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.
Bibliographical noteFunding Information:
We thank Missy Rudeen, Alexis Rogers, Alisha Fischer, Taelor Haase, and many undergraduate students for help in the lab. We also thank Marty Dekkers for sharing expertise and advice on methods for virus detection. We received support from the NSF program in Ecology and Evolution of Infectious Disease (grant DEB-1015805) to E.T.B. and E.W.S. and from the DOE Office of Science, BER, Great Lakes Bioenergy Center (DEFC02-07ER64494), USDA NIFA Sustainable Biofuels Program award 2011-67009-30137, and MSU AgBioResearch funding to C.M.M. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication, DOE Office of Science -Great Lakes Bioenergy Center provided funding to Carolyn M. Malmstrom under grant number BER DE-FC02-07ER64494. USDA NIFA Sustainable Biofuel Program Award provided funding to Carolyn M. Malmstrom under grant number 2011-67009-30137. MSU AgBioResearch provided funding to Carolyn M. Malmstrom. National Science Foundation (NSF) provided funding to Eric W. Seabloom and Elizabeth T. Borer under grant number DEB-1015805.