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

被引:34
|
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
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
收藏
页码:193 / 201
页数:9
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