Zhang Ning, McCarthy Meaghan L, Smart Christine D
Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Geneva, NY 14456.
Department of Biology, Hobart and William Smith Colleges, Geneva, NY 14456.
Plant Dis. 2008 Jun;92(6):953-960. doi: 10.1094/PDIS-92-6-0953.
There are numerous fungal and oomycete pathogens that cause severe damage to solanaceous crops. Rapid and accurate detection and identification of these pathogens is critical for plant disease management. Recently, DNA array technology has been successfully applied for simultaneous detection of multiple microorganisms from various habitats. The goal of this project was to develop a multiplex detection and identification system for major fungal and oomycete pathogens of solanaceous crops. To facilitate this goal, we used a membrane-based macroarray technology that included at least two specific oligonucleotides per pathogen. Based on the internal transcribed spacer sequences of the rRNA genes, 105 oligonucleotides (17 to 27 bases long) specific for 25 pathogens of solanaceous crops were designed and spotted on a nylon membrane. The array was tested against the 25 target pathogen species, 46 infected field samples, and a number of nontarget species. Our results indicate that the oligonucleotide-based macroarray detection system is a reliable and effective method for pathogen detection and identification even when multiple pathogens are present in a field sample.
有许多真菌和卵菌病原体对茄科作物造成严重损害。快速准确地检测和鉴定这些病原体对于植物病害管理至关重要。最近，DNA芯片技术已成功应用于同时检测来自各种生境的多种微生物。本项目的目标是开发一种用于茄科作物主要真菌和卵菌病原体的多重检测和鉴定系统。为实现这一目标，我们使用了基于膜的宏阵列技术，每个病原体至少包含两个特异性寡核苷酸。基于rRNA基因的内部转录间隔区序列，设计了105个针对25种茄科作物病原体的寡核苷酸（长度为17至27个碱基），并点样在尼龙膜上。该阵列针对25种目标病原体物种、46个感染的田间样本以及一些非目标物种进行了测试。我们的结果表明，基于寡核苷酸的宏阵列检测系统是一种可靠且有效的病原体检测和鉴定方法，即使田间样本中存在多种病原体。
