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Molecular approach for analysis of model fungal genes encoding ligninolytic peroxidases in wood‐decaying soil systems
Author(s) -
Stuardo M,
Vásquez M,
Vicuña R,
González B
Publication year - 2004
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2003.01442.x
Subject(s) - phanerochaete , chrysosporium , polymerase chain reaction , biology , white rot , lignin peroxidase , gene , soil microbiology , dna extraction , microbiology and biotechnology , inoculation , botany , peroxidase , enzyme , bacteria , horticulture , genetics , lignin , biochemistry
Aims: 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. Methods and 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. Significance and Impact of the Study: This is the first assessment of the use of consensus PCR primers for direct detection of ligninolytic peroxidase genes in wood‐decaying soil systems.