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Title: Methodological Guidelines for Accurate Detection of Viruses in Wild Plant Species

Abstract

Here 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 amplificationmore » 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.« less

Authors:
ORCiD logo [1];  [2];  [3];  [2];  [2];  [4]
  1. Univ. of Minnesota, Saint Paul, MN (United States). Dept of Ecology, Evolution, and Behavior; Planat Pathology, Montfavet, France (Europe)
  2. Univ. of Minnesota, Saint Paul, MN (United States). Dept of Ecology, Evolution, and Behavior
  3. Loyola Marymount Univ., Los Angeles, CA (United States)
  4. Michigan State Univ., East Lansing, MI (United States)
Publication Date:
Research Org.:
Univ. of Wisconsin, Madison, WI (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1470727
Grant/Contract Number:  
FC02-07ER64494
Resource Type:
Accepted Manuscript
Journal Name:
Applied and Environmental Microbiology
Additional Journal Information:
Journal Volume: 82; Journal Issue: 6; Journal ID: ISSN 0099-2240
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Lacroix, Christelle, Renner, Kurra, Cole, Ellen, Seabloom, Eric W., Borer, Elizabeth T., and Malmstrom, Carolyn M. Methodological Guidelines for Accurate Detection of Viruses in Wild Plant Species. United States: N. p., 2016. Web. doi:10.1128/AEM.03538-15.
Lacroix, Christelle, Renner, Kurra, Cole, Ellen, Seabloom, Eric W., Borer, Elizabeth T., & Malmstrom, Carolyn M. Methodological Guidelines for Accurate Detection of Viruses in Wild Plant Species. United States. doi:10.1128/AEM.03538-15.
Lacroix, Christelle, Renner, Kurra, Cole, Ellen, Seabloom, Eric W., Borer, Elizabeth T., and Malmstrom, Carolyn M. Fri . "Methodological Guidelines for Accurate Detection of Viruses in Wild Plant Species". United States. doi:10.1128/AEM.03538-15. https://www.osti.gov/servlets/purl/1470727.
@article{osti_1470727,
title = {Methodological Guidelines for Accurate Detection of Viruses in Wild Plant Species},
author = {Lacroix, Christelle and Renner, Kurra and Cole, Ellen and Seabloom, Eric W. and Borer, Elizabeth T. and Malmstrom, Carolyn M.},
abstractNote = {Here 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.},
doi = {10.1128/AEM.03538-15},
journal = {Applied and Environmental Microbiology},
number = 6,
volume = 82,
place = {United States},
year = {2016},
month = {1}
}

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