Development of a Real-Time RT-PCR SYBR Green Assay forTomato ring spot virusin Grape

Autor:	Barrie E. Overton, Deborah S. Grove, Elwin L. Stewart, Nancy G. Wenner, Xinshun Qu, F. E. Gildow
Rok vydání:	2007
Předmět:	Plant Science Coat protein Biology Virology Virus law.invention Economic risk Real-time polymerase chain reaction law Primer dimer Primer (molecular biology) Vitis vinifera Agronomy and Crop Science Polymerase chain reaction
Zdroj:	Plant Disease. 91:1083-1088
ISSN:	1943-7692 0191-2917 1083-1088
DOI:	10.1094/pdis-91-9-1083
Popis:	Stewart, E. L., Qu, X. S., Overton, B. E., Gildow, F. E., Wenner, N. G., and Grove, D. S. 2007. Development of a real-time RT-PCR SYBR Green assay for Tomato ring spot virus in grape. Plant Dis. 91:1083-1088. 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. Additional keywords: Grape yellow vein virus, Vitis vinifera
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2bc2e3bf494f5a87db19c28620f5dd7a https://doi.org/10.1094/pdis-91-9-1083 Zobrazit plný text záznamu