207 results on '"Azevedo JL"'
Search Results
2. Effects of IGF-1 onIn VitroCulture of Bovine Preantral Follicles are Dose-Dependent
- Author
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Jimenez, CR, primary, de Azevedo, JL, additional, Silveira, RG, additional, Penitente-Filho, J, additional, Carrascal-Triana, EL, additional, Zolini, AM, additional, Araujo, VR, additional, Torres, CAA, additional, and Gonçalves, WG, additional
- Published
- 2016
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3. Potential of Metarhizium anisopliae (Metsch.) Sorokin (Ascomycetes, hypocreales) in the control of Bonagota salubricola (Meyrick) (Lepidoptera, Tortricidae) and its compatibility with chemical insecticides
- Author
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Anhalt, FA., Azevedo, JL., Sugayama, RL., Specht, A., and Barros, NM.
- Subjects
lagarta enroladeira da macieira ,apple ,biological control ,entomopathogenic fungi ,fungos entomopatogênicos ,maçã ,leafroller ,controle biológico - Abstract
Several insects are harmful to apples grown in Brazil, especially the leafroller Bonagota salubricola (Lepidoptera, Tortricidae), usually controlled with chemical insecticides. The purpose of the present study was to investigate the use of Metarhizium anisopliae strains in the control of the apple leafroller, by assessing their virulence to B. salubricola larvae in bioassays with suspensions of 2 x 10(5) to 2 x 10(9) conidia/mL as well as their relationship with protease expression. The most effective strain underwent a compatibility test with chemical insecticides. The M. anisopliae E6 strain showed a good performance, with up to 88% mortality and a LT50 of 1.66 days. The virulence was positively correlated with a higher enzymatic activity. The E6 strain was compatible with tebufenozide, evidencing its potential to control B. salubricola. Diversos insetos são nocivos à macieira no Brasil, destacando-se a lagarta-enroladeira Bonagota salubricola (Lepidoptera: Tortricidae), controlada com inseticidas químicos. Este trabalho objetivou avaliar linhagens de Metarhizium anisopliae passíveis de serem utilizadas no controle desta praga, determinando-se a virulência sobre larvas de B. salubricola em bioensaios com suspensões de 2 x 10(5) a 2 x 10(9) conídios/mL considerando-se mortalidade larval e pupal, bem como a sua relação com a expressão de proteases. A linhagem mais efetiva foi submetida a teste de compatibilidade com inseticidas químicos. A linhagem E6 de M. anisopliae destacou-se por proporcionar mortalidade de 88% com TL50 de 1,66 dias; tal virulência relacionou-se positivamente com maior produção enzimática. Esta linhagem foi compatível com tebufenozide, evidenciado seu potencial no controle de B. salubricola.
- Published
- 2010
4. Pathogenicity of four strains of entomopathogenic fungi against the bovine tick Boophilus microplus
- Author
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de Barros Nm, Onofre Sb, Azevedo Jl, and Miniuk Cm
- Subjects
Entomopathogenic fungi ,Male ,General Veterinary ,Metarhizium anisopliae var. anisopliae ,fungi ,Body Weight ,Fungi ,General Medicine ,Biology ,Tick ,Pathogenicity ,biology.organism_classification ,Metarhizium flavoviride ,Microbiology ,Conidium ,Ticks ,parasitic diseases ,Fungal strain ,Bioassay ,Animals ,Cattle ,Female ,Pest Control, Biological - Abstract
Objective—To evaluate the pathogenicity of 4 strains of the entomopathogenic fungi Metarhizium flavoviride var flavoviride and Metarhizium anisopliae var anisopliae in vitro against the bovine tick Boophilus microplus. Sample Population—300 B microplus females were used for each series of 5 conidial suspensions of the 4 fungal strains; a control group of 60 females was treated with sterilized water. Procedure—For each fungal strain, sporulating cultures were used to produce conidial suspensions containing 104, 105, 106, 107, and 108 conidia/ml. A bioassay was performed by immersing B microplus females in conidial suspensions; the procedure was repeated with a new group of 60 females for each dilution (a total of 300). A control group was immersed in the same volume of sterilized distilled water. Ticks were maintained at 27 C and 80% relative humidity for 7 days, and the following variables were evaluated: weight before egging, egging weight, percentage eclosion, reproductive efficiency, and percentage control. Results—A low index of eclosion was found in the treated groups, with a progressive decrease in percentage eclosion (control = 100%) with increased conidial concentration. The highest level of effectiveness of biocontrol was detected in groups treated with concentrations of 107 and 108 conidia/ml. Conclusions and Clinical Relevance—Strains of the entomopathogenic fungi M anisopliae var anisopliae and M flavoviride var flavoviride were effective as biological control agents against B microplus, with M flavoviridestrain CG-291 being the most effective. The pathogenicity of these strains and the development of an application method for this fungus for control of ticks may result in improved animal productivity and a reduction in environmental pollution through the use of fewer chemical insecticides. (Am J Vet Res 2001;62:1478–1480)
- Published
- 2001
5. Effects of Growth Hormone on In Situ Culture of Bovine Preantral Follicles are Dose Dependent.
- Author
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Jimenez, CR, Azevedo, JL, Silveira, RG, Penitente‐Filho, J, Carrascal‐Triana, EL, Zolini, AM, Araújo, VR, and Torres, CAA
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BOS , *SOMATOTROPIN , *OVARIAN atresia , *OVUM , *OVARIES , *REPRODUCTION - Abstract
Content The objective of this study was to evaluate different concentrations of growth hormone ( GH) on the development of bovine preantral follicles cultured included in the ovarian tissue ( in situ) on the rates of morphologically normal, viable, primordial and developing follicles, as well as the oocyte and follicle diameter and ultrastructural analysis. Ovarian fragments collected from cows with no cross-breeds defined were cultured in situ for 1 and 7 days in minimal essential medium ( α- MEM+) supplemented with different concentrations of recombinant human GH (0, 10, 25, 50 ng/ml). The ovarian fragments non-cultured (control) and cultured were processed for classic histology, mechanical isolation and electron transmission microscopy ( MET). The parameters underwent anova (Tukey′s and Dunnett′s tests) and chi-square test ( χ2). After 7 days of culture, the treatment with 50 ng/ml GH showed no differences with fresh control (p > 0.05) and had greater effectiveness than in the 0, 10 and 25 ng/ml GH concentrations of the morphologically normal follicles. Regarding the primordial follicles, a reduction was observed in the 50 ng/ml GH concentration concomitant with the significant increase in developing follicles, differing from both the fresh control and the other GH concentrations tested. In addition, 50 ng/ml GH showed a larger follicle and oocyte diameter when compared to the other treatments cultured. Similar structures were ultrastructurally observed in the control group, 50 ng/ml GH. Follicles cultured in 10 ng/ml GH showed nuclear invagination, vacuoles and lesioned basal membrane. Hence, it is concluded that 50 ng/ml GH is the most effective concentration for the development of preantral follicles cultured in situ. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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6. Effects of IGF-1 on In Vitro Culture of Bovine Preantral Follicles are Dose-Dependent.
- Author
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Jimenez, CR, Azevedo, JL, Silveira, RG, Penitente‐Filho, J, Carrascal‐Triana, EL, Zolini, AM, Araujo, VR, Torres, CAA, and Gonçalves, WG
- Subjects
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SOMATOMEDIN C , *BOS , *GROWTH factors , *ULTRASTRUCTURE (Biology) , *QUANTITATIVE research , *IN vitro studies , *REPRODUCTION - Abstract
Contents This study aimed at assessing the effect of different concentrations of the growth factor similar to insulin 1 ( IGF-1) in the development, survival and ultrastructure of the bovine preantral follicles cultured in situ. Fragments of bovine ovarian cortical tissue were cultured during 1 and 7 days in 1 ml of α- MEM+, supplemented with different concentrations of human recombinant IGF-1 (0, 30, 70 and 100 ng/ml), in an incubator at 37°C and 5% of CO2 in 24-well plates with total replacement of the medium every 2 days. Non-cultured ovarian fragments (control) and ovarian fragments cultured during 1 and 7 days were processed for classic histology, mechanical isolation and electron transmission microscopy ( ETM). Parameters such as normality, viability, activation, development, diameter and ultrastructure were evaluated. All statistical analyses were carried out using sas Version 9.2. The results showed that the percentage of follicles morphologically normal in the IGF-1 30 ng/ml treatment was similar to the fresh control (p > 0.05) both on the day 1 and on the day 7 of in vitro culture. In the viability analysis, the cultured treatments maintained the percentage of viable follicles during the entire culture period (p > 0.05). After 7 days of culture, the IGF-1 30 ng/ml treatment showed higher percentages of developing follicles (48.33%) than those of the fresh control (22.22%) and the cultured treatments (p < 0.05). Also, after 7 days of culture, IGF-1 30 ng/ml presented a higher follicular diameter when compared to the control and other concentrations of IGF-1 tested. Ultrastructurally, the non-cultured control and IGF-1 30 ng/ml, after 7 days of culture, showed conserved oocytes, nuclei and organelles. Hence, it is concluded that IGF-1 30 ng/ml was the most efficient concentration for the development of bovine preantral follicles cultured in vitro. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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7. Potential of Metarhizium anisopliae (Metsch.) Sorokin (Ascomycetes, hypocreales) in the control of Bonagota salubricola (Meyrick) (Lepidoptera, Tortricidae) and its compatibility with chemical insecticides
- Author
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Anhalt, FA., primary, Azevedo, JL., additional, Sugayama, RL., additional, Specht, A., additional, and Barros, NM., additional
- Published
- 2010
- Full Text
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8. Endophytic fungi from Musa acuminata and their reintroduction into axenic plants
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Pereira, Jo, Maria Lucia Carneiro Vieira, and Azevedo, Jl
9. Genetic and molecular analysis of Aspergillus nidulans transformants obtained by the biolistic process
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Maria Helena Fungaro, Poli, Rc, Pizziranikleiner, Aa, Besin, E., Lejeune, B., and Azevedo, Jl
10. Radiotolerant endophytic bacteria and analysis of the effects of 137 Cesium on the metabolome of Pantoea sp.
- Author
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Dos Santos Oliveira JA, Polli AD, Ferreira AP, Lopes NB, Mangolim CA, Vicentini VEP, Polonio JC, Ramos AVG, Baldoqui DC, Pamphile JA, and Azevedo JL
- Abstract
Some bacteria have developed mechanisms to withstand the stress caused by ionizing radiation. The ability of these radioresistant microorganisms to survive high levels of radiation is primarily attributed to their DNA repair mechanisms and the production of protective metabolites. To determine the effect of irradiation on bacterial growth, we propose to compare the metabolites produced by the irradiated isolates to those of the control (non-irradiated isolates) using mass spectrometry, molecular networking, and chemometric analysis. We identified the secondary metabolites produced by these bacteria and observed variations in growth following irradiation. Notably, after 48 h of exposure to radiation, Pantoea sp. bacterial cells exhibited a significant 6-log increase compared to non-irradiated cells. Non-irradiated cells produce exclusively Pyridindolol, 1-hydroxy-4-methylcarbostyril, N-alkyl, and N-2-alkoxyethyl diethanolamine, while 5'-methylthioadenosine was detected only in irradiated cells. These findings suggest that the metabolic profile of Pantoea sp. remained relatively stable. The results obtained from this study have the potential to facilitate the development of innovative strategies for harnessing the capabilities of endophytic bacteria in radiological protection and bioremediation of radionuclides., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)
- Published
- 2024
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11. Synthesis, characterization, and reusability of novel nanobiocomposite of endophytic fungus Aspergillus flavus and magnetic nanoparticles (Fe 3 O 4 ) with dye bioremediation potential.
- Author
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Polli AD, Oliveira Junior VA, Ribeiro MADS, Polonio JC, Rosini B, Oliveira JADS, Bini RD, Golias HC, Fávaro-Polonio CZ, Orlandelli RC, Vicentini VEP, Cotica LF, Peralta RM, Pamphile JA, and Azevedo JL
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- Biodegradation, Environmental, Fungi, Coloring Agents, Endophytes, Aspergillus flavus, Magnetite Nanoparticles
- Abstract
The incorrect disposal of textile dyes, such as Reactive Black 5 (RB5), causes several problems for living beings and the quality of the environment. Nanobiocomposites (NBC) produced from endophytic fungi (potentially remediation dyes-agents) and magnetic nanoparticles have high biotechnological potential due to their superparamagnetic behavior, which would allow their recovery through the magnetic field after the bioremediation process. This work aimed to obtain a new nanobiocomposite from the interaction of magnetite nanoparticles (Fe
3 O4 ) with the endophyte Aspergillus flavus (Af-CL-7) to evaluate its bioremediation capacity and to reduce the toxicity of RB5 and its reuse. Before obtaining the NBC, Af-CL-7 showed discoloration of RB5 and it was tolerant to all tested concentrations of this dye. The discovery of the nanobiocomposite textile dye bioremediator product presents a significant environmental advantage by addressing the issue of water pollution caused by textile dyes. The NBC called Af-Fe3 O4 was successfully obtained with the magnetized endophyte, and their magnetic properties were verified by VSM analysis and by action of magnetic fields generated by Nd-Fe-B magnets SEM analyzes showed that the nanoparticles did not cause any damage to the hypha morphology, and TEM analyzes confirmed the presence of nanoparticles in the fungus wall and also inside the cell. The NBC Af-Fe3 O4 and Af-CL-7 showed, respectively, 96.1% and 92.2% of RB5 discoloration in the first use, 91.1% e 86.2% of discoloration in the validation test, and 89.0% in NBC reuse. In the toxicological bioassay with Lactuca sativa seeds, NBC showed a positive reduction in the toxicity of RB5 after treatment, allowing the hypocotyl growth to be statistically similar to the control with water. Thus, we highlight the promising obtaining process of NBC that could be applied in bioremediation of contaminated waters, wherein the industrial economic cost will depend on the fermentation efficiency, biomass production and nanoparticle synthesis., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests Veronica Elisa Pimenta Vicentini reports financial support was provided by National Council for Scientific and Technological Development. Andressa Domingos Polli reports financial support was provided by Coordination of Higher Education Personnel Improvement. Julio Cesar Polonio has patent #BR102020014930-0 issued to Universidade Estadual de Maringá. The co-authors has relationships with Universidade Estadual de Maringá as employers or students., (Copyright © 2023 Elsevier Ltd. All rights reserved.)- Published
- 2023
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12. Mechanisms of Mucor sp. CM3 isolated from the aquatic macrophyte Eichhornia crassipes (Mart.) Solms to increase cadmium bioremediation.
- Author
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de Almeida TT, Tschoeke BAP, Quecine MC, Tezzoto T, Gaziola SA, Azevedo RA, Piotto FA, Orlandelli RC, Dourado MN, and Azevedo JL
- Subjects
- Cadmium toxicity, Antioxidants pharmacology, Mucor, Biodegradation, Environmental, Metals pharmacology, Endophytes, Eichhornia, Soil Pollutants analysis
- Abstract
Bioremediation of toxic metals is a feasible and low-cost remediation tool to reduce metal contamination. Plant-fungus interactions can improve this technique. Eichhornia crassipes (Mart.) Solms is a macrophyte reported to bioremediate contaminated water. Thus, the present study aimed to isolate endophytic fungi from E. crassipes, select a highly cadmium (Cd) tolerant isolate and evaluate its bioremediation potential. This was evaluated by (1) the fungus tolerance and capacity to accumulate Cd; (2) Cd effects on cell morphology (using SEM and TEM) and on the fungal antioxidant defense system, as well as (3) the effect on model plant Solanum lycopersicum L. cultivar Calabash Rouge, inoculated with the endophyte fungus and exposed to Cd. Our results selected the endophyte Mucor sp. CM3, which was able to tolerate up to 1000 g/L of Cd and to accumulate 900 mg of Cd/g of biomass. Significant changes in Mucor sp. CM3 morphology were observed when exposed to high Cd concentrations, retaining this metal both in its cytoplasm and in its cell wall, which may be linked to detoxification and metal sequestration mechanisms related to the formation of Cd-GSH complexes. In addition, Cd stress induced the activation of all tested antioxidant enzymes - superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GR) - in this endophytic fungus. Moreover, when inoculated in tomato plants, this fungus promoted plant growth (in treatments without Cd) and induced an increased metal translocation to plant shoot, showing its potential to increase metal bioremediation. Therefore, this study indicates that the isolated endophyte Mucor sp. CM3 can be applied as a tool in different plant conditions, improving plant bioremediation and reducing the environmental damage caused by Cd, while also promoting plant growth in the absence of contaminants., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
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- 2023
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13. Meta-omics integration approach reveals the effect of soil native microbiome diversity in the performance of inoculant Azospirillum brasilense .
- Author
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Ferrarezi JA, Defant H, de Souza LF, Azevedo JL, Hungria M, and Quecine MC
- Abstract
Plant growth promoting bacteria (PGPB) have been used as integrative inputs to minimize the use of chemical fertilizers. However, a holistic comprehension about PGPB-plant-microbiome interactions is still incipient. Furthermore, the interaction among PGPB and the holobiont (host-microbiome association) represent a new frontier to plant breeding programs. We aimed to characterize maize bulk soil and rhizosphere microbiomes in irradiated soil (IS) and a native soil (NS) microbial community gradient (dilution-to-extinction) with Azospirillum brasilense Ab-V5, a PGPB commercial inoculant. Our hypothesis was that plant growth promotion efficiency is a result of PGPB niche occupation and persistence according to the holobiont conditions. The effects of Ab-V5 and NS microbial communities were evaluated in microcosms by a combined approach of microbiomics (species-specific qPCR, 16S rRNA metataxonomics and metagenomics) and plant phenomics (conventional and high-throughput methods). Our results revealed a weak maize growth promoting effect of Ab-V5 inoculation in undiluted NS, contrasting the positive effects of NS dilutions 10
-3 , 10-6 , 10-9 and IS with Ab-V5. Alpha diversity in NS + Ab-V5 soil samples was higher than in all other treatments in a time course of 25 days after sowing (DAS). At 15 DAS, alpha diversity indexes were different between NS and IS, but similar in all NS dilutions in rhizospheric samples. These differences were not persistent at 25 DAS, demonstrating a stabilization process in the rhizobiomes. In NS 10-3 +Ab-V5 and NS 10-6 Ab-V5, Ab-V5 persisted in the maize rhizosphere until 15 DAS in higher abundances compared to NS. In NS + Ab-V5, abundance of six taxa were positively correlated with response to (a)biotic stresses in plant-soil interface. Genes involved in bacterial metabolism of riboses and amino acids, and cresol degradation were abundant on NS 10-3 + Ab-V5, indicating that these pathways can contribute to plant growth promotion and might be a result of Ab-V5 performance as a microbial recruiter of beneficial functions to the plant. Our results demonstrated the effects of holobiont on Ab-V5 performance. The meta-omics integration supported by plant phenomics opens new perspectives to better understanding of inoculants-holobiont interaction and for developing better strategies for optimization in the use of microbial products., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ferrarezi, Defant, de Souza, Azevedo, Hungria and Quecine.)- Published
- 2023
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14. The key role of indole-3-acetic acid biosynthesis by Bacillus thuringiensis RZ2MS9 in promoting maize growth revealed by the ipdC gene knockout mediated by the CRISPR-Cas9 system.
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Figueredo EF, Cruz TAD, Almeida JR, Batista BD, Marcon J, Andrade PAM, Hayashibara CAA, Rosa MS, Azevedo JL, and Quecine MC
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- Plant Growth Regulators metabolism, Tryptophan metabolism, Gene Knockout Techniques, CRISPR-Cas Systems, Indoleacetic Acids metabolism, Zea mays genetics, Zea mays metabolism, Bacillus thuringiensis genetics, Bacillus thuringiensis metabolism
- Abstract
The bacterial biosynthesis of indole-3-acetic acid (IAA) is often related to the beneficial effects of plant growth-promoting rhizobacteria (PGPR) on plant development. In PGPR belonging to the Bacillus genus, the synthesis of IAA may occur through different metabolic pathways that are still poorly understood. B. thuringiensis (Bt) is well known for its insecticidal properties; however, its beneficial features are not limited to pest control. Our group has been studed the beneficial effects of Bt strain RZ2MS9 as growth promoter in a range of plant crops, including soybean, tomato, and maize. We recently demonstrated that bacterial IAA biosynthesis plays an important role in the ability of RZ2MS9 to benefit plant development. However, the molecular involved mechanisms in the IAA biosynthesis by this bacterium in the beneficial interaction with plants remain unclear. Here, we investigated the genetic basis of IAA biosynthesis by RZ2MS9. We knocked out the ipdC gene, involved in IAA biosynthesis via the tryptophan-dependent IPyA pathway, using the CRISPR-Cas9 system. Our results showed that, by disrupting the IPyA pathway, the amount of IAA synthesized by the mutant RZ2MS9 (ΔipdC) in the presence of tryptophan drops 57%. The gene knockout did not affect the bacterial growth, but it did affect its ability to colonize maize. Moreover, deactivating the ipdC gene in RZ2MS9 significantly reduces its ability to promote maize growth. ΔipdC performed worse than RZ2MS9 in almost all evaluated plant parameters, including total root length, projected root area, lateral roots, aerial part dry matter, and germination speed index. Therefore, we demonstrated that tryptophan-dependent IAA biosynthesis via the IPyA pathway by RZ2MS9 is strongly influenced by the ipdC gene. Furthermore, IAA biosynthesis by RZ2MS9 is a major mechanism used by this PGPR to promote maize growth., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier GmbH. All rights reserved.)
- Published
- 2023
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15. Differential Chemical Profile of Metabolite Extracts Produced by the Diaporthe citri (G-01) Endophyte Mediated by Varying the Fermented Broth pH.
- Author
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Polonio JC, Ribeiro MADS, Fávaro-Polonio CZ, Meurer EC, Azevedo JL, Golias HC, and Pamphile JA
- Abstract
Endophytic microorganisms show great potential for biotechnological exploitation because they are able to produce a wide range of secondary compounds involved in endophyte−plant adaptation, and their interactions with other living organisms that share the same microhabitat. Techniques used to chemically extract these compounds often neglect the intrinsic chemical characteristics of the molecules involved, such as the ability to form conjugate acids or bases and how they influence the solubilities of these molecules in organic solvents. Therefore, in this study, we aimed to evaluate how the pH of the fermented broth affects the process used to extract the secondary metabolites of the Diaporthe citri strain G-01 endophyte with ethyl acetate as the organic solvent. The analyzed samples, conducted by direct-infusion electrospray-ionization mass spectrometry, were grouped according to the pH of the fermented broth (i.e., <7 and ≥7). A more extreme pH (i.e., 2 or 12) was found to affect the chemical profile of the sample. Moreover, statistical analysis enabled us to determine the presence or absence of ions of high importance; for example, ions at 390.7 and 456.5 m/z were observed mainly at acidic pH, while 226.5, 298.3, and 430.1 m/z ions were observed at pH ≥ 7. Extraction at a pH between 4 and 9 may be of interest for exploring the differential secondary metabolites produced by endophytes. Furthermore, pH influences the chemical phenotype of the fungal metabolic extract.
- Published
- 2022
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16. Antimicrobial and antioxidant activities of secondary metabolites from endophytic fungus Botryosphaeria fabicerciana (MGN23-3) associated to Morus nigra L.
- Author
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Silva AAD, Polonio JC, Bulla AM, Polli AD, Castro JC, Soares LC, Oliveira-Junior VA, Vicentini VEP, Oliveira AJB, Gonçalves JE, Gonçalves RAC, Azevedo JL, Abreu-Filho BA, and Pamphile JA
- Subjects
- Anti-Bacterial Agents chemistry, Antioxidants chemistry, Bacteria metabolism, Microbial Sensitivity Tests, Plant Extracts chemistry, Staphylococcus aureus metabolism, Anti-Infective Agents pharmacology, Ascomycota metabolism, Morus metabolism
- Abstract
This study was to evaluate the biological activity of the extract of Botryosphaeria fabicerciana isolated from leaves of Morus nigra . The volatile compounds from the crude extract were analysed by GC-MS which demonstrate that mellein and β-orcinaldehyde were are the major compounds. The best minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract was observed against Gram-positive bacteria, with a MIC of 15.6 μg/mL towards B. cereus and MIC of 62.5 μg/mL towards S. aureus and B. subtilis . MBC values of 31.25 μg/mL, 62.5 μg/mL, and 250 μg/mL were observed towards B. cereus , B. subtilis , and S. aureus , respectively. The cytotoxicity analyses showed CC
50 of 115 μg/mL. The crude extract showed antioxidant activity by the DPPH, ABTS, and FRAP assays. Therefore, the extract of the endophytic fungus presented biotechnological potential as an antibacterial and antioxidant agent.- Published
- 2022
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17. Influence of plant growth-promoting endophytes Colletotrichum siamense and Diaporthe masirevici on tomato plants ( Lycopersicon esculentum Mill.).
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da Silva Santos SDS, da da Silva AA, Polonio JC, Polli AD, Orlandelli RC, Dos Santos Oliveira JADS, Brandão Filho JUT, Azevedo JL, and Pamphile JA
- Abstract
The protective and growth-promoting activities of Colletrotrichum and Diaporthe endophytes on tomato plants ( Lycopersicon esculentum Mill.) are underexplored. We screened 40 endophytic fungi associated with Mexican shrimp plant ( Justicia brandegeana ) using an in vitro dual culture assay for Fusarium oxysporum , one of the most important phytopathogens of tomato plants. The three best antagonists, Colletotrichum siamense (JB224.g1), C. siamense (JB252.g1), and Diaporthe masirevicii (JB270), were identified based on multilocus sequence analysis. They were assessed in vitro for their inhibition of F. oxysporum and phosphate solubilisation capacity, and for the production of indole acetic acid. Greenhouse experiments verified the growth-promoting effects of these endophytes and the suppression of F. oxysporum symptoms in tomato plants. Under greenhouse conditions, the JB252.g1 and JB270 isolates showed positive results for seedling emergence speed. The radicular system depth of plants inoculated with JB270 was greater than that in uninoculated plants (27.21 vs 21.95 cm). The soil plant analysis development chlorophyll metre (SPAD) index showed statistically significant results, especially for the endophyte JB224.g1 (36.99) compared to the control plants (30.90) and plants infected solely with F. oxysporum (33.64)., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)
- Published
- 2022
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18. Retrotransposons and multilocus sequence analysis reveals diversity and genetic variability in endophytic fungi-associated with Serjania laruotteana Cambess.
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da Silva Ribeiro A, Polonio JC, Dos Santos Oliveira JA, Ferreira AP, Alves LH, Mateus NJ, Mangolin CA, de Azevedo JL, and Pamphile JA
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- Multilocus Sequence Typing, Phylogeny, Retroelements genetics, Biodiversity, Fungi classification, Fungi genetics, Sapindaceae microbiology
- Abstract
The composition of endophytic communities is dynamic and demonstrates host specificity; besides, they have great intra- and interspecific genetic variability. In this work, we isolated leaf endophytic fungi from Serjania laruotteana, identify them using multilocus analysis, and evaluate the genetic variability using IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microssatellite amplified polymorphism). A total of 261 fungi were isolated and 58 were identified. Multilocus phylogenetic analysis using the partial sequences from the ITS1-5.8S-ITS2 regions, elongation factor 1-alpha, β-tubulin, actin, glyceraldehyde-3-phosphate dehydrogenase, and calmodulin genes identify that most strains belonged to the Colletotrichum and Diaporthe genera, other isolated genera were Xylaria, Phyllosticta, Muyocopron, Fusarium, Nemania, Plectosphaerella, Corynespora, Bipolaris, and Curvularia. The IRAP and REMAP analyzes were performed with Colletotrichum and Diaporthe genera and showed 100% of polymorphism and high intra- and interspecific variability. This is the first report of the diversity of endophytic fungi from S. laruotteana. In addition, it demonstrated that the IRAP and REMAP can be used to distinguish morphologically similar lineages, revealing differences even strains of the same species., (© 2021. Sociedade Brasileira de Microbiologia.)
- Published
- 2021
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19. Bacillus thuringiensis RZ2MS9, a tropical plant growth-promoting rhizobacterium, colonizes maize endophytically and alters the plant's production of volatile organic compounds during co-inoculation with Azospirillum brasilense Ab-V5.
- Author
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de Almeida JR, Bonatelli ML, Batista BD, Teixeira-Silva NS, Mondin M, Dos Santos RC, Bento JMS, de Almeida Hayashibara CA, Azevedo JL, and Quecine MC
- Subjects
- Plant Roots microbiology, Zea mays metabolism, Zea mays microbiology, Azospirillum brasilense genetics, Azospirillum brasilense metabolism, Bacillus thuringiensis genetics, Volatile Organic Compounds metabolism
- Abstract
The beneficial features of Bacillus thuringiensis (Bt) are not limited to its role as an insecticide; it is also able to promote plant growth interacting with plants and other plant growth-promoting rhizobacterium (PGPR). The PGPR Bt strain RZ2MS9 is a multi-trait maize growth promoter. We obtained a stable mutant of RZ2MS9 labelled with green fluorescent protein (RZ2MS9-GFP). We demonstrated that the Bt RZ2MS9-GFP successfully colonizes maize's roots and leaves endophytically. We evaluated whether RZ2MS9 has an additive effect on plant growth promotion when co-inoculated with Azospirillum brasilense Ab-V5. The two strains combined enhanced maize's roots and shoots dry weight around 50% and 80%, respectively, when compared to the non-inoculated control. However, non-differences were observed comparing RZ2MS9 alone and when co-inoculated with Ab-V5, In addition, we used co-inoculation experiments in glass chambers to analyse the plant's volatile organic compounds (VOCs) production during the maize-RZ2MS9 and maize-RZ2MS9-Ab-V5 interaction. We found that the single and co-inoculation altered maize's VOCs emission profile, with an increase in the production of indoles in the co-inoculation. Collectively, these results increase our knowledge about the interaction between the Bt and maize, and provide a new possibility of combined application with the commercial inoculant A. brasilense Ab-V5., (© 2021 Society for Applied Microbiology and John Wiley & Sons Ltd.)
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- 2021
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20. Secondary metabolites of Curvularia sp. G6-32, an endophyte of Sapindus saponaria , with antioxidant and anticholinesterasic properties.
- Author
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Polli AD, Ribeiro MADS, Garcia A, Polonio JC, Santos CM, Silva AA, Orlandelli RC, Castro JC, Abreu-Filho BA, Cabral MRP, Sarragiotto MH, Pamphile JA, and Azevedo JL
- Subjects
- Butyrylcholinesterase, Endophytes chemistry, Secondary Metabolism, Antioxidants pharmacology, Cholinesterase Inhibitors pharmacology, Curvularia chemistry, Sapindus microbiology
- Abstract
In the present study, the biological activity of an extract of the secondary metabolites (E-G6-32) produced by the Curvularia sp. G6-32 endophyte (isolated from the medicinal plant Sapindus saponaria L.) was investigated. The antioxidant potential was confirmed by the DPPH (22.5%) and ABTS (62.7%) assays, and the total phenolic compound content was 40 µg gallic acid equivalents/mg. The extract E-G6-32 displayed good inhibitory activity toward butyrylcholinesterase (BuChE; IC
50 = 110 ± 0.05 µg mL-1 ). The extract E-G6-32 was subjected to spectroscopic and mass spectrometry analyses. Comparison with the literature data confirmed that (-)-asperpentyn ( 1 ) was a major component. Asperpentyn belongs to the epoxyquinone family, which has attractive structural complexity, diverse functional groups, and a broad range of biological activities, including specific enzyme inhibitory activity. Our results suggest that Curvularia sp. G6-32 is a promising source of bioactive secondary metabolites and contains (-)-asperpentyn, which has potential pharmaceutical interest.[Figure: see text].- Published
- 2021
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21. On the genetic architecture in a public tropical maize panel of the symbiosis between corn and plant growth-promoting bacteria aiming to improve plant resilience.
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Yassue RM, Carvalho HF, Gevartosky R, Sabadin F, Souza PH, Bonatelli ML, Azevedo JL, Quecine MC, and Fritsche-Neto R
- Abstract
Exploring the symbiosis between plants and plant growth-promoting bacteria (PGPB) is a new challenge for sustainable agriculture. Even though many works have reported the beneficial effects of PGPB in increasing plant resilience for several stresses, its potential is not yet widely explored. One of the many reasons is the differential symbiosis performance depending on the host genotype. This opens doors to plant breeding programs to explore the genetic variability and develop new cultivars with higher responses to PGPB interaction and, therefore, have higher resilience to stress. Hence, we aimed to study the genetic architecture of the symbiosis between PGPB and tropical maize germplasm, using a public association panel and its impact on plant resilience. Our findings reveal that the synthetic PGPB population can modulate and impact root architecture traits and improve resilience to nitrogen stress, and 37 regions were significant for controlling the symbiosis between PGPB and tropical maize. In addition, we found two overlapping SNPs in the GWAS analysis indicating strong candidates for further investigations. Furthermore, genomic prediction analysis with genomic relationship matrix computed using only significant SNPs obtained from GWAS analysis substantially increased the predictive ability for several traits endorsing the importance of these genomic regions for the response of PGPB. Finally, the public tropical panel reveals a significant genetic variability to the symbiosis with the PGPB and can be a source of alleles to improve plant resilience., Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-021-01257-6., Competing Interests: Conflict of interestThe authors declare no competing interests., (© The Author(s), under exclusive licence to Springer Nature B.V. 2021.)
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- 2021
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22. Mycoviruses infecting Colletotrichum spp.: A comprehensive review.
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Casas LL, Azevedo JL, Almeida LN, Costa-Neto PQ, Bianco RA, and Pereira JO
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- Humans, Phylogeny, Spores, Fungal, Virulence, Colletotrichum, Fungal Viruses genetics, RNA Viruses
- Abstract
Colletotrichum is one of the most economically important fungal genera, which affects a wide range of hosts, specifically tropical and subtropical crops. Thus far, there have been several records of mycovirus infection in Colletotrichum spp., primarily by viruses of the Partitiviridae family. There have also been records of infections by mycoviruses of the Chrysoviridae family. Mycoviruses are (+)ssRNA and dsRNA genome viruses, which may or may not be enveloped. To date, no mycovirus with a DNA genome has been isolated from Colletotrichum spp. Typically, mycoviruses cause latent infections, although hypo- and hypervirulence have also been reported in Colletotrichum spp. In addition to its effects on pathogenic behavior, mycovirus infection can lead to important physiological changes, such as altered morphological characteristics, reduced vegetative growth, and suppressed conidia production. Therefore, research on mycoviruses infecting phytopathogenic fungi can help develop alternative methods to chemical control, which can cause irreversible damage to humans and the environment. From an agricultural perspective, mycoviruses can contribute to sustainable agriculture as biological control agents via changes in fungal physiology, ultimately resulting in the total loss of or reduction in the virulence of these pathogens.
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- 2021
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23. The auxin-producing Bacillus thuringiensis RZ2MS9 promotes the growth and modifies the root architecture of tomato (Solanum lycopersicum cv. Micro-Tom).
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Batista BD, Dourado MN, Figueredo EF, Hortencio RO, Marques JPR, Piotto FA, Bonatelli ML, Settles ML, Azevedo JL, and Quecine MC
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- Plant Roots growth & development, Plant Roots microbiology, Bacillus thuringiensis physiology, Indoleacetic Acids metabolism, Solanum lycopersicum growth & development, Solanum lycopersicum microbiology
- Abstract
Strains of Bacillus thuringiensis (Bt) are commonly commercialized as bioinoculants for insect pest control, but their benefits go beyond their insecticidal property: they can act as plant growth-promoters. Auxins play a major role in the plant growth promotion. However, the mechanism of auxin production by the Bacilli group, and more specifically by Bt strains, is unclear. In previous work, the plant growth-promoting rhizobacterium (PGPR) B. thuringiensis strain RZ2MS9 increased the corn roots. This drew our attention to the strain's auxin production trait, earlier detected in vitro. Here, we demonstrate that in its genome, RZ2MS9 harbours the complete set of genes required in two pathways that are used for Indole acetic acid (IAA) production. We also detected that the strain produces almost five times more IAA during the stationary phase. The bacterial application increased the shoot dry weight of the Micro-Tom (MT) tomato by 24%. The application also modified MT root architecture, with an increase of 26% in the average lateral root length and inhibition of the axial root. At the cellular level, RZ2MS9-treated MT plants presented elongated root cortical cells with intensified mitotic activity. Altogether, these are the best characterized auxin-associated phenotypes. Besides that, no growth alteration was detected in the auxin-insensitive diageotropic (dgt) plants either with or without the RZ2MS9 inoculation. Our results suggest that auxins play an important role in the ability of B. thuringiensis RZ2MS9 to promote MT growth and provide a better understanding of the auxin production mechanism by a Bt strain., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
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- 2021
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24. Chromolaena laevigata (Asteraceae) as a source of endophytic non-aflatoxigenic Aspergillus flavus: chemical profile in different culture conditions and biological applications.
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Balbinot RB, de Oliveira JAM, Bernardi DI, Polli AD, Polonio JC, Cabral MRP, Zanqueta ÉB, Endo EH, Meneguello JE, Cardoso RF, Azevedo JL, Dias Filho BP, Nakamura TU, do Carmo MRB, Sarragiotto MH, Pamphile JA, and Baldoqui DC
- Subjects
- Aflatoxins, Aspergillus flavus genetics, Biological Products pharmacology, Endophytes, Aspergillus flavus chemistry, Biological Products chemistry, Chromolaena microbiology
- Abstract
Endophytes are microorganisms that form symbiotic relationships with their host. These microorganisms can produce a variety of secondary metabolites, some of which have inhibitory effects on pests and pathogens or even act to promote plant growth. Due to these characteristics, these microorganisms are used as sources of biologically active substances for a wide range of biotechnological applications. Based on that, the aim of this study was to evaluate the production of metabolites of the endophytic Aspergillus flavus CL7 isolated from Chromolaena laevigata, in four different cultivation conditions, and to determine the antimicrobial, cytotoxic, antiviral, and antioxidant potential of these extracts. The multiphasic approach used to identify this strain was based on morphology and ITS gene sequence analysis. The chemical investigation of A. flavus using potato dextrose and minimal medium, using both stationary and agitated methods, resulted in the isolation of kojic acid, α-cyclopiazonic acid, and 20,25-dihydroxyaflavinine. Another 18 compounds in these extracts were identified by UHPLC-HRMS/MS, of which dideacetyl parasiticolide A has been described for the first time from A. flavus. Aflatoxins, important chemomarkers of A. flavus, were not detected in any of the extracts, thus indicating that the CL7 strain is non-aflatoxigenic. The biological potential of all extracts was evaluated, and the best results were observed for the extract obtained using minimal medium against Trichophyton rubrum and Mycobacterium tuberculosis., (© 2021. Sociedade Brasileira de Microbiologia.)
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- 2021
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25. Endophytic Colletotrichum siamense for Biocontrol and Resistance Induction in Guarana Seedlings.
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Casas LL, Pereira JO, Costa-Neto PQ, Silva JF, Almeida LN, Bianco RA, and Azevedo JL
- Abstract
Paullinia cupana var. sorbilis , known in Brazil as guarana plant, is an important plant and a major traditional crop in the State of Amazonas. It is a native Brazilian species of great economic and social importance, particularly in the Amazon region. Anthracnose caused by Colletotrichum spp. is the main challenge for this crop. Therefore, the present study verified whether C. siamense , an endophytic fungus infected with a mycovirus, could protect the seedlings and reduce or eliminate the characteristic symptoms. Total proteins and enzymatic activities of pathogenesis-related proteins (PRPs), including peroxidase (POX), chitinase (CHI), and phenylalanine ammonia lyase (PAL), were quantified. Guarana seedlings of cultivar Maués were sprayed with a C. siamense conidia suspension (5.0 × 10
3 conidia/mL). After ten days, the seedlings were sprayed with a suspension of the phytopathogen's conidia (1.0 × 106 conidia/mL). One group of these seedlings received the fungicide indicated for this crop. The fungicide was applied twice with an interval of 15 days between applications. Negative control seedlings did not receive any treatment (except water and fertilization), and positive control seedlings were treated only with the phytopathogen. The experiment was conducted between December 2019 and February 2020 in a greenhouse. The treatments were applied at an average temperature of 25°C and 85% relative humidity. Leaflets were randomly collected from each treatment group at 0, 48, 72, and 96 hours after pathogen inoculation and analyzed for total protein and enzyme production (POX, PAL, and CHI). After 28 days, the percentage of leaf lesions on the seedlings was evaluated. C. siamense inoculation reduced lesions. There were differences in total proteins and PRPs at different timepoints after inoculation, except for CHI activity, among treatments. To the best of our knowledge, this is the first record of resistance induction in guarana plants., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this paper., (Copyright © 2021 Luana L. Casas et al.)- Published
- 2021
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26. Metabolic extract of the endophytic fungus Flavodon flavus isolated from Justicia brandegeana in the control of Alicyclobacillus acidoterrestris in commercial orange juice.
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Silva AA, Polonio JC, Polli AD, Oliveira JADS, Soares LC, Oliveira Junior VA, Vicentini VEP, Oliveira AJB, Gonçalves JE, Gonçalves RAC, Azevedo JL, Pamphile JA, and Abreu Filho BA
- Subjects
- Anti-Bacterial Agents pharmacology, Microbial Sensitivity Tests, Nisin pharmacology, Alicyclobacillus drug effects, Citrus sinensis microbiology, Food Microbiology methods, Fruit and Vegetable Juices microbiology, Justicia chemistry, Justicia microbiology, Polyporales chemistry
- Abstract
In this work, the antibacterial activity of a crude extract of the endophytic fungus Flavodon flavus (JB257), isolated from leaves of Justicia brandegeana, was evaluated against both the vegetative and sporulated forms of Alicyclobacillus acidoterrestris. The microdilution technique was performed in order to determine the antibacterial activity of the crude extract alone as well as in combination with the bacteriocin, nisin. The minimum inhibitory concentration (MIC) of the crude extract and nisin alone against A. acidoterrestris vegetative forms were 250 μg/mL and 31.5 μg/mL, respectively, while the minimum bactericidal concentrations (MBC) were 1000 μg/mL and 62.5 μg/mL,respectively. For A. acidoterrestris spores, treatment with the crude extract at a concentration of 500 μg/mL caused a 47% reduction in growth, while nisin at 62.5 μg/mL could reduce 100% of the growth. The in vitro evaluation of the crude extract combined with nisin against A. acidoterrestris by the Checkerboard method showed a synergistic interaction between the two compounds. In addition, greater selectivity towards bacterial cells over host cells, a human hepatocyte cell line, was achieved when the crude extract was combined with nisin, Using scanning electron microscopy, interferences in the cell membrane of A. acidoterrestris could be observed after treatment with the crude extract. The results presented in this study indicate that the crude extract of the endophyte F. flavus has biotechnological potential in the food industry, especially for the treatment of orange juices through the control of A. acidoterrestris., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2021
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27. Tibouchina granulosa (Vell.) Cogn (Melastomataceae) as source of endophytic fungi: isolation, identification, and antiprotozoal activity of metabolites from Phyllosticta capitalensis.
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Golias HC, Polonio JC, Dos Santos Ribeiro MA, Polli AD, da Silva AA, Bulla AM, Volpato H, Nakamura CV, Meurer EC, Azevedo JL, and Pamphile JA
- Subjects
- Animals, Antiprotozoal Agents isolation & purification, Ascomycota classification, Endophytes chemistry, Endophytes classification, Endophytes isolation & purification, Leishmania drug effects, Microbial Sensitivity Tests, Secondary Metabolism, Trypanosoma drug effects, Antiprotozoal Agents pharmacology, Ascomycota chemistry, Ascomycota isolation & purification, Melastomataceae microbiology
- Abstract
Endophytes are microorganisms that form symbiotic relationships with their own host. Included in this group are the species Phyllosticta capitalensis, a group of fungi that include saprobes that produce bioactive metabolites. The present study aimed to identify the cultivable endophytic fungal microbiota present in healthy leaves of Tibouchina granulosa (Desr.) Cogn. (Melastomataceae) and investigate secondary metabolites produced by a strain of P. capitalensis and their effects against both Leishmania species and Trypanossoma cruzi. Identification of the strains was accomplished through multilocus sequencing analysis (MLSA), followed by phylogenetic analysis. The frequency of colonization was 73.66% and identified fungi belonged to the genus Diaporthe, Colletotrichum, Phyllosticta, Xylaria, Hypoxylon, Fusarium, Nigrospora, and Cercospora. A total of 18 compounds were identified by high-resolution mass spectrum analysis (UHPLC-HRMS), including fatty acids based on linoleic acid and derivatives, from P. capitalensis. Crude extracts had activity against Leishmania amazonensis, L. infantum, and Trypanosoma cruzi, with inhibitory concentration (IC
50 ) values of 17.2 μg/mL, 82.0 μg/mL, and 50.13 μg/mL, respectively. This is the first report of the production of these compounds by the endophytic P. capitalensis isolated from T. granulosa.- Published
- 2020
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28. Limitation of nitrogen source facilitated the production of nonmeiotic recombinants in Aspergillus nidulans.
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Sybuia PA, Pereira TS, Esquissato GNM, de Castro-Prado G, de Oliveira HCG, Azevedo JL, Pamphile JA, Pereira OCN, de Carvalho Nunes WM, Zanutto CA, and de Castro-Prado MAA
- Subjects
- Benomyl chemistry, Culture Media chemistry, Diploidy, Haploidy, Nitrates chemistry, Aspergillus nidulans genetics, Mitosis, Nitrogen chemistry, Recombination, Genetic
- Abstract
Aspergillus nidulans is a fungal model organism extensively used in genetic approaches. It may reproduce sexually and asexually, with a well-defined parasexual cycle. The current paper demonstrates that the limitation of nitrogen source facilitates the production of A. nidulans's nonmeiotic recombinants directly from heterokaryons, without the recovery of the diploid phase. Heterokaryons formed between master strains were inoculated in sodium nitrate-low (basal medium [BM]) and sodium nitrate-rich media (minimal medium [MM]). All mitotic segregants produced by the heterokaryons were tested for their mitotic stability in the presence of benomyl, the haploidizing agent. Only mitotically stable haploid segregants were selected for subsequent analysis. Phenotypic analyses of such haploids favored the characterization of nonmeiotic recombinants. As the number of such recombinants was higher in BM than in MM, nitrogen limitation may have facilitated the isolation of nonmeiotic recombinants from heterokaryons by stimulating nuclear fusion still inside the heterokaryotic mycelium as a survival strategy., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2020
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29. Bacterial communities associated with anthracnose symptomatic and asymptomatic leaves of guarana, an endogenous tropical crop, and their pathogen antagonistic effects.
- Author
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Bonatelli ML, Tsui S, Batista BD, Dourado MN, Kitajima EW, Andreote FD, Pylro VS, Pereira JO, de Azevedo JL, and Quecine MC
- Subjects
- Acinetobacter classification, Acinetobacter genetics, Acinetobacter isolation & purification, Amylases metabolism, Anthracosis microbiology, Bacillus classification, Bacillus genetics, Bacillus isolation & purification, Klebsiella classification, Klebsiella genetics, Klebsiella isolation & purification, Microbiota, Plant Diseases microbiology, Plant Leaves microbiology, Polygalacturonase metabolism, Proteobacteria classification, Proteobacteria genetics, Pseudomonas classification, Pseudomonas genetics, Pseudomonas isolation & purification, RNA, Ribosomal, 16S genetics, Rainforest, Siderophores metabolism, Antibiosis physiology, Biological Control Agents isolation & purification, Colletotrichum growth & development, Paullinia microbiology, Proteobacteria isolation & purification
- Abstract
Plants are colonized by diverse microorganisms that can substantially impact their health and growth. Understanding bacterial diversity and the relationships between bacteria and phytopathogens may be key to finding effective biocontrol agents. We evaluated the bacterial community associated with anthracnose symptomatic and asymptomatic leaves of guarana, a typical tropical crop. Bacterial communities were assessed through culture-independent techniques based on extensive 16S rRNA sequencing, and cultured bacterial strains were evaluated for their ability to inhibit the growth of Colletotrichum sp. as well as for enzyme and siderophore production. The culture-independent method revealed that Proteobacteria was the most abundant phylum, but many sequences were unclassified. The emergence of anthracnose disease did not significantly affect the bacterial community, but the abundance of the genera Acinetobacter, Pseudomonas and Klebsiella were significantly higher in the symptomatic leaves. In vitro growth of Colletotrichum sp. was inhibited by 11.38% of the cultured bacterial strains, and bacteria with the highest inhibition rates were isolated from symptomatic leaves, while asymptomatic leaves hosted significantly more bacteria that produced amylase and polygalacturonase. The bacterial isolate Bacillus sp. EpD2-5 demonstrated the highest inhibition rate against Colletotrichum sp., whereas the isolates EpD2-12 and FD5-12 from the same genus also had high inhibition rates. These isolates were also able to produce several hydrolytic enzymes and siderophores, indicating that they may be good candidates for the biocontrol of anthracnose. Our work demonstrated the importance of using a polyphasic approach to study microbial communities from plant diseases, and future work should focus on elucidating the roles of culture-independent bacterial communities in guarana anthracnose disease.
- Published
- 2019
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30. Additive and heterozygous (dis)advantage GWAS models reveal candidate genes involved in the genotypic variation of maize hybrids to Azospirillum brasilense.
- Author
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Vidotti MS, Lyra DH, Morosini JS, Granato ÍSC, Quecine MC, Azevedo JL, and Fritsche-Neto R
- Subjects
- Azospirillum brasilense physiology, Genotype, Heterozygote, Host-Pathogen Interactions genetics, Hybridization, Genetic, Nitrogen metabolism, Phenotype, Plant Diseases genetics, Plant Diseases microbiology, Plant Roots genetics, Plant Roots microbiology, Stress, Physiological, Zea mays microbiology, Genes, Plant genetics, Genome-Wide Association Study methods, Polymorphism, Single Nucleotide, Zea mays genetics
- Abstract
Maize genotypes can show different responsiveness to inoculation with Azospirillum brasilense and an intriguing issue is which genes of the plant are involved in the recognition and growth promotion by these Plant Growth-Promoting Bacteria (PGPB). We conducted Genome-Wide Association Studies (GWAS) using additive and heterozygous (dis)advantage models to find candidate genes for root and shoot traits under nitrogen (N) stress and N stress plus A. brasilense. A total of 52,215 Single Nucleotide Polymorphism (SNP) markers were used for GWAS analyses. For the six root traits with significant inoculation effect, the GWAS analyses revealed 25 significant SNPs for the N stress plus A. brasilense treatment, in which only two were overlapped with the 22 found for N stress only. Most were found by the heterozygous (dis)advantage model and were more related to exclusive gene ontology terms. Interestingly, the candidate genes around the significant SNPs found for the maize-A. brasilense association were involved in different functions previously described for PGPB in plants (e.g. signaling pathways of the plant's defense system and phytohormone biosynthesis). Our findings are a benchmark in the understanding of the genetic variation among maize hybrids for the association with A. brasilense and reveal the potential for further enhancement of maize through this association., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2019
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31. Gloeosporiocide, a new antifungal cyclic peptide from Streptomyces morookaense AM25 isolated from the Amazon bulk soil.
- Author
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Vicente Dos Reis G, Abraham WR, Grigoletto DF, de Campos JB, Marcon J, da Silva JA, Quecine MC, de Azevedo JL, Ferreira AG, and de Lira SP
- Subjects
- Antifungal Agents chemistry, Chromatography, High Pressure Liquid, Magnetic Resonance Spectroscopy, Microbial Sensitivity Tests, Peptides, Cyclic chemistry, Phylogeny, RNA, Ribosomal, 16S genetics, Tandem Mass Spectrometry, Antibiosis, Antifungal Agents isolation & purification, Antifungal Agents pharmacology, Peptides, Cyclic isolation & purification, Peptides, Cyclic pharmacology, Soil Microbiology, Streptomyces metabolism
- Abstract
Actinobacteria are known by their ability to produce several antimicrobial compounds of biotechnological interest. Thus, in this study, we isolated and identified by partial 16S RNA sequencing ∼100 actinobacteria isolates from guarana (Paullinia cupana) bulk soil. Besides, we isolated from the actinobacteria Streptomyces morookaense AM25 a novel cyclic peptide, named gloeosporiocide, molecular formula C44H48N11O7S3 (calculated 938.2901), and characterized by the presence of cyclized cysteins to form three thiazols. The novel compound had activity against the plant pathogen Colletotrichum gloeosporioides, assayed by the paper disk diffusion method (42.7% inhibition, 0.1 mg disk-1) and by the microdilution assay (1.25 g L-1). Our results reveal the potential of the actinobacteria from the Amazon rhizospheric soils as biocontrol agents as well as producers of new compounds with antifungal activity. Thus, this work constitutes a step forward in the development of the biotechnology of actinobacteria in the production of compounds of agronomic interest., (© FEMS 2019.)
- Published
- 2019
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32. Agrobacterium-Mediated Transformation of Diaporthe schini Endophytes Associated with Vitis labrusca L. and Its Antagonistic Activity Against Grapevine Phytopathogens.
- Author
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Felber AC, Polonio JC, Orlandelli RC, Costa AT, Ambrosio-Albuquerque EP, Castro RA, Quecine-Verdi MC, de Azevedo JL, Pamphile JA, and Mourão KSM
- Abstract
Fungus-caused diseases are among the greatest losses in grapevine culture. Biological control of pathogens by endophytes may be used to decrease fungicide application rates and environmental impacts. Previously, Diaporthe sp. B46-64 and C27-07 were highlighted as antagonists of grapevine phytopathogens. Herein, molecular multigene (ITS-TUB-TEF1) identification and phylogenetic analysis allowed the identification of these endophytes as belonging to Diaporthe schini species. Agrobacterium tumefaciens -mediated transformation was employed for obtaining 14 stable and traceable gfp - or DsRed -expressing transformants, with high transformation efficiency: 96% for the pFAT-GFP plasmid and 98% for pCAM-DsRed plasmid. Transformants were resistant to hygromycin B with gene hph confirmed by polymerase chain reaction and proved to be mitotically stable, expressing the fluorescent phenotype, with morphological differences in the colonies when compared with wild strains. In vitro antagonism tests revealed an increased antagonistic activity of some transformant strains. The current genetic transformation of D. schini mediated by A. tumefaciens proved to be an efficient technique within the randomized insertion of reporter genes for the monitoring of the strain in the environment.
- Published
- 2019
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33. Curvulin and spirostaphylotrichins R and U from extracts produced by two endophytic Bipolaris sp. associated to aquatic macrophytes with antileishmanial activity.
- Author
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de Almeida TT, Ribeiro MADS, Polonio JC, Garcia FP, Nakamura CV, Meurer EC, Sarragiotto MH, Baldoqui DC, Azevedo JL, and Pamphile JA
- Subjects
- Ascomycota, Eichhornia microbiology, Endophytes, Inhibitory Concentration 50, Leishmania drug effects, Magnetic Resonance Spectroscopy, Mitosporic Fungi, Plant Extracts chemistry, Plant Extracts toxicity, Tandem Mass Spectrometry, Eichhornia chemistry
- Abstract
In the present study, biological activity and chemical composition of two crude extracts of endophytic fungal strains of Bipolaris genera isolated from two species of aquatic macrophytes: Eichhornia azurea (Kunth) and Eichhornia crassipes (Mart.) were investigated. The nuclear magnetic resonance and mass spectrometry data provided the identification of three main compounds: curvulin ( 1 ), spirostaphylotrichin R ( 2 ) and U ( 3 ). The fragmentation mechanism of the precursor ions towards collision induced dissociation (CID) tandem mass spectrometry experiment (MS/MS) is also proposed. Furthermore, biological screening of the crude extracts displayed antileishmanial activity with IC
50 values ranging from 70-84.2 μg.mL-1 .- Published
- 2018
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34. Sequential medium with GH and IGF-1 improved in vitro development of bovine preantral follicles enclosed in ovarian tissue.
- Author
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Jimenez CR, de Azevedo JL, Ciro Alexandre Alves T, Penitente-Filho JM, and Gonçalves WG
- Subjects
- Animals, Cattle, Female, Oocytes drug effects, Ovarian Follicle drug effects, Tissue Culture Techniques veterinary, Growth Hormone pharmacology, Insulin-Like Growth Factor I pharmacology, Oocytes physiology, Ovarian Follicle physiology
- Abstract
The growth hormone (GH) and growth insulin-like factor-1 (IGF-1) act directly upon the regulation and growth in the different phases of preantral follicles. Thus, it is necessary to define their sequentiality until the in vitro preovulatory development. Therefore, the study aimed to assess the effects of a sequential medium containing GH and/or IGF-1 in the long-duration in vitro culture of preantral ovarian follicles. Ovarian fragments were cultivated: first half (days 1-7), second half (days 7-14) or during 14 culture days. Treatments were identified as: αMEM+; GH → IGF-1; IGF-1 → GH and GH + IGF-1. The culture was designed in 24-well plates, in an incubator at 37°C and 5% CO
2 . The parameters of normality, viability, follicles (primordial/in developing) and follicle diameter were evaluated. In addition, the ultrastructure was confirmed with electron transmission microscopy. The results showed that the culture treated with GH → IGF-1 kept the follicular normality and the viability until the 14th day of culture and increased both in the follicular development until 7th day and in the follicular diameter until 14th day, when compared to the control. The treatments IGF-1 → GH and GH + IGF-1 were not effective in the developing and follicular diameter after 7 days of culture, and also reduced the percentage of viability. It is concluded that the bovine preantral follicles cultured in the sequential medium treated with GH → IGF-1 improved the follicular development until the first half of the culture and kept these parameters with normality, viability and ultrastructure until the second half of the in vitro culture., (© 2018 Blackwell Verlag GmbH.)- Published
- 2018
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35. The potential of compounds isolated from Xylaria spp. as antifungal agents against anthracnose.
- Author
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Elias LM, Fortkamp D, Sartori SB, Ferreira MC, Gomes LH, Azevedo JL, Montoya QV, Rodrigues A, Ferreira AG, and Lira SP
- Subjects
- Colletotrichum drug effects, Colletotrichum physiology, Endophytes genetics, Endophytes isolation & purification, Endophytes metabolism, Fungicides, Industrial chemistry, Fungicides, Industrial isolation & purification, Mass Spectrometry, Molecular Structure, Phylogeny, Plant Diseases microbiology, Xylariales genetics, Xylariales isolation & purification, Xylariales metabolism, Endophytes chemistry, Fungicides, Industrial pharmacology, Paullinia microbiology, Plant Diseases prevention & control, Xylariales chemistry
- Abstract
Anthracnose is a crop disease usually caused by fungi in the genus Colletotrichum or Gloeosporium. These are considered one of the main pathogens, causing significant economic losses, such as in peppers and guarana. The current forms of control include the use of resistant cultivars, sanitary pruning and fungicides. However, even with the use of some methods of controlling these cultures, the crops are not free of anthracnose. Additionally, excessive application of fungicides increases the resistance of pathogens to agrochemicals and cause harm to human health and the environment. In order to find natural antifungal agents against guarana anthracnose, endophytic fungi were isolated from Amazon guarana. The compounds piliformic acid and cytochalasin D were isolated by chromatographic techniques from two Xylaria spp., guided by assays with Colletotrichum gloeosporioides. The isolated compounds were identified by spectrometric techniques, as NMR and mass spectrometry. This is the first report that piliformic acid and cytochalasin D have antifungal activity against C. gloeosporioides with MIC 2.92 and 2.46μmolmL
-1 respectively. Captan and difenoconazole were included as positive controls (MIC 16.63 and 0.02μmolmL-1 , respectively). Thus, Xylaria species presented a biotechnological potential and production of different active compounds which might be promising against anthracnose disease., (Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)- Published
- 2018
- Full Text
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36. Biocontrol of Botrytis cinerea and Calonectria gracilis by eucalypts growth promoters Bacillus spp.
- Author
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Paz ICP, Santin RCM, Guimarães AM, Rosa OPPD, Quecine MC, Silva MCPE, Azevedo JL, and Matsumura ATS
- Subjects
- Biological Assay, Plant Diseases microbiology, Volatile Organic Compounds chemistry, Bacillus, Biological Control Agents, Botrytis growth & development, Eucalyptus microbiology, Hypocreales growth & development
- Abstract
The clonal Eucalyptus plants are commonly obtained by vegetative propagation under a protected environment. This system improves the Botrytis cinerea and Calonectria spp infection on the young eucalypts plantings, resulting gray mold and cutting rot respectively. Currently, the unique available control method is based on chemicals. As alternative, novel methods to manage plant diseases, endophytic microorganisms could be an interesting alternative. Thus, we aimed to evaluate endophytic Bacillus isolated from eucalypts as a biocontrol agent against Botrytis cinerea and Calonectria gracilis, important fungal pathogens in the greenhouse, using clonal plantlets of E. urograndis. Eight endophytic strains of Bacillus, previously described as eucalyptus growth promoters, were evaluated in vitro and in vivo against Botrytis cinerea and Calonectria gracilis. The diffusible metabolites assay showed the potential of endophytic Bacillus to decrease the growth of both pathogens. Differences in the susceptibility of the pathogens to bacterial volatile metabolites were observed, B. cinerea showed more susceptible than Calonectria gracilis. In vivo assays, Bacillus amyloliquefaciens EUCB 10 demonstrated better overall reductions in these diseases. Based on the results obtained from the in vitro and in vivo analyses, we suggest that the endophytic B. amyloliquefaciens strain EUCB 10 constitutes a promising biocontrol agent against B. cinerea and Calonectria gracilis. Furthermore, this is the first reporting of B. amyloliquefaciens previously describe as plant growth promoter and also as potential control agent of B. cinerea and Calonectria gracilis to eucalyptus., (Copyright © 2018 Elsevier Ltd. All rights reserved.)
- Published
- 2018
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37. Bioprospection of Culturable Endophytic Fungi Associated with the Ornamental Plant Pachystachys lutea.
- Author
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da Silva Ribeiro A, Polonio JC, Costa AT, Dos Santos CM, Rhoden SA, Azevedo JL, and Pamphile JA
- Subjects
- Antibiosis, Colletotrichum physiology, Endophytes classification, Endophytes genetics, Fungi classification, Fungi genetics, Fusarium physiology, Magnoliopsida growth & development, Phylogeny, Plant Diseases microbiology, Plant Leaves microbiology, Endophytes isolation & purification, Endophytes physiology, Fungi isolation & purification, Fungi physiology, Magnoliopsida microbiology, Plant Diseases prevention & control
- Abstract
Endophytes are fungi and bacteria that inhabit plant tissues without causing disease. Endophytes have characteristics that are important for the health of the plant and have been isolated from several plants of economic and medicinal interest but rarely from ornamental plants. The current study isolates and identifies endophytic fungi from the leaves of Pachystachys lutea and evaluates the antagonistic activity of these endophytes as well as cellulase production by the endophytes. Fungi were isolated by fragmentation from surface-disinfected leaves and were identified by the sequencing of the ITS gene and the genes coding for EF 1-α and β-tubulin followed by multilocus sequence analysis. Molecular taxonomic analysis revealed that 78% of the identified fungi belonged to the genus Diaporthe. We also identified strains belonging to the genera Colletotrichum, Phyllosticta, Xylaria, Nemania, and Alternaria. Most of the strains tested were able to inhibit the growth of pathogenic fungi, especially PL09 (Diaporthe sp.), which inhibited the growth of Colletotrichum sp., and PL03 (Diaporthe sp.), which inhibited the growth of Fusarium oxysporum. The production of cellulase ranged from 0.87 to 1.60 μmol/min. Foliar endophytic fungal isolates from P. lutea showed promising results for the in vitro control of plant pathogens and for cellulase production. This paper is the first report on culturable endophytic fungi isolated from the ornamental plant P. lutea.
- Published
- 2018
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38. Endophytic bacterial microbiome associated with leaves of genetically modified (AtAREB1) and conventional (BR 16) soybean plants.
- Author
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Montanari-Coelho KK, Costa AT, Polonio JC, Azevedo JL, Marin SRR, Fuganti-Pagliarini R, Fujita Y, Yamaguchi-Shinozaki K, Nakashima K, Pamphile JA, and Nepomuceno AL
- Subjects
- Arabidopsis genetics, Bacteria genetics, Bacteria isolation & purification, Biodiversity, DNA, Bacterial genetics, Droughts, Endophytes genetics, Endophytes isolation & purification, Fabaceae genetics, Fabaceae microbiology, Phylogeny, RNA, Ribosomal, 16S genetics, Soil Microbiology, Arabidopsis Proteins genetics, Bacteria classification, Basic-Leucine Zipper Transcription Factors genetics, Endophytes classification, Microbiota genetics, Plant Leaves microbiology, Plants, Genetically Modified genetics, Glycine max genetics, Glycine max microbiology
- Abstract
Plant leaves (phyllosphere) have a great potential for colonization and microbial growth, consisting of a dynamic environment in which several factors can interfere with the microbial population structure. The use of genetically modified (GM) plants has introduced several traits in agriculture, such as the improvement of plant drought tolerance, as observed in the AtAREB1 transcription factor overexpression in soybean (Glycine max L. Merrill). The present study aimed at investigating the taxonomic and functional profile of the leaf microbial community of bacteria found in GM (drought-tolerant event 1Ea2939) and conventional (BR 16) soybean plants. Bacterial DNA was extracted from leaf samples collected from each genotype and used for microbial diversity and richness analysis through the MiSeq Illumina platform. Functional prediction was performed using the PICRUSt tool and the STAMP v 2.1.3 software. The obtainment of the GM event 1Ea2939 showed minimum effects on the microbial community and in the potential for chemical-genetic communication, i.e. in the potential for symbiotic and/or mutualistic interaction between plants and their natural microbiota.
- Published
- 2018
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39. Mangrove endophyte promotes reforestation tree (Acacia polyphylla) growth.
- Author
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Castro RA, Dourado MN, Almeida JR, Lacava PT, Nave A, Melo IS, Azevedo JL, and Quecine MC
- Subjects
- Acacia growth & development, Bacteria classification, Bacteria genetics, Bacteria metabolism, Brazil, Endophytes classification, Endophytes genetics, Endophytes metabolism, Indoleacetic Acids metabolism, Phylogeny, Trees growth & development, Wetlands, Acacia microbiology, Bacteria isolation & purification, Endophytes isolation & purification, Trees microbiology
- Abstract
Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000km
2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their ability to fix nitrogen and solubilize phosphorous. Bacteria that tested positive for both of these tests were examined further to determine their level of indole acetic acid production. Two strains with high indole acetic acid production were selected for use as inoculants for reforestation trees, and then the growth of the plants was evaluated under field conditions. The bacterium Pseudomonas fluorescens (strain MCR1.10) had a low phosphorus solubilization index, while this index was higher in the other strain used, Enterobacter sp. (strain MCR1.48). We used the reforestation tree Acacia polyphylla. The results indicate that inoculation with the MCR1.48 endophyte increases Acacia polyphylla shoot dry mass, demonstrating that this strain effectively promotes the plant's growth and fitness, which can be used in the seedling production of this tree. Therefore, we successfully screened the biotechnological potential of endophyte isolates from mangrove, with a focus on plant growth promotion, and selected a strain able to provide limited nutrients and hormones for in plant growth., (Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)- Published
- 2018
- Full Text
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40. Screening of tropically derived, multi-trait plant growth- promoting rhizobacteria and evaluation of corn and soybean colonization ability.
- Author
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Batista BD, Lacava PT, Ferrari A, Teixeira-Silva NS, Bonatelli ML, Tsui S, Mondin M, Kitajima EW, Pereira JO, Azevedo JL, and Quecine MC
- Subjects
- Bacillus isolation & purification, Bacillus physiology, Bacteria classification, Bacteria genetics, Bacteria growth & development, Bacteria isolation & purification, Brazil, Burkholderia isolation & purification, Burkholderia physiology, Crops, Agricultural, DNA, Ribosomal genetics, Indoleacetic Acids metabolism, Nitrogen metabolism, Nitrogen Fixation, Phenotype, Phosphates metabolism, Phylogeny, Plant Roots growth & development, Plant Roots microbiology, RNA, Ribosomal, 16S genetics, Rhizosphere, Seeds growth & development, Seeds microbiology, Siderophores metabolism, Soil Microbiology, Bacterial Physiological Phenomena, Plant Development, Glycine max growth & development, Glycine max microbiology, Zea mays growth & development, Zea mays microbiology
- Abstract
The present study assessed the plant growth-promoting (PGP) traits and diversity of culturable rhizobacteria associated with guarana (Paullinia cupana), a typical tropical plant. Ninety-six bacteria were isolated, subjected to biochemical tests, and identified by partial or total 16S rDNA sequencing. Proteobacteria and Firmicutes were the dominant rhizospheric phyla found, and Burkholderia and Bacillus were the most abundant genera. Thirteen strains exhibited the four PGP traits evaluated, and most of them belonged to the genus Burkholderia. Two multi-trait PGP strains, RZ2MS9 (Bacillus sp.) and RZ2MS16 (Burkholderia ambifaria), expressively promoted corn and soybean growth under greenhouse conditions. Compared to the non-inoculated control, increases in corn root dry weight of 247.8 and 136.9% were obtained with RZ2MS9 and RZ2MS16 inoculation, respectively, at 60days after seeding. The dry weights of corn and soybean shoots were significantly higher than those of non-inoculated plants, showing increases of more than 47% for both strains and crops. However, soybean root dry weight did not increased after bacterial inoculation with either strain. The colonization behavior of RZ2MS16 was assessed using GFP-labeling combined with fluorescence microscopy and a cultivation-based approach for quantification. RZ2MS16:gfp was able to colonize the roots and shoots of corn and soybean, revealing an endophytic behavior., (Copyright © 2017 Elsevier GmbH. All rights reserved.)
- Published
- 2018
- Full Text
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41. A Novel Multifunctional β-N-Acetylhexosaminidase Revealed through Metagenomics of an Oil-Spilled Mangrove.
- Author
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Soares FL, Marcon J, Pereira E Silva MC, Khakhum N, Cerdeira LT, Ottoni JR, Domingos DF, Taketani RG, de Oliveira VM, Lima AOS, Azevedo JL, Rodrigues JLM, and Andreote FD
- Abstract
The use of culture-independent approaches, such as metagenomics, provides complementary access to environmental microbial diversity. Mangrove environments represent a highly complex system with plenty of opportunities for finding singular functions. In this study we performed a functional screening of fosmid libraries obtained from an oil contaminated mangrove site, with the purpose of identifying clones expressing hydrolytic activities. A novel gene coding for a β- N -acetylhexosaminidase with 355 amino acids and 43KDa was retrieved and characterized. The translated sequence showed only 38% similarity to a β- N -acetylhexosaminidase gene in the genome of Veillonella sp. CAG:933, suggesting that it might constitute a novel enzyme. The enzyme was expressed, purified, and characterized for its enzymatic activity on carboxymethyl cellulose, p-Nitrophenyl-2acetamide-2deoxy-β-d-glucopyranoside, p-Nitrophenyl-2acetamide-2deoxy-β-d-galactopyranoside, and 4-Nitrophenyl β-d-glucopyranoside, presenting β- N -acetylglucosaminidase, β-glucosidase, and β-1,4-endoglucanase activities. The enzyme showed optimum activity at 30 °C and pH 5.5. The characterization of the putative novel β- N -acetylglucosaminidase enzyme reflects similarities to characteristics of the environment explored, which differs from milder conditions environments. This work exemplifies the application of cultivation-independent molecular techniques to the mangrove microbiome for obtaining a novel biotechnological product., Competing Interests: The authors declare no conflict of interest. The founding sponsors had role in the collection of samples and support in the publication of the results.
- Published
- 2017
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42. Activity of the endophytic fungi Phlebia sp. and Paecilomyces formosus in decolourisation and the reduction of reactive dyes' cytotoxicity in fish erythrocytes.
- Author
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Bulla LM, Polonio JC, Portela-Castro AL, Kava V, Azevedo JL, and Pamphile JA
- Subjects
- Animals, Anthraquinones adverse effects, Anthraquinones metabolism, Coloring Agents adverse effects, Endophytes metabolism, Erythrocytes drug effects, Fishes, Fungal Proteins metabolism, Laccase metabolism, Micronucleus Tests veterinary, Naphthalenesulfonates adverse effects, Naphthalenesulfonates metabolism, Textile Industry, Waste Disposal, Fluid, Basidiomycota metabolism, Biodegradation, Environmental, Coloring Agents metabolism, Environmental Monitoring, Paecilomyces metabolism
- Abstract
The current study investigates the potential for discolouration and degradation of Reactive Blue 19 and Reactive Black 5 textile dyes by endophytic fungi Phlebia sp. and Paecilomyces formosus as well as the potential cytotoxicity of products or by-products generated by the treatments in fish erythrocytes. It was observed at 30 days that both endophytes showed biodegradation activity with 0.1 g mL
-1 of dyes. P. formosus showed highest extracellular and intracellular protein content levels after the 15th day, and Phlebia sp. stands out for production of extracellular laccase, indicating that this enzyme may be associated with the decolouration capacity. The dyes showed toxic effects in fishes at 0.01 g mL-1 concentration, resulting in the appearance of micronuclei in erythrocyte cells. When degraded dyes treated by endophytes were tested, the frequency of micronuclei reduced approximately 20%, indicating the effectiveness of these endophytic in the treatment of textile dyes with less environmental impact, thus indicating a potential for application of these fungi in bioremediation process.- Published
- 2017
- Full Text
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43. β-(1→3,1→6)-d-glucans produced by Diaporthe sp. endophytes: Purification, chemical characterization and antiproliferative activity against MCF-7 and HepG2-C3A cells.
- Author
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Orlandelli RC, Corradi da Silva ML, Vasconcelos AFD, Almeida IV, Vicentini VEP, Prieto A, Hernandez MDD, Azevedo JL, and Pamphile JA
- Subjects
- Antineoplastic Agents isolation & purification, Antineoplastic Agents pharmacology, Ascomycota chemistry, Carbohydrate Conformation, Cell Proliferation drug effects, Drug Screening Assays, Antitumor, Endophytes chemistry, Fungal Polysaccharides isolation & purification, Fungal Polysaccharides pharmacology, Hep G2 Cells, Humans, Inhibitory Concentration 50, MCF-7 Cells, Piper microbiology, Antineoplastic Agents chemistry, Fungal Polysaccharides chemistry
- Abstract
This study reports the characterization and antiproliferative activity of exopolysaccharides (EPS) produced by submerged cultures of the endophytes Diaporthe sp. JF766998 and Diaporthe sp. JF767007 isolated from the medicinal plant Piper hispidum Sw. Both strains secreted a crude EPS that, upon size exclusion chromatography, showed to contain a heteropolysaccharide (galactose, glucose and mannose) and a high-molecular weight glucan. Data from methylation analysis, FTIR and NMR spectroscopy (
1 H, COSY, TOCSY and HSQC-DEPT) indicated that the purified glucan consisted of a main chain of glucopyranosyl β-(1→3) linkages substituted at O-6 by glucosyl residues. According to MTT assay, some treatments of both β-glucans have antiproliferative activity against human breast carcinoma (MCF-7) and hepatocellular carcinoma (HepG2-C3A) cells after 24 and 48h of treatment, exhibiting a degree of inhibition ratio that reached the highest values at 400μg/mL: 58.0% (24h) and 74.6% (48h) for MCF-7 cells, and 61.0% (24h) and 83.3% (48h) for HepG2-C3A cells. These results represent the first reports on the characterization and antiproliferative effect of β-glucans from Diaporthe species and also expand the knowledge about bioactive polysaccharides from endophytic sources., (Copyright © 2016 Elsevier B.V. All rights reserved.)- Published
- 2017
- Full Text
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44. Draft Genome Sequence of Multitrait Plant Growth-Promoting Bacillus sp. Strain RZ2MS9.
- Author
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Batista BD, Taniguti LM, Almeida JR, Azevedo JL, and Quecine MC
- Abstract
Bacillus sp. strain RZ2MS9 is a multitrait soybean and maize growth-promoting bacterium isolated in Brazil from guarana's rhizosphere. Here, we present the draft genome sequence of RZ2MS9 and its genes involved in many features related to plant growth promotion., (Copyright © 2016 Batista et al.)
- Published
- 2016
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45. 3-Nitropropionic acid production by the endophytic Diaporthe citri: Molecular taxonomy, chemical characterization, and quantification under pH variation.
- Author
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Polonio JC, Ribeiro MA, Rhoden SA, Sarragiotto MH, Azevedo JL, and Pamphile JA
- Subjects
- Ascomycota genetics, Ascomycota isolation & purification, Culture Media chemistry, Endophytes genetics, Endophytes isolation & purification, Hydrogen-Ion Concentration, Multilocus Sequence Typing, Plants microbiology, Ascomycota classification, Ascomycota metabolism, Endophytes classification, Endophytes metabolism, Nitro Compounds metabolism, Phylogeny, Propionates metabolism
- Abstract
3-nitropropionic acid (3-NPA) is a nitrogenated compound produced by plants and fungi and has been associated with poisoning episodes in humans, animals, and to induction of Huntington disease symptoms in rats. The production of 3-NPA by endophytes has been reported, but the function and biosynthesis are not well-defined. The specie of endophytic strain G-01 was confirmed as Diaporthe citri using a multilocus sequence analysis, and was verified different concentrations of 3-NPA produced at different initial pHs by these strain. The chemical analysis indicated that 3-NPA was the majority compound present in the crude extracts. The better extraction condition was at an initial pH of 7.0 for 22 d, yielding about 80 % of 3-NPA per mg of extract. It was observed that the concentration of 3-NPA increased after the initial consumption of reduction sugars, indicating that the compound is produced after the high energetic production phase of the fungus. These and other studies demonstrate the production of this compound by plants and endophytic fungi, indicating that 3-NPA may be involved in defence and nutrition systems of endophytes and host plants, and they also might participate in the biogeochemical nitrogen cycle., (Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
46. Extracellular enzymatic profiles and taxonomic identification of endophytic fungi isolated from four plant species.
- Author
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Alberto RN, Costa AT, Polonio JC, Santos MS, Rhoden SA, Azevedo JL, and Pamphile JA
- Subjects
- DNA, Ribosomal Spacer genetics, Endophytes classification, Fungi enzymology, Phylogeny, Plants microbiology, Sequence Analysis, DNA, Species Specificity, Endophytes enzymology, Endophytes isolation & purification, Enzymes metabolism, Extracellular Space enzymology, Fungi classification, Fungi isolation & purification
- Abstract
Plants of medicinal and economic importance have been studied to investigate the presence of enzyme-producing endophytic fungi. The characterization of isolates with distinct enzyme production potential may identify suitable alternatives for specialized industry. At Universidade Estadual de Maringá Laboratory of Microbial Biotechnology, approximately 500 isolates of endophytic fungi have been studied over the last decade from various host plants, including medicinally and economically important species, such as Luehea divaricata (Martius et Zuccarini), Trichilia elegans A. Juss, Sapindus saponaria L., Piper hispidum Swartz, and Saccharum spp. However, only a fraction of these endophytes have been identified and evaluated for their biotechnological application, having been initially grouped by morphological characteristics, with at least one representative of each morphogroup tested. In the current study, several fungal strains from four plants (L. divaricata, T. elegans, S. saponaria, and Saccharum spp) were identified by ribosomal DNA typing and evaluated semi-quantitatively for their enzymatic properties, including amylase, cellulase, pectinase, and protease activity. Phylogenetic analysis revealed the presence of four genera of endophytic fungi (Diaporthe, Saccharicola, Bipolaris, and Phoma) in the plants examined. According to enzymatic tests, 62% of the isolates exhibited amylase, approximately 93% cellulase, 50% pectinase, and 64% protease activity. Our results verified that the composition and abundance of endophytic fungi differed between the plants tested, and that these endophytes are a potential enzyme production resource of commercial and biotechnological value.
- Published
- 2016
- Full Text
- View/download PDF
47. Rhizosphere bacteriome of the medicinal plant Sapindus saponaria L. revealed by pyrosequencing.
- Author
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Garcia A, Polonio JC, Polli AD, Santos CM, Rhoden SA, Quecine MC, Azevedo JL, and Pamphile JA
- Subjects
- Bacteria classification, Biodiversity, Phylogeny, Plants, Medicinal genetics, RNA, Ribosomal, 16S genetics, Soil Microbiology, Microbiota, Plants, Medicinal microbiology, Rhizosphere, Sapindus genetics, Sapindus microbiology, Sequence Analysis, DNA methods, Temperature
- Abstract
Sapindus saponaria L. of Sapindaceae family is popularly known as soldier soap and is found in Central and South America. A study of such medicinal plants might reveal a more complex diversity of microorganisms as compared to non-medicinal plants, considering their metabolic potential and the chemical communication between their natural microbiota. Rhizosphere is a highly diverse microbial habitat with respect to both the diversity of species and the size of the community. Rhizosphere bacteriome associated with medicinal plant S. saponaria is still poorly known. The objective of this study was to assess the rhizosphere microbiome of the medicinal plant S. saponaria using pyrosequencing, a culture-independent approach that is increasingly being used to estimate the number of bacterial species present in different environments. In their rhizosphere microbiome, 26 phyla were identified from 5089 sequences of 16S rRNA gene, with a predominance of Actinobacteria (33.54%), Acidobacteria (22.62%), and Proteobacteria (24.72%). The rarefaction curve showed a linear increase, with 2660 operational taxonomic units at 3% distance sequence dissimilarity, indicating that the rhizosphere microbiome associated with S. saponaria was highly diverse with groups of bacteria important for soil management, which could be further exploited for agricultural and biotechnological purposes.
- Published
- 2016
- Full Text
- View/download PDF
48. The diversity of citrus endophytic bacteria and their interactions with Xylella fastidiosa and host plants.
- Author
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Azevedo JL, Araújo WL, and Lacava PT
- Abstract
The bacterium Xylella fastidiosa is the causal agent of citrus variegated chlorosis (CVC) and has been associated with important losses in commercial orchards of all sweet orange [Citrus sinensis (L.)] cultivars. The development of this disease depends on the environmental conditions, including the endophytic microbial community associated with the host plant. Previous studies have shown that X. fastidiosa interacts with the endophytic community in xylem vessels as well as in the insect vector, resulting in a lower bacterial population and reduced CVC symptoms. The citrus endophytic bacterium Methylobacterium mesophilicum can trigger X. fastidiosa response in vitro, which results in reduced growth and induction of genes associated with energy production, stress, transport, and motility, indicating that X. fastidiosa has an adaptive response to M. mesophilicum. Although this response may result in reduced CVC symptoms, the colonization rate of the endophytic bacteria should be considered in studies that intend to use this endophyte to suppress CVC disease. Symbiotic control is a new strategy that uses symbiotic endophytes as biological control agents to antagonize or displace pathogens. Candidate endophytes for symbiotic control of CVC must occupy the xylem of host plants and attach to the precibarium of sharpshooter insects to access the pathogen. In the present review, we focus on interactions between endophytic bacteria from sweet orange plants and X. fastidiosa, especially those that may be candidates for control of CVC.
- Published
- 2016
- Full Text
- View/download PDF
49. RNA applications for endophytic research.
- Author
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Polonio JC, Polli AD, Azevedo JL, and Pamphile JA
- Subjects
- Bacteria genetics, Fungi genetics, Phylogeny, RNA metabolism, RNA, Small Interfering metabolism, RNA, Small Nuclear metabolism, Endophytes classification, Endophytes genetics
- Abstract
Endophytic microorganisms, mainly bacteria and fungi, have intrinsic relationships with the host plants, involving complex chemical and genetic communication networks. The relationship among these organisms involves the development of regulatory mechanisms of gene expression that control their development and response to different interactions. Although RNA molecules are already being used in studies of microorganism diversity and taxonomy, for example, using comparisons of rRNA regions, they may also be useful tools in the exploration of gene regulation and modeling of other molecules, such as the analysis of microRNA and small interfering RNAs. Transcriptional profile analyses are capable of providing robust information on biosynthetic pathways, genes involved in the interaction and differential production of metabolites by endophytes, using RNA-seq approaches. In-depth studies of RNA types and their functions in endophytes may provide valuable information that can be used for biotechnological manipulation of microorganisms to produce metabolites, bioremediation, biological control of pathogens, and decrease plant diseases, among other economically important applications. Our study highlights the present state of knowledge of studies involving endophytes, RNA molecules, and future perspectives., Competing Interests: The authors declare no conflict of interest.
- Published
- 2016
- Full Text
- View/download PDF
50. Endophytic cultivable bacterial community obtained from the Paullinia cupana seed in Amazonas and Bahia regions and its antagonistic effects against Colletotrichum gloeosporioides.
- Author
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Silva MC, Polonio JC, Quecine MC, Almeida TT, Bogas AC, Pamphile JA, Pereira JO, Astolfi-Filho S, and Azevedo JL
- Subjects
- Bacteria genetics, Bacterial Load, Biodiversity, Brazil, Cluster Analysis, Colony Count, Microbial, DNA, Bacterial chemistry, DNA, Bacterial genetics, DNA, Ribosomal chemistry, DNA, Ribosomal genetics, Endophytes genetics, Endophytes physiology, Enzymes analysis, Pest Control, Biological methods, Phylogeny, Plant Diseases prevention & control, RNA, Ribosomal, 16S genetics, Seeds microbiology, Sequence Analysis, DNA, Antibiosis, Bacteria classification, Bacteria isolation & purification, Colletotrichum growth & development, Endophytes classification, Endophytes isolation & purification, Paullinia microbiology
- Abstract
Guarana (Paullinia cupana var. sorbilis) is a plant from the Amazonas region with socio-economic importance. However, guarana production has been increasingly affected by unfavorable conditions resulting from anthracnose, caused by the Colletotrichum fungal genus, which primarily affects mainly the Amazonas region. The aim of the present study was to isolate bacterial endophytes from the seeds of guarana plants obtained from Amazonas region and the Northeast state of Bahia, a region where this disease is not a problem for guarana plantations. The number of bacterial Colony Forming Units (CFU/g seeds) was 2.4 × 10(4) from the Bahia and 2.9 × 10(4) from the Amazonas region. One hundred and two isolated bacteria were evaluated in vitro against the phytopathogenic strain Colletotrichum gloeosporioides L1. These isolates were also analyzed for the enzymatic production of amylase, cellulase, protease, pectinase, lipase and esterase. Approximately 15% of isolates, showing high antagonistic activity, and the production of at least one enzyme were identified through the partial sequencing of 16S rDNA. The genus Bacillus was the most frequently observed, followed by Paenibacillus, Ochrobactrum, Microbacterium and Stenotrophomonas. Proteolytic activity was observed in 24 isolates followed by amylolytic, pectinolytic and cellulolytic activities. No esterase and lipase production was detected. Most of the isolates, showing antagonistic effects against C. gloeosporioides and high enzymatic activities, were isolated from the anthracnose-affected region. A biocontrol method using the endophytes from guarana seeds could be applied in the future, as these bacteria are vertically transferred to guarana seedlings., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
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