Department of Plant Biosecurity, College of Plant Protection, College of Plant Protection, China Agricultural University, Beijing 100193, China.
National Agro-Tech Extension and Service Center, Beijing 100125, China.
J Econ Entomol. 2023 Jun 13;116(3):956-962. doi: 10.1093/jee/toad014.
Bactrocera tsuneonis and Bactrocera minax are the most destructive pests that damage citrus in China. These key pests hinder the citrus trade, cause significant financial losses, drastically lower citrus production and quality, and decrease farmer enthusiasm for citrus planting. Bactrocera minax and B. tsuneonis are very similar in all life stages. There are limited morphological characteristics to differentiate the adult species, and it is nearly impossible to differentiate these two species in the egg and larval stages. Loop-mediated isothermal amplification (LAMP) is a rapid and robust diagnostic tool used to identify these two species accurately. We designed two sets of primers to distinguish B. minax and B. tsuneonis using DNA barcoding region of the COI gene. Only 50 min was needed under a constant temperature of 65ºC to determine the species of the two flies. The reaction system has high specificity and sensitivity, in which these two species can be accurately distinguished between different geographical populations and 1.0 ng/μL was the lowest DNA concentration that could be detected. Our primers can quickly identify these key pests without knowing their morphology, which could facilitate plant protection workers at the primary level to solve problems in plant quarantine.
实蝇科中的桔小实蝇和瓜实蝇是中国危害柑橘的最具破坏性的害虫。这些关键害虫阻碍了柑橘贸易，造成了巨大的经济损失，大幅降低了柑橘的产量和质量，降低了农民种植柑橘的积极性。桔小实蝇和瓜实蝇在所有生命阶段都非常相似。成虫的形态特征有限，几乎不可能在卵和幼虫阶段区分这两个物种。环介导等温扩增（LAMP）是一种快速而强大的诊断工具，可用于准确识别这两种物种。我们使用 COI 基因的 DNA 条形码区域设计了两套引物来区分瓜实蝇和桔小实蝇。在 65°C 的恒温下，仅需 50 分钟即可确定两种蝇的物种。该反应体系具有高度的特异性和灵敏度，可在不同地理种群之间准确区分这两个物种，检测的最低 DNA 浓度为 1.0ng/μL。我们的引物可以在不知道形态的情况下快速识别这些关键害虫，这有助于基层植保工作者解决植物检疫中的问题。
