BACKGROUNDGrapholitha molesta and Carposina sasakii are notorious pests that damage fruits, and are ranked among the most high‐priority control targets in orchards. Chronic mixed infestation of these two economically destructive species not only pose formidable challenges to integrated pest management efforts, but also have substantial impacts on global fruit trade. Therefore, a non‐destructive, rapid and efficient identification technology is urgently needed. In comparison with conventional detection methods, the environmental DNA (eDNA) approach does not require direct contact or capture of organisms and can monitor multiple species simultaneously. In this study, an innovative eDNA‐based methodology was developed and optimized for precise detection of G. molesta and Carposina sasakii, and was finally applied to assess their infestation levels across different orchards.RESULTSThe results demonstrated that eDNA with the highest concentration and quality was obtained through a filtration process combining a 10 μm mixed cellulose ester (MCE) membrane and a 0.45 μm cellulose acetate (CA) membrane, after soaking the victim pulp for 15 min. The primers developed for these two species of fruit moths exhibited high sensitivity, with minimum detection limits of 0.1 and 0.01 ng/ul, respectively.CONCLUSIONThe eDNA‐based technology successfully detected the target moth species in orchards, regardless of whether the fruit was damaged by one or more moths. This study provided a critical theoretical foundation for the development of non‐destructive fruit pest testing technology, and also offered valuable references for eDNA research on other species, which will be vital to the field monitoring of fruit‐boring moths. © 2025 Society of Chemical Industry.

中文翻译：

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基于环境 DNA 的 Grapholitha molesta 和 Carposina sasakii 检测及其田间应用

背景 Grapholitha molesta 和 Carposina sasakii 是臭名昭著的损害水果的害虫，是果园中优先级最高的控制目标之一。这两种具有经济破坏性的物种的长期混合侵扰不仅对有害生物的综合治理工作构成了巨大挑战，而且对全球水果贸易也产生了重大影响。因此，迫切需要一种无损、快速、高效的识别技术。与传统检测方法相比，环境 DNA （eDNA） 方法不需要直接接触或捕获生物体，并且可以同时监测多个物种。在这项研究中，开发了一种基于 eDNA 的创新方法并进行了优化，用于精确检测 G. molesta 和 Carposina sasakii，并最终应用于评估它们在不同果园中的侵染水平。结果结果表明，在浸泡受害果肉 15 分钟后，通过 10 μm 混合纤维素酯 （MCE） 膜和 0.45 μm 醋酸纤维素 （CA） 膜的组合过滤过程获得最高浓度和质量的 eDNA。为这两种果蛾开发的引物表现出高灵敏度，最小检出限分别为 0.1 ng/μL 和 0.01 ng/ul。结论基于 eDNA 的技术成功检测了果园中的目标飞蛾物种，无论果实是否被一种或多种飞蛾破坏。本研究为无损果害虫检测技术的发展提供了重要的理论基础，也为其他物种的 eDNA 研究提供了有价值的参考，这对蛀果蛾的野外监测至关重要。© 2025 化工学会.