Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response

Detalhes bibliográficos
Autor(a) principal: Hurtado, Fernando Manuel Matias
Data de Publicação: 2020
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
Texto Completo: https://www.teses.usp.br/teses/disponiveis/11/11144/tde-13082020-184821/
Resumo: NAC proteins are one of the largest families of plant-specific transcription factors (TFs). They regulate diverse biological processes, including secondary cell wall formation (SCW). SCW is the main component of wood and source of raw material in forestry industry, Tectona grandis (teak) specie is recognized for presenting one of the most valuable woods in the world due to its extraordinary qualities of color, density and durability. Studies of teak genomics and transcriptomics were recently unveiled, producing information that allowed to identify teak NAC transcription factors and to analyse their evolution and expression profile during wood formation. It was identified 130 NAC proteins that were coded by 107 gene loci. These proteins were classified into 23 clades of the NAC family, together with Populus, Eucalyptus, and Arabidopsis. Data on transcription expression revealed specific temporal and spatial expression patterns for the majority of teak NAC genes. The qRT-PCR indicated expression of VASCULAR NAC DOMAIN proteins involved in SCW formation in xylem vessels of 16-year-old juvenile trees. In addition, it was characterized by heterologous expression, the novel salt stress-responsive TgNAC01 gene. Three independent transgenic tobacco lines overexpressing TgNAC01 were generated. It was demonstrated that the TgNAC01 protein is localized only in the nucleus. Analyses of TgNAC01 relative expression, stem length and chlorophyll content were evaluated in two months old transgenic plants growing in greenhouse. Tobacco plants overexpressing TgNAC01 showed higher stem length, an increase in leaf senescence, and tolerance to 300 mM of NaCl when compared to wild type (WT) plants. Water potential, gas exchange and chlorophyll fluorescence were used to determine the salt stress tolerance. In this research, it was demonstrated that TgNAC01 transcription factor acts as a positive regulator in xylem development and as a transcriptional activator mediated by salt stress and leaf senescence. These results open a way to further understanding of NAC transcription factor genes in T. grandis wood formation and stress tolerance, potentially being useful for future studies aiming to improve productivity and wood quality using biotechnological approaches.
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spelling Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress responseCaracterização da família de fatôres de transcrição NAC de Tectona grandis envolvidos na deposição de parede celular secundaria e reposta à estresseEstresse salinoFormação de madeiraNAC proteinProteína NACSalt stressSecondary xylemWood formationXilema secundárioNAC proteins are one of the largest families of plant-specific transcription factors (TFs). They regulate diverse biological processes, including secondary cell wall formation (SCW). SCW is the main component of wood and source of raw material in forestry industry, Tectona grandis (teak) specie is recognized for presenting one of the most valuable woods in the world due to its extraordinary qualities of color, density and durability. Studies of teak genomics and transcriptomics were recently unveiled, producing information that allowed to identify teak NAC transcription factors and to analyse their evolution and expression profile during wood formation. It was identified 130 NAC proteins that were coded by 107 gene loci. These proteins were classified into 23 clades of the NAC family, together with Populus, Eucalyptus, and Arabidopsis. Data on transcription expression revealed specific temporal and spatial expression patterns for the majority of teak NAC genes. The qRT-PCR indicated expression of VASCULAR NAC DOMAIN proteins involved in SCW formation in xylem vessels of 16-year-old juvenile trees. In addition, it was characterized by heterologous expression, the novel salt stress-responsive TgNAC01 gene. Three independent transgenic tobacco lines overexpressing TgNAC01 were generated. It was demonstrated that the TgNAC01 protein is localized only in the nucleus. Analyses of TgNAC01 relative expression, stem length and chlorophyll content were evaluated in two months old transgenic plants growing in greenhouse. Tobacco plants overexpressing TgNAC01 showed higher stem length, an increase in leaf senescence, and tolerance to 300 mM of NaCl when compared to wild type (WT) plants. Water potential, gas exchange and chlorophyll fluorescence were used to determine the salt stress tolerance. In this research, it was demonstrated that TgNAC01 transcription factor acts as a positive regulator in xylem development and as a transcriptional activator mediated by salt stress and leaf senescence. These results open a way to further understanding of NAC transcription factor genes in T. grandis wood formation and stress tolerance, potentially being useful for future studies aiming to improve productivity and wood quality using biotechnological approaches.As proteínas NAC são uma das maiores famílias de fatôres de transcrição específicos de plantas (TFs). Eles regulam diversos processos biológicos, incluindo a formação de parede celular secundária (SWC). A SCW é a principal componente da madeira e fonte de matéria-prima na indústria florestal. A espécie Tectona grandis (teca) é reconhecida por apresentar uma das madeiras mais valiosas do mundo devido às suas extraordinárias qualidades de côr, densidade e durabilidade. Estudos de genômica e transcriptômica de teca foram recentemente disponibilizados, essas informações permitiram identificar os fatores de transcrição NAC e analisá-los em relação a sua evolução e seu perfil de expressão durante a formação da madeira. Foram identificadas 130 proteínas NAC que são codificadas por 107 loci de genes. Essas proteínas foram classificadas em 23 clados da família NAC, juntamente com Populus, Eucalyptus e Arabidopsis. Os dados de expressão dos transcritos revelaram padrões de expressão temporal e espacial específicos para a maioria dos genes NAC de teca. O qRT-PCR indicou expressão de proteínas de Domínio Vascular NAC (VASCULAR NAC DOMAIN) envolvidas na formação de SCW em vasos do xilema de árvores jovens de 16 anos de idade. Ainda, foi caracterizado por expressão heteróloga o novo gene TgNAC01 responsivo ao estresse salino. Foram geradas três linhas de tabaco transgênicas independentes que superexpressaram TgNAC01. A proteína TgNAC01 foi demonstrada estar localizada no núcleo. A expressão relativa do gene TgNAC01, comprimento do caule e o teor de clorofila foram analisados nas plantas transgênicas com dois meses de crescimento em casa de vegetação. As plantas de tabaco que superexpressam TgNAC01 mostraram maior crescimento, aumento na senescência foliar e maior tolerância ao estresse salino na concentração de 300 mM de NaCl em comparação com plantas controle (WT). O potencial hídrico, trocas gasosas e a fluorescência da clorofila foram utilizados para determinar a tolerância ao estresse salino. Neste trabalho de pesquisa, foi demonstrado que o fator de transcrição TgNAC01 atua como um regulador positivo no desenvolvimento do xilema e como um ativador transcricional da regulação mediada pelo estresse salino e senescência foliar. Estes resultados abrem caminho para uma maior compreensão sobre os genes do fator de transcrição NAC na formação da madeira e na tolerância à estresse em teca, sendo potencialmente úteis para estudos futuros que visam melhorar a produtividade e a qualidade da madeira usando abordagens biotecnológicas.Biblioteca Digitais de Teses e Dissertações da USPCarrer, HelaineHurtado, Fernando Manuel Matias2020-06-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/11/11144/tde-13082020-184821/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2020-08-17T17:40:02Zoai:teses.usp.br:tde-13082020-184821Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212020-08-17T17:40:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
Caracterização da família de fatôres de transcrição NAC de Tectona grandis envolvidos na deposição de parede celular secundaria e reposta à estresse
title Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
spellingShingle Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
Hurtado, Fernando Manuel Matias
Estresse salino
Formação de madeira
NAC protein
Proteína NAC
Salt stress
Secondary xylem
Wood formation
Xilema secundário
title_short Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
title_full Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
title_fullStr Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
title_full_unstemmed Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
title_sort Characterization of NAC transcription factor family of Tectona grandis involved in secondary cell wall deposition and stress response
author Hurtado, Fernando Manuel Matias
author_facet Hurtado, Fernando Manuel Matias
author_role author
dc.contributor.none.fl_str_mv Carrer, Helaine
dc.contributor.author.fl_str_mv Hurtado, Fernando Manuel Matias
dc.subject.por.fl_str_mv Estresse salino
Formação de madeira
NAC protein
Proteína NAC
Salt stress
Secondary xylem
Wood formation
Xilema secundário
topic Estresse salino
Formação de madeira
NAC protein
Proteína NAC
Salt stress
Secondary xylem
Wood formation
Xilema secundário
description NAC proteins are one of the largest families of plant-specific transcription factors (TFs). They regulate diverse biological processes, including secondary cell wall formation (SCW). SCW is the main component of wood and source of raw material in forestry industry, Tectona grandis (teak) specie is recognized for presenting one of the most valuable woods in the world due to its extraordinary qualities of color, density and durability. Studies of teak genomics and transcriptomics were recently unveiled, producing information that allowed to identify teak NAC transcription factors and to analyse their evolution and expression profile during wood formation. It was identified 130 NAC proteins that were coded by 107 gene loci. These proteins were classified into 23 clades of the NAC family, together with Populus, Eucalyptus, and Arabidopsis. Data on transcription expression revealed specific temporal and spatial expression patterns for the majority of teak NAC genes. The qRT-PCR indicated expression of VASCULAR NAC DOMAIN proteins involved in SCW formation in xylem vessels of 16-year-old juvenile trees. In addition, it was characterized by heterologous expression, the novel salt stress-responsive TgNAC01 gene. Three independent transgenic tobacco lines overexpressing TgNAC01 were generated. It was demonstrated that the TgNAC01 protein is localized only in the nucleus. Analyses of TgNAC01 relative expression, stem length and chlorophyll content were evaluated in two months old transgenic plants growing in greenhouse. Tobacco plants overexpressing TgNAC01 showed higher stem length, an increase in leaf senescence, and tolerance to 300 mM of NaCl when compared to wild type (WT) plants. Water potential, gas exchange and chlorophyll fluorescence were used to determine the salt stress tolerance. In this research, it was demonstrated that TgNAC01 transcription factor acts as a positive regulator in xylem development and as a transcriptional activator mediated by salt stress and leaf senescence. These results open a way to further understanding of NAC transcription factor genes in T. grandis wood formation and stress tolerance, potentially being useful for future studies aiming to improve productivity and wood quality using biotechnological approaches.
publishDate 2020
dc.date.none.fl_str_mv 2020-06-03
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