Microbial Community Analysis Using Terminal Restriction Fragment Length Polymorphism (T-RFLP) Analysis: Field Study Results

IRG/WP 08-20377

G T Kirker, S V Diehl, M L Prewitt, W J Diehl

The effects of chlorothalonil (CTN) and butylated hydroxytoluene (BHT) on the fungal community in southern yellow pine (SYP) were assessed using terminal restriction fragment length polymorphism (T-RFLP) analysis. Field stakes treated with 0.25% and 0.37% ammoniacal copper quat (ACQ-C), 0.1 and 0.25% CTN, and 0.1 and 0.25% CTN combined with 2% BHT were installed with untreated controls in field sites in Mississippi. Stakes were sampled at 90 days intervals and rated for decay damage. Microbial DNA was extracted from wood and amplified using non-specific (bacteria and fungi) and specific (basidiomycete) primers. Alpha diversity (richness and diversity), species composition, and beta diversity, were all calculated using T-RFLP data. Results indicate the presence of wood preservatives containing biocides: (1) increases initial colonization by bacteria that decreases over time, (2) slows the initial colonization of field stakes by fungi resulting in lower richness and diversity that increased over time, and (3) increased richness and diversity of basidiomycetes. Preservative treatment also changed the community composition of bacteria, fungi, and basidiomycetes found in wood, that became more similar over time to untreated controls for fungi and basidiomycetes, but not bacteria. The beta diversity of treated samples was less similar in early stages of exposure (3-9 months), but coalesced over time into stable populations that were similar to fungal and basidiomycete communities in untreated controls, but still bacteria remained different. Correlations were found between depletion of 0.1% CTN and increasing bacterial and fungal diversity.


Keywords: microbial community, T-RFLP, field sites, wood decay, colonization

Conference: 08-05-25/29, Istanbul, Turkey


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