RESUMO
Fungi have been recently recognized as organisms able to grow in presence of high salt concentration with halophilic and halotolerance properties and their ligninolytic enzyme complex have an unspecific action enabling their use to degradation of a number of xenobiotic compounds. In this work, both the effect of salt and polyols on growth of the basidiomycetes strains, on their ability to produce ligninolytic enzyme and diuron degradation were evaluated. Results showed that the presence of NaCl in the culture medium affected fungal specimens in different ways. Seven out of ten tested strains had growth inhibited by salt while Dacryopinax elegans SXS323, Polyporus sp MCA128 and Datronia stereoides MCA167 fungi exhibited higher biomass production in medium containing 0.5 and 0.6 mol.L-1 of NaCl, suggesting to be halotolerant. Polyols such as glycerol and mannitol added into the culture media improved the biomass and ligninases production by D. elegans but the fungus did not reveal consumption of these polyols from media. This fungus degraded diuron in medium control, in presence of NaCl as well as polyols, produced MnP, LiP and laccase.
Assuntos
Basidiomycota/enzimologia , Basidiomycota/metabolismo , Herbicidas/metabolismo , Oxigenases/metabolismo , Cloreto de Sódio/metabolismo , Biomassa , Biotransformação , Basidiomycota/efeitos dos fármacos , Basidiomycota/crescimento & desenvolvimento , Meios de Cultura/química , Diurona/metabolismo , Inibidores do Crescimento/metabolismo , Inibidores do Crescimento/toxicidade , Polímeros/metabolismo , Polímeros/toxicidade , Cloreto de Sódio/toxicidadeRESUMO
Environments contaminated with heavy metals negatively impact the living organisms. Ectomycorrhizal fungi have shown important role in these impacted sites. Thus, this study aimed to evaluate the copper-resistance of ectomycorrhizal fungi isolates Pisolithus microcarpus - UFSC-Pt116; Pisolithus sp. - UFSC-PT24, Suillus sp. - UFSM RA 2.8 and Scleroderma sp. - UFSC-Sc124 to different copper doses in solid and liquid media. The copper doses tested were: 0.00, 0.25, 0.5, 0.75, 1.0 and 1.25 mmol L-1 in the solid medium and 0.00, 0.32, 0.64 and 0.96 mmol L-1 in the liquid medium. Copper was amended as copper sulphate in order to supplement the culture medium MNM at pH 4.8, with seven replicates to each fungus-dose combination. The fungal isolates were incubated for 30 days at 28 °C. UFSC-Pt116 showed high copper-resistance such as accessed by CL50 determinations (concentration to reduce 50% of the growth) as while as UFSC-PT24 displayed copper-resistance mechanism at 0.50 mmol L-1 in solid medium. The UFSC-PT24 and UFSC-Sc124 isolates have increased copper-resistance in liquid medium. The higher production of extracellular pigment was detected in UFSC-Pt116 cultures. The UFSC-Pt116 and UFSC-PT24 isolates showed higher resistance for copper and produced higher mycelium biomass than the other isolates. In this way, the isolates UFSG-Pt116 and UFSC-PT24 can be important candidates to survive in copper-contaminated areas, and can show important role in plants symbiosis in these contaminated sites.
Assuntos
Basidiomycota/efeitos dos fármacos , Cobre/toxicidade , Farmacorresistência Fúngica , Micorrizas/efeitos dos fármacos , Meios de Cultura/química , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Temperatura , Fatores de TempoRESUMO
The model ectomycorrhizal fungus Pisolithus microcarpus isolate 441 was transformed by using Agrobacterium tumefaciens LBA1100 and AGL-1. The selection marker was the Shble gene of Streptoallotecius hidustanus, conferring resistance to phleomycin, under the control of the gpd gene promoter and terminator of Schizophyllum commune. Transformation resulted in phleomycin resistant clones which were confirmed by PCR to contain the resistance cassette. A. tumefaciens-mediated gene transfer would allow the development of RNA interference technology in P. microcarpus.
El hongo ectomicorrícico modelo Pisolithus microcarpus aislamiento 441 fue transformado utilizando Agrobacterium tumefaciens LBA 1100 y AGL-1. El marcador de selección fue el gen Shble de Streptoallotecius hidustanus, el cual confiere resistencia a fleomicina, bajo el control del promotor y terminador del gen gpd de Schizophyllum commune. La transformación resultó en clones resistentes a fleomicina comprobándose por PCR la presencia del transgen. La transferencia génica mediada por Agrobacterium podría permitir el desarrollo de la tecnología de interferencia por ARN en P. microcarpus.
Assuntos
Agrobacterium tumefaciens/genética , Proteínas de Bactérias/genética , Basidiomycota/genética , DNA Bacteriano/genética , Transformação Genética , Agrobacterium tumefaciens/fisiologia , Basidiomycota/efeitos dos fármacos , Farmacorresistência Bacteriana/genética , Farmacorresistência Fúngica/genética , Genes Sintéticos , Reação em Cadeia da Polimerase , Fleomicinas/farmacologia , Regiões Promotoras Genéticas/genética , Seleção Genética , Schizophyllum/genéticaRESUMO
El desarrollo de dos cepas de Pycnoporus cinnabarinus (Jacq. ex Fr.) Karts, fue inhibido durante 72 horas al agregar al medio de cultivo clorpromacina, ketoconazol o cicloheximida respectivamente. La cepa Pc 58 fue inhibida por 33 ug/ml de clorpromacina y la cepa Pc470.6 por 40 ug/ml. Cicloheximida inhibió el crecimiento de la cepa Pc58 a una concentración de 3 ug/ml y a la cepa Pc470.6 a 5 ug/ml. El agente más eficaz para inhibir el crecimiento de estas cepas de P. cinnabarinus fue el Ketoconazol, que actuó a concentraciones menores de 0,5 ug/ml en la cepa Pc58 y a 0,9 ug/ml en la cepa Pc470.6