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利用定量PCR技术早期检测花生田孢子捕捉样品中的空气传播接种体

Early Detection of Airborne Inoculum of in Spore Trap Samples from Peanut Fields Using Quantitative PCR.

作者信息

Munir Misbakhul, Wang Hehe, Dufault Nicholas S, Anco Daniel J

机构信息

Department of Plant and Environment Science, Clemson University, Edisto Research and Education Center, Blackville, SC 29817, USA.

Department of Plant Pathology, University of Florida, Gainesville, FL 32611, USA.

出版信息

Plants (Basel). 2020 Oct 9;9(10):1327. doi: 10.3390/plants9101327.

Abstract

A quantitative PCR (qPCR)-assay was developed to detect airborne inoculum of , causal agent of late leaf spot (LLS) on peanut, collected with a modified impaction spore trap. The qPCR assay was able to consistently detect as few as 10 spores with purified DNA and 25 spores based on crude DNA extraction from rods. In 2019, two spore traps were placed in two peanut fields with a history of LLS. Sampling units were replaced every 2 to 4 days and tested with the developed qPCR assay, while plots were monitored for symptom development. The system detected inoculum 35 to 56 days before visual symptoms developed in the field, with detection related to environmental parameters affecting pathogen life-cycle and disease development. This study develops the framework of the qPCR spore trap system and represents the initial steps towards validation of the performance of the system for use as a decision support tool to complement integrated management of LLS.

摘要

开发了一种定量PCR(qPCR)检测方法,用于检测用改良撞击式孢子捕捉器收集的花生晚期叶斑病(LLS)病原体的空气传播接种体。该qPCR检测方法能够基于从杆中提取的粗DNA一致地检测到低至10个纯化DNA孢子和25个孢子。2019年,在两个有LLS病史的花生田中放置了两个孢子捕捉器。每2至4天更换一次采样单元,并用开发的qPCR检测方法进行检测,同时监测地块的症状发展情况。该系统在田间出现可见症状前35至56天检测到接种体,检测结果与影响病原体生命周期和病害发展的环境参数有关。本研究建立了qPCR孢子捕捉器系统的框架,并代表了验证该系统作为决策支持工具以补充LLS综合管理性能的初步步骤。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e2/7650562/baf7bd615729/plants-09-01327-g001.jpg

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