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评估 DNA 检测平台与被动风力孢子捕捉器结合在加拿大对马铃薯和番茄晚疫病的早期监测中的应用。

Assessing the Use of DNA Detection Platforms Combined with Passive Wind-Powered Spore Traps for Early Surveillance of Potato and Tomato Late Blight in Canada.

机构信息

Sporometrics Inc., Toronto, Ontario M6K 3J1, Canada.

Department of Plant Agriculture, University of Guelph, Ridgetown, Ontario N0P 2C0, Canada.

出版信息

Plant Dis. 2021 Nov;105(11):3610-3622. doi: 10.1094/PDIS-12-20-2695-RE. Epub 2021 Nov 7.

Abstract

Quantitative PCR (qPCR), loop-mediated amplification (LAMP), and lateral flow strip-based recombinase polymerase amplification (RPA-LFS) assays were assessed for early detection of , the global causal agent of potato and tomato late blight, on passive wind-powered spore traps known as Spornados. Spore traps were deployed in potato and tomato fields during the 2018, 2019, and 2020 growing seasons in the provinces of Alberta, British Columbia, Manitoba, Prince Edward Island, and Ontario. All assays used DNA extracts from Spornado cassette membranes targeting the nuclear ribosomal internal transcribed spacer. A total of 1,003 Spornado samples were qPCR tested, yielding 115 positive samples for spores. In further assessment of these samples, LAMP detected in 108 (93.9%) of 115 qPCR positive samples, and RPA-LFS detected it in 103 (89.6%). None of the assays showed cross-reaction with other species or pathogenic fungi known to infect potato and tomato. The qPCR detected ≤1 fg of DNA, and LAMP and RPA-LFS amplified 10 fg in as little as 10 min. All assays detected before the first report of late blight symptoms in commercial potato or tomato fields within each region or province. The combination of Spornado passive samplers with qPCR, LAMP, or RPA-LFS proved a valuable spore trapping system for early surveillance of late blight in potato and tomato. Both LAMP and RPA-LFS showed potential as alternative approaches to qPCR for in-field monitoring of .

摘要

定量聚合酶链式反应(qPCR)、环介导扩增(LAMP)和基于侧向流条的重组酶聚合酶扩增(RPA-LFS)检测法被用于早期检测马铃薯和番茄晚疫病的全球病原体——致病疫霉,这些检测法是在称为 Spornados 的被动风力孢子捕捉器上进行的。2018 年、2019 年和 2020 年生长季节期间,在艾伯塔省、不列颠哥伦比亚省、马尼托巴省、爱德华王子岛省和安大略省的马铃薯和番茄田中部署了孢子捕捉器。所有的检测方法都使用来自 Spornado 盒式膜的 DNA 提取物,针对的是核核糖体内部转录间隔区。对总共 1003 个 Spornado 样本进行了 qPCR 检测,有 115 个样本对 孢子呈阳性。在对这些样本的进一步评估中,LAMP 在 115 个 qPCR 阳性样本中的 108 个(93.9%)中检测到了它,而 RPA-LFS 在 103 个(89.6%)中检测到了它。这些检测方法均未与其他已知感染马铃薯和番茄的 种或病原真菌发生交叉反应。qPCR 检测到的 DNA 量低至≤1 fg,而 LAMP 和 RPA-LFS 在 10 分钟内即可扩增 10 fg。所有的检测方法都在每个地区或省份的商业马铃薯或番茄田中首次报告晚疫病症状之前检测到了 。Spornado 被动采样器与 qPCR、LAMP 或 RPA-LFS 的结合证明是一种用于早期监测马铃薯和番茄晚疫病的有价值的孢子捕捉系统。LAMP 和 RPA-LFS 均显示出作为 qPCR 的替代方法,用于现场监测 的潜力。

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