HR: 1330h
AN: B22A-0807    [PDF]
TI: Use of a Microarray to Detect Expression of Genes for Lignin-Degrading Enzymes in Soil Fungi
AU: * Bailey, V L
EM: vanessa.bailey@pnl.gov
AF: Pacific Northwest National Lab, 902 Battelle Blvd, Richland, WA 99352 United States
AU: Smith, J L
EM: jlsmith@mail.wsu.edu
AF: USDA-ARS, 215 Johnson Hall, Pullman, WA 99164 United States
AU: Bolton, H
EM: harvey.bolton@pnl.gov
AF: Pacific Northwest National Lab, 902 Battelle Blvd, Richland, WA 99352 United States
AB: Lignin is a complex biopolymer that is degraded by fungi. Several extracellular enzymes have been implicated in degradation and include lignin peroxidases, laccases, manganese peroxidases, and glyoxal oxidases. Versions of these enzymes are produced by multiple species of fungi, and in some cases, multiple versions of a single enzyme may be produced by the same species of fungus. Previous research has indicated changes in fungal activity and diversity along a tallgrass prairie restoration chronosequence (Fermi National Lab, IL). A cDNA microarray was designed to interrogate the expression and microbial source of these lignin degrading enzymes in the chronosequence soils. We hypothesized that less diversity in gene expression would be detected in a farmed soil than in a restored prairie soil. The array had 46 oligonucleotides (15-25mer) that represent each of the enzymes listed above. Messenger RNA was extracted from 32 one-gram subsamples of the target soils then all of the extracts were pooled prior to RNA precipitation and mRNA purification. Aminoallyl modified dUTPs were incorporated during reverse transcription, after which the cDNA was labeled with Alexa-555 dye. The labeled cDNA was hybridized with the microarray for 24 hours and then imaged. Preliminary results support the hypothesis that fewer genes were expressed in the farmed soil than in the restored soil.
DE: 4805 Biogeochemical cycles (1615)
DE: 4806 Carbon cycling
DE: 4815 Ecosystems, structure and dynamics
SC: Biogeosciences [B]
MN: 2003 Fall Meeting