摘要
【目的】快速鉴定我国进境植物检疫性害虫南瓜实蝇Bactrocera tau(Walker),防止该虫传入扩散。【方法】基于物种特异性PCR(SS-PCR)技术,选取20种形态近似的常见实蝇,将南瓜实蝇作为阳性对照,提取基因组DNA模板,选用mt DNA COⅠ序列,检查同源系列,设计筛选1对可快速准确鉴定南瓜实蝇的种特异性引物NF404和NR610,并进行PCR检测和验证,建立一种利用种特异性引物的快速鉴定方法。【结果】南瓜实蝇在207 bp的位置扩增出一条清晰且单一的目标条带,其余果实蝇未见条带。将截获的南瓜实蝇与其同亚属的近缘种具条实蝇、瓜实蝇的不同虫态和成虫部分虫体组织的虫样,采用本实验室建立的SS-PCR鉴定方法进行验证,结果一致。【结论】建立的南瓜实蝇的SS-PCR鉴定方法种特异性和稳定性强,能快速鉴定目标种类,解决口岸进境果蔬中截获实蝇幼虫、蛹等不同虫态和残体难于快速鉴定问题。
【Objective】A rapid method to detect Bactrocera tau(Walker)was established for inspection on produce to prevent potential introduction and spread of the invasive pest.【Method】Based on the species-specific PCR(SS-PCR)technology,B.tau as control and 20 common fruit flies of similar morphology were used to extract the genomic DNA templates,and the homology of the specimens checked with the COI mt DNA sequence.A rapid and accurate identification method applying the B.tau-specific primers NF404 and NR610 was designed and verified by PCR amplification and electrophoresis analysis.【Result】The newly developed SS-PCR detection method clearly amplified a single 207 bp band only on the target B.tau,not on any other fruit flies.Comparisons on the test results on the insects or body parts of all fruit flies with homogenous band sequence to B.tau validated the new methodology.【Conclusion】The newly established SS-PCR method exhibited high repeatability,specificity,and accuracy in detecting B.tau.It was considered applicable for the entry/exit and quarantine inspection on the invasive insect,larvae or dead body parts found on vegetables and fruits.
作者
黄振
侯有明
郭琼霞
陈韶萍
HUANG Zhen;HOU Youming;GUO Qiongxia;CHEN Shaoping(College of Plant Protection,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;Fuzhou Changle Airport Customs,Fuzhou,Fujian 350209,China;Fuzhou Customs Technology Center,Fuzhou,Fujian 350001,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2021年第2期215-220,共6页
Fujian Journal of Agricultural Sciences
基金
福建省自然科学基金项目(2011J01066、2012J01061)。
作者简介
黄振(1985−),男,硕士,高级农艺师,研究方向:农业昆虫与害虫防治(E-mail:HZ15880010585@126.com);通信作者:侯有明(1966−),男,博士,教授,研究方向:农业昆虫与害虫防治(E-mail:ymhou@fjau.edu.cn);通信作者:郭琼霞(1954−),女,研究员,研究方向:植物检疫(E-mail:gqxfjciq@126.com)。
