"PathoScout” - portable DNA analysis for Precision Agriculture

在疾病流行可能大大降低产品适销性和产量的情况下，种植高价值园艺作物和大面积大田作物的农民面临着控制真菌和卵菌病原体的挑战。及早发现导致重要疾病的病原体将使作物保护产品仅在需要时才能使用，这将最大化控制效果并减少不必要的使用，从而降低成本和降低生产的碳足迹。该项目将使用Rothamsted的培养收集和存档的孢子样本，与项目合作伙伴一起，研究为特定病原体创建诊断灯分析试剂盒的可行性，开发和测试从孢子中释放DNA的通用方法，包括捕获在轮状孢子陷阱的采样棒上的孢子，测试纯化DNA的测试方法，并测试一个工程流体系统，以自动化处理样本以释放DNA，纯化它并执行灯测试。我们将重点关注保护和户外作物的重要病原体：灰霉病菌(许多园艺作物的灰霉病)、致病疫霉(马铃薯疫病)、白粉病(葡萄白粉病)和条锈菌(小麦黄锈病)。稳定的LAMP检测试剂将由Polygenyn有限公司生产，并由Rothamsted进行测试。该诊断测试本身将被设计用于一种轻型、便携式、电池供电的设备，该设备将由Agi-Samplers Ltd生产，使用Microbritt有限公司开发的流体盒。该设备将与对研磨的叶片或水果样本以及使用孢子捕捉器捕获空气中的空气颗粒物样本进行的测试兼容。总体目标是探索创造一种便携式、用户友好和低成本的诊断工具来支持作物保护的可行性。基于LAMP的新型真菌孢子检测技术，用户可以使用捕获的空气传播孢子或叶或水果样本进行当天测试。成功的可行性成果将提供一个自动化DNA分析平台，其应用范围超出农业领域。它将改变作物保护做法，并提高英国农业对气候变化导致的更零星疾病的适应能力。

Farmers of high value horticultural crops and large acre field crops face challenges to control fungal and oomycete pathogens in conditions where disease epidemics can substantially reduce marketability of produce and yield. Early detection of pathogens that cause important diseases will allow crop protection products to be applied only when needed, which will maximise control efficacy and reduce unnecessary applications thereby reducing costs and lowering the carbon footprint of production.This project will use Rothamsted's culture collection and archived spore samples, along with project partners, to research the feasibility of creating kits of diagnostic LAMP assays for specific pathogens, develop and test generic methods to release DNA from spores, including spores trapped onto sampling rods of rotorod spore traps, test methods to purify the DNA and test an engineered fluidic system to automate the handling of samples to release DNA, purify it and perform a LAMP assay. We will focus on important pathogens of protected and outdoor crops; Botrytis cinerea (grey mould of many horticultural crops), Phytophthora infestans (potato blight), Erysiphe necator (grape powdery mildew) and Puccinia striiformis (wheat yellow rust).Stabilised LAMP assay reagents will be produced by Polygenyn Ltd and tested by Rothamsted. The diagnostic test itself will be designed for use in a lightweight, portable, battery-powered device, which will be produced by Agri-Samplers Ltd using a fluidic cassette developed by Microbritt Ltd. The device will be compatible with tests performed on ground-up leaf or fruit samples and also samples of airborne particulates captured in air using a spore trap.The overall aim is to explore the feasibility of creating a portable user-friendly and low-cost diagnostics tool to support crop protection, based on novel fungal spore detection by LAMP technology for same-day testing by the user with samples of either captured airborne spores or leaf or fruit samples.A successful feasibility outcome will deliver an automated DNA analysis platform with broad applications beyond agriculture. It will transform crop protection practices and improve the resilience of UK agriculture to diseases made more sporadic by climate change.