【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。更多还原

【Abstract】 The technique of molecular detection of Callosobruchus analis(Fabricius) and Callosobruchus maculatus(Fabricius) was studied using the methods of polymerase chain reaction and enzyme digestion.According to the alignments of mtDNA COⅠ 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.maculatus 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.更多还原