应用SS-PCR技术设计种特异性引物快速鉴定瓜实蝇  

Rapid Identification of Bactrocera cucurbitae(Coquillett)by Using Species-specific PCR Technique

作　　者：黄振 郭琼霞[3] 陈韶萍 HUANG Zhen;GUO Qiong-xia;CHEN Shao-ping(Fuzhou Changle Airport Customs,Fuzhou,Fujian 350209;College of Plant Protection,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002;Fuzhou Customs Technology Center,Fuzhou,Fujian 350001)

机构地区：[1]福州长乐机场海关,福建福州350209 [2]福建农林大学植物保护学院,福建福州350002 [3]福州海关技术中心,福建福州350001

出　　处：《安徽农业科学》2020年第6期83-86,96,共5页Journal of Anhui Agricultural Sciences

基　　金：福建省自然科学基金项目(2011J01066、2012J01061);福建出入境检验检疫局科技项目(FK2010-27、FK2011-56)。

摘　　要：[目的]为了促进果蔬进出口贸易和快速通关,防止瓜实蝇[Bactrocera cucurbitae(Coquillett)]进一步传入扩散,迫切需要开展适合实蝇快速鉴定技术的研究。[方法]应用SS-PCR技术,基于mtDNA COⅠ序列,设计了1对能够准确鉴定瓜实蝇的种特异性引物GF85和GR531。选用瓜实蝇作为阳性对照,南瓜实蝇[B.tau(Walker)]、具条实蝇[B.scutellata(Hendel)]等其他19种待测实蝇作为阴性对照,提取供试虫样基因组DNA,并进行PCR扩增和电泳检测。[结果]仅虫样瓜实蝇能在约447 bp位置扩增出一条清晰且单一的条带,其余的虫样未出现条带。[结论]将该试验建立的SS-PCR鉴定方法应用于实际进出境果蔬截获和疫情监测的虫样的鉴定,并得到验证,表明该方法具特异性、稳定性强,可在实际的实蝇鉴定工作中应用。[Objective]In order to promote the import and export trade of fruits and vegetables and rapid customs clearance,prevent further damage from Bactrocera cucurbitae(Coquillett),there is an urgent need to develop a rapid identification technology suitable for the fly.[Method]A pair of primers,GF85 and GR531,was designed and synthesized based on the mtDNA CO I sequence.B.cucurbitae(Coquillett)was chosen as the positive control and other 19 species of fruit flies including B.tau(Walker),B.scutellata(Hendel)and so on were used as the negative controls,for the PCR amplification and gel electrophoresis.[Result]Clear and single 447 bp band was amplified only in B.cucurbitae(Coquillett),but not in the other 19 fruit flies.[Conclusion]The SS-PCR identification method established in this experiment has been applied and verified in inspection and quarantine,indicating that this method has high species specificity and can be applied in infestation monitoring and quarantine surveillance of fruit flies in ports.

关 键 词：瓜实蝇 MTDNA COⅠ 种特异性引物 SS-PCR 快速鉴定 

分 类 号：S433[农业科学—农业昆虫与害虫防治]