Abstract
Grapevine red blotch virus (GRBV) is an emerging virus of significant viticultural importance throughout North America. Here, we report the development of a simple protocol for point-of-use detection of GRBV. Extraction of nucleic acids is not required; instead, the whole intact plant can simply be pricked with a sterile pipette tip, which is then incubated in sterile distilled water to provide the sample template in a loop-mediated isothermal amplification (LAMP) reaction. This method is 10,000 times more sensitive than conventional PCR, costs under a dollar per sample, and can be completed from sampling to readout in just over half an hour.
References
Blanco-Ulate B, Hopfer H, Figueroa-Balderas R, Ye Z, Rivero RM, Albacete A, Pérez-Alfocea F, Koyama R, Anderson MM, Smith RJ, Ebeler SE, Cantu D (2017) Red blotch disease alters grape berry development and metabolism by interfering with the transcriptional and hormonal regulation of ripening. J Exp Bot 68:1225–1238
Cieniewicz EJ, Pethybridge SJ, Gorny A, Madden LV, McLane H, Perry KL, Fuchs M (2017) Spatiotemporal spread of grapevine red blotch-associated virus in a California vineyard. Virus Res 241:156–162
Gambino G, Perrone I, Gribaudo I (2008) A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants. Phytochem Anal 19:520–525
Gasperin-Bulbarela J, Licea-Navarro AF, Pino-Villar C, Hernandez-Martinez R, Carrillo-Tripp J (2018) First report of grapevine red blotch virus in Mexico. Plant Dis. https://doi.org/10.1094/pdis-07-18-1227-pdn
Krenz B, Thompson JR, McLane HL, Fuchs M, Perry KL (2014) Grapevine red blotch-associated virus Is Widespread in the United States. Phytopathology 104:1232–1240
Li R, Fuchs MF, Perry KL, Mekuria T, Zhang S (2017) Development of a fast Amplifyrp Acceler8 diagnostic assay for grapevine red blotch virus. J Plant Pathol 99:657–662
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:e63
Poojari S, Lowery DT, Rott M, Schmidt AM, Úrbez-Torres JR (2017) Incidence, distribution and genetic diversity of Grapevine red blotch virus in British Columbia. Can J Plant Pathol 39:201–211
Reynard J-S, Brodard J, Dubuis N, Zufferey V, Schumpp O, Schaerer S, Gugerli P (2018) Grapevine red blotch virus: absence in Swiss vineyards and analysis of potential detrimental effect on viticultural performance. Plant Dis 102:651–655
Setiono FJ, Chatterjee D, Fuchs M, Perry KL, Thompson JR (2018) The distribution and detection of grapevine red blotch virus in its host depend on time of sampling and tissue type. Plant Dis 102:2187–2193
Sudarshana MR, Perry KL, Fuchs MF (2015) Grapevine red blotch-associated virus, an emerging threat to the grapevine industry. Phytopathology 105:1026–1032
Vargas-Asencio J, Wojciechowska K, Raskerville M, Gomez AL, Perry KL, Thompson JR (2017) The complete nucleotide sequence and genomic characterization of grapevine asteroid mosaic associated virus. Virus Res 227:82–87
Yepes LM, Cieniewicz E, Krenz B, McLane H, Thompson JR, Perry KL, Fuchs M (2018) Causative role of grapevine red blotch virus in red blotch disease. Phytopathology 108:902–909
Acknowledgements
Many thanks to the Cornell Latin American Studies Program Fellowship for funding JLR2 while at Cornell. We are also grateful to John Keeton and colleagues at the USDA-ARS Plant Genetic Resources Unit for providing us with dormant field cane material from the USDA grapevine germplasm repository, and to Heather McLane, Aisha Younas and Alex Cha for their technical and greenhouse support. Thanks also to Marc Fuchs for critically reading the manuscript. This work was supported by USDA-NIFA Hatch project number 1013513, and the College of Agriculture and Life Sciences, Cornell University.
Funding
This study was funded by a Cornell Latin American Studies Program Fellowship, USDA-NIFA Hatch project number 1013513 and the College of Agriculture and Life Sciences, Cornell University.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
All authors declare they have no conflict of interest.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Handling Editor: Elvira Fiallo-Olivé.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Romero Romero, J.L., Carver, G.D., Arce Johnson, P. et al. A rapid, sensitive and inexpensive method for detection of grapevine red blotch virus without tissue extraction using loop-mediated isothermal amplification. Arch Virol 164, 1453–1457 (2019). https://doi.org/10.1007/s00705-019-04207-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00705-019-04207-y