Molecular approach for analysis of model fungal genes encoding ligninolytic peroxidases in wood-decaying soil systems.
OBJECTIVE
Test the use of nondegenerated consensus polymerase chain reaction (PCR) primers targeting lip and mnp sequences to detect ligninolytic fungi in wood-decaying soil systems, avoiding the need for enrichment or isolation on traditional fungal media culture.


RESULTS
The PCR primers were tested with total DNA isolated from incubations of wood-soil systems inoculated or not with the white-rot fungi Phanerochaete chrysosporium, or a white-rot sample obtained from a Nothofagus forest. The PCR products for lip and mnp sequences were only obtained in soil with P. chrysosporium-colonized wood chips. In these soil samples, reverse transcription-PCR analysis of lip and mnp PCR products indicated expression of LipA, LipB, LipJ and MnP isoenzymes.


CONCLUSIONS
This is the first assessment of the use of consensus PCR primers for direct detection of ligninolytic peroxidase genes in wood-decaying soil systems.
Stuardo
M
M
Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, P. Universidad Católica de Chile and Millennium Institute for Fundamental and Applied Biology, Santiago, Chile.
Vásquez
M
M
Vicuña
R
R
González
B
B
D001487:Basidiomycota / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D004271:DNA, Fungal / Q000737:chemistry / Q000302:isolation & purification; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal; D010544:Peroxidases / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology
D004271:DNA, Fungal; D005656:Fungal Proteins; D010544:Peroxidases; C042858:lignin peroxidase; C051129:manganese peroxidase