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Q-RT-PCR for Assessing Archaea, Bacteria, and Fungi During Leaf Decomposition in a Stream

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Abstract

Leaf disks of Tilia cordata were exposed for up to 5 weeks in a first-order stream in Nova Scotia, Canada. The exponential decay rate k was 0.008 day−1. Ergosterol levels increased linearly to a maximum of 134 μg g−1 dry leaf mass. Release of conidia peaked at 700 day−1 mg−1 on leaves that had been exposed for 3 weeks; after 5 weeks, it declined to 15 mg−1. In total, 23 taxa of aquatic hyphomycetes were distinguished. Anguillospora filiformis contributed over 76% of the conidia during weeks 1, 2, and 3, and 16.5% in week 5. Three sets of primers specific for Bacteria, Archaea, and Fungi were applied in quantitative real-time polymerase chain reaction (Q-RT-PCR) to estimate relative DNA amounts. Archaeal DNA was consistently present at low levels. Bacterial and fungal DNA peaked between weeks 2 and 3, and declined in week 5. With the exception of week 1, fungal DNA exceeded bacterial DNA by between 12 and 110%.

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Acknowledgements

Financial support by an NSERC Discovery grant to FB is gratefully acknowledged. We thank Amanda Cockshutt and Chris Brown for introducing us to RT-PCR. Meredith Hullar provided valuable advice concerning suitable primers for Archaea.

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  1. Department of Biology, Mount Allison University, Sackville, New Brunswick, E4L 1G7, Canada

    Mayura A. Manerkar, S. Seena & Felix Bärlocher

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  1. Mayura A. Manerkar
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  2. S. Seena
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  3. Felix Bärlocher
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Correspondence to Felix Bärlocher.

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Manerkar, M.A., Seena, S. & Bärlocher, F. Q-RT-PCR for Assessing Archaea, Bacteria, and Fungi During Leaf Decomposition in a Stream. Microb Ecol 56, 467–473 (2008). https://doi.org/10.1007/s00248-008-9365-z

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