首页> 外文期刊>Journal of Applied Entomology >A rapid diagnostic tool for two species of Tetradacus (Diptera: Tephritidae: Bactrocera) based on species-specific PCR. (Special Issue: Biology, ecology, behavior, host status and management of invasive and native fruit flies of Europe, Africa and the Middle East.)
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A rapid diagnostic tool for two species of Tetradacus (Diptera: Tephritidae: Bactrocera) based on species-specific PCR. (Special Issue: Biology, ecology, behavior, host status and management of invasive and native fruit flies of Europe, Africa and the Middle East.)

机译:基于物种特异性PCR的两种四足纲(Diptera:Tephritidae:Bactrocera)的快速诊断工具。 (特刊:欧洲,非洲和中东的入侵性和本土果蝇的生物学,生态学,行为,寄主状况和管理。)

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摘要

Bactrocera minax and Bactrocera tsuneonis are two sibling species of the Bactrocera subgenus Tetradacus with significant quarantine importance in Asia. It is difficult to identify or distinguish them only based on morphological characteristics. In this study, microscopic observations showed that morphological features of adult samples were congruent with the diagnosis of the two species. In this study, we describe species-specific PCR for the molecular identification of B. minax and B. tsuneonis. Species-specific primer pairs were designed on the basis of variations in the mtDNA COI barcode sequences among Bactrocera spp. A 499 bp-specific fragment of B. minax and a 337 bp absolute product of B. tsuneonis were generated by their respective primer pairs, whilst no such diagnostic bands were present in any of the other fruit fly species tested. The results of the sensitivity test demonstrated that the detection limit of DNA template concentration was 1 ng/ micro l for B. minax and 0.1 ng/ micro l for B. tsuneonis. This assay exhibited high specificity, reliability, rapid and low cost for all ontogenetic stages. Our data show that species-specific PCR is a powerful tool that can be used for the identification of B. minax and B. tsuneonis in quarantine work.
机译:Bactrocera minax和Bactrocera tsuneonis是Bactrocera亚属Tetradacus的两个兄弟种,在亚洲具有重要的检疫重要性。仅基于形态特征很难识别或区分它们。在这项研究中,显微镜观察表明,成年样品的形态特征与这两种物种的诊断是一致的。在这项研究中,我们描述了物种特异性PCR用于B. minax和B. tsuneonis的分子鉴定。根据Bactrocera spp的mtDNA COI条码序列的差异设计物种特异性引物对。它们各自的引物对产生了499 bp的B. minax特异片段和337 bp的B. tsuneonis绝对产物,而在所测试的任何其他果蝇中均未出现这样的诊断带。敏感性测试的结果表明,DNA模板浓度的检出限对于米氏芽孢杆菌为1 ng /μl,对于通花芽孢杆菌为0.1 ng /μl。该测定法对所有个体发育阶段均显示出高特异性,可靠性,快速和低成本。我们的数据表明,物种特异性PCR是一种强大的工具,可用于在检疫工作中鉴定B. Minax和B. tsuneonis。

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