Abstract

Traditional methods to identify aquatic hyphomycetes rely on the morphology of released conidia, which can lead to misidentifications or underestimates of species richness due to convergent morphological evolution and the presence of non-sporulating mycelia. Molecular methods allow fungal identification irrespective of the presence of conidia or their morphology. As a proof-of-concept, we established a quantitative real-time polymerase chain reaction (qPCR) assay to accurately quantify the amount of DNA as a proxy for the biomass of an aquatic hyphomycete species (Alatospora pulchella). Our study showed discrimination even among genetically closely-related species, with a high sensitivity and a reliable quantification down to 9.9 fg DNA (3 PCR forming units; LoD) and 155.0 fg DNA (47 PCR forming units; LoQ), respectively. The assay's specificity was validated for environmental samples that harboured diverse microbial communities and likely contained PCR-inhibiting substances. This makes qPCR a promising tool to gain deeper insights into the ecological roles of aquatic hyphomycetes and other microorganisms.

Published in

PLoS ONE
2017, Volume: 12, number: 4
Publisher: PUBLIC LIBRARY SCIENCE

SLU Authors

• Feckler, Alexander

  • Department of Aquatic Sciences and Assessment, Swedish University of Agricultural Sciences
    
  • University of Koblenz-Landau

• Bundschuh, Mirco

  • Department of Aquatic Sciences and Assessment, Swedish University of Agricultural Sciences
    
  • University of Koblenz-Landau

UKÄ Subject classification

Microbiology


Publication identifier

DOI: https://doi.org/10.1371/journal.pone.0174634

Permanent link to this page (URI)

https://res.slu.se/id/publ/82745