研究旨在准确、高效鉴定识别叶蝉科昆虫,克服传统分类方法的不足,为有效防治提供依据。以新疆4种叶蝉若虫和2种叶蝉成虫为试材,基于mt DNA COI基因,通过PCR扩增、测序技术获得6种叶蝉的基因序列。结果表明:6个样本的COI序列A+T含量高于G...研究旨在准确、高效鉴定识别叶蝉科昆虫,克服传统分类方法的不足,为有效防治提供依据。以新疆4种叶蝉若虫和2种叶蝉成虫为试材,基于mt DNA COI基因,通过PCR扩增、测序技术获得6种叶蝉的基因序列。结果表明:6个样本的COI序列A+T含量高于G+C,A/T碱基偏移明显,符合昆虫mt DNA碱基构成特点;6种叶蝉种间平均遗传距离为0.232,样本A、B、E种内平均遗传距离为0.0031,符合“10倍阈值定律”;样本A、B、E与数据库中相关叶蝉的序列相似度高于99%,与形态分类结果一致;6种叶蝉在系统发育树上位置清晰,分类明确,在各分类阶元间形成独立单系。获得6条COI基因序列可作为叶蝉快速鉴定的DNA条形码,其中4种若虫的基因序列为叶蝉科昆虫首次报道。展开更多
[Objective]The paper was to establish a rapid identification method of Bactrocera cilifera(Hendel)with species-specific primers(SS-COI).[Method]Using B.cilifera(Hendel)as the positive control,and 19 species of fruit f...[Objective]The paper was to establish a rapid identification method of Bactrocera cilifera(Hendel)with species-specific primers(SS-COI).[Method]Using B.cilifera(Hendel)as the positive control,and 19 species of fruit flies such as B.diaphora(Coquillett)and B.dorsalis(Hendel)as the negative controls,a pair of species-specific primers,YF290 and YR511,were designed and screened for accurate identification of B.cilifera,based on mitochondrial DNA COI sequence.[Result]The PCR products were amplified and detected by electrophoresis.Only a clear and single band was observed at about 222 bp in the positive control,while no bands were found in the other negative controls.[Conclusion]The established rapid identification method with species-specific primers(SS-COI)is of great practical significance for rapid identification of fruit flies intercepted from import and export fruits and vegetables at ports,and for rapid clearance and early warning of import fruits and vegetables at ports.展开更多
【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...【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。展开更多
文摘研究旨在准确、高效鉴定识别叶蝉科昆虫,克服传统分类方法的不足,为有效防治提供依据。以新疆4种叶蝉若虫和2种叶蝉成虫为试材,基于mt DNA COI基因,通过PCR扩增、测序技术获得6种叶蝉的基因序列。结果表明:6个样本的COI序列A+T含量高于G+C,A/T碱基偏移明显,符合昆虫mt DNA碱基构成特点;6种叶蝉种间平均遗传距离为0.232,样本A、B、E种内平均遗传距离为0.0031,符合“10倍阈值定律”;样本A、B、E与数据库中相关叶蝉的序列相似度高于99%,与形态分类结果一致;6种叶蝉在系统发育树上位置清晰,分类明确,在各分类阶元间形成独立单系。获得6条COI基因序列可作为叶蝉快速鉴定的DNA条形码,其中4种若虫的基因序列为叶蝉科昆虫首次报道。
基金Supported by Natural Science Foundation of Fujian Province (2011J01066, 2012JO1061)。
文摘[Objective]The paper was to establish a rapid identification method of Bactrocera cilifera(Hendel)with species-specific primers(SS-COI).[Method]Using B.cilifera(Hendel)as the positive control,and 19 species of fruit flies such as B.diaphora(Coquillett)and B.dorsalis(Hendel)as the negative controls,a pair of species-specific primers,YF290 and YR511,were designed and screened for accurate identification of B.cilifera,based on mitochondrial DNA COI sequence.[Result]The PCR products were amplified and detected by electrophoresis.Only a clear and single band was observed at about 222 bp in the positive control,while no bands were found in the other negative controls.[Conclusion]The established rapid identification method with species-specific primers(SS-COI)is of great practical significance for rapid identification of fruit flies intercepted from import and export fruits and vegetables at ports,and for rapid clearance and early warning of import fruits and vegetables at ports.
文摘【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。
