Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.
J Microbiol Biotechnol. 2010 Jan;20(1):193-201.
A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in the horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of 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 102 CFU/g tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with filed 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.
本研究开发了一种用于检测园艺土壤和植物组织中青枯雷尔氏菌的特异性和快速实时 PCR 检测方法。根据青枯雷尔氏菌 UDP-3-O-酰基-GlcNAc 去乙酰化酶基因上游区设计了特异性引物 RSF/RSR，可特异性扩增来自 28 株青枯雷尔氏菌的 159bpPCR 产物，这些菌株代表了所有遗传多样性的 AluI 型和所有 6 个生物型，但不能从任何其他非目标物种中扩增出来。在该实时 PCR 检测中，可检测到 102CFU/g 番茄茎和园艺土壤中的下限。田间样本和人工感染样本的高灵敏度和特异性表明，该方法可能是一种用于准确预测和诊断青枯雷尔氏菌检测和定量的有用工具。
