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SRX22754758: Pyrosequencing sequencing of ITS1 region of fungal hyphae: isolated from mesh bags buried in a spruce and fir forest
1 LS454 (454 GS FLX) run: 1 spots, 171 bases, 55,962b downloads

Design: Every sample from each plot was amplified individually with a primer unique to that plot, and all amplicons (nine successful samples from each plot) were pooled in an equimolar mixture by combining equal amounts of each PCR amplicon at a standard concentration. The final 20 ng L1 mixture, containing six unique pyrosequencing primers, each representing one of six different plots, was amplified in a 1/8 plate Next Generation pyrosequencing reaction on a Roche GS-FLX at the McGill University and Genome Quebec Innovation Center.
Submitted by: University of British Columbia Okanagan Campus
Study: Representative sequences from each OTU yielded from pyrosequencing of fungi in sand-filled hyphae-trapping mesh bags within fifteen years of clearcut harvest
show Abstracthide Abstract
Shifts in ectomycorrhizal (ECM) fungal community composition occur after clearcut logging, resulting inthe loss of some dominant forest fungi. Because decaying wood is a remnant of the original forest andan important habitat for ECM fungal species, we examined ECM spruce roots and hyphae in 1-ha coarsewoody debris (CWD) retention and removal plots (N = 3) at a high elevation spruce forest 12 and 13 yearsafter clearcut harvesting to test for a medium-term effect of CWD retention. Root tips from ten Piceaengelmannii (Parry ex. Engelm.) saplings per plot were grouped morphologically, and the ECM fungalsymbiont identified by Sanger sequencing. Sand-filled hyphae-trapping mesh bags were buried amongstthe roots of saplings for one year. PCR product from all bags per plot was pooled and submitted for GS-FLXTitanium sequencing. Forty-six of 89 operational taxonomic units (OTUs) from root tips and 50 OTUs fromhyphae were identified as ECM taxa. The most abundant taxa amongst root tip OTUs were Thelephoraterrestris, Alloclavaria purpurea, Amphinema byssoides, and Tylospora asterophora. The most abundant OTUs from mesh bags were ectomycorrhizal taxa, and the most abundant of these were A. byssoides, T. terrestris, Wilcoxina mikolae, and T. asterophora. The retention of CWD had no detectable effect on taxon richness, evenness or diversity of ECM fungi on root tips or in mesh bags; however, there was a detectable shift in species composition. The relative abundance of A. byssoides root tips was significantly higher at removal plots while A. purpurea was a significant indicator species for CWD removal. Our results suggest that the retention of CWD at the time of harvest has affected ECM habitat at this site, and has resulted in altered ECM species composition, even though the logs are still hard and intact.
Library:
Name: CP07H4KUX
Instrument: 454 GS FLX
Strategy: AMPLICON
Source: GENOMIC
Selection: PCR
Layout: SINGLE
Runs: 1 run, 1 spots, 171 bases, 55,962b
Run# of Spots# of BasesSizePublished
SRR27065426117155,962b2025-01-01

ID:
30791980

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