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用便携式实时 PCR 系统进行土壤载体 Phytopathogens 的现场分子检测
On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System
JoVE 杂志
环境科学
Author Produced
This content is Free Access.
JoVE 杂志 环境科学
On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System

用便携式实时 PCR 系统进行土壤载体 Phytopathogens 的现场分子检测

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16,616 Views

14:15 min

February 23, 2018

DOI:

14:15 min
February 23, 2018

16588 Views

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Hi my name’s Joseph DeShields, I’m with the Department of Plant Pathology at Washington State University and today I’ll be showing you an onsite detection protocol for detecting soil-borne diseases in soil.Onsite diagnosis of plant diseases in the field is pivotal for effective disease management and can ultimately reduce secondary spread of pathogens in the field.Soil-borne pathogens are particularly difficult to diagnose due to the large and complex soil environments and this can make understanding soil-borne diseases difficult.Polymerase chain reaction, otherwise known as PCR, is currently the most widely-accepted sensitive and accurate approach to detecting plant pathogens in the field.The following protocol demonstrates a sensitive and onsite method for detecting soil-borne pathogens comprised of a magnetic bead-based DNA extraction, PCR, which can be done using a portable PCR instrument, and data analysis, which can be done remotely, using a laptop computer.Magnetic bead-based DNA extraction.The DNA and RNA extraction kit was prepared and used per the manufacturer’s instructions with slight modifications.All reagents were stored at room temperature and the prepared Proteinase K was stored at a 20

Summary

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快速准确地检测植物病原体, 特别是土壤传播病原体, 对于防止田间植物病害进一步接种生产和增殖至关重要。采用便携式实时 PCR 检测系统开发的方法, 能够在野外条件下进行现场诊断。

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