摘要
本研究以在检疫中经常被截获的5种豆象为研究对象,通过对GenBank中5种豆象mtDNA COⅠ序列的查询,并进行序列比对,共设计了30对引物,通过筛选获得5对能分别鉴定5种豆象的特异引物,并用50、25、10、5、1、0.5、0.1、0.05ng/μL 8个不同浓度系列的豆象DNA模板来检测灵敏度,结果表明,豌豆象和四纹豆象特异引物对的灵敏度为0.1ng/μL,蚕豆象、菜豆象和绿豆象特异引物对的灵敏度为0.5ng/μL。该方法可用于快速准确地鉴定5种豆象,对于豆象类害虫的检测监测具有重要意义。
Molecular identification of five bean weevils that were frequently intercepted from customs was studied in this paper.Based on the mtDNA COⅠ gene sequences of Callosobruchus chinensis,Bruchus rufimanus,B.pisorum,Acanthoscelides obtectus and C.maculatus from GenBank,30 pairs of primers were designed,and the five bean weevils can be specifically identified by PCR amplification with five pairs of specific primers.Moreover,the sensitivity of PCR was detected at 50,25,10,5,1,0.5,0.1 and 0.05 ng/μL genomic DNA of C.chinensis,B.rufimanus,B.pisorum,A.obtectus and C.maculatus,respectively.The results showed that the sensitivity for B.pisorum and C.maculatus was 0.1 ng/μL,while 0.5 ng/μL for the other three.The method provides a rapid and reliable tool for identification of the five bean weevils,which could be applied in detecting and monitoring of bean weevil pests.
作者
刘昌燕
李莉
焦春海
万正煌
Liu Changyan Li Li Jiao Chunhai Wan Zhenghuang(Institute of Food Crops, Hubei Academy of Agricultural Sciences, Hubei Key Laboratory of Food Crop Germplasm and Genetic Improvement, Wuhan 430064, Chin)
出处
《植物保护》
CAS
CSCD
北大核心
2017年第4期120-124,共5页
Plant Protection
基金
国家现代农业产业技术体系建设专项(CARS-09)
科技部国际合作重点项目(2011DFB31620)
湖北省农业科技创新中心资助项目(2016-620-000-001-006)
