Stewart Elwin L, Qu Xinshun, Overton Barrie E, Gildow Fred E, Wenner Nancy G, Grove Deborah S
Department of Plant Pathology, The Pennsylvania State University, University Park 16802.
Huck Institute, Wartik Laboratory, The Pennsylvania State University.
Plant Dis. 2007 Sep;91(9):1083-1088. doi: 10.1094/PDIS-91-9-1083.
Grapevines infected with Tomato ring spot virus (ToRSV) pose an economic risk for growers in the northeastern United States. This study describes a one-step real-time reverse-transcription polymerase chain reaction (RT-PCR) SYBR Green assay for detecting ToRSV in grapevines. Two newly designed primer pairs based on the ToRSV coat protein gene sequence were evaluated for specificity and optimized for a SYBR Green assay. The primer pair ToRSV1f/1r yielded a 130-bp product with strong primer-dimer products, whereas the primer pair ToRSV2f/2r yielded a 330-bp product with weak primer dimer products. Real-time RT-PCR detected ToRSV in more naturally infected grapevines maintained in the greenhouse than did enzyme-linked immunosorbent assay. The nucleotide sequences of the fragments amplified from grapevine growing in Pennsylvania using real-time PCR were divergent from previously published sequences.
感染番茄环斑病毒（ToRSV）的葡萄藤给美国东北部的种植者带来了经济风险。本研究描述了一种用于检测葡萄藤中ToRSV的一步法实时逆转录聚合酶链反应（RT-PCR）SYBR Green检测方法。基于ToRSV外壳蛋白基因序列新设计的两对引物，对其特异性进行了评估，并针对SYBR Green检测进行了优化。引物对ToRSV1f/1r产生了一个130 bp的产物，引物二聚体产物较强，而引物对ToRSV2f/2r产生了一个330 bp的产物，引物二聚体产物较弱。实时RT-PCR检测到温室中更多自然感染的葡萄藤中的ToRSV，比酶联免疫吸附测定法检测到的更多。使用实时PCR从宾夕法尼亚州种植的葡萄藤中扩增的片段的核苷酸序列与先前发表的序列不同。
