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1.
Biomolecules ; 9(12)2019 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-31779176

RESUMO

Trichoderma species are known for their ability to produce lytic enzymes, such as exoglucanases, endoglucanases, chitinases, and proteases, which play important roles in cell wall degradation of phytopathogens. ß-glucanases play crucial roles in the morphogenetic-morphological process during the development and differentiation processes in Trichoderma species, which have ß-glucans as the primary components of their cell walls. Despite the importance of glucanases in the mycoparasitism of Trichoderma spp., only a few functional analysis studies have been conducted on glucanases. In the present study, we used a functional genomics approach to investigate the functional role of the gluc31 gene, which encodes an endo-ß-1,3-glucanase belonging to the GH16 family in Trichoderma harzianum ALL42. We demonstrated that the absence of the gluc31 gene did not affect the in vivo mycoparasitism ability of mutant T. harzianum ALL42; however, gluc31 evidently influenced cell wall organization. Polymer measurements and fluorescence microscopy analyses indicated that the lack of the gluc31 gene induced a compensatory response by increasing the production of chitin and glucan polymers on the cell walls of the mutant hyphae. The mutant strain became more resistant to the fungicide benomyl compared to the parental strain. Furthermore, qRT-PCR analysis showed that the absence of gluc31 in T. harzianum resulted in the differential expression of other glycosyl hydrolases belonging to the GH16 family, because of functional redundancy among the glucanases.


Assuntos
Antibiose/genética , Parede Celular/enzimologia , Parede Celular/metabolismo , Endo-1,3(4)-beta-Glucanase/metabolismo , Trichoderma/enzimologia , Trichoderma/metabolismo , Ascomicetos/metabolismo , Benomilo/farmacologia , Parede Celular/química , Parede Celular/efeitos dos fármacos , Quitina/metabolismo , Endo-1,3(4)-beta-Glucanase/genética , Fusarium/metabolismo , Regulação Fúngica da Expressão Gênica/genética , Genômica , Microscopia de Fluorescência , Filogenia , Rhizoctonia/metabolismo , Trichoderma/efeitos dos fármacos , Trichoderma/patogenicidade , beta-Glucanas/metabolismo
2.
PLoS One ; 11(11): e0167135, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27875592

RESUMO

Inducing apoptosis is an interesting therapeutic approach to develop drugs that act against helminthic parasites. Researchers have investigated how curcumin (CUR), a biologically active compound extracted from rhizomes of Curcuma longa, affects Schistosoma mansoni and several cancer cell lines. This study evaluates how CUR influences the induction of apoptosis and oxidative stress in couples of adult S. mansoni worms. CUR decreased the viability of adult worms and killed them. The tegument of the parasite suffered morphological changes, the mitochondria underwent alterations, and chromatin condensed. Different apoptotic parameters were determined in an attempt to understand how CUR affected adult S. mansoni worms. CUR induced DNA damage and fragmentation and increased the expression of SmCASP3/7 transcripts and the activity of Caspase 3 in female and male worms. However, CUR did not intensify the activity of Caspase 8 in female or male worms. Evaluation of the superoxide anion and different antioxidant enzymes helped to explore the mechanism of parasite death further. The level of superoxide anion and the activity of Superoxide Dismutase (SOD) increased, whereas the activity of Glutathione-S-Transferase (GST), Glutathione reductase (GR), and Glutathione peroxidase (GPX) decreased, which culminated in the oxidation of proteins in adult female and male worms incubated with CUR. In conclusion, CUR generated oxidative stress followed by apoptotic-like-events in both adult female and male S. mansoni worms, ultimately killing them.


Assuntos
Apoptose/efeitos dos fármacos , Curcumina/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Schistosoma mansoni/metabolismo , Animais , Caspase 3/metabolismo , Caspase 8/metabolismo , Feminino , Proteínas de Helminto/metabolismo , Masculino , Oxirredutases/metabolismo , Superóxidos/metabolismo
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