginatus Williams and Granara de Willink is an invasive pest with strong diffusion and fecundity.It has caused serious damage to the papaya industry in Central America,Florida(USA),Guam(USA)and India.Pa.marginatus was first discovered in Fujian Province(Fuzhou and Zhangzhou)in 2017,showing great potential risks to papaya and other fruit crops,as well as flower industry in Fujian province.Because of the small body size and similar morphological characteristics,the morphological identification of mealybugs was inefficient.Rapid molecular identification of different species could be achieved through the use of DNA barcoding technology.Therefore,a technology for rapid molecular identification of Pa.marginatus was established based on the species-specific PCR method.【Methods】A species-specific PCR method based on ribosomal DNA-28S gene fragment(28S rDNA)was exploited to establish one technology for rapid detection and identification of Pa.marginatus.The additional 10 species of mealybugs(Phenacoccus solenopsis,Dysmicoccus boninsis,Nipaecoccus viridis,Phenacoccus solani,Pseudococcus comstocki,Pseudococcus cryptus,Planococcus lilacinus,Pseudococcus odermatti,Planococcus minor and Phenacoccus madeirensi)were collected in the fields as the contrast.In order to ensure the uniqueness of the source of DNA,the DNA templates were all extracted from one single female adult of these 11 species of mealybugs,respectively.28S rDNA of the 11 species was amplified by a pair of universal primers(S3660/A335).The obtained partial fragments of 28S rDNA were sequenced.And the phylogenetic tree was established by using a Neighbor-joining(NJ)method.According to the obtained 28S rDNA gene partial sequence of the 11 species and 28S rDNA gene sequences of Paracoccus galzerae in GeneBank database,the sequence alignment and analysis were performed on DNAMAN.28S rDNA species-specific primers(28S-ParF/28S-MarR)for Pa.marginatus were designed by selecting the sites with large differences in the sequence.And then,the

关 键 词：木瓜秀粉蚧 粉蚧 28S种特异引物 分子检测 快速鉴定 

分 类 号：S667.9[农业科学—果树学]