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1.
Phytopathology ; 112(10): 2072-2083, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35522048

ABSTRACT

Bacterial wilt, caused by the Ralstonia solanacearum species complex (RSSC), is the most destructive potato disease in Kenya. Studies were conducted to (i) determine the molecular diversity of RSSC strains associated with bacterial wilt of potato in Kenya, (ii) generate an RSSC distribution map for epidemiological inference, and (iii) determine whether phylotype II sequevar 1 strains exhibit epidemic clonality. Surveys were conducted in 2018 and 2019, in which tubers from wilting potato plants and stem samples of potential alternative hosts were collected for pathogen isolation. The pathogen was phylotyped by multiplex PCR and 536 RSSC strains typed at a sequevar level. Two RSSC phylotypes were identified, phylotype II (98.4%, n = 506 [sequevar 1 (n = 505) and sequevar 2 (n = 1)]) and phylotype I (1.6%, n = 30 [sequevar 13 (n = 9) and a new sequevar (n = 21)]). The phylotype II sequevar 1 strains were haplotyped using multilocus tandem repeat sequence typing (TRST) schemes. The TRST scheme identified 51 TRST profiles within the phylotype II sequevar 1 strains with a modest diversity index (HGDI = 0.87), confirming the epidemic clonality of RSSC phylotype II sequevar 1 strains in Kenya. A minimum spanning tree and mapping of the TRST profiles revealed that TRST27 '8-5-12-7-5' is the primary founder of the clonal complex of RSSC phylotype II sequevar 1 and is widely distributed via latently infected seed tubers. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Subject(s)
Ralstonia solanacearum , Solanum tuberosum , Kenya/epidemiology , Phylogeny , Plant Diseases/microbiology , Ralstonia , Ralstonia solanacearum/genetics , Solanum tuberosum/microbiology
2.
Methods Mol Biol ; 2354: 375-385, 2021.
Article in English | MEDLINE | ID: mdl-34448170

ABSTRACT

Potato bacterial wilt is caused by the devastating bacterial pathogen Ralstonia solanacearum. Quantitative resistance to this disease has been and is currently introgressed from a number of wild relatives into cultivated varieties through laborious breeding programs. Here, we present two methods that we have developed to facilitate the screening for resistance to bacterial wilt in potato. The first one uses R. solanacearum reporter strains constitutively expressing the luxCDABE operon or the green fluorescent protein (gfp) to follow pathogen colonization in potato germplasm. Luminescent strains are used for nondestructive live imaging, while fluorescent ones enable precise pathogen visualization inside the plant tissues through confocal microscopy. The second method is a BIO-multiplex-PCR assay that is useful for sensitive and specific detection of viable R. solanacearum (IIB-1) cells in latently infected potato plants. This BIO-multiplex-PCR assay can specifically detect IIB-1 sequevar strains as well as strains belonging to all four R. solanacearum phylotypes and is sensitive enough to detect without DNA extraction ten bacterial cells per mL in complex samples.The described methods allow the detection of latent infections in roots and stems of asymptomatic plants and were shown to be efficient tools to assist potato breeding programs.


Subject(s)
Ralstonia solanacearum , Solanum tuberosum , Multiplex Polymerase Chain Reaction , Operon , Plant Diseases , Ralstonia solanacearum/genetics
3.
Biotechnol Biofuels ; 14(1): 96, 2021 Apr 17.
Article in English | MEDLINE | ID: mdl-33865436

ABSTRACT

BACKGROUND: Wood-decay basidiomycetes are effective for the degradation of highly lignified and recalcitrant plant substrates. The degradation of lignocellulosic materials by brown-rot strains is carried out by carbohydrate-active enzymes and non-enzymatic Fenton mechanism. Differences in the lignocellulose catabolism among closely related brown rots are not completely understood. Here, a multi-omics approach provided a global understanding of the strategies employed by L. sulphureus ATCC 52600 for lignocellulose degradation. RESULTS: The genome of Laetiporus sulphureus ATCC 52600 was sequenced and phylogenomic analysis supported monophyletic clades for the Order Polyporales and classification of this species within the family Laetiporaceae. Additionally, the plasticity of its metabolism was revealed in growth analysis on mono- and disaccharides, and polysaccharides such as cellulose, hemicelluloses, and polygalacturonic acid. The response of this fungus to the presence of lignocellulosic substrates was analyzed by transcriptomics and proteomics and evidenced the occurrence of an integrated oxidative-hydrolytic metabolism. The transcriptomic profile in response to a short cultivation period on sugarcane bagasse revealed 125 upregulated transcripts, which included CAZymes (redox enzymes and hemicellulases) as well as non-CAZy redox enzymes and genes related to the synthesis of low-molecular-weight compounds. The exoproteome produced in response to extended cultivation time on Avicel, and steam-exploded sugarcane bagasse, sugarcane straw, and Eucalyptus revealed 112 proteins. Contrasting with the mainly oxidative profile observed in the transcriptome, the secretomes showed a diverse hydrolytic repertoire including constitutive cellulases and hemicellulases, in addition to 19 upregulated CAZymes. The secretome induced for 7 days on sugarcane bagasse, representative of the late response, was applied in the saccharification of hydrothermally pretreated grass (sugarcane straw) and softwood (pine) by supplementing a commercial cocktail. CONCLUSION: This study shows the singularity of L. sulphureus ATCC 52600 compared to other Polyporales brown rots, regarding the presence of cellobiohydrolase and peroxidase class II. The multi-omics analysis reinforces the oxidative-hydrolytic metabolism involved in lignocellulose deconstruction, providing insights into the overall mechanisms as well as specific proteins of each step.

4.
Plant Dis ; 105(4): 958-964, 2021 Apr.
Article in English | MEDLINE | ID: mdl-32886041

ABSTRACT

Resistance to sterol demethylation inhibitor (DMI) fungicides in Monilinia fructicola, causal agent of brown rot of stone fruit, has been reported in the southeastern and eastern United States and in Brazil. DMI resistance of some M. fructicola isolates, in particular those recovered from the southeastern United States, is associated with a sequence element termed "Mona" that causes overexpression of the cytochrome demethylase target gene MfCYP51. In this study, we conducted statewide surveys of Michigan stone fruit orchards from 2009 to 2011 and in 2019, and we determined the sensitivity to propiconazole of a total of 813 isolates of M. fructicola. A total of 80.7% of Michigan isolates were characterized as resistant to propiconazole by relative growth assays, but the Mona insert was not uniformly detected and was present in some isolates that were not characterized as DMI resistant. Gene expression assays indicated that elevated expression of MfCYP51 was only weakly correlated with DMI resistance in M. fructicola isolates from Michigan, and there was no obvious correlation between the presence of the Mona element and elevated expression of MfCYP51. However, sequence analysis of MfCYP51 from 25 DMI-resistant isolates did not reveal any point mutations that could be correlated with resistance. Amplification and sequencing upstream of MfCYP51 resulted in detection of DNA insertions in a wide range of isolates typed by DMI phenotype and the presence of Mona or other unique sequences. The function of these unique sequences or their presence upstream of MfCYP51 cannot be correlated to a DMI-resistant genotype at this time. Our results indicate that DMI resistance was established in Michigan populations of M. fructicola by 2009 to 2011, and that relative resistance levels have continued to increase to the point that practical resistance is present in most orchards. In addition, the presence of the Mona insert is not a marker for identifying DMI-resistant isolates of M. fructicola in Michigan.


Subject(s)
Fungicides, Industrial , Ascomycota , Brazil , Demethylation , Drug Resistance, Fungal/genetics , Fungicides, Industrial/pharmacology , Michigan , Southeastern United States
5.
Plant Dis ; 2020 Aug 25.
Article in English | MEDLINE | ID: mdl-32896216

ABSTRACT

Eggplant (Solanum melongena L.) is an important solanaceous crop that is produced mainly in tropical and subtropical regions and is widely consumed worldwide. In 2018, eggplant production in Mexico was approximately 80000 t, and Sinaloa State contributed 96% of this production; however, this crop suffers significant losses from plant pathogens. In December 2019, fruits from commercial orchards (geographical coordinates: 24°45'39.39''N, 107°26'57.30''O) with visible brown soft rot and profuse white mycelia were analysed. On V8 medium, pieces of tissue obtained from the border of lesions were plated and incubated between 25°C and 36°C. After five days, a dense cottony mycelium with a slightly petaloid pattern was observed at 25°C and did not grow at 36°C. Isolates of that pathogen were heterothallic, and microscopic preparations showed development of coenocytic mycelium and spheroid sporangia that were noncaducous and papillate, measuring 35.6 ± 5.8 x 27.1 ± 4.4. Based on morphological characteristics, the eggplant soft rot causal agent was identified as Phytophthora nicotianae Breda de Haan (Erwin and Ribeiro, 1996). From a representative isolate denominated PhySm01, two DNA regions (internal transcribed spacer (ITS) and the large subunit ribosomal (28S)) were amplified and sequenced with ITS1-ITS4 and NL1-LR3 primers, respectively. The obtained ITS sequence (GenBank accession number MT508842) showed 100% identity with several P. nicotianae sequences (Access MT065840, MH290435 and MG570057) with 100% query coverage and 740 matching nucleotides. For the 28S sequence (accession number MT508843), the identity with strains N° Access EU080889 and EU080508 of P. nicotianae was 99.86%, with 100 query coverage and 729 matching nucleotides. Further, phylogenetic analysis from P. nicotianae strain PhySm01 and GenBank reference sequences was carried out by Maximum Likelihood method with Mega 7 software based on the ITS sequences, which verified the species identification. To fulfill Koch's postulates, a suspension containing 1 x 104 zoospores/mL of the oomycete isolated from the original diseased eggplant fruit was used to inoculate ten healthy and disinfested fruits. Sterile water was used as a control. Three wounds per fruit were made with a sterile needle, and 20 µL of the zoospore suspension (or water) was placed on each. All fruits were placed into plastic bags with moistened paper and incubated at 25°C for three days. Thisest was repeated twice with similar results. Initial symptoms developed 24 hours after inoculation with brown soft tissue forming around the inoculated area, and profuse soft rot accompanied by white mycelium was observed two days after inoculation. No symptoms developed on the control fruits during this time. Pieces of necrotic tissue were plated on V8 medium and incubated as described previously. The reisolated pathogen was compared to the original isolate and had the same morphological characteristics. Phytophthora nicotianae has a worldwide distribution and can infect multiple solanaceous crops, including tobacco (Gallup et al., 2018). In addition, it has been reported on other hosts outside of the Solanaceae family, causing economically important losses in citrus and strawberry. In India, it causes necrosis of citrus fruits and roots, leading to tree decline (Das et al., 2016), and in the United States, it causes crown rot of strawberry (Marin et al., 2018). In Mexico, P. nicotianae has been reported on vinca (Alvarez-Rodriguez et al., 2013); however, this is the first report of P. nicotianae causing fruit soft rot of eggplant, and this pathogen represents a new threat when the environment is favourable for disease outbreaks. References Alvarez-Rodriguez, B. et al. 2013. Plant Dis. 97: 1257. https://doi.org/10.1094/PDIS-04-13-0400-PDN Das, A. K. et al. 2016. J. Plant Pathol. 98: 55. DOI: 10.4454/JPP.V98I1.038 Erwin, D. C. and Ribeiro, O. K. 1996. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN. 562 Pp. Gallup, C. A. et al. 2018. Plan Dis. 102: 1108. https://doi.org/10.1094/PDIS-02-17-0295-RE Marin, M. V. et al. 2018. Plant Dis. 102:1463. https://doi.org/10.1094/PDIS-08-17-1333-PDN.

6.
Environ Sci Pollut Res Int ; 27(8): 8467-8480, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31902077

ABSTRACT

The native state of lignocellulosic biomass is highly resistant to enzymatic hydrolysis and the fermentation process of biofuel production. Brown-rot fungi use an extracellular Fenton system to degrade lignocellulosic biomass in the initial stages of decay. In this work, the combined effects of Mn2+, Fe2+, and NO3- inducers were evaluated based on the activities of hydrolytic enzymes and Fe3+ reduction as well as the catechol-type compound production during wheat straw pretreatment by the brown-rot fungus Gloeophyllum trabeum. Weight loss and chemical changes were evaluated to establish the culture conditions for stimulating wheat straw degradation using a central composite design. The results showed that weight loss and the Fe3+-reducing activity were promoted at the highest concentrations of Fe2+. A positive effect on catechol compound production by the addition of Mn2+ and NO3- was observed. Cellulase activity was increased at the highest concentration of NO3-. The multiple optimizations of G. trabeum culture conditions in wheat straw resulted in 11.3% weight loss and 0.47 total crystallinity index at 0.24 M NO3-, 0.95 mM Fe2+, and 0.85 mM Mn2+ after 40 days. The wheat straw pretreatment by G. trabeum for 10 days increased glucose recovery. The results indicated that the wheat straw pretreatment using G. trabeum with biodegradation inducers could be a complementary step to physicochemical pretreatment of lignocellulosic biomass for production of second-generation ethanol.


Subject(s)
Basidiomycota , Ethanol/chemistry , Triticum , Basidiomycota/chemistry , Hydrolysis , Lignin , Triticum/chemistry
7.
Rev. cient. (Guatem.) ; 28(1): [32]-[43], 20181107.
Article in Spanish | LILACS-Express | LILACS | ID: biblio-963803

ABSTRACT

Neolentinus ponderosus y N. lepideus son dos especies de hongos saprobios que poseen uso alimenticio tradicional en comunidades de los departamentos de Huehuetenango y Totonicapán, Guatemala. La capacidad degradadora de ambas especies les confiere potencial para el estudio de la producción de cuerpos fructíferos a nivel de sustrato, por lo que en esta investigación se evaluó el crecimiento de una cepa nativa de N. ponderosus y una de N. lepideus en el medio de cultivo agar extracto de malta (EMA) a diferente pH y también se estudió la degradación de la madera de dos especies de pino (Pinus tecunumanii y P. ayacahuite) en cámaras de podredumbre durante 12 meses. El mayor diámetro de crecimiento miceliar de la cepa N. ponderosus se observó a pH 7.0, en tanto que para N. lepideus se obtuvo a pH 5.0 y 5.6. Las colonias de ambas cepas evidenciaron textura afelpada, borde regular a irregular, color blanco, con o sin pigmento difusible, olor frutal, hifas de 1.0-5.0 µm de diámetro, con clamidosporas y fíbulas. La madera de P. tecunumanii degradada por la cepa de N. ponderosus y la madera de P. ayacahuite por la cepa N. lepideus, presentaron porcentajes de pérdida de peso de 8.76 (±5.58) y 12.07 (±5.66), respectivamente y en ambos casos se alcanzó solamente el estadio temprano de degradación. Los resultados de este estudio sientan las bases para investigaciones posteriores en los que se evalúe el cultivo en troncos de ambas especies para la producción de cuerpos fructíferos con fines alimenticios y comerciales


Neolentinus ponderosus and N. lepideus are two saprophytic fungi species used traditionally in Huehuetenango and Totonicapán, Guatemala. The degradative capacity of both species confers them potential for fruiting bodies production. This study evaluated the mycelial growth of two native strains of N. ponderosus and N. lepideus in malt extract agar (EMA) at different pH and the degradation of wood from two pine species in rot chambers during 12 months. pH 7.0 was the most appropriate for the mycelial growth of N. ponderosus and for N. lepideus were 5.0 and 5.6. The colonies of both strains showed fruity odor, velvety texture, regular to irregular edge, white color, with or without diffusible pigment, hyphae with 1-5 µm width, chlamydospores and clamp connections. Wood from Pinus tecunumanii and P. ayacahuite exhibit weight-loss percentages between 8.76 ± 5.58 and 12.07 ± 5.66, with N. ponderosus 145.2003 and N. lepideus 90.2002, respectively. In both cases reached the early stage of brown-rot decay. These results could be useful for future research that evaluate the fruiting bodies production in logs for food and commercial purposes

8.
Waste Manag ; 79: 240-250, 2018 Sep.
Article in English | MEDLINE | ID: mdl-30343752

ABSTRACT

White-rot and brown-rot fungi have complementary mechanisms to selectively degrade lignin and holocellullose, respectively. Thereby, a fungal co-culture of a white-rot and a brown-rot fungal could result in efficient strategy for a mild lignocellulosic biomass pretreatment. In this work, single, sequential and co-inoculation of the selective-lignin degrading white-rot fungus Ganoderma lobatum and the brown-rot fungus Gloeophyllum trabeum were evaluated as biological pretreatments of wheat straw to enhance enzymatic hydrolysis of cellulose. The single cultures of G. lobatum and G. trabeum exhibited preferential degradation of lignin and hemicellulose, respectively. The total crystallinity index decreased in samples pretreated with G. trabeum but not with G. lobatum. The pretreatment with single cultures of G. lobatum or G. trabeum increased glucose yields by 43.6% and 26.1% respectively compared to untreated straw. Although co-inoculation resulted in higher yields of glucose when compared with single cultures, only a slight synergistic effect between fungi was observed. Contrary, the sequential inoculation of G. lobatum incubated for 10 days followed by G. trabeum incubated for 10 days more showed a strong synergic effect on enzymatic hydrolysis. This sequential culture showed the highest glucose yield (191.5 mg g-1 wheat straw), 2.8-fold higher than untreated wheat straw, and 140-150% higher than the single-cultures of G. lobatum and G. trabeum, respectively.


Subject(s)
Basidiomycota , Triticum , Fungi , Hydrolysis , Lignin
9.
Sci. agric ; 74(6): 489-491, Nov.-Dec.2017. graf, tab
Article in English | VETINDEX | ID: biblio-1497670

ABSTRACT

Healthy peach fruit is not considered a preferred host for Drosophila suzukii (Matsumura); however, it becomes a more preferable host when damaged. Thus, damaged peach fruit is of importance not only in terms of economic losses, but also because it serves as a reservoir for this fly species. This work aimed to evaluate the suitability of peaches mechanically damaged or harboring the brown rot disease as hosts for D. suzukii compared to hosts for which they have a low (undamaged peach) and high (undamaged strawberry) preference. Damaged peaches were as susceptible as undamaged strawberries. Fungus infection alone did not increase the susceptibility of peaches compared to undamaged peaches. This information should be considered when an Integrated Pest Management program is defined in a scenario of different species of fruit production.


Subject(s)
Animals , Pest Control/history , Pest Control/trends , Drosophila/classification , Prunus persica/adverse effects
10.
Sci. agric. ; 74(6): 489-491, Nov.-Dec.2017. graf, tab
Article in English | VETINDEX | ID: vti-15603

ABSTRACT

Healthy peach fruit is not considered a preferred host for Drosophila suzukii (Matsumura); however, it becomes a more preferable host when damaged. Thus, damaged peach fruit is of importance not only in terms of economic losses, but also because it serves as a reservoir for this fly species. This work aimed to evaluate the suitability of peaches mechanically damaged or harboring the brown rot disease as hosts for D. suzukii compared to hosts for which they have a low (undamaged peach) and high (undamaged strawberry) preference. Damaged peaches were as susceptible as undamaged strawberries. Fungus infection alone did not increase the susceptibility of peaches compared to undamaged peaches. This information should be considered when an Integrated Pest Management program is defined in a scenario of different species of fruit production.(AU)


Subject(s)
Animals , Drosophila/classification , Prunus persica/adverse effects , Pest Control/history , Pest Control/trends
11.
Front Plant Sci ; 8: 1221, 2017.
Article in English | MEDLINE | ID: mdl-28769944

ABSTRACT

The current bacterial wilt infestation level in the potato fields in the Peruvian Andes was investigated by collecting stem samples from wilted plants and detecting Ralstonia solanacearum. In total 39 farmers' fields located in the central and northern Peru between the altitudes 2111 and 3742 m above sea level were sampled. R. solanacearum was detected in 19 fields, and in 153 out of the 358 samples analyzed. Phylogenetic analysis using the partial sequence of the endoglucanase gene on strains collected in Peru between 1966 and 2016 from potato, pepper, tomato, plantain or soil, divided the strains in phylotypes I, IIA, and IIB. The Phylotype IIB isolates formed seven sequevar groups including the previously identified sequevars 1, 2, 3, 4, and 25. In addition to this, three new sequevars of phylotype IIB were identified. Phylotype IIA isolates from Peru clustered together with reference strains previously assigned to sequevars 5, 39, 41, and 50, and additionally one new sequevar was identified. The Phylotype I strain was similar to the sequevar 18. Most of the Peruvian R. solanacearum isolates were IIB-1 strains. In the old collection sampled between 1966 and 2013, 72% were IIB-1 and in the new collection at 2016 no other strains were found. The pathogenicity of 25 isolates representing the IIA and IIB sequevar groups was tested on potato, tomato, eggplant and tobacco. All were highly aggressive on potato, but differed in pathogenicity on the other hosts, especially on tobacco. All IIA strains caused latent infection on tobacco and some strains also caused wilting, while IIB strains caused only few latent infections on this species. In conclusion, high molecular diversity was found among the R. solanacearum strains in Peru. Most of the variability was found in areas that are no longer used for potato cultivation and thus these strains do not pose a real threat for potato production in the country. Compared to the previous data from the 1990s, the incidence of bacterial wilt has decreased in Peru. The epidemics are likely caused by infected seed tubers carrying the clonal brown rot strain IIB-1.

12.
J Agric Food Chem ; 64(20): 4029-47, 2016 May 25.
Article in English | MEDLINE | ID: mdl-27133976

ABSTRACT

Brown rot (BR) caused by Monilinia spp., has been an economic problem for the stone fruit market due to dramatic losses, mainly during the postharvest period. There is much literature about basic aspects of Monilinia spp. infection, which indicates that environment significantly influences its occurrence in the orchard. However, progress is needed to sustainably limit this disease: the pathogen is able to develop resistance to pesticides, and most of BR resistance research programs in plant models perish. Solving this problem becomes important due to the need to decrease chemical treatments and reduce residues on fruit. Thus, research has recently increased, exploring a wide range of disease control strategies (e.g., genetic, chemical, physical). Summarizing this information is difficult, as studies evaluate different Monilinia and Prunus model species, with diverse strategies and protocols. Thus, the purpose of this review is to present the diversity and distribution of agents causing BR, focusing on the biochemical mechanisms of Monilinia spp. infection both of the fungi and of the fruit, and report on the resistance sources in Prunus germplasm. This review comprehensively compiles the information currently available to better understand mechanisms related to BR resistance.


Subject(s)
Ascomycota/physiology , Plant Diseases/microbiology , Prunus/microbiology , Ascomycota/genetics , Ascomycota/isolation & purification , Fruit/microbiology , Plant Diseases/prevention & control
13.
Can J Microbiol ; 62(5): 442-8, 2016 May.
Article in English | MEDLINE | ID: mdl-26991236

ABSTRACT

Bacterial wilt is a devastating plant disease caused by the bacterial pathogen Ralstonia solanacearum species complex and affects different crops. Bacterial wilt infecting potato is also known as brown rot (BR) and is responsible for significant economic losses in potato production, especially in developing countries. In Bolivia, BR affects up to 75% of the potato crop in areas with high incidence and 100% of stored potatoes. The disease has disseminated since its introduction to the country in the mid-1980s mostly through contaminated seed tubers. To avoid this, local farmers multiply seed tubers in highlands because the strain infecting potatoes cannot survive near-freezing temperatures that are typical in the high mountains. Past disease surveys have shown an increase in seed tubers with latent infection in areas at altitudes lower than 3000 m a.s.l. Since global warming is increasing in the Andes Mountains, in this work, we explored the incidence of BR in areas at altitudes above 3000 m a.s.l. Results showed BR presence in the majority of these areas, suggesting a correlation between the increase in disease incidence and the increase in temperature and the number of irregular weather events resulting from climate change. However, it cannot be excluded that the increasing availability of latently infected seed tubers has boosted the spread of BR.


Subject(s)
Climate Change , Plant Diseases/microbiology , Ralstonia solanacearum , Solanum tuberosum/microbiology , Bolivia , Cold Temperature , Temperature
14.
Electron. j. biotechnol ; Electron. j. biotechnol;18(6): 451-458, Nov. 2015. ilus, graf, tab
Article in English | LILACS | ID: lil-772290

ABSTRACT

Background Laccase has been considered important for the degradation of lignocellulose by wood rot fungi. The properties and functions of laccase in white rot fungi have been investigated extensively, but those from brown rot fungi remain largely unknown. In this paper, a laccase isoform Pplcc2 from the brown rot fungus Postia placenta MAD-698-R was expressed heterologously in Pichia pastoris GS115, purified and the properties of the enzyme were determined. Results The molecular weight of the protein was determined to be 67 kDa using SDS-PAGE. It cannot oxidize syringaldazine (SGZ), but it can oxidize 2,2'-azino-di-(3-ethylbenzothialozin-6-Sulfonic acid) (ABTS) and 2,6-dimethoxyphenol (DMP). Specific activity for ABTS was 1960 ± 19 Unit/mg. The catalytic constant (k cat) was 1213 ± 18.3 s-1 for ABTS and 293.2 ± 21.9 s-1 for DMP. Km was 22.08 µM for ABTS and 11.62 µM for DMP. The optimal pH for the oxidation of ABTS and DMP was 3.5 and 5.0 respectively. The optimal temperature for the oxidation of ABTS and DMP was 60°C. Conclusions This is the first identified thermo activated and thermostable laccase in brown rot fungi. This investigation will contribute to understanding the roles played by laccases in brown rot fungi.


Subject(s)
Wood/microbiology , Laccase , Coriolaceae/enzymology , Pichia , Temperature , Enzyme Stability , Electrophoresis, Polyacrylamide Gel
15.
Acta amaz ; Acta amaz;43(4): 429-438, Dec. 2013. tab, ilus
Article in Portuguese | LILACS-Express | LILACS, VETINDEX | ID: biblio-1455163

ABSTRACT

This work aimed to evaluate the use of colorimetry in the characterization of biodeterioration marupá, jequitibá and cumaru wood, submitted to white and brown rot. For this purpose, specimens of each Amazonian species were submitted to accelerated decay tests, according to American Society for Testing and Materials - ASTM D2017, during 20 weeks. With a colorimeter, the colorimetric parameters L* (lightness), a* (green-red coordinate), b* (blue-yellow coordinate) and total color change (ΔE) were determined weekly until the fourth week, and then every two weeks until the twentieth week. Additionally, it was determined the mass loss. The results indicated that the colorimetry was effective in monitoring of biodeterioration of wood, as well as to differentiate white and brown rot. The best predictions of natural resistance to white and brown rot were obtained with the parameters b* and L*, respectively. We also emphasizes the use of ΔE to estimate the biodeterioration of wood, because of their significant correlations with weight loss.


Objetivou-se avaliar o emprego da colorimetria na caracterização da biodeterioração das madeiras de marupá, jequitibá e cumaru submetidas ao ataque de fungos de podridão branca e parda. Para tanto, corpos de prova de cada espécie amazônica foram submetidos a ensaios de apodrecimento acelerado, de acordo com a American Society for Testing and Materials - ASTM D2017, durante 20 semanas. Os parâmetros colorimétricos L* (luminosidade), a* (coordenada verde-vermelho), b* (coordenada amarelo-azul) e a variação total da cor (ΔE) foram determinados semanalmente até a quarta semana, e posteriormente a cada duas semanas até a vigésima semana, com auxílio de um espectrofotocolorímetro. Adicionalmente, foram determinadas as perdas de massa dos corpos de prova. Os resultados permitiram destacar que a colorimetria foi eficaz no monitoramento da biodeterioração da madeira, bem como para diferenciação da podridão branca e parda. As melhores predições da resistência natural aos fungos de podridão branca e parda foram obtidas com os parâmetros b* e L*, respectivamente. Enfatiza-se também, o emprego da ΔE como parâmetro auxiliar na estimativa da biodeterioração da madeira, em razão de suas correlações significativas com a perda de massa.

16.
Acta amaz. ; 43(4): 429-438, dez. 2013. tab, ilus
Article in Portuguese | VETINDEX | ID: vti-20321

ABSTRACT

This work aimed to evaluate the use of colorimetry in the characterization of biodeterioration marupá, jequitibá and cumaru wood, submitted to white and brown rot. For this purpose, specimens of each Amazonian species were submitted to accelerated decay tests, according to American Society for Testing and Materials - ASTM D2017, during 20 weeks. With a colorimeter, the colorimetric parameters L* (lightness), a* (green-red coordinate), b* (blue-yellow coordinate) and total color change (ΔE) were determined weekly until the fourth week, and then every two weeks until the twentieth week. Additionally, it was determined the mass loss. The results indicated that the colorimetry was effective in monitoring of biodeterioration of wood, as well as to differentiate white and brown rot. The best predictions of natural resistance to white and brown rot were obtained with the parameters b* and L*, respectively. We also emphasizes the use of ΔE to estimate the biodeterioration of wood, because of their significant correlations with weight loss.(AU)


Objetivou-se avaliar o emprego da colorimetria na caracterização da biodeterioração das madeiras de marupá, jequitibá e cumaru submetidas ao ataque de fungos de podridão branca e parda. Para tanto, corpos de prova de cada espécie amazônica foram submetidos a ensaios de apodrecimento acelerado, de acordo com a American Society for Testing and Materials - ASTM D2017, durante 20 semanas. Os parâmetros colorimétricos L* (luminosidade), a* (coordenada verde-vermelho), b* (coordenada amarelo-azul) e a variação total da cor (ΔE) foram determinados semanalmente até a quarta semana, e posteriormente a cada duas semanas até a vigésima semana, com auxílio de um espectrofotocolorímetro. Adicionalmente, foram determinadas as perdas de massa dos corpos de prova. Os resultados permitiram destacar que a colorimetria foi eficaz no monitoramento da biodeterioração da madeira, bem como para diferenciação da podridão branca e parda. As melhores predições da resistência natural aos fungos de podridão branca e parda foram obtidas com os parâmetros b* e L*, respectivamente. Enfatiza-se também, o emprego da ΔE como parâmetro auxiliar na estimativa da biodeterioração da madeira, em razão de suas correlações significativas com a perda de massa.(AU)

17.
Bioresour Technol ; 148: 624-7, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24080442

ABSTRACT

Dioxins are a class of extremely hazardous molecules that might pose a threat to the environment. This work evaluated the microbial degradation of 1,2,3,4-tetrachlorodibenzo-p-dioxin (1,2,3,4-TCDD), in liquid broth using three brown-rot fungi and one white-rot fungi as control. A fast and reliable extraction method with recoveries of over 98% together with a validated GC-MS method was developed, and applied to quantify 1,2,3,4-TCDD in liquid broth, mycelia and reaction flask, with detection limits of 10 ppb. Among the four strains tested, brown-rot fungus Aspergillus aculeatus showed best results, removing up to 21% of dioxin after 30-day incubation. The results open both a path for biotechnological interest in bioremediation purposes and environmental behavior studies by using brown-rot fungus.


Subject(s)
Basidiomycota/metabolism , Culture Media/metabolism , Polychlorinated Dibenzodioxins/analogs & derivatives , Analysis of Variance , Biodegradation, Environmental , Biomass , Gas Chromatography-Mass Spectrometry , Linear Models , Polychlorinated Dibenzodioxins/chemistry , Polychlorinated Dibenzodioxins/metabolism , Reproducibility of Results
18.
Ciênc. rural ; Ciênc. rural (Online);43(4): 623-630, abr. 2013. ilus, tab
Article in Portuguese | LILACS | ID: lil-669376

ABSTRACT

Objetivou-se avaliar o emprego de ensaios de dureza Rockwell na caracterização da biodeterioração das madeiras de Simarouba amara (marupá), Cariniana micrantha (jequitibá) e Dipteryx odorata (cumaru), submetidas ao ataque dos fungos apodrecedores Trametes versicolor (podridão branca) e Gloeophyllum trabeum (podridão parda). Para tanto, corpos de prova de cada espécie amazônica foram submetidos a ensaios de apodrecimento acelerado, de acordo com a ASTM D2017 (2005), durante 20 semanas. A caracterização da dureza Rockwell foi realizada semanalmente até a 4ª semana e, posteriormente, a cada duas semanas, até a 20ª semana. Adicionalmente, foram determinadas as perdas de massa, conforme critérios estabelecidos pela ASTM D2017 (2005). Quanto aos resultados, destaca-se que a dureza Rockwell pode ser empregada como ferramenta principal na caracterização da biodeterioração da madeira, sendo sensível na predição dos estágios iniciais. Dentre as madeiras amazônicas, o cumaru apresentou a maior resistência natural aos fungos apodrecedores. Com relação aos fungos, a podridão parda atacou mais intensamente as três madeiras amazônicas.


This study aimed to evaluate the use of Rockwell hardness in the characterization of the biodeterioration of Simarouba amara (marupá), Cariniana micrantha (jequitibá) e Dipteryx odorata (cumaru) woods, which were submitted to decay fungi Trametes versicolor (white rot) and Gloeophyllum trabeum (brown rot). For this purpose, specimens of each Amazon species were submitted to decay fungi, according to ASTM D2017 (2005). The characterization of the Rockwell hardness was performed weekly until the fourth week, and then every two weeks until the twentieth week. Additionally, it was determined the mass loss, according to criteria established by ASTM D2017 (2005). The results indicated that the Rockwell hardness can be used as a primary tool in the characterization of the biodeterioration of wood, being sensitive in predicting the early stages. Among the Amazon woods, cumaru wood showed a greater natural resistance to decay fungi. With respect to decay fungi, brown rot more intensely attacked the three Amazon woods.

19.
Ci. Rural ; 43(4)2013.
Article in Portuguese | VETINDEX | ID: vti-708574

ABSTRACT

This study aimed to evaluate the use of Rockwell hardness in the characterization of the biodeterioration of Simarouba amara (marupá), Cariniana micrantha (jequitibá) e Dipteryx odorata (cumaru) woods, which were submitted to decay fungi Trametes versicolor (white rot) and Gloeophyllum trabeum (brown rot). For this purpose, specimens of each Amazon species were submitted to decay fungi, according to ASTM D2017 (2005). The characterization of the Rockwell hardness was performed weekly until the fourth week, and then every two weeks until the twentieth week. Additionally, it was determined the mass loss, according to criteria established by ASTM D2017 (2005). The results indicated that the Rockwell hardness can be used as a primary tool in the characterization of the biodeterioration of wood, being sensitive in predicting the early stages. Among the Amazon woods, cumaru wood showed a greater natural resistance to decay fungi. With respect to decay fungi, brown rot more intensely attacked the three Amazon woods.


Objetivou-se avaliar o emprego de ensaios de dureza Rockwell na caracterização da biodeterioração das madeiras de Simarouba amara (marupá), Cariniana micrantha (jequitibá) e Dipteryx odorata (cumaru), submetidas ao ataque dos fungos apodrecedores Trametes versicolor (podridão branca) e Gloeophyllum trabeum (podridão parda). Para tanto, corpos de prova de cada espécie amazônica foram submetidos a ensaios de apodrecimento acelerado, de acordo com a ASTM D2017 (2005), durante 20 semanas. A caracterização da dureza Rockwell foi realizada semanalmente até a 4ª semana e, posteriormente, a cada duas semanas, até a 20ª semana. Adicionalmente, foram determinadas as perdas de massa, conforme critérios estabelecidos pela ASTM D2017 (2005). Quanto aos resultados, destaca-se que a dureza Rockwell pode ser empregada como ferramenta principal na caracterização da biodeterioração da madeira, sendo sensível na predição dos estágios iniciais. Dentre as madeiras amazônicas, o cumaru apresentou a maior resistência natural aos fungos apodrecedores. Com relação aos fungos, a podridão parda atacou mais intensamente as três madeiras amazônicas.

20.
Ci. Rural ; 43(4)2013.
Article in Portuguese | VETINDEX | ID: vti-708288

ABSTRACT

This study aimed to evaluate the use of Rockwell hardness in the characterization of the biodeterioration of Simarouba amara (marupá), Cariniana micrantha (jequitibá) e Dipteryx odorata (cumaru) woods, which were submitted to decay fungi Trametes versicolor (white rot) and Gloeophyllum trabeum (brown rot). For this purpose, specimens of each Amazon species were submitted to decay fungi, according to ASTM D2017 (2005). The characterization of the Rockwell hardness was performed weekly until the fourth week, and then every two weeks until the twentieth week. Additionally, it was determined the mass loss, according to criteria established by ASTM D2017 (2005). The results indicated that the Rockwell hardness can be used as a primary tool in the characterization of the biodeterioration of wood, being sensitive in predicting the early stages. Among the Amazon woods, cumaru wood showed a greater natural resistance to decay fungi. With respect to decay fungi, brown rot more intensely attacked the three Amazon woods.


Objetivou-se avaliar o emprego de ensaios de dureza Rockwell na caracterização da biodeterioração das madeiras de Simarouba amara (marupá), Cariniana micrantha (jequitibá) e Dipteryx odorata (cumaru), submetidas ao ataque dos fungos apodrecedores Trametes versicolor (podridão branca) e Gloeophyllum trabeum (podridão parda). Para tanto, corpos de prova de cada espécie amazônica foram submetidos a ensaios de apodrecimento acelerado, de acordo com a ASTM D2017 (2005), durante 20 semanas. A caracterização da dureza Rockwell foi realizada semanalmente até a 4ª semana e, posteriormente, a cada duas semanas, até a 20ª semana. Adicionalmente, foram determinadas as perdas de massa, conforme critérios estabelecidos pela ASTM D2017 (2005). Quanto aos resultados, destaca-se que a dureza Rockwell pode ser empregada como ferramenta principal na caracterização da biodeterioração da madeira, sendo sensível na predição dos estágios iniciais. Dentre as madeiras amazônicas, o cumaru apresentou a maior resistência natural aos fungos apodrecedores. Com relação aos fungos, a podridão parda atacou mais intensamente as três madeiras amazônicas.

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