Quantitation of fungal mRNAs in complex substrates by reverse transcription PCR and its application to Phanerochaete chrysosporium-colonized soil.
Thorough analysis of fungi in complex substrates has been hampered by inadequate experimental tools for assessing physiological activity and estimating biomass. We report a method for the quantitative assessment of specific fungal mRNAs in soil. The method was applied to complex gene families of Phanerochaete chrysosporium, a white-rot fungus widely used in studies of organopollutant degradation. Among the genes implicated in pollutant degradation, two closely related lignin peroxidase transcripts were detected in soil. The pattern of lignin peroxidase gene expression was unexpected; certain transcripts abundant in defined cultures were not detected in soil cultures. Transcripts encoding cellobiohydrolases and beta-tubulin were also detected. The method will aid in defining the roles of specific genes in complex biological processes such as organopollutant degradation, developing strategies for strain improvement, and identifying specific fungi in environmental samples.
Lamar
R T
RT
Forest Products Laboratory, U.S. Department of Agriculture, Madison, Wisconsin 53705, USA.
Schoenike
B
B
Vanden Wymelenberg
A
A
Stewart
P
P
Dietrich
D M
DM
Cullen
D
D
D001483:Base Sequence; D001487:Basidiomycota / Q000302:isolation & purification*; D043366:Cellulose 1,4-beta-Cellobiosidase; D017931:DNA Primers; D006026:Glycoside Hydrolases / Q000235:genetics*; D008969:Molecular Sequence Data; D010416:Pentachlorophenol; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D012988:Soil Microbiology; D014404:Tubulin / Q000235:genetics
D017931:DNA Primers; D012333:RNA, Messenger; D014404:Tubulin; D010416:Pentachlorophenol; D006026:Glycoside Hydrolases; D043366:Cellulose 1,4-beta-Cellobiosidase