曹学仁　周益林

摘要 植物病原菌的菌源量是病害发生和流行的重要因子之一，对其精准的定量测定或检测可大大提高植物病害预测的准确性，本文对实时荧光定量PCR （qPCR）与数字PCR在植物病原菌定量检测、以及基于RNA水平的real-time PCR和基于核酸染料（EMA/PMA）与qPCR相结合的技术在植物病原菌活体定量检测中的应用进行了综述，并展望其在植物病害流行和预测中的应用前景。

关键词 植物病原菌; 定量检测; 实时定量PCR; 数字PCR; 活体检测

中图分类号： S 432

文献标识码： A

DOI： 10.16688/j.zwbh.2019463

Research progress in quantitative detection of plant pathogens using PCR technique

CAO Xueren1， ZHOU Yilin2*

（1. Key Laboratory of Integrated Pest Management on Tropical Crops， Ministry of Agriculture and Rural Affairs， Institute of

Environment and Plant Protection， Chinese Academy of Tropical Agricultural Sciences， Haikou 571101， China; 2. State

Key Laboratory for Biology of Plant Diseases and Insect Pests， Institute of Plant Protection，

Chinese Academy of Agricultural Sciences， Beijing 100193， China）

Abstract

The inoculum of plant pathogens is an important factor related with disease epidemics. Precise quantification of inoculum greatly help the prediction of diseases. This review summarized the applications of real-time quantitative PCR （qPCR） and digital PCR in the quantification of plant pathogens. The use of real-time reverse-transcriptase PCR and the combination of viability dyes and qPCR in viability detection of plant pathogens were also reviewed. The future perspectives of molecular quantitative detection of the pathogens in plant disease epidemics were discussed.

Key words

plant pathogen; quantitative detection; real-time quantitative PCR; digital PCR; viability detection

在植物病害流行過程中，病原菌的菌源量是一种重要的驱动因子，是病害预测的一个重要参数，如土壤中芸薹根肿菌Plasmodiophora brassicae的含量与病害发生程度显著相关，小麦条锈菌Puccinia striiformis f.sp. tritici和白粉菌Blumeria graminis f.sp. tritici的越冬菌量和第二年早春的病情也存在显著的相关性等[1-3]，因此准确获得病原菌的菌源量对于一些病害的预测和治理有十分重要的作用。

传统病原菌菌源量数据的获取主要依靠田间调查，但是一方面仅根据症状来判断病情容易出现误判，如苗期小麦条锈病与叶锈病容易混淆;另一方面对在寄主组织内潜伏侵染或未显症或隐症病原菌来说，由于此时尚未显症，无法准确估计病原菌的菌源量，从而影响对病害的预测。近年来，快速发展的分子生物学技术如实时荧光定量PCR、数字PCR等为植物病原菌的定量检测提供了新工具。

1 基于实时荧光定量PCR（qPCR）的植物病原菌定量检测

实时荧光定量PCR（real-time quantitative PCR，qPCR）技术通过在PCR反应体系中加入特定的……