Development of a real-time RT-PCR SYBR green assay for Tomato ring spot virus in grape

Elwin L. Stewart; Xinshun Qu; Barrie E. Overton; Fred E. Gildow; Nancy G. Wenner; Deborah S. Grove

doi:10.1094/PDIS-91-9-1083

Development of a real-time RT-PCR SYBR green assay for Tomato ring spot virus in grape

Elwin L. Stewart, Xinshun Qu, Barrie E. Overton, Fred E. Gildow, Nancy G. Wenner, Deborah S. Grove

Plant Pathology and Environmental Microbiology

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	1083-1088
Number of pages	6
Journal	Plant disease
Volume	91
Issue number	9
DOIs	https://doi.org/10.1094/PDIS-91-9-1083
State	Published - Sep 2007

All Science Journal Classification (ASJC) codes

Agronomy and Crop Science
Plant Science

Access to Document

10.1094/PDIS-91-9-1083

Cite this

@article{25a7035f7b254a7db5eac30f16bbb260,

title = "Development of a real-time RT-PCR SYBR green assay for Tomato ring spot virus in grape",

abstract = "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.",

author = "Stewart, {Elwin L.} and Xinshun Qu and Overton, {Barrie E.} and Gildow, {Fred E.} and Wenner, {Nancy G.} and Grove, {Deborah S.}",

year = "2007",

month = sep,

doi = "10.1094/PDIS-91-9-1083",

language = "English (US)",

volume = "91",

pages = "1083--1088",

journal = "Plant disease",

issn = "0191-2917",

publisher = "American Phytopathological Society",

number = "9",

}

TY - JOUR

T1 - Development of a real-time RT-PCR SYBR green assay for Tomato ring spot virus in grape

AU - Stewart, Elwin L.

AU - Qu, Xinshun

AU - Overton, Barrie E.

AU - Gildow, Fred E.

AU - Wenner, Nancy G.

AU - Grove, Deborah S.

PY - 2007/9

Y1 - 2007/9

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=34548248170&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34548248170&partnerID=8YFLogxK

U2 - 10.1094/PDIS-91-9-1083

DO - 10.1094/PDIS-91-9-1083

M3 - Article

AN - SCOPUS:34548248170

SN - 0191-2917

VL - 91

SP - 1083

EP - 1088

JO - Plant disease

JF - Plant disease

IS - 9

ER -

Development of a real-time RT-PCR SYBR green assay for Tomato ring spot virus in grape

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this