Imperial College London
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ProfessorAlfriedVogler

Faculty of Natural Sciences, Department of Life Sciences (Silwood Park)

Professor of Molecular Systematics
 
 
 
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Contact

 

+44 (0)20 7942 5613a.vogler

 
 
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Location

 

Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Miller:2016:10.1002/ece3.1925,
author = {Miller, K and Vogler, A and Hopkins, K and Inward, D},
doi = {10.1002/ece3.1925},
journal = {Ecology and Evolution},
pages = {1590--1600},
title = {Metabarcoding of fungal communities associated with bark beetles},
url = {http://dx.doi.org/10.1002/ece3.1925},
volume = {6},
year = {2016}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Many species of fungi are closely allied with bark beetles, including many treepathogens, but their species richness and patterns of distribution remain largelyunknown. We established a protocol for metabarcoding of fungal communitiesdirectly from total genomic DNA extracted from individual beetles, showingthat the ITS3/4 primer pair selectively amplifies the fungal ITS. Using threespecimens of bark beetle from different species, we assess the fungal diversityassociated with these specimens and the repeatability of these estimates in PCRsconducted with different primer tags. The combined replicates produced 727fungal Operational Taxonomic Units (OTUs) for the specimen of Hylastes ater,435 OTUs for Tomicus piniperda, and 294 OTUs for Trypodendron lineatum,while individual PCR reactions produced on average only 229, 54, and 31OTUs for the three specimens, respectively. Yet, communities from PCR replicateswere very similar in pairwise comparisons, in particular when consideringspecies abundance, but differed greatly among the three beetle specimens. Differentprimer tags or the inclusion of amplicons in separate libraries did notimpact the species composition. The ITS2 sequences were identified with theLowest Common Ancestor approach and correspond to diverse lineages offungi, including Ophiostomaceae and Leotiomycetes widely found to be treepathogens. We conclude that Illumina MiSeq metabarcoding reliably capturesfungal diversity associated with bark beetles, although numerous PCR replicatesare recommended for an exhaustive sample. Direct PCR from beetle DNAextractions provides a rapid method for future surveys of fungal species diversityand their associations with bark beetles and environmental variables.
AU  - Miller,K
AU  - Vogler,A
AU  - Hopkins,K
AU  - Inward,D
DO  - 10.1002/ece3.1925
EP  - 1600
PY  - 2016///
SN  - 2045-7758
SP  - 1590
TI  - Metabarcoding of fungal communities associated with bark beetles
T2  - Ecology and Evolution
UR  - http://dx.doi.org/10.1002/ece3.1925
UR  - http://hdl.handle.net/10044/1/28355
VL  - 6
ER  -
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