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Abstract Detail


MSA - Ecology/Pathology

Amend, Anthony S [1], Seifert, Keith A. [2], Samson, Robert A. [3], Bruns , Thomas [1].

Spores indoors: a pyrosequencing look at what fungi the cat dragged in.

Changes in building practices stimulated largely by increased oil prices have led to construction of tightly sealed buildings which are prone to developing moisture problems that promote fungal growth. Concurrently, increased use of immunosuppressant drugs and the advent of AIDS have led to an increased frequency of human fungal infections, including infections caused by species not previously considered pathogenic. For these reasons it is important to begin to characterize the presence, prevalence and roles of the indoor fungi with whom we interact daily. While real-time PCR assays have been developed to detect the most common and demonstratively pathogenic taxa, most studies of indoor fungi use culturing methods to characterize indoor fungal assemblages. Both of these methodologies inherently restrict the detectable diversity to a limited and phylogenetically narrow set of taxa. Conversely, indoor fungal diversity detected by direct DNA sampling techniques is only beginning to be explored. Here, we characterize the fungal diversity in settled indoor dust from nearly 100 samples from every inhabited continent and Micronesia. Nucleotide sequence data from the ITS1, ITS2 and D1-D2 regions of the large subunit rDNA were sequenced using the Roche 454 Titanium pyrosequencing platform. This multilocus bar-coding approach enables analyses of taxonomic and phylogenetic diversity of indoor fungal assemblages at a sampling depth of thousands of reads per locus per sample. In parallel, high-throughput culturing efforts enable isolation of voucher specimens for subsequent barcode referencing. We compare the taxonomic and phylogenetic diversity detected among loci and analyze global patterns of distance-decay of assemblage similarity. Furthermore, we test the hypothesis that fungal species distribution is scaled to substrate. Methods used to determine assay sensitivity and detection thresholds of various spore morphologies are discussed.


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1 - University of California, Berkeley, Department of Plant Biology, 321 Koshland Hall, Berkeley, CA, 94720, USA
2 - Biodiversity (Mycology & Botany), Agriculture & Agri-Food Canada, Eastern Cereal and Oilseed Research Centre, 960 Carling Ave., Ottawa, Ontario, K1A 0C6, Canada
3 - CBS Fungal Biodiversity Centre, P.O. Box 85167, 3508 AD , Utrecht, The Netherlands

Keywords:
microbial biogepgraphy
indoor fungi
pyrosequencing
bioinformatics.

Presentation Type: Oral Paper:Papers for Topics
Session: 18
Location: Cottonwood A/Snowbird Center
Date: Monday, July 27th, 2009
Time: 11:30 AM
Number: 18006
Abstract ID:716