鹰嘴豆象与四纹豆象的分子检测技术  

Technique of molecular detection of Callosobruchus analis and Callosobruchus maculatus(Coleoptera:Bruchidae)

作　　者：顾杰[1,2] 王莉萍[1] 毛雅琴[1] 顾忠盈[2] 朱光耀[2] 杜予州[1] 

机构地区：[1]扬州大学应用昆虫研究所,扬州225009 [2]江苏出入境检验检疫局,南京210001

出　　处：《植物保护学报》2010年第3期211-216,共6页Journal of Plant Protection

基　　金：江苏出入境检验检疫局科技项目(2006KJ36);江苏省科技攻关项目(BE2005348)

摘　　要：结合聚合酶链反应(polymerase chain reaction,PCR)和酶切方法,对鹰嘴豆象Callosobruchus analis(Fabricius)和四纹豆象Callosobruchus maculatus(Fabricius)的分子检测技术进行了研究。根据两种豆象m tDNACOⅠ基因序列的比对结果设计了2对特异性PCR引物,并筛选到2个特异性内切酶SnaⅠ和MboⅠ,通过PCR反应(最适退火温度为55℃)和PCR产物的酶切反应检测2个近缘种。可靠性检测表明,2对引物针对不同地理种群和个体以及不同DNA浓度的鹰嘴豆象和四纹豆象均能扩增出相应的目的条带。限制性内切酶验证结果表明,PCR技术具有较高的准确性和灵敏度,弥补了形态鉴定的不足。The technique of molecular detection of Callosobruchus analis （Fabricius） and CaUosobruchus maculatus （Fabricius） was studied using the methods of polymerase chain reaction and enzyme digestion. According to the alignments of mtDNA CO I gene sequences of bean weevils, two specific primers were designed that the optimal annealing temperature for PCR was 55 ℃ and two restriction enzymes were screened out for molecular detection. The results showed that different populations, individuals and consistence of C. analis and C. mactdatus could obtain corresponding amplificatory bands by the specific primers and the results were testified by the enzyme digestion experiment. The PCR detection and enzyme digestion with high sensitivity and accuracy could make＇ up for a lack of morphology.

关 键 词：鹰嘴豆象 四纹豆象 PCR 酶切 检测 

分 类 号：S433[农业科学—农业昆虫与害虫防治]