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1.
DNA Res ; 27(2)2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32531032

RESUMO

White-rot (WR) fungi are pivotal decomposers of dead organic matter in forest ecosystems and typically use a large array of hydrolytic and oxidative enzymes to deconstruct lignocellulose. However, the extent of lignin and cellulose degradation may vary between species and wood type. Here, we combined comparative genomics, transcriptomics and secretome proteomics to identify conserved enzymatic signatures at the onset of wood-decaying activity within the Basidiomycota genus Pycnoporus. We observed a strong conservation in the genome structures and the repertoires of protein-coding genes across the four Pycnoporus species described to date, despite the species having distinct geographic distributions. We further analysed the early response of P. cinnabarinus, P. coccineus and P. sanguineus to diverse (ligno)-cellulosic substrates. We identified a conserved set of enzymes mobilized by the three species for breaking down cellulose, hemicellulose and pectin. The co-occurrence in the exo-proteomes of H2O2-producing enzymes with H2O2-consuming enzymes was a common feature of the three species, although each enzymatic partner displayed independent transcriptional regulation. Finally, cellobiose dehydrogenase-coding genes were systematically co-regulated with at least one AA9 lytic polysaccharide monooxygenase gene, indicative of enzymatic synergy in vivo. This study highlights a conserved core white-rot fungal enzymatic mechanism behind the wood-decaying process.


Assuntos
Desidrogenases de Carboidrato/genética , Proteínas Fúngicas/genética , Lignina/genética , Pycnoporus/enzimologia , Desidrogenases de Carboidrato/metabolismo , Celulose/metabolismo , Proteínas Fúngicas/metabolismo , Genoma Fúngico , Lignina/metabolismo , Filogenia , Pycnoporus/classificação , Pycnoporus/genética , Madeira/metabolismo , Madeira/microbiologia
2.
Science ; 333(6043): 762-5, 2011 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-21764756

RESUMO

Brown rot decay removes cellulose and hemicellulose from wood--residual lignin contributing up to 30% of forest soil carbon--and is derived from an ancestral white rot saprotrophy in which both lignin and cellulose are decomposed. Comparative and functional genomics of the "dry rot" fungus Serpula lacrymans, derived from forest ancestors, demonstrated that the evolution of both ectomycorrhizal biotrophy and brown rot saprotrophy were accompanied by reductions and losses in specific protein families, suggesting adaptation to an intercellular interaction with plant tissue. Transcriptome and proteome analysis also identified differences in wood decomposition in S. lacrymans relative to the brown rot Postia placenta. Furthermore, fungal nutritional mode diversification suggests that the boreal forest biome originated via genetic coevolution of above- and below-ground biota.


Assuntos
Basidiomycota/genética , Biodiversidade , Parede Celular/metabolismo , Micorrizas/genética , Árvores/microbiologia , Madeira/microbiologia , Basidiomycota/classificação , Basidiomycota/enzimologia , Basidiomycota/fisiologia , Evolução Biológica , Biota , Coriolaceae/enzimologia , Coriolaceae/genética , Coriolaceae/fisiologia , Perfilação da Expressão Gênica , Genes Fúngicos , Genômica , Lignina/metabolismo , Magnoliopsida/microbiologia , Micorrizas/enzimologia , Micorrizas/fisiologia , Oxirredutases/genética , Oxirredutases/metabolismo , Peroxidases/genética , Peroxidases/metabolismo , Filogenia , Proteoma , Simbiose , Traqueófitas/microbiologia , Madeira/metabolismo
3.
PLoS Genet ; 6(9): e1001135, 2010 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-20885784

RESUMO

Coprinopsis cinerea (also known as Coprinus cinereus) is a multicellular basidiomycete mushroom particularly suited to the study of meiosis due to its synchronous meiotic development and prolonged prophase. We examined the 15-hour meiotic transcriptional program of C. cinerea, encompassing time points prior to haploid nuclear fusion though tetrad formation, using a 70-mer oligonucleotide microarray. As with other organisms, a large proportion (∼20%) of genes are differentially regulated during this developmental process, with successive waves of transcription apparent in nine transcriptional clusters, including one enriched for meiotic functions. C. cinerea and the fungi Saccharomyces cerevisiae and Schizosaccharomyces pombe diverged ∼500-900 million years ago, permitting a comparison of transcriptional programs across a broad evolutionary time scale. Previous studies of S. cerevisiae and S. pombe compared genes that were induced upon entry into meiosis; inclusion of C. cinerea data indicates that meiotic genes are more conserved in their patterns of induction across species than genes not known to be meiotic. In addition, we found that meiotic genes are significantly more conserved in their transcript profiles than genes not known to be meiotic, which indicates a remarkable conservation of the meiotic process across evolutionarily distant organisms. Overall, meiotic function genes are more conserved in both induction and transcript profile than genes not known to be meiotic. However, of 50 meiotic function genes that were co-induced in all three species, 41 transcript profiles were well-correlated in at least two of the three species, but only a single gene (rad50) exhibited coordinated induction and well-correlated transcript profiles in all three species, indicating that co-induction does not necessarily predict correlated expression or vice versa. Differences may reflect differences in meiotic mechanisms or new roles for paralogs. Similarities in induction, transcript profiles, or both, should contribute to gene discovery for orthologs without currently characterized meiotic roles.


Assuntos
Basidiomycota/citologia , Basidiomycota/genética , Sequência Conservada/genética , Evolução Molecular , Regulação Fúngica da Expressão Gênica , Genes Fúngicos/genética , Meiose/genética , Núcleo Celular/genética , Análise por Conglomerados , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Perfilação da Expressão Gênica , História Antiga , Família Multigênica/genética , Saccharomyces cerevisiae/genética , Schizosaccharomyces/genética , Fatores de Tempo , Transcrição Gênica
4.
Nat Biotechnol ; 28(9): 957-63, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20622885

RESUMO

Much remains to be learned about the biology of mushroom-forming fungi, which are an important source of food, secondary metabolites and industrial enzymes. The wood-degrading fungus Schizophyllum commune is both a genetically tractable model for studying mushroom development and a likely source of enzymes capable of efficient degradation of lignocellulosic biomass. Comparative analyses of its 38.5-megabase genome, which encodes 13,210 predicted genes, reveal the species's unique wood-degrading machinery. One-third of the 471 genes predicted to encode transcription factors are differentially expressed during sexual development of S. commune. Whereas inactivation of one of these, fst4, prevented mushroom formation, inactivation of another, fst3, resulted in more, albeit smaller, mushrooms than in the wild-type fungus. Antisense transcripts may also have a role in the formation of fruiting bodies. Better insight into the mechanisms underlying mushroom formation should affect commercial production of mushrooms and their industrial use for producing enzymes and pharmaceuticals.


Assuntos
Sequência de Bases , Genoma Fúngico/genética , Modelos Biológicos , Schizophyllum/genética , Carpóforos/genética , Carpóforos/crescimento & desenvolvimento , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Genes Fúngicos Tipo Acasalamento , Loci Gênicos/genética , Schizophyllum/crescimento & desenvolvimento , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Madeira/microbiologia
5.
Proc Natl Acad Sci U S A ; 107(26): 11889-94, 2010 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-20547848

RESUMO

The mushroom Coprinopsis cinerea is a classic experimental model for multicellular development in fungi because it grows on defined media, completes its life cycle in 2 weeks, produces some 10(8) synchronized meiocytes, and can be manipulated at all stages in development by mutation and transformation. The 37-megabase genome of C. cinerea was sequenced and assembled into 13 chromosomes. Meiotic recombination rates vary greatly along the chromosomes, and retrotransposons are absent in large regions of the genome with low levels of meiotic recombination. Single-copy genes with identifiable orthologs in other basidiomycetes are predominant in low-recombination regions of the chromosome. In contrast, paralogous multicopy genes are found in the highly recombining regions, including a large family of protein kinases (FunK1) unique to multicellular fungi. Analyses of P450 and hydrophobin gene families confirmed that local gene duplications drive the expansions of paralogous copies and the expansions occur in independent lineages of Agaricomycotina fungi. Gene-expression patterns from microarrays were used to dissect the transcriptional program of dikaryon formation (mating). Several members of the FunK1 kinase family are differentially regulated during sexual morphogenesis, and coordinate regulation of adjacent duplications is rare. The genomes of C. cinerea and Laccaria bicolor, a symbiotic basidiomycete, share extensive regions of synteny. The largest syntenic blocks occur in regions with low meiotic recombination rates, no transposable elements, and tight gene spacing, where orthologous single-copy genes are overrepresented. The chromosome assembly of C. cinerea is an essential resource in understanding the evolution of multicellularity in the fungi.


Assuntos
Cromossomos Fúngicos/genética , Coprinus/genética , Evolução Molecular , Sequência de Bases , Mapeamento Cromossômico , Coprinus/citologia , Coprinus/crescimento & desenvolvimento , Sistema Enzimático do Citocromo P-450/genética , Primers do DNA/genética , Proteínas Fúngicas/genética , Duplicação Gênica , Genoma Fúngico , Meiose/genética , Dados de Sequência Molecular , Família Multigênica , Filogenia , Proteínas Quinases/genética , RNA Fúngico/genética , Recombinação Genética , Retroelementos/genética
6.
Mycol Res ; 112(Pt 3): 389-98, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18313909

RESUMO

Proteolytic enzymes, particularly secreted proteases of fungal origin, are among the most important of industrial enzymes, yet the biochemical properties and substrate specificities of these proteins have been difficult to characterize. Genomic sequencing offers a powerful tool to identify potentially novel proteases. The genome of the model basidiomycete Coprinopsis cinereus was found to have an unusually high number of metalloproteases that closely match the M36 peptidase family known as fungalysins. The eight predicted C. cinereus fungalysins divide into two groups upon comparison with fungalysins from other fungi. One member, CcMEP1, is most similar to the single representative fungalysins from the basidiomycetes Phanerochaete chrysosporium, Cryptococcus neoformans, and Ustilago maydis, and to the fungalysin type-protein from Aspergillus fumigatus. The remaining seven C. cinereus predicted fungalysins form a group with similarity to three predicted M36 peptidases of Laccaria bicolor. All eight of the C. cinereus enzymes contain both the signature M36 Pfam domain and the FTP propeptide domain. All contain large propeptides with considerable sequence conservation near a proposed cleavage site. The predicted mature enzymes range in size from 37-46 kDa and have isoelectric points that are mildly acidic to neutral. The proximity of these genes to telomeres and/or to transposable elements may have contributed to the expansion of this gene family in C. cinereus.


Assuntos
Agaricales/enzimologia , Metaloproteases/química , Metaloproteases/genética , Família Multigênica , Agaricales/classificação , Agaricales/genética , Sequência de Aminoácidos , Aspergillus fumigatus/enzimologia , Cryptococcus neoformans/enzimologia , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Genes Fúngicos , Dados de Sequência Molecular , Phanerochaete/enzimologia , Regiões Promotoras Genéticas , Alinhamento de Sequência , Ustilago/enzimologia
7.
Can J Microbiol ; 53(11): 1278-81, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18026222

RESUMO

The leaf-litter fungus Coprinus cinereus maintains a pool of free amino acid in its mycelium. When the organism is grown under conditions of high nitrogen availability with 13.2 mmol.L-1 L-asparagine as the nitrogen source, the primary constituents of this pool are glutamine, alanine, and glutamic acid. Together these 3 amino acids comprise approximately 70% of the pool. Nitrogen deprivation reduces the size of the free amino acid pool by 75%, and neither a high concentration of ammonium nor a protein nitrogen source support a similar pool size as L-asparagine. Nitrogen deprivation also reduces the concentration of glutamine to the pool while increasing glutamate. Concomitant with this shift is a marked increase in mycelial ammonium.


Assuntos
Aminoácidos/análise , Coprinus/química , Micélio/química , Asparagina/análise , Coprinus/metabolismo , Nitrogênio/análise
8.
Curr Microbiol ; 47(5): 444-9, 2003 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-14669925

RESUMO

Wood-decay fungi depend upon recycling of nitrogen-containing molecules to maintain growth in nitrogen-deficient environments. One of the pools that can support growth in these organisms is the pool of free amino acids. The free amino acid (AA) composition of Schizophyllum commune mycelium grown on the surface of nitrogen-rich (M = 6.6 mM L-asparagine) and nitrogen-poor medium (M01 = 0.06 mM L-asparagine) has been examined: When mycelium is grown on M, alanine, glutamate, and asparagine account for almost 2/3 of the amino acid pool. The free amino acid concentration is reduced by 75% for mycelium grown on the M01 medium, with alanine and glutamate predominating. In addition, free NH4+ increases by 60% in nitrogen-deprived mycelia. Except for asparagine, which is absorbed by the apices, the concentration of all free amino acids is higher in the centers of M-grown, 4-day-old mycelia than in the apices. Hyphae grown to exponential growth on M and transferred to M01 for 12 h show greater free amino acid and NH4+ concentrations in the apices, most likely indicating increased translocation to the apices.


Assuntos
Aminoácidos/metabolismo , Sulfato de Amônio/metabolismo , Nitrogênio/metabolismo , Schizophyllum/metabolismo , Alanina/análise , Asparagina/metabolismo , Meios de Cultura/química , Glutamina/análise , Glutamina/metabolismo , Micélio/química , Schizophyllum/crescimento & desenvolvimento
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