Methodological Guidelines for Accurate Detection of Viruses in Wild Plant Species

Ecological understanding of disease risk, emergence, and dynamics and of the efficacy of control strategies relies heavily on efficient tools for microorganism identification and characterization. Misdetection, such as the misclassification of infected hosts as healthy, can strongly bias estimates o...

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Published in:	Applied and environmental microbiology Vol. 82; no. 6; pp. 1966 - 1975
Main Authors:	Lacroix, Christelle, Renner, Kurra, Cole, Ellen, Seabloom, Eric W, Borer, Elizabeth T, Malmstrom, Carolyn M
Format:	Journal Article
Language:	English
Published:	United States American Society for Microbiology 01-03-2016
Subjects:	Agricultural sciences Barley yellow dwarf virus BASIC BIOLOGICAL SCIENCES Flowers & plants Immunoassay Immunologic Techniques - methods Immunologic Techniques - standards Life Sciences Methods Microbiology Microbiology and Parasitology Microorganisms Molecular Biology - methods Molecular Biology - standards Phytopathology and phytopharmacy Plant Diseases - virology Plant Viruses - isolation & purification Plants - virology Serologic Tests - methods Vegetal Biology Virology Virology - methods Virology - standards Viruses inhibitors Infectious disease detection false-negative RT-PCR extraction pathogen distribution ELISA
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Summary:	Ecological understanding of disease risk, emergence, and dynamics and of the efficacy of control strategies relies heavily on efficient tools for microorganism identification and characterization. Misdetection, such as the misclassification of infected hosts as healthy, can strongly bias estimates of disease prevalence and lead to inaccurate conclusions. In natural plant ecosystems, interest in assessing microbial dynamics is increasing exponentially, but guidelines for detection of microorganisms in wild plants remain limited, particularly so for plant viruses. To address this gap, we explored issues and solutions associated with virus detection by serological and molecular methods in noncrop plant species as applied to the globally important Barley yellow dwarf virus PAV (Luteoviridae), which infects wild native plants as well as crops. With enzyme-linked immunosorbent assays (ELISA), we demonstrate how virus detection in a perennial wild plant species may be much greater in stems than in leaves, although leaves are most commonly sampled, and may also vary among tillers within an individual, thereby highlighting the importance of designing effective sampling strategies. With reverse transcription-PCR (RT-PCR), we demonstrate how inhibitors in tissues of perennial wild hosts can suppress virus detection but can be overcome with methods and products that improve isolation and amplification of nucleic acids. These examples demonstrate the paramount importance of testing and validating survey designs and virus detection methods for noncrop plant communities to ensure accurate ecological surveys and reliable assumptions about virus dynamics in wild hosts.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 PMCID: PMC4784055 USDOE FC02-07ER64494 Citation Lacroix C, Renner K, Cole E, Seabloom EW, Borer ET, Malmstrom CM. 2016. Methodological guidelines for accurate detection of viruses in wild plant species. Appl Environ Microbiol 82:1966–1975. doi:10.1128/AEM.03538-15.
ISSN:	0099-2240 1098-5336
DOI:	10.1128/AEM.03538-15