Transcriptomic Responses of the Softwood-Degrading White-Rot Fungus Phanerochaete carnosa during Growth on Coniferous and Deciduous Wood

ABSTRACT To identify enzymes that could be developed to reduce the recalcitrance of softwood resources, the transcriptomes of the softwood-degrading white-rot fungus Phanerochaete carnosa were evaluated after growth on lodgepole pine, white spruce, balsam fir, and sugar maple and compared to the transcriptome of P. carnosa after growth on liquid nutrient medium. One hundred fifty-two million paired-end reads were obtained, and 63% of these reads were mapped to 10,257 gene models from P. carnosa. Five-hundred thirty-three of these genes had transcripts that were at least four times more abundant during growth on at least one wood medium than on nutrient medium. The 30 transcripts that were on average over 100 times more abundant during growth on wood than on nutrient medium included 6 manganese peroxidases, 5 cellulases, 2 hemicellulases, a lignin peroxidase, glyoxal oxidase, and a P450 monooxygenase. Notably, among the genes encoding putative cellulases, one encoding a glycosyl hydrolase family 61 protein had the highest relative transcript abundance during growth on wood. Overall, transcripts predicted to encode lignin-degrading activities were more abundant than those predicted to encode carbohydrate-active enzymes. Transcripts predicted to encode three MnPs represented the most highly abundant transcripts in wood-grown cultivations compared to nutrient medium cultivations. Gene set enrichment analyses did not distinguish transcriptomes resulting from softwood and hardwood cultivations, suggesting that similar sets of enzyme activities are elicited by P. carnosa grown on different wood substrates, albeit to different expression levels.

[1]  Igor Grigoriev,et al.  Comparative Transcriptome and Secretome Analysis of Wood Decay Fungi Postia placenta and Phanerochaete chrysosporium , 2010, Applied and Environmental Microbiology.

[2]  D. Cullen,et al.  Role of fungal peroxidases in biological ligninolysis. , 2008, Current opinion in plant biology.

[3]  A. Salamov,et al.  Genome, transcriptome, and secretome analysis of wood decay fungus Postia placenta supports unique mechanisms of lignocellulose conversion , 2009, Proceedings of the National Academy of Sciences.

[4]  F. Feltus,et al.  The first genome-level transcriptome of the wood-degrading fungus Phanerochaete chrysosporium grown on red oak , 2009, Current Genetics.

[5]  M. Galbe,et al.  A review of the production of ethanol from softwood , 2002, Applied Microbiology and Biotechnology.

[6]  Roger M. Rowell,et al.  The Chemistry of solid wood , 1984 .

[7]  L. Lo Leggio,et al.  Stimulation of lignocellulosic biomass hydrolysis by proteins of glycoside hydrolase family 61: structure and function of a large, enigmatic family. , 2010, Biochemistry.

[8]  J. Y. Zhu,et al.  Woody biomass pretreatment for cellulosic ethanol production: Technology and energy consumption evaluation. , 2010, Bioresource technology.

[9]  Sofia M. C. Robb,et al.  MAKER: an easy-to-use annotation pipeline designed for emerging model organism genomes. , 2007, Genome research.

[10]  Katherine H. Huang,et al.  Genome sequence of the lignocellulose degrading fungus Phanerochaete chrysosporium strain RP78 , 2004, Nature Biotechnology.

[11]  J. Ralph,et al.  Evidence for cleavage of lignin by a brown rot basidiomycete. , 2008, Environmental microbiology.

[12]  H. Doddapaneni,et al.  Microarray-based global differential expression profiling of P450 monooxygenases and regulatory proteins for signal transduction pathways in the white rot fungus Phanerochaete chrysosporium , 2005, Molecular Genetics and Genomics.

[13]  A. Salamov,et al.  Computational analysis of the Phanerochaete chrysosporium v2.0 genome database and mass spectrometry identification of peptides in ligninolytic cultures reveal complex mixtures of secreted proteins. , 2006, Fungal genetics and biology : FG & B.

[14]  T. Kirk,et al.  Degradation of Gymnosperm (Guaiacyl) vs. Angiosperm (Syringyl/Guaiacyl) Lignins by Phanerochaete chrysosporium , 1985 .

[15]  B. Henrissat,et al.  Synergism of Cellulases from Trichoderma reesei in the Degradation of Cellulose , 1985, Bio/Technology.

[16]  W. Kenealy,et al.  Growth and fermentation responses of Phanerochaete chrysosporium to O2 limitation , 2004 .

[17]  T. M. Wood,et al.  Fungal cellulases. , 1992, Biochemical Society transactions.

[18]  K. Igarashi,et al.  Quantitative transcriptional analysis of the genes encoding glycoside hydrolase family 7 cellulase isozymes in the basidiomycete Phanerochaete chrysosporium. , 2009, FEMS microbiology letters.

[19]  H. Doddapaneni,et al.  Genome-wide structural and evolutionary analysis of the P450 monooxygenase genes (P450ome) in the white rot fungus Phanerochaete chrysosporium : Evidence for gene duplications and extensive gene clustering , 2005, BMC Genomics.

[20]  Ichael,et al.  Analysis of Character Correlations Among Wood Decay Mechanisms , Mating Systems , and Substrate Ranges in Homobasidiomycetes , 2001 .

[21]  Jonathan P. Bollback,et al.  Bayesian Inference of Phylogeny and Its Impact on Evolutionary Biology , 2001, Science.

[22]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[23]  R. Pettersen,et al.  The chemical composition of wood , 1984 .

[24]  Yan Lin,et al.  Ethanol fermentation from biomass resources: current state and prospects , 2006, Applied Microbiology and Biotechnology.

[25]  K. Jensen,et al.  Pathways for Extracellular Fenton Chemistry in the Brown Rot Basidiomycete Gloeophyllum trabeum , 2001, Applied and Environmental Microbiology.

[26]  John P. Huelsenbeck,et al.  MrBayes 3: Bayesian phylogenetic inference under mixed models , 2003, Bioinform..

[27]  M. Gerstein,et al.  RNA-Seq: a revolutionary tool for transcriptomics , 2009, Nature Reviews Genetics.

[28]  B. Cathala,et al.  Organization Behavior of Guaiacyl and Guaiacyl/Syringyl Dehydrogenation Polymers (Lignin Model Compounds) at the Air/Water Interface , 2000 .

[29]  E. Sjöström,et al.  Wood Chemistry: Fundamentals and Applications , 1981 .

[30]  C. Felby,et al.  Effect of storage on extractives from particle surfaces of softwood and hardwood raw materials for wood pellets , 2009, European Journal of Wood and Wood Products.

[31]  Jill Gaskell,et al.  Transcriptome and Secretome Analyses of Phanerochaete chrysosporium Reveal Complex Patterns of Gene Expression , 2009, Applied and Environmental Microbiology.

[32]  Liisa Viikari,et al.  Evaluation of wet oxidation pretreatment for enzymatic hydrolysis of softwood , 2004, Applied biochemistry and biotechnology.

[33]  T. Wood Fungal cellulases. , 1992, Biochemical Society transactions.

[34]  Zhaolei Zhang,et al.  An atlas of chaperone–protein interactions in Saccharomyces cerevisiae: implications to protein folding pathways in the cell , 2009, Molecular systems biology.

[35]  E. Master,et al.  Proteomic characterization of lignocellulose-degrading enzymes secreted by Phanerochaete carnosa grown on spruce and microcrystalline cellulose , 2010, Applied Microbiology and Biotechnology.

[36]  B. Morgenstern,et al.  AUGUSTUS at EGASP: using EST, protein and genomic alignments for improved gene prediction in the human genome , 2006, Genome Biology.

[37]  G. F. Leatham,et al.  Lignin Distribution in Wood Delignified by White-Rot Fungi: X-Ray Microanalysis of Decayed Wood Treated with Bromine , 1988 .

[38]  D. Hibbett,et al.  Molecular Evolution and Diversity of Lignin Degrading Heme Peroxidases in the Agaricomycetes , 2008, Journal of Molecular Evolution.

[39]  R. Blanchette,et al.  Structure, Organization, and Transcriptional Regulation of a Family of Copper Radical Oxidase Genes in the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium , 2006, Applied and Environmental Microbiology.

[40]  B. W. Bogan,et al.  Expression of lip genes during growth in soil and oxidation of anthracene by Phanerochaete chrysosporium , 1996, Applied and environmental microbiology.

[41]  K. Welinder Superfamily of plant, fungal and bacterial peroxidases , 1992 .

[42]  H. H. Burdsall A contribution to the taxonomy of the genus Phanerochaete (Corticiaceae, Aphyllophorales) , 1985 .