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1.
Food Chem ; 339: 127884, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32858387

RESUMO

The differences in Dictyophora indusiata fruiting bodies between peach-shaped and mature stage during the postharvest were systematically investigated through quantitative proteomic and metabolomic analyses. A total of 951 differentially expressed proteins were identified, 571 upregulated and 380 downregulated in the mature fruiting body; additionally, 173 upregulated and 165 downregulated differential abundance metabolites were screened. Integrated proteome and metabolome analyses showed that, during the maturation of D. indusiata fruiting bodies, glycerophospholipids were hydrolyzed and drastically decreased, the degradation of glucan was upregulated, the degradation and synthesis of chitin were simultaneously enhanced, and proteins were dominated via catabolism. Along with vigorous material metabolism, energy production was enhanced through the upregulated TCA-cycles and oxidative phosphorylation. In addition, the synthesis of antioxidant substances and the decomposition of peroxides were enhanced in mature fruiting bodies. These omics analyses of D. indusiata provide high-throughput data and reveal the changes in the post-harvest morphological development.


Assuntos
Basidiomycota/metabolismo , Metabolômica/métodos , Proteômica/métodos , Antioxidantes/metabolismo , Basidiomycota/química , Metabolismo dos Carboidratos/genética , Análise por Conglomerados , Regulação para Baixo , Metabolismo Energético/genética , Carpóforos/metabolismo , Proteínas Fúngicas/metabolismo , Metaboloma , Regulação para Cima
2.
Plant Mol Biol ; 104(1-2): 113-136, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32627097

RESUMO

KEY MESSAGE: Present study revealed a complex relationship among histone H3 methylation (examined using H3K4/K27me3 marks), cytosine DNA methylation and differential gene expression during Lr28 mediated leaf rust resistance in wheat. During the present study, genome-wide histone modifications were examined in a pair of near isogenic lines (NILs) (with and without Lr28 in the background of cv. HD2329). The two histone marks used included H3K4me3 (an activation mark) and H3K27me3 (a repression mark). The results were compared with levels of expression (using RNA-seq) and DNA methylation (MeDIP) data obtained using the same pair of NILs. Some of the salient features of the present study include the following: (i) large scale differential binding sites (DBS) were available for only H3K4me3 in the susceptible cultivar, but for both H3K4me3 and H3K27me3 in its resistant NIL; (ii) DBSs for H3K27me3 mark were more abundant (> 80%) in intergenic regions, whereas DBSs for H3K4me3 were distributed in all genomic regions including exons, introns, intergenic, TTS (transcription termination sites) and promoters; (iii) fourteen (14) genes associated with DBSs showed co-localization for both the marks; (iv) only a small fraction (7% for H3K4me3 and 12% for H3K27me3) of genes associated with DBSs matched with the levels of gene expression inferred from RNA-seq data; (v) validation studies using qRT-PCR were conducted on 26 selected representative genes; results for only 11 genes could be validated. The proteins encoded by important genes involved in promoting infection included domains generally carried by R gene proteins such as Mlo like protein, protein kinases and purple acid phosphatase. Similarly, proteins encoded by genes involved in resistance included those carrying domains for lectin kinase, R gene, aspartyl protease, etc. Overall, the results suggest a very complex network of downstream genes that are expressed during compatible and incompatible interactions; some of the genes identified during the present study may be used in future validation studies involving RNAi/overexpression approaches.


Assuntos
Basidiomycota/metabolismo , Resistência à Doença/genética , Genes de Plantas/genética , Genoma de Planta/genética , Histonas/genética , Doenças das Plantas/genética , Triticum/genética , Triticum/metabolismo , Imunoprecipitação da Cromatina , Metilação de DNA , Regulação da Expressão Gênica de Plantas , Ligação Genética , Histonas/metabolismo , Anotação de Sequência Molecular , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/microbiologia , Regiões Promotoras Genéticas , Reprodutibilidade dos Testes , Alinhamento de Sequência , Análise de Sequência , Análise de Sequência de RNA , Transcrição Genética , Triticum/microbiologia
3.
Biochem Biophys Res Commun ; 526(4): 1138-1142, 2020 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-32317185

RESUMO

Brown macroalgae is a promising marine biomass for the production of bioethanol and biodiesel fuels. Here we investigate the biochemical processes used by marine oleaginous yeast for assimilating the major carbohydrate found in brown macroalgae. Briefly, yeast Rhodosporidiobolus fluvialis strain Y2 was isolated from seawater and grown in minimal medium containing reduced sugar alcohol mannitol as the sole carbon source with a salinity comparable to seawater. Conditions limiting nitrogen were used to facilitate lipid synthesis. R. fluvialis Y2 yielded 55.1% (w/w) and 39.1% (w/w) of lipids, per dry cell weight, from mannitol in the absence and presence of salinity, respectively. Furthermore, mannitol, as a sugar source, led to an increase in the composition of polyunsaturated fatty acids, linoleic acid (C18:2) and linolenic acid (C18:3), compared to glucose. This suggests that oxidation of mannitol leads to the activation of NADH-dependent fatty acid desaturases in R. fluvialis Y2. Such fatty acid composition may contribute to the cold-flow properties of biodiesel fuels. Our results identified a salt-tolerant oleaginous yeast species with unique metabolic traits, demonstrating a key role as a decomposer in the global carbon cycle through marine ecosystems. This is the first study on mannitol-induced synthesis of lipids enriched with polyunsaturated fatty acids by marine yeast.


Assuntos
Organismos Aquáticos/metabolismo , Basidiomycota/metabolismo , Ácidos Graxos Insaturados/metabolismo , Manitol/metabolismo , Organismos Aquáticos/ultraestrutura , Basidiomycota/efeitos dos fármacos , Basidiomycota/isolamento & purificação , Basidiomycota/ultraestrutura , Ácidos Graxos Insaturados/biossíntese , Nitrogênio/farmacologia , Oxirredução
4.
Chemosphere ; 253: 126585, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32278187

RESUMO

This pioneering study aimed to determine the activity concentrations of 210Po, 210Pb and uranium (234U, 235U, 238U) radionuclides in fruit bodies of wild bolete Boletus bainiugan Dentinger and to estimate its edible safety, which may give scientific evidence for the consumption of this species. The analyses were performed using alpha spectrometer after digestion, exchange resins separation and deposition. Measurement data were analysed and interpolation maps reflecting 210Po, 210Pb and uranium (234U, 235U, 238U) geographical distribution in Yunnan province (China) were presented. In addition, from the perspective of food safety, the possible related effective radiation dose to mushrooms consumers were estimated. The results indicated that 210Po, 210Pb and uranium (234U, 235U, 238U) radionuclides contents in B. bainiugan were significantly different with respect to geographical distribution, and their possible intake in a part of the region was considerably higher. A very interesting observation was done according to the values of 235U/238U activity ratio indicating the occurrence of uranium faction from the global fallout of nuclear weapon tests.


Assuntos
Basidiomycota/metabolismo , Monitoramento de Radiação/métodos , Poluentes Radioativos do Solo/metabolismo , Urânio/metabolismo , Agaricales , China , Frutas/química , Chumbo/análise , Radioisótopos de Chumbo/metabolismo , Polônio/metabolismo , Radioisótopos/análise , Poluentes Radioativos do Solo/análise , Urânio/análise
5.
Appl Environ Microbiol ; 86(10)2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32144102

RESUMO

Vibralactone, a hybrid compound derived from phenols and a prenyl group, is a strong pancreatic lipase inhibitor with a rare fused bicyclic ß-lactone skeleton. Recently, a researcher reported a vibralactone derivative (compound C1) that caused inhibition of pancreatic lipase with a half-maximal inhibitory concentration of 14 nM determined by structure-based optimization, suggesting a potential candidate as a new antiobesity treatment. In the present study, we sought to identify the main gene encoding prenyltransferase in Stereum vibrans, which is responsible for the prenylation of phenol leading to vibralactone synthesis. Two RNA silencing transformants of the identified gene (vib-PT) were obtained through Agrobacterium tumefaciens-mediated transformation. Compared to wild-type strains, the transformants showed a decrease in vib-PT expression ranging from 11.0 to 56.0% at 5, 10, and 15 days in reverse transcription-quantitative PCR analysis, along with a reduction in primary vibralactone production of 37 to 64% at 15 and 21 days, respectively, as determined using ultra-high-performance liquid chromatography-mass spectrometry analysis. A soluble and enzymatically active fusion Vib-PT protein was obtained by expressing vib-PT in Escherichia coli, and the enzyme's optimal reaction conditions and catalytic efficiency (Km /k cat) were determined. In vitro experiments established that Vib-PT catalyzed the C-prenylation at C-3 of 4-hydroxy-benzaldehyde and the O-prenylation at the 4-hydroxy of 4-hydroxy-benzenemethanol in the presence of dimethylallyl diphosphate. Moreover, Vib-PT shows promiscuity toward aromatic compounds and prenyl donors.IMPORTANCE Vibralactone is a lead compound with a novel skeleton structure that shows strong inhibitory activity against pancreatic lipase. Vibralactone is not encoded by the genome directly but rather is synthesized from phenol, followed by prenylation and other enzyme reactions. Here, we used an RNA silencing approach to identify and characterize a prenyltransferase in a basidiomycete species that is responsible for the synthesis of vibralactone. The identified gene, vib-PT, was expressed in Escherichia coli to obtain a soluble and enzymatically active fusion Vib-PT protein. In vitro characterization of the enzyme demonstrated the catalytic mechanism of prenylation and broad substrate range for different aromatic acceptors and prenyl donors. These characteristics highlight the possibility of Vib-PT to generate prenylated derivatives of aromatics and other compounds as improved bioactive agents or potential prodrugs.


Assuntos
Basidiomycota/metabolismo , Dimetilaliltranstransferase/metabolismo , Proteínas Fúngicas/metabolismo , Lactonas/metabolismo , Basidiomycota/enzimologia , Escherichia coli/metabolismo , Microrganismos Geneticamente Modificados/metabolismo , Interferência de RNA , Reação em Cadeia da Polimerase Via Transcriptase Reversa
6.
An Acad Bras Cienc ; 92(1): e20181174, 2020 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-32187277

RESUMO

Grape pomace from Vitis labrusca is an important sub-product of the "American table wine" industry. It is recalcitrant to degradation, and its accumulation is a serious problem with negative environmental impacts. We analyzed the ability of five white-rot fungi to transform this residue in-vitro. Mass loss and phenol removal in grape pomace treated with each fungus were compared after 30-day solid-state fermentation. Since Peniophora albobadia isolate LPSC 285 was the fungus that showed the highest degradative ability and the lowest free phenol levels in the residue transformed, we selected this fungus to monitor its effect on this residue after 30, 60, and 90 days of incubation. We analyzed mass loss of the residue caused by the fungus activity and its chemical changes using Fourier transform infrared spectroscopy. After 90 days of incubation, Peniophora albobadia isolate LPSC 285 reduced grape pomace mass by 20.48%, which was associated with degradation of polysaccharides and aromatic structures. We concluded that Peniophora albobadia LPSC # 285 isolate is a promising fungus to transform grape pomace from Vitis labrusca under solid-state fermentation conditions.


Assuntos
Basidiomycota/metabolismo , Biotransformação , Fermentação , Vitis/química , Resíduos Industriais , Fenóis/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo , Vitis/metabolismo
7.
BMC Genomics ; 21(1): 181, 2020 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-32093624

RESUMO

BACKGROUND: Sporobolomyces pararoseus is regarded as an oleaginous red yeast, which synthesizes numerous valuable compounds with wide industrial usages. This species hold biotechnological interests in biodiesel, food and cosmetics industries. Moreover, the ballistospores-shooting promotes the colonizing of S. pararoseus in most terrestrial and marine ecosystems. However, very little is known about the basic genomic features of S. pararoseus. To assess the biotechnological potential and ballistospores-shooting mechanism of S. pararoseus on genome-scale, the whole genome sequencing was performed by next-generation sequencing technology. RESULTS: Here, we used Illumina Hiseq platform to firstly assemble S. pararoseus genome into 20.9 Mb containing 54 scaffolds and 5963 predicted genes with a N50 length of 2,038,020 bp and GC content of 47.59%. Genome completeness (BUSCO alignment: 95.4%) and RNA-seq analysis (expressed genes: 98.68%) indicated the high-quality features of the current genome. Through the annotation information of the genome, we screened many key genes involved in carotenoids, lipids, carbohydrate metabolism and signal transduction pathways. A phylogenetic assessment suggested that the evolutionary trajectory of the order Sporidiobolales species was evolved from genus Sporobolomyces to Rhodotorula through the mediator Rhodosporidiobolus. Compared to the lacking ballistospores Rhodotorula toruloides and Saccharomyces cerevisiae, we found genes enriched for spore germination and sugar metabolism. These genes might be responsible for the ballistospores-shooting in S. pararoseus NGR. CONCLUSION: These results greatly advance our understanding of S. pararoseus NGR in biotechnological potential and ballistospores-shooting, which help further research of genetic manipulation, metabolic engineering as well as its evolutionary direction.


Assuntos
Basidiomycota/genética , Genoma Fúngico , Filogenia , Sequenciamento Completo do Genoma , Basidiomycota/metabolismo , Biotecnologia , Metabolismo dos Carboidratos , Carotenoides/metabolismo , Genômica , Metabolismo dos Lipídeos , Análise de Sequência de RNA
8.
Cell Biochem Biophys ; 78(1): 111-119, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32062829

RESUMO

There are different varieties of mushrooms not yet studied spread all over the planet. The objective of this study was to evaluate biochemical properties and effects on mitochondrial respiration of eight Basidiomycete mushrooms: Flaviporus venustus EF30, Hydnopolyporus fimbriatus EF41 and EF44, Inonotus splitgerberi EF46, Oudemansiella canarii EF72, Perenniporia sp. EF79, Phellinus linteus EF81, and Pleurotus albidus EF84. Total phenols, ABTS, TEAC, FRAP, and ORAC were measured in order to determine the antioxidant capacity. Antimicrobial potential was studied by disc-diffusion and microdilution method. Cytotoxicity was determined in murine peritoneal macrophages. The bioenergetic aspects were evaluated by the uncoupling of the oxidative phosphorylation in mitochondrias. The H. fimbriatus mushroom was the one that presented the most significant results for the antioxidant assays. Three mushrooms presented antimicrobial activity, indicating a potential for formulation of drugs. The results suggest that I. spligerberi has an uncoupling activity, even at the lowest concentration tested, dissipating the mitochondrial electrochemical gradient. On the other hand, P. albidus has effect only on succinate-oxidase activity without influencing mitochondrial respiratory efficiency. Therefore, both interfere negatively in mitochondrial respiration. In relation with the cytotoxicity in peritoneal macrophages, O. canarii and F. venustus were cytotoxic in this type of cells.


Assuntos
Basidiomycota/química , Mitocôndrias/efeitos dos fármacos , Fenóis/química , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Antioxidantes/química , Antioxidantes/metabolismo , Basidiomycota/metabolismo , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/metabolismo , Mitocôndrias/metabolismo , Oxirredução , Fosforilação Oxidativa/efeitos dos fármacos , Consumo de Oxigênio/efeitos dos fármacos , Fenóis/isolamento & purificação , Fenóis/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Ácido Succínico/química
9.
Carbohydr Polym ; 230: 115680, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31887951

RESUMO

Three main fractions, one neutral polysaccharide (ACPN-1a) and two acidic polysaccharides (ACPA-1a and ACPA-2a), were isolated from crude polysaccharides of Auricularia cornea var. Li. (ACP). The physicochemical properties and structural characteristics of these fractions were investigated using High Performance Gel Permeation Chromatography (HPGPC), High Performance Liquid Chromatography (HPLC), Scanning Electron Microscopy (SEM), Fourier Transform Infrared (FTIR), and Nuclear Magnetic Resonance (NMR) spectra. The results showed that ACPN-1a with a higher molecular weight (∼2.18 × 106 Da) was a ß-(1→3)-glucan with (1→6)-linked ß-Glcp residues for side chains. ACPA-1a was a heteropolysaccharide and had the backbone of (1→3)-linked ß-Glcp and (1→6)-linked α-Manp and side chains with ß-Xylp or ß-GlcpA residues. ACPA-2a had the average molecular weight of 8.5 × 105 Da and was composed of a (1→6)-linked α-Manp backbone and side chains with ß-Xylp or ß-GlcpA residues. The data provided useful information for further investigation on the structure-activity relationship of Auricularia cornea var. Li. polysaccharides.


Assuntos
Basidiomycota/metabolismo , Polissacarídeos/química , Polissacarídeos/isolamento & purificação
10.
PLoS One ; 15(1): e0227478, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31899917

RESUMO

Indole-3-acetic acid (IAA) is an imperative phytohormone for plant growth and development. Ectomycorrhizal fungi (ECM) are able to produce IAA. However, only a few studies on IAA biosynthesis pathways in ECM fungi have been reported. This study aimed to investigate the IAA biosynthesis pathway of six ECM cultures including Astraeus odoratus, Gyrodon suthepensis, Phlebopus portentosus, Pisolithus albus, Pisolithus orientalis and Scleroderma suthepense. The results showed that all ECM fungi produced IAA in liquid medium that had been supplemented with L-tryptophan. Notably, fungal IAA levels vary for different fungal species. The detection of indole-3-lactic acid and indole-3-ethanol in the crude culture extracts of all ECM fungi indicated an enzymatic reduction of indole-3-pyruvic acid and indole-3-acetaldehyde, respectively in the IAA biosynthesis via the indole-3-pyruvic acid pathway. Moreover, the tryptophan aminotransferase activity confirmed that all ECM fungi synthesize IAA through the indole-3-pyruvic acid pathway. Additionally, the elongation of rice and oat coleoptiles was stimulated by crude culture extract. This is the first report of the biosynthesis pathway of IAA in the tested ECM fungi.


Assuntos
Basidiomycota/metabolismo , Ácidos Indolacéticos/metabolismo , Micorrizas/metabolismo , Reguladores de Crescimento de Planta/metabolismo , Cromatografia Líquida de Alta Pressão , Colorimetria , Proteínas Fúngicas/metabolismo , Ácidos Indolacéticos/análise , Reguladores de Crescimento de Planta/análise , Triptofano/metabolismo , Triptofano Transaminase/metabolismo
11.
Cryobiology ; 92: 26-33, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-31580830

RESUMO

A number of living creatures in the Antarctic region have developed characteristic adaptation of cold weather by producing antifreeze proteins (AFP). Antifreeze peptide (Afp1m) fragment have been designed in the sequence of strings from native proteins. The objectives of this study were to assess the properties of Afp1m to cryopreserve skin graft at the temperature of -10 °C and -20 °C and to assess sub-zero injuries in Afp1m cryopreserved skin graft using light microscopic techniques. In the present study, a process was developed to cryopreserve Sprague-Dawley (SD) rat skin grafts with antifreeze peptide, Afp1m, α-helix peptide fragment derived from Glaciozyma antractica yeast. Its viability assessed by different microscopic techniques. This study also described the damages caused by subzero temperatures (-10 and -20 °C) on tissue cryopreserved in different concentrations of Afp1m (0.5, 1, 2, 5 and 10 mg/mL) for 72 h. Histological scores of epidermis, dermis and hypodermis of cryopreserved skin grafts showed highly significant difference (p < 0.01) among the different concentrations at -10 and -20 °C. In conclusion, the integrity of cryopreserved skin grafts with lower concentrations of Afp1m (0.5, 1 and 2 mg/mL) or at -20 °C was not maintained. The present study attested that Afp1m is a good cryoprotective agent for the cryopreservation of skin graft. Higher Afp1m concentrations (5 and 10 mg/mL) at -10 °C found to be suitable for the future in vivo study using (SD) rat skin grafts.


Assuntos
Proteínas Anticongelantes/farmacologia , Criopreservação/métodos , Crioprotetores/farmacologia , Transplante de Pele/métodos , Aclimatação/fisiologia , Animais , Regiões Antárticas , Basidiomycota/metabolismo , Congelamento , Masculino , Conformação Proteica em alfa-Hélice , Ratos , Ratos Sprague-Dawley , Pele
12.
J Ind Microbiol Biotechnol ; 47(1): 145-154, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31734813

RESUMO

Putative iron-reductase (IR) genes from Serpula lacrymans with similarity to the conserved iron-binding domains of cellobiose dehydrogenase (CDH) enzymes have been identified. These genes were cloned and expressed to functionally characterize their activity and role in the decomposition of lignocellulose. The results show that IR1 and IR2 recombinant enzymes have the ability to depolymerize both lignin and cellulose, are capable of the reduction of ferric iron to the ferrous form, and are capable of the degradation of nitrated lignin. Expression of these genes during wheat straw solid-state fermentation was shown to correlate with the release of compounds associated with lignin decomposition. The results suggest that both IR enzymes mediate a non-enzymatic depolymerisation of lignocellulose and highlight the potential of chelator-mediated Fenton systems in the industrial pre-treatment of biomass.


Assuntos
Basidiomycota/metabolismo , FMN Redutase/metabolismo , Lignina/metabolismo , Basidiomycota/genética , Fenômenos Bioquímicos , Fermentação , Triticum/metabolismo
13.
Prep Biochem Biotechnol ; 50(1): 66-73, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31502910

RESUMO

Response surface methodology was applied to maximize the yield and production of carotenoids by Sporidiobolus pararoseus WZ012 using citrus juice. A high concentration of yeast extract and citrus juice favored carotenoid production and biomass concentration, respectively. Under optimal conditions, a more than 51 percent (from 860 to 1300 µg/g) and 62 percent (from 17.05 to 27.66 mg/L) respective enhancement in intracellular and total carotenoid production was achieved. Finally, this process was successfully upscaled in a 5-L fermentor. A comparison of the carotenoid distributions revealed that torulene (61.3%) was the dominant carotenoid when using the citrus based medium, while the main carotenoid was ß-carotene (62.5%) when using the glucose medium. The present work provides an alternative method to produce high-value products derived from waste and low-grade citrus.


Assuntos
Basidiomycota/metabolismo , Citrus/metabolismo , Microbiologia Industrial/métodos , Biomassa , Reatores Biológicos , Carotenoides/metabolismo , Fermentação , Glucose/metabolismo , Microbiologia Industrial/instrumentação
14.
New Phytol ; 225(2): 880-895, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31529497

RESUMO

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), poses a tremendous threat to the production of wheat worldwide. The molecular mechanisms of Pst effectors that regulate wheat immunity are poorly understood. In this study, we identified an effector Pst18363 from Pst that suppresses plant cell death in Nicotiana benthamiana and in wheat. Knocking down Pst18363 expression by virus-mediated host-induced gene silencing significantly decreased the number of rust pustules, indicating that Pst18363 functions as an important pathogenicity factor in Pst. Pst18363 was proven to interact with wheat Nudix hydrolase 23 TaNUDX23. In wheat, silencing of TaNUDX23 by virus-induced gene silencing increased reactive oxygen species (ROS) accumulation induced by the avirulent Pst race CYR23, whereas overexpression of TaNUDX23 suppressed ROS accumulation induced by flg22 in Arabidopsis. In addition, TaNUDX23 suppressed Pst candidate effector Pst322-trigged cell death by decreasing ROS accumulation in N. benthamiana. Knocking down of TaNUDX23 expression attenuated Pst infection, indicating that TaNUDX23 is a negative regulator of defence. In N. benthamiana, Pst18363 stabilises TaNUDX23. Overall, our data suggest that Pst18363 stabilises TaNUDX23, which suppresses ROS accumulation to facilitate Pst infection.


Assuntos
Basidiomycota/metabolismo , Proteínas Fúngicas/metabolismo , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Triticum/microbiologia , Basidiomycota/patogenicidade , Morte Celular , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Interações Hospedeiro-Patógeno , Doenças das Plantas/imunologia , Imunidade Vegetal , Proteínas de Plantas/genética , Ligação Proteica , Estabilidade Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Tabaco/citologia , Triticum/citologia , Triticum/genética , Triticum/imunologia , Regulação para Cima/genética
15.
Int J Biol Macromol ; 146: 875-886, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31726131

RESUMO

De novo transcriptome assembly and shotgun proteome analysis of Dictyophora indusiata fruiting bodies were performed. A total of 19,704 unigenes were sequenced, and 4380 proteins were identified. Annotation and functional analysis of the identified proteins were significantly enriched in small molecule synthetic and metabolic processes, protein modification regulation (phosphorylation and ubiquitination), and vesicle transport. Furthermore, quantitative developmental transcriptome analysis was performed between the peach-shaped and mature fruiting bodies, and the results revealed that the metabolism and transport activities were upregulated in the mature stage, while protein translation was downregulated; this regulation is likely the main reason for the significant changes in the nutrients of fruiting bodies. Furthermore, the cell wall stress-dependent MAPK sub-pathway was activated in the mature stage, and fungal cell wall degradation-related genes were upregulated, which could promote reconstruction of the cell wall and might play a key role in the morphological development of D. indusiata fruiting bodies.


Assuntos
Basidiomycota/crescimento & desenvolvimento , Basidiomycota/genética , Carpóforos/crescimento & desenvolvimento , Carpóforos/genética , Perfilação da Expressão Gênica , Proteoma/metabolismo , Transcriptoma/genética , Basidiomycota/enzimologia , Basidiomycota/metabolismo , Carpóforos/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Ontologia Genética , Genes Fúngicos , Sistema de Sinalização das MAP Quinases/genética , Anotação de Sequência Molecular
16.
Biosci Biotechnol Biochem ; 84(3): 433-444, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31738699

RESUMO

Ascomycota and basidiomycota fungi are prolific sources of biologically active natural products. Recent genomic data and bioinformatic analysis indicate that fungi possess a large number of biosynthetic gene clusters for bioactive natural products but more than 90% are silent. Heterologous expression in the filamentous fungi as hosts is one of the powerful tools to expression of the silent gene clusters. This review introduces recent studies on the total biosynthesis of representative family members via common platform intermediates, genome mining of novel di- and sesterterpenoids including detailed cyclization pathway, and development of expression host for basidiomycota genes with efficient genome editing method. In addition, this review will discuss the several strategies, for the generation of structural diversity, which are found through these studies.


Assuntos
Ascomicetos/metabolismo , Basidiomycota/metabolismo , Produtos Biológicos/metabolismo , Ascomicetos/genética , Basidiomycota/genética , Vias Biossintéticas/genética , Genes Fúngicos , Família Multigênica
17.
J Med Food ; 23(2): 153-160, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31747350

RESUMO

Oxidative stress triggered by hyperglycemia is thought to be a major factor in the development of liver disease during diabetes mellitus (DM). The aim of this study was to determine the antioxidant capacity of aqueous extracts of selenium-enriched Auricularia auricular (AESA) and further investigate the hepatic protection and potential mechanism of AESA in a mouse model of diabetes. An in vitro antioxidant assay confirmed that AESA exhibited better antioxidant capacity characterized by increased reducing power and scavenging capacity of free radicals, such as diphenyl picrylhydrazyl radical, hydroxyl radical, and superoxide radical. The diabetic model was induced by high-fat diet combined with a single injection of streptozotocin in C57BL/6 mice. Our results showed that AESA treatment improved diabetes-induced disorders of lipid metabolisms and alleviated liver damage in diabetic mice. Furthermore, the antioxidant enzyme activities of glutathione peroxidase and catalase in liver were increased and malondialdehyde level was decreased with AESA treatment compared with those in the DM group. In parallel, AESA significantly reduced the contents of tumor necrosis factor-alpha and interleukin-1beta in liver in comparison with the DM group. In addition, western blot results showed that the expression of receptor for advanced glycation end products (RAGE), phospho-c-Jun NH2-terminal kinase, phospho-extracellular signal-regulated kinase, and phospho-p38 kinase were remarkably decreased in AESA treatment group compared with DM group. Taken together, supplementation of AESA may effectively attenuate diabetic hepatopathology by exerting antioxidant function through the RAGE/mitogen-activated protein kinase pathway.


Assuntos
Antioxidantes/farmacologia , Basidiomycota/metabolismo , Dieta Hiperlipídica/efeitos adversos , Hepatopatias/tratamento farmacológico , Selênio/farmacologia , Animais , Diabetes Mellitus Experimental , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Teste de Tolerância a Glucose , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Hepatopatias/etiologia , Masculino , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Estreptozocina/efeitos adversos , Fator de Necrose Tumoral alfa/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
18.
Anal Bioanal Chem ; 412(2): 449-462, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31797019

RESUMO

In the last decades, microbial oils have been extensively investigated as a renewable platform for biofuel and oleochemical production. Offering a potent alternative to plant-based oils, oleaginous microorganisms have been the target of ongoing metabolic engineering aimed at increasing growth and lipid yields, in addition to specialty fatty acids. Discovery proteomics is an attractive tool for elucidating lipogenesis and identifying metabolic bottlenecks, feedback regulation, and competing biosynthetic pathways. One prominent microbial oil producer is Cutaneotrichosporon oleaginosus, due to its broad feedstock catabolism and high lipid yield. However, this yeast has a recalcitrant cell wall and high cell lipid content, which complicates efficient and unbiased protein extraction for downstream proteomic analysis. Optimization efforts of protein sample preparation from C. oleaginosus in the present study encompasses the comparison of 8 lysis methods, 13 extraction buffers, and 17 purification methods with respect to protein abundance, proteome coverage, applicability, and physiochemical properties (pI, MW, hydrophobicity in addition to COG, and GO analysis). The optimized protocol presented in this work entails a one-step extraction method utilizing an optimal lysis method (liquid homogenization), which is augmented with a superior extraction buffer (50 mM Tris, 8/2 M Urea/Thiourea, and 1% C7BzO), followed by either of 2 advantageous purification methods (hexane/ethanol or TCA/acetone), depending on subsequent applications and target studies. This work presents a significant step forward towards implementation of efficient C. oleaginosus proteome mining for the identification of potential targets for genetic optimization of this yeast to improve lipogenesis and production of specialty lipids. Graphical abstract.


Assuntos
Basidiomycota/metabolismo , Proteínas Fúngicas/metabolismo , Proteômica , Proteínas Fúngicas/isolamento & purificação , Lipídeos/isolamento & purificação , Proteólise , Solubilidade
19.
Int J Biol Macromol ; 144: 231-241, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31843615

RESUMO

Genome data mining of the Antarctic yeast, Glaciozyma antarctica PI12 revealed an expansin-like protein encoding sequence (GaEXLX1). The GaEXLX1 protein is 24.8 kDa with a high alkaline pI of 9.81. Homology modeling of GaEXLX1 showed complete D1 and D2 domains of a conventional expansin. The protein exhibited 36% sequence similarity to Clavibacter michiganensis EXLX1 (PDB: 4JCW). Subsequently, a recombinant GaEXLX1 protein was produced using Escherichia coli expression system. Incubation with Avicel, filter paper and cotton fiber showed that the protein can disrupt the surface of crystalline and pure cellulose, suggesting a cell wall modification activity usually exhibited by expansin-like proteins. Binding assays displayed that GaEXLX1 can bind to polymeric substrates, including those postulated to be present in the sea ice ecosystem such as crab chitin and moss lichenan. GaEXLX1 may assist in the recognition and loosening of these substrates in the sea ice prior to hydrolysis by other extracellular enzymes. Similar loosening mechanism to classical expansin-like protein has been postulated for this psychrophilic protein based on several conserved residues of GaEXLX1 involved in binding interaction identified by docking analyses.


Assuntos
Basidiomycota/metabolismo , Proteínas Fúngicas/metabolismo , Simulação de Acoplamento Molecular , Nutrientes/metabolismo , Sequência de Aminoácidos , Regiões Antárticas , Parede Celular/metabolismo , Celulose/química , Ecossistema , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Fúngicas/química , Camada de Gelo , Simulação de Dinâmica Molecular , Filogenia , Proteínas Recombinantes , Alinhamento de Sequência
20.
Ecotoxicol Environ Saf ; 188: 109935, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31740233

RESUMO

Chromium (Cr) is one of the most toxic heavy metals and a health hazard to millions of people worldwide. Ectomycorrhizal (ECM) fungi can assist plants in phytoremediation of heavy metal contaminated soil. Cr tolerance differs among ECM fungal varieties, but the underlying molecular mechanisms of Cr tolerance in ECM fungi are not clear. This study identified, analysed and compared the Cr(VI)-induced transcriptional changes between Cr(VI)-tolerant strain (Pisolithus sp. 1 LS-2017) and Cr(VI)-sensitive strain (Pisolithus sp. 2 LS-2017) by de novo transcriptomic analysis. The results showed that 93,642 assembled unique transcripts representing the 22,353 (46.76%) unigenes matched the proteins we have known in the Nr database and 47,801 unigenes were got from the Pisolithus spp. For DEGs between the control and 10 mg/L Cr(VI) treatment, cyanoamino acid metabolic, type I diabetes mellitus metabolism, nitrogen metabolism and beta-Alanine metabolism pathways were significantly enriched (p < 0.05) in Pisolithus sp. 1 LS-2017. Two nitrate reductase family genes (nidD, niiA) provide Cr(VI) tolerance for Pisolithus sp. 1 LS-2017 by regulating Cr(VI) reduction. In addition, NO produced by nidD, niiA regulated denitrification can alleviate Cr(VI) induced oxidative stress. In Pisolithus sp. 2 LS-2017, the alcC, aldA and lcf2 gene may alleviate Cr(VI) induced oxidative stress by protecting SH groups and increasing secondary metabolism, reducing detoxify aldehydes to carboxylic acids and producing LCPUFAs respectively; .T gene regulate Cr(VI) induced wound healing by pigmentation and stability of melanin in spore; MKP2 gene accelerate Cr(VI) induced cell death and gpmA gene regulated Cr(VI) induced energy emergency.


Assuntos
Basidiomycota/genética , Cromo/toxicidade , Genes Fúngicos , Micorrizas/genética , Poluentes do Solo/toxicidade , Transcriptoma/efeitos dos fármacos , Adaptação Fisiológica , Basidiomycota/metabolismo , Biodegradação Ambiental , Desnitrificação , Perfilação da Expressão Gênica , Micorrizas/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Solo/química , Microbiologia do Solo/normas
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