Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo

Detalhes bibliográficos
Autor(a) principal: Mota, Paulo Roberto da
Data de Publicação: 2015
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
dARK ID: ark:/38995/0013000006fq0
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/5000
Resumo: Fungi of the genus Trichoderma have emerged as biological control agents, especially those belonging to the species Trichoderma harzianum with various mechanisms involved include production of antibiotics, mycoparasitism, competition for nutrients and host plant resistance induction. Mycoparasitism is a sequential process that involves recognition of the host contact, adhesion, expression of specific genes and secretion of secondary metabolites. Several hydrolytic enzymes are involved in this process among them chitinases, beta-glucanases and proteases. Recent studies of T. harzianum interaction with some pathogens have shown that other enzymes may also be involved in this mechanism, among which alphamannosidase. Alpha-mannosidase has an essential role in the recycling of glycoproteins in filamentous fungi, but little is known about the role of these enzymes in mycoparasitism. This study aims to characterize the expression of T. alphamannosidase gene harzianum after growth in cell wall of Sclerotinia sclerotiorum (SS), Fusarium solani (FS) and Fusarium oxysporum (FO) as well as during the interaction in vivo . It was used as a control cell wall T. harzianum (TH). Also, jobs obtaining mutants of this gene in T. harzianum were performed. Our data show that alpha-mannosidase gene was highly expressed as T. harzianum grown in the presence of cell wall SS (4.5X), FS (18X) and FO (75X) compared to the control TH (1X). In comparison situation it was noticed a 75X greater expression in TH x FO state during contact as compared with the control (TH x TH). In other situations of confrontation did not observe a significant increase in expression. The promoter region, the selection mark and the terminator region of the alpha-mannosidase of T. harzianum gene was cloned into the pRS426 vector, and the deletion cassette was assembled by the recombination technique in Saccharomyces cerevisiae. Then, these vectors were used for transformation of T. harzianum and transformants were obtained by selection on plates containing hygromycin b. Approximately 50 transformants were obtained, but none of the alpha-mannosidase gene was successfully deleted. Obtaining more transformants is necessary for us to have a mutant alpha-mannosidase gene deleted.
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spelling Ulhoa, Cirano Joséhttp://lattes.cnpq.br/8368469162867277Silva, Roberto do Nascimentohttp://lattes.cnpq.br/5771309284349109Ulhoa, Cirano JoséVieira, Pabline MarinhoGeorg, Raphaela de Castrohttp://lattes.cnpq.br/7089252809031211Mota, Paulo Roberto da2015-12-04T10:27:57Z2015-08-28MOTA, P. R. Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo. 2015. 54 f. Dissertação (Mestrado em Biologia) - Universidade Federal de Goiás, Goiânia, 2015.http://repositorio.bc.ufg.br/tede/handle/tede/5000ark:/38995/0013000006fq0Fungi of the genus Trichoderma have emerged as biological control agents, especially those belonging to the species Trichoderma harzianum with various mechanisms involved include production of antibiotics, mycoparasitism, competition for nutrients and host plant resistance induction. Mycoparasitism is a sequential process that involves recognition of the host contact, adhesion, expression of specific genes and secretion of secondary metabolites. Several hydrolytic enzymes are involved in this process among them chitinases, beta-glucanases and proteases. Recent studies of T. harzianum interaction with some pathogens have shown that other enzymes may also be involved in this mechanism, among which alphamannosidase. Alpha-mannosidase has an essential role in the recycling of glycoproteins in filamentous fungi, but little is known about the role of these enzymes in mycoparasitism. This study aims to characterize the expression of T. alphamannosidase gene harzianum after growth in cell wall of Sclerotinia sclerotiorum (SS), Fusarium solani (FS) and Fusarium oxysporum (FO) as well as during the interaction in vivo . It was used as a control cell wall T. harzianum (TH). Also, jobs obtaining mutants of this gene in T. harzianum were performed. Our data show that alpha-mannosidase gene was highly expressed as T. harzianum grown in the presence of cell wall SS (4.5X), FS (18X) and FO (75X) compared to the control TH (1X). In comparison situation it was noticed a 75X greater expression in TH x FO state during contact as compared with the control (TH x TH). In other situations of confrontation did not observe a significant increase in expression. The promoter region, the selection mark and the terminator region of the alpha-mannosidase of T. harzianum gene was cloned into the pRS426 vector, and the deletion cassette was assembled by the recombination technique in Saccharomyces cerevisiae. Then, these vectors were used for transformation of T. harzianum and transformants were obtained by selection on plates containing hygromycin b. Approximately 50 transformants were obtained, but none of the alpha-mannosidase gene was successfully deleted. Obtaining more transformants is necessary for us to have a mutant alpha-mannosidase gene deleted.Fungos do gênero Trichoderma têm se destacado como agentes de controle biológico, principalmente aqueles pertencentes à espécie Trichoderma harzianumcom vários mecanismos envolvidos incluem produção de antibióticos, micoparasitismo, competição por nutrientes e indução de resistência da planta hospedeira. Micoparasitismo é um processo sequencial que envolve reconhecimento do hospedeiro, contato, adesão, expressão de genes específicos e secreção de metabólitos secundários. Várias enzimas hidrolíticas estão envolvidas neste processo dentre elas quitinases, beta-glucanases e proteases. Estudos recentes de interação de T. harzianum com alguns fitopatógenos tem demonstrado que outras enzimas podem também estar envolvidas neste mecanismo, dentre elas as alfamanosidases. As alfa-manosidases tem um papel fundamental na reciclagem de glicoproteínas em fungos filamentosos, mas pouco se sabe sobre o papel destas enzimas no micoparasitismo. Este trabalho tem como objetivo a caracterização da expressão do gene de alfa-manosidase de T. harzianumapós crescimento em parede celular de Sclerotinia sclerotiorum (SS), Fusarium solani (FS) e Fusarium oxysporum (FO), bem como durante a interação in vivo. Como controle foi utilizado parede celular de T. harzianum (TH). Além disto, trabalhos de obtenção de mutantes deste gene em T. harzianum foram realizados. Nossos dados mostraram que o gene de alfa-manosidase foi altamente expresso quando T. harzianum cresceu na presença de parede celular de SS (4.5X), FS (18X) e FO (75X) quando comparado com o controle TH (1X). Em situação de confronto foi observado uma expressão de 75X maior na situação TH x FO durante o contato, quando comparado com o controle (TH x TH). Na outras situações de confronto não observamos um aumento significativo de expressão. A região promotora, a marca de seleção e a região terminadora do gene da alfa-manosidase de T. harzianumfoi clonado no vetor pRS426 e o cassete de deleção foi montado através da técnica de recombinação em Saccharomyces cerevisiae . Em seguida, estes vetores foram utilizados para a transformação de T. harzianume os transformantes foram obtidos por seleção em placas contendo higromicina b. Aproximadamente 50 transformantes foram obtidos, mas em nenhum deles o gene da alfa-manosidase foi deletado com sucesso. A obtenção de mais transformantes se faz necessário para que possamos ter um mutante com o gene da alfa-manosidase deletado.Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2015-12-04T10:24:50Z No. of bitstreams: 2 Dissertação - Paulo Roberto da Mota - 2015.pdf: 1806908 bytes, checksum: d6798b75ceeef2f456ddde6fbf387e77 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-12-04T10:27:57Z (GMT) No. of bitstreams: 2 Dissertação - Paulo Roberto da Mota - 2015.pdf: 1806908 bytes, checksum: d6798b75ceeef2f456ddde6fbf387e77 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Made available in DSpace on 2015-12-04T10:27:57Z (GMT). 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dc.title.por.fl_str_mv Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
title Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
spellingShingle Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
Mota, Paulo Roberto da
Fitopatógenos
Micoparasitismo
Competição
Parede celular e expressão
Plant pathogens
Mycoparasitism
Competition
Ccellular wall and expression
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
title_short Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
title_full Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
title_fullStr Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
title_full_unstemmed Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
title_sort Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo
author Mota, Paulo Roberto da
author_facet Mota, Paulo Roberto da
author_role author
dc.contributor.advisor1.fl_str_mv Ulhoa, Cirano José
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/8368469162867277
dc.contributor.advisor-co1.fl_str_mv Silva, Roberto do Nascimento
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/5771309284349109
dc.contributor.referee1.fl_str_mv Ulhoa, Cirano José
dc.contributor.referee2.fl_str_mv Vieira, Pabline Marinho
dc.contributor.referee3.fl_str_mv Georg, Raphaela de Castro
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/7089252809031211
dc.contributor.author.fl_str_mv Mota, Paulo Roberto da
contributor_str_mv Ulhoa, Cirano José
Silva, Roberto do Nascimento
Ulhoa, Cirano José
Vieira, Pabline Marinho
Georg, Raphaela de Castro
dc.subject.por.fl_str_mv Fitopatógenos
Micoparasitismo
Competição
Parede celular e expressão
topic Fitopatógenos
Micoparasitismo
Competição
Parede celular e expressão
Plant pathogens
Mycoparasitism
Competition
Ccellular wall and expression
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
dc.subject.eng.fl_str_mv Plant pathogens
Mycoparasitism
Competition
Ccellular wall and expression
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
description Fungi of the genus Trichoderma have emerged as biological control agents, especially those belonging to the species Trichoderma harzianum with various mechanisms involved include production of antibiotics, mycoparasitism, competition for nutrients and host plant resistance induction. Mycoparasitism is a sequential process that involves recognition of the host contact, adhesion, expression of specific genes and secretion of secondary metabolites. Several hydrolytic enzymes are involved in this process among them chitinases, beta-glucanases and proteases. Recent studies of T. harzianum interaction with some pathogens have shown that other enzymes may also be involved in this mechanism, among which alphamannosidase. Alpha-mannosidase has an essential role in the recycling of glycoproteins in filamentous fungi, but little is known about the role of these enzymes in mycoparasitism. This study aims to characterize the expression of T. alphamannosidase gene harzianum after growth in cell wall of Sclerotinia sclerotiorum (SS), Fusarium solani (FS) and Fusarium oxysporum (FO) as well as during the interaction in vivo . It was used as a control cell wall T. harzianum (TH). Also, jobs obtaining mutants of this gene in T. harzianum were performed. Our data show that alpha-mannosidase gene was highly expressed as T. harzianum grown in the presence of cell wall SS (4.5X), FS (18X) and FO (75X) compared to the control TH (1X). In comparison situation it was noticed a 75X greater expression in TH x FO state during contact as compared with the control (TH x TH). In other situations of confrontation did not observe a significant increase in expression. The promoter region, the selection mark and the terminator region of the alpha-mannosidase of T. harzianum gene was cloned into the pRS426 vector, and the deletion cassette was assembled by the recombination technique in Saccharomyces cerevisiae. Then, these vectors were used for transformation of T. harzianum and transformants were obtained by selection on plates containing hygromycin b. Approximately 50 transformants were obtained, but none of the alpha-mannosidase gene was successfully deleted. Obtaining more transformants is necessary for us to have a mutant alpha-mannosidase gene deleted.
publishDate 2015
dc.date.accessioned.fl_str_mv 2015-12-04T10:27:57Z
dc.date.issued.fl_str_mv 2015-08-28
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dc.identifier.citation.fl_str_mv MOTA, P. R. Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo. 2015. 54 f. Dissertação (Mestrado em Biologia) - Universidade Federal de Goiás, Goiânia, 2015.
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identifier_str_mv MOTA, P. R. Caracterizaçãoda alfa-manosidase produzida por Trichoderma harzianum durante o micoparasitismo. 2015. 54 f. Dissertação (Mestrado em Biologia) - Universidade Federal de Goiás, Goiânia, 2015.
ark:/38995/0013000006fq0
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dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Instituto de Ciências Biológicas - ICB (RG)
publisher.none.fl_str_mv Universidade Federal de Goiás
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