RESUMO
At present, 25 species are accepted in Haploporus and are distributed in Asia, Europe, North America, South America, Australia, and Africa. In this study, two new species, Haploporus ecuadorensis from Ecuador and H. monomitica from China, are described and illustrated based on morphological examination and phylogenetic analyses. H. ecuadorensis is characterized by annual, resupinate basidiomata with pinkish buff to honey yellow hymenophore when dry, round to angular pores of 2-4 per mm, a dimitic hyphal structure with generative hyphae bearing clamp connections, hyphae at dissepiment edge usually with one or two simple septa, the presence of dendrohyphidia and cystidioles, and oblong to ellipsoid basidiospores measuring 14.9-17.9 × 6.9-8.8 µm. Haploporus monomitica differs from other Haploporus species in that it has a monomitic hyphal system and strongly dextrinoid basidiospores. The differences between the new species and morphologically similar and phylogenetically related species are discussed. In addition, an updated key to 27 species of Haploporus is provided.
Assuntos
Basidiomycota , Polyporales , Polyporales/genética , Filogenia , DNA Espaçador Ribossômico/genética , DNA Espaçador Ribossômico/química , Equador , Basidiomycota/genética , China , Esporos Fúngicos/genéticaRESUMO
The family Steccherinaceae includes genera with smooth, hydnoid, and poroid hymenophores, monomitic to dimitic hyphal systems, and generative hyphae with clamps or simple septa. Steccherinum is the largest genus in the family, with a worldwide distribution, and is characterized mainly by a dimitic hyphal system and presence of thick-walled encrusted cystidia. Species traditionally included in Steccherinum, however, have been transferred to other genera based on results of molecular phylogenetic analyses. Even though knowledge of Steccherinaceae has increased in the past few years, very little is known about the hydnoid species of the family, especially from the Neotropics. In this study, we present morphological and phylogenetic analyses on hydnoid specimens of Steccherinaceae collected in the Neotropics. Molecular data of nuc internal transcribed spacer region ITS1-5.8S-ITS rDNA (ITS) and portions of nuc 28S rDNA (28S), translation elongation factor 1-α (tef1), and the largest subunit of RNA polymerase II (rpb1) were obtained from Brazilian collections. Types and original collections were studied for morphological comparison. Samples we studied grouped in four different genera of Steccherinaceae: Cabalodontia, Etheirodon, Metuloidea, and Steccherinum. Three new neotropical species, Cabalodontia delicata, Etheirodon purpureum, and Steccherinum larssonii, are described. In addition, the new combinations Cabalodontia albofibrillosa and Metuloidea reniformis are proposed. The four genera presented in this study are compared and discussed in detail.
Assuntos
Polyporales , DNA Fúngico/genética , DNA Ribossômico/genética , DNA Espaçador Ribossômico/genética , Filogenia , Polyporales/genética , RNA Ribossômico 28S , Análise de Sequência de DNARESUMO
Ceriporiopsis subvermispora is a white-rot fungus with a high specificity towards lignin mineralization when colonizing dead wood or lignocellulosic compounds. Its lignocellulose degrading system is formed by cellulose hydrolytic enzymes, manganese peroxidases, and laccases that catalyze the efficient depolymerization and mineralization of lignocellulose. To determine if this metabolic specialization has modified codon usage of the lignocellulolytic system, improving its adaptation to the fungal translational machine, we analyzed the adaptation to host codon usage (CAI), tRNA pool (tAI, and AAtAI), codon pair bias (CPB), and the number of effective codons (Nc). These indexes were correlated with gene expression of C. subvermispora, in the presence of glucose and Aspen wood. General gene expression was not correlated with the index values. However, in media containing Aspen wood, the induction of expression of lignocellulose-degrading genes, showed significantly (p < 0.001) higher values of CAI, AAtAI, CPB, tAI, and lower values of Nc than non-induced genes. Cellulose-binding proteins and manganese peroxidases presented the highest adaptation values. We also identified an expansion of genes encoding glycine and glutamic acid tRNAs. Our results suggest that the metabolic specialization to use wood as the sole carbon source has introduced a bias in the codon usage of genes involved in lignocellulose degradation. This bias reduces codon diversity and increases codon usage adaptation to the tRNA pool available in C. subvermispora. To our knowledge, this is the first study showing that codon usage is modified to improve the translation efficiency of a group of genes involved in a particular metabolic process.
Assuntos
Uso do Códon , Lacase/metabolismo , Lignina/metabolismo , Peroxidases/metabolismo , Polyporales/metabolismo , RNA de Transferência/genética , Catálise , Hidrólise , Lacase/genética , Peroxidases/genética , Filogenia , Polyporales/genética , Polyporales/crescimento & desenvolvimentoRESUMO
The genus Antrodiella includes resupinate and pileate species of polypores with a dimitic hyphal system, small, globose to cylindrical basidiospores, absence of cystidia, tetrapolar mating system, and haplo-dikaryotic nuclear behavior. Recent studies, however, indicate that Antrodiella is highly polyphyletic, so many of its species have been transferred to other genera. This study reviews the systematic status and diversity of Antrodiella from the Neotropics based, in part, on studies of type specimens. Collections from Brazil were used for molecular analysis of nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS), nuc 28S rDNA (28S), and portions of genes encoding translation elongation factor 1-α (tef1) and the second largest subunit of RNA polymerase II (rpb2). Eight genera are confirmed to include Neotropical species treated as Antrodiella in a broad sense: Aegis, Antrodiella s. str., Flaviporus, Metuloidea, Mycorrhaphium, Rickiopora, Trametopsis, and Trullella. Molecular data reveal the occurrence of two new species, described as Antrodiella trivialis, the only Neotropical species of Antrodiella s. str. known so far, and Mycorrhaphium hispidum. In addition, Antrodiella luteocontexta was found to nest in the genus Aegis, close to the Grifolaceae and Polyporaceae; therefore, the new combination Aegis luteocontexta is proposed. Comments on the eight Antrodiella-related genera as well as species with uncertain taxonomic position are provided, together with a key to their identification.
Assuntos
Variação Genética , Filogenia , Polyporales/classificação , Polyporales/genética , Brasil , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Fator 1 de Elongação de Peptídeos/genética , Polyporales/isolamento & purificação , RNA Polimerase II/genética , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Clima TropicalRESUMO
AIMS: Isolate and characterize a laccase-encoding gene (lac I) of Phlebia brevispora BAFC 633, as well as cloning and expressing cDNA of lac I in Pichia pastoris. And to obtain a purified and characterized recombinant laccase to analyse the biotechnological application potential. METHODS AND RESULTS: Lac I was cloned and sequenced, it contains 2447 pb obtained by PCR and long-distance inverse PCR. Upstream of the structural region of the laccase gene, response elements such as metals, antioxidants, copper, nitrogen and heat shock were found. The coding region consisted of a 1563-pb ORF encoding 521 amino acids. Lac I was functionally expressed in P. pastoris and it was shown that the gene cloned using the α-factor signal peptide was more efficient than the native signal sequence, in directing the secretion of the recombinant protein. Km and highest kcat /Km values towards ABTS, followed by 2,6-dimethylphenol, were similar to other laccases. Lac I showed tolerance to NaCl and solvents, and nine synthetic dyes could be degraded to different degrees. CONCLUSIONS: Lac I-encoding gene could be successfully sequenced having cis-acting elements located at the regulatory region. It was found that lac I cDNA expressed in P. pastoris using the α-factor signal peptide was more efficient than the native signal sequence. The purified Lac I exhibited high tolerance towards NaCl and various solvents and degraded some recalcitrant synthetic dyes. SIGNIFICANCE AND IMPACT OF THE STUDY: The cis-acting elements may be involved in the transcriptional regulation of laccase gene expression. These results may provide a further insight into potential ways of optimizing fermentation process and also open new frontiers for engineering strong promoters for laccase production. The Lac I stability in chloride and solvents and broad decolorization of synthetic dyes are important for its use in organic synthesis work and degradation of dyes from textile effluents respectively.
Assuntos
Proteínas Fúngicas/genética , Lacase/genética , Lignina/metabolismo , Polyporales/enzimologia , Clonagem Molecular , Estabilidade Enzimática , Proteínas Fúngicas/química , Proteínas Fúngicas/isolamento & purificação , Proteínas Fúngicas/metabolismo , Expressão Gênica , Cinética , Lacase/química , Lacase/isolamento & purificação , Lacase/metabolismo , Pichia/genética , Pichia/metabolismo , Reação em Cadeia da Polimerase , Polyporales/química , Polyporales/genética , Sinais Direcionadores de Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
Rigidoporus microporus (Polyporales, Basidiomycota) syn. Rigidoporus lignosus is the most destructive root pathogen of rubber plantations distributed in tropical and sub-tropical regions. Our primary objective was to characterize Nigerian isolates from rubber tree and compare them with other West African, Southeast Asian and American isolates. To characterize the 20 isolates from Nigeria, we used sequence data of the nuclear ribosomal DNA ITS and LSU, ß-tubulin and translation elongation factor 1-α (tef1) gene sequences. Altogether, 40 isolates of R. microporus were included in the analyses. Isolates from Africa, Asia and South/Central America formed three distinctive clades corresponding to at least three species. No phylogeographic pattern was detected among R. microporus collected from West and Central African rubber plantations suggesting continuous gene flow among these populations. Our molecular phylogenetic analysis suggests the presence of two distinctive species associated with the white rot disease. Phylogenetic analyses placed R. microporus in the Hymenochaetales in the vicinity of Oxyporus. This is the first study to characterize R. microporus isolates from Nigeria through molecular phylogenetic techniques, and also the first to compare isolates from rubber plantations in Africa and Asia.
Assuntos
Hevea/microbiologia , Filogenia , Doenças das Plantas/microbiologia , Polyporales/classificação , Polyporales/isolamento & purificação , Dados de Sequência Molecular , Polyporales/genéticaRESUMO
We investigated the phylogenetic relationships of Postia species from Patagonia with rDNA ITS and LSU sequences, together with morphological, cultural and biological features. All species in the genus were included in a "Postia clade" irrespective of whether their spores were thin- or thick-walled. This clade is characterized by tetrapolar mating, a normal nuclear behavior, metachromatic generative hyphae and absence of fiber hyphae in culture. One subclade merged the austral taxa P. pelliculosa and P. punctata, but otherwise no distinct relationships were found regarding spore shape, spore wall thickness and geographical distribution of taxa. The austral P. venata and the endemic P. carbophila, with thin-walled basidiospores, occupied variable positions in both analyses. Postia caesia from Patagonia grouped within the P. caesia species complex but on a separate branch. In contrast, P. rennyi and P. balsamea from Patagonia corresponded well with strains from other geographic areas. The two austral species in Ryvardenia, R. cretacea and R. campyla, characterized by non-metachromatic hyphae, bipolar mating and an astatocoenocytic nuclear behavior, formed an independent subclade among the dimitic genera of the "Antrodia clade", far from other Postia taxa within which they had been placed previously, supporting their inclusion in a genus of their own. Postia carbophila grouped with other Postia species and not with Postia (Rhodonia) placenta as suggested previously on the basis of morphological comparisons. Instead, the latter species grouped with taxa in the dimitic genus Amyloporia with which it shares heterocytic nuclear behavior. A separation between specimens of Postia pelliculosa and Ryvardenia cretacea from either side of the Pacific (i.e. SE Australia/New Zealand and S Argentina/S Chile) suggests they could be considered different at the species level from a phylogenetic point of view.
Assuntos
Polyporales/classificação , Argentina , Sequência de Bases , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Hifas , Dados de Sequência Molecular , Filogenia , Polyporales/genética , Polyporales/isolamento & purificação , Análise de Sequência de DNA , Especificidade da Espécie , Esporos FúngicosRESUMO
An α-galactosidase was isolated from a culture filtrate of Lenzites elegans (Spreng.) ex Pat. MB445947 grown on citric pectin as carbon source. It was purified to electrophoretic homogeneity by ammonium sulfate precipitation, gel filtration chromatography and anion-exchange chromatography. The relative molecular mass of the native purified enzyme was 158 kDa determined by gel filtration and it is a homodimer (Mr subunits = 61 kDa). The optimal temperature for enzyme activity was in the range 60-80 °C. This α-galactosidase showed a high thermostability, retaining 94 % of its activity after preincubation at 60 °C for 2 h. The optimal pH for the enzyme was 4.5 and it was stable from pH 3 to 7.5 when the preincubation took place at 60 °C for 2 h. It was active against several α-galactosides such as p-nitrophenyl-α-D-galactopyranoside, α-D-melibiose, raffinose and stachyose. The α-galactosidase is a glycoprotein with 26 % of structural sugars. Galactose was a non-competitive inhibitor with a Ki = 22 mM versus p-nitrophenyl-α-D-galactoside and 12 mM versus α-D-melibiose as substrates. Glucose was a simple competitive inhibitor with a Ki = 10 mM. Cations such as Hg(2+) and p-chloromercuribenzoate were also inhibitors of this activity, suggesting the presence of -SH groups in the active site of the enzyme. On the basis of the sequence of the N-terminus (SPDTIVLDGTNFALN) the studied α-galactosidase would be a member of glycosyl hydrolase family 36 (GH 36). Given the high optimum temperature and heat stability of L. elegans α-galactosidase, this fungus may become a useful source of α-galactosidase production for multiple applications.
Assuntos
Proteínas Fúngicas/química , Proteínas Fúngicas/isolamento & purificação , Polyporales/enzimologia , alfa-Galactosidase/química , alfa-Galactosidase/isolamento & purificação , Sequência de Aminoácidos , Estabilidade Enzimática , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Temperatura Alta , Concentração de Íons de Hidrogênio , Cinética , Dados de Sequência Molecular , Peso Molecular , Polyporales/química , Polyporales/genética , Polyporales/metabolismo , Alinhamento de Sequência , Especificidade por Substrato , alfa-Galactosidase/genética , alfa-Galactosidase/metabolismoRESUMO
This is a review of the available knowledge on nuclear behavior of the mycelium within polypore genera (Agaricomycetes, Basidiomycota). Information on 68 genera showed that nuclear behavior is a distinct and consistent feature at genus level and can be coupled with phylogenetic differentiation. The sole exception was found in Polyporus, where different species with normal, heterocytic and astatocoenocytic nuclear behaviors were found. Of the 68 genera treated 41 (60.3%) displayed a normal nuclear behavior, nine (13.2%) were heterocytic, nine (13.2%) were astatocoenocytic and another eight (11.8%) were holocoenocytic. In 95% of the genera a unique compatibility system was found, with the exceptions of Antrodia, which includes both homothallic and bipolar species all associated with a normal nuclear behavior, and Spongipellis, in which bipolar and tetrapolar species are found, all displaying an astatocoenocytic nuclear behavior. Normal and heterocytic nuclear behaviors were associated mostly with tetrapolarity, astatocoenocity was associated mostly with bipolarity, and holocoenocity was associated with either bipolarity or purported homothallism. The combination of nuclear behavior with mating system and brown or white rot capability appeared as a strong feature characterizing and distinguishing the genera of polypores, each combination being valuable to differentiate between apparently related genera, as is supported by phylogenetic studies. Several examples are presented to support this idea, as well as the cases of species that are problematic to this concept. Poroid genera of Hymenochaetaceae were treated apart because of the lack of knowledge regarding their nuclear behavior. In addition new information on the sexuality and/or nuclear behavior of 15 polyporoid taxa is given.
Assuntos
Núcleo Celular/fisiologia , Polyporales/fisiologia , Núcleo Celular/genética , DNA Fúngico/genética , DNA Ribossômico/genética , DNA Espaçador Ribossômico/genética , Genes Fúngicos Tipo Acasalamento , Micélio/genética , Micélio/fisiologia , Filogenia , Polyporales/classificação , Polyporales/genéticaRESUMO
Two laccase isoenzyme genes (lcc2 and lcc3) from the white-rot fungus Coriolopsis rigida were cloned, and together with the previously described lcc1, their transcript levels were analysed by Quantitative RT-PCR in order to study their expression patterns under a range of putative inducers (Cu(2+), Mn(2+), Fe(3+), 2,6-dimethoxy-1,4-benzoquinone, H(2)O(2,) caffeine, amphotericin B and syringic acid). The highest induction was observed for lcc1 in presence of copper, and thus, a kinetic study was performed to analyze its effect on temporary lcc1 gene expression. Our results showed that upregulation due to copper was linked to growth stage, being highest during the trophophase and decreasing during the idiophase. Amphotericin B increased levels of transcripts of lcc1 and lcc2, syringic acid upregulated lcc1 and lcc3 and 2,6-dimethoxy-1,4-benzoquinone induced lcc2 and lcc3. Possible reasons for why laccase genes from C. rigida are differentially regulated at the transcriptional level are discussed.