Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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A Real-Time PCR Assay for the Quantitative Detection of Ralstonia solanacearum in Horticultural Soil and Plant Tissues

  • Chen, Yun (Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Engineering Center of Bioresource Pesticide in Jiangsu Province, and Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture) ;
  • Zhang, Wen-Zhi (Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Engineering Center of Bioresource Pesticide in Jiangsu Province, and Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture) ;
  • Liu, Xin (Institute of Biotechnology, Zhejiang University) ;
  • Ma, Zhong-Hua (Institute of Biotechnology, Zhejiang University) ;
  • Li, Bo (Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Engineering Center of Bioresource Pesticide in Jiangsu Province, and Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture) ;
  • Allen, Caitilyn (Department of Plant Pathology, University of Wisconsin) ;
  • Guo, Jian-Hua (Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Engineering Center of Bioresource Pesticide in Jiangsu Province, and Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture)
  • Published : 2010.01.31
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Abstract

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.

Keywords

References

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