Sholberg Peter, O'Gorman Dan, Bedford Karen, Lévesque C André
Agriculture and Agri-Food Canada, PARC, Summerland BC.
Agriculture and Agri-Food Canada, ECORC, Ottawa ON.
Plant Dis. 2005 Nov;89(11):1143-1150. doi: 10.1094/PD-89-1143.
Short DNA gene sequences (oligonucleotides) from the ribosomal spacer regions of bacterial and fungal pathogens were used to identify and monitor economically important apple diseases. The oligonucleotides or probes were attached to a nylon membrane by an amine modified linker arm and arranged in a precise pattern to form an array for detecting five pathogens corresponding to five apple diseases. Initially the specificity of the probes was determined by hybridizing pure cultures of the pathogens to the probes. The DNA array correctly identified Botrytis cinerea, Penicillium expansum, Podosphaera leucotricha, Venturia inaequalis, and Erwinia amylovora and eliminated closely related species. When the array was used to monitor V. inaequalis ascospores collected from spore traps located in orchards, it confirmed the presence of ascospores as predicted by the disease forecasting model. Preliminary tests to quantify P. leucotricha populations using grayscale values was effective to 20 conidia per leaf disk. The DNA array is a promising new detection system for accurate identification of several pathogens in a single test with the potential for being a new tool for epidemiological studies.
利用细菌和真菌病原体核糖体间隔区的短DNA基因序列（寡核苷酸）来鉴定和监测具有经济重要性的苹果病害。寡核苷酸或探针通过胺修饰的连接臂连接到尼龙膜上，并以精确的模式排列形成一个阵列，用于检测与五种苹果病害相对应的五种病原体。最初，通过将病原体的纯培养物与探针杂交来确定探针的特异性。该DNA阵列正确地鉴定了灰葡萄孢、扩展青霉、白叉丝单囊壳、苹果黑星菌和梨火疫病菌，并排除了亲缘关系较近的物种。当该阵列用于监测从果园孢子捕捉器收集的苹果黑星菌子囊孢子时，它证实了病害预测模型所预测的子囊孢子的存在。使用灰度值对白叉丝单囊壳种群进行定量的初步试验在每片叶盘20个分生孢子时是有效的。该DNA阵列是一种很有前景的新型检测系统，能够在一次测试中准确鉴定多种病原体，有可能成为流行病学研究的新工具。
