A Real-Time PCR Assay for the Quantitative Detection of Ralstonia solanacearum in Horticultural Soil and Plant Tissues

被引：35

作者：

Chen Yun ^{[2
,3
,4
]}

Zhang, Wen-Zhi ^{[2
,3
,4
]}

Liu, Xin ^{[1
]}

Ma, Zhong-Hua ^{[1
]}

Li, Bo ^{[2
,3
,4
]}

Allen, Caitilyn ^{[5
]}

Guo, Jian-Hua ^{[2
,3
,4
]}

机构：

[1] Zhejiang Univ, Inst Biotechnol, Hangzhou 310029, Zhejiang, Peoples R China

[2] Nanjing Agr Univ, Coll Plant Protect, Dept Plant Pathol, Nanjing 210095, Peoples R China

[3] Engn Ctr Bioresource Pesticide Jiangsu Prov, Nanjing 210095, Peoples R China

[4] Minist Agr, Key Lab Monitoring & Management Crop Dis & Pest I, Nanjing 210095, Peoples R China

[5] Univ Wisconsin, Dept Plant Pathol, Madison, WI 53706 USA

来源：

JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY | 2010年 / 20卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Detection; quantification; Ralstonia solanacearum; real-time PCR; PHYLOGENETIC-RELATIONSHIPS; BURKHOLDERIA SOLANACEARUM; SENSITIVE DETECTION; SEQUENCE-ANALYSIS; STRAINS; AMPLIFICATION; DIVERSITY; SURVIVAL; MULTIPLEX; BIOVAR-2;

D O I：

10.4014/jmb.0906.06019

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of the UDP-3-O-acyl-GlcNAc deacetylase gene from R solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of 10(2) CFU/g tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with field samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.

引用

页码：193 / 201

页数：9

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