RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia

Detalhes bibliográficos
Autor(a) principal: Fagundes, Lucas França
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
dARK ID: ark:/38995/001300000dshf
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/8948
Resumo: Due in most part to the rapid growth and adaptability of the Eucalyptus in Brazil, the country's forestry sector is one of the most competitive of the world. Drought is one of the main obstacles of the culture, which has been expanding to the country’s interior regions such as the Cerrado biome. The objective of this work was to identify genes and biological processes involved in the responses of the Eucalyptus roots to drought. The experiment was conducted in a completely randomized design with 2 treatments (control and deficit) x 3 root RNA gathering time (3, 6 and 12h after start of treatment) and three biological replicates (commercial clones 1528, I144, VM1). The plants grew in hydroponics for 50 days, before the experiment start by using PEG8000 to induce water deficit (osmotic potential of -0.9 MPa). The root RNA of the seedlings was extracted using the RNeasy Plant Mini® kit (Quiagen). RNA samples were sequenced on the HiSeq 2000 platform (Illumina), yielding approximately 626 million 50 bp sequences in single end mode. After quality control, the sequences were aligned in the reference genome of Eucalyptus grandis. After counting reads aligned on each gene, differential expression analyses were performed with edgeR package of R/Bioconductor. A total of 6169 differentially expressed genes (DEG) (FDR ≤ 5% and Log2 FoldChange > |2|) were identified. Of these, 3350 were up-regulated and 2819 were repressed under osmotic stress, considering all the time-series evaluations. At three hours after treatment the highest number of DEG was obtained, with 3150, followed by six hours (2494 DEG) and 12h (525 DEG). Functional enrichment analysis was performed with the Fisher exact test, by software Blast2GO, with the DEG associated with their respective Gene Ontology (GO) terms. As a total of 90 GO terms, enriched and non-redundant, was verified that our results indicate a rapid initial response of Eucalyptus roots induced by PEG 8000 in hydroponics. The most remarkable characteristics of this response under hydroponic conditions are: increased energy consumption via carbohydrate catabolism, activation of the glycolytic pathway, repression of the respiratory chain in the mitochondria, increase of the fermentation pathway, partial execution of the citric acid cycle and repression of the primary metabolism (DNA replication and RNA biosynthesis, development and growth). These enriched functional terms point to the predominance of hypoxia stress, induced by the high viscosity of PEG in hydroponics, as compared to the osmotic stress.
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spelling Novaes, Evandrohttp://lattes.cnpq.br/0568272239145336Novaes, EvandroAntunes, Adriana MariaMoraes, Moemy Gomes dehttp://lattes.cnpq.br/1960687146765217Fagundes, Lucas França2018-10-05T14:14:11Z2017-12-18FAGUNDES, L. F. RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia. 2017. 85 f. Dissertação (Mestrado em Genética e Melhoramento de Plantas) - Universidade Federal de Goiás, Goiânia, 2017.http://repositorio.bc.ufg.br/tede/handle/tede/8948ark:/38995/001300000dshfDue in most part to the rapid growth and adaptability of the Eucalyptus in Brazil, the country's forestry sector is one of the most competitive of the world. Drought is one of the main obstacles of the culture, which has been expanding to the country’s interior regions such as the Cerrado biome. The objective of this work was to identify genes and biological processes involved in the responses of the Eucalyptus roots to drought. The experiment was conducted in a completely randomized design with 2 treatments (control and deficit) x 3 root RNA gathering time (3, 6 and 12h after start of treatment) and three biological replicates (commercial clones 1528, I144, VM1). The plants grew in hydroponics for 50 days, before the experiment start by using PEG8000 to induce water deficit (osmotic potential of -0.9 MPa). The root RNA of the seedlings was extracted using the RNeasy Plant Mini® kit (Quiagen). RNA samples were sequenced on the HiSeq 2000 platform (Illumina), yielding approximately 626 million 50 bp sequences in single end mode. After quality control, the sequences were aligned in the reference genome of Eucalyptus grandis. After counting reads aligned on each gene, differential expression analyses were performed with edgeR package of R/Bioconductor. A total of 6169 differentially expressed genes (DEG) (FDR ≤ 5% and Log2 FoldChange > |2|) were identified. Of these, 3350 were up-regulated and 2819 were repressed under osmotic stress, considering all the time-series evaluations. At three hours after treatment the highest number of DEG was obtained, with 3150, followed by six hours (2494 DEG) and 12h (525 DEG). Functional enrichment analysis was performed with the Fisher exact test, by software Blast2GO, with the DEG associated with their respective Gene Ontology (GO) terms. As a total of 90 GO terms, enriched and non-redundant, was verified that our results indicate a rapid initial response of Eucalyptus roots induced by PEG 8000 in hydroponics. The most remarkable characteristics of this response under hydroponic conditions are: increased energy consumption via carbohydrate catabolism, activation of the glycolytic pathway, repression of the respiratory chain in the mitochondria, increase of the fermentation pathway, partial execution of the citric acid cycle and repression of the primary metabolism (DNA replication and RNA biosynthesis, development and growth). These enriched functional terms point to the predominance of hypoxia stress, induced by the high viscosity of PEG in hydroponics, as compared to the osmotic stress.Devido em grande parte ao rápido crescimento e adaptabilidade do Eucalyptus no Brasil, o setor florestal do país é um dos mais competitivos do mundo. O déficit hídrico é um dos principais entraves atuais da cultura, que vem se expandindo para as regiões interioranas do país, como o Cerrado. O presente trabalho teve como objetivos identificar os genes e os processos biológicos relacionados à resposta das raízes do Eucalyptus ao déficit hídrico. O experimento foi conduzido em delineamento inteiramente casualizados com 2 tratamentos (controle e déficit) x 3 tempos de coleta (3, 6 e 12h após início do tratamento) e três réplicas biológicas (clones comerciais 1528, I144, VM1). As plantas cresceram em hidroponia por 50 dias, quando foi iniciado o experimento de déficit hídrico induzido com PEG8000 (potencial osmótico de -0,9 MPa). O RNA das raízes das mudas foi extraído utilizando o kit RNeasy Plant Mini® (Quiagen). As amostras de RNA foram sequenciadas na plataforma HiSeq 2000 (Illumina), produzindo aproximadamente 626 milhões de sequências de 50 pb no modo single end. Após controle qualidade, as sequências foram alinhadas no genoma de referência de Eucalyptus grandis. Após contagem das sequências alinhadas em cada gene, realizou-se análises de expressão diferencial com o pacote edgeR do R/Bioconductor. Foram identificados 6169 genes diferencialmente expressos (GDE) (FDR ≤ 5% e Log2 FoldChange > |2|). Desses, 3350 foram ativados e 2819 reprimidos sob estresse osmótico, considerando todos os tempos avaliados. O tempo de três horas foi o que mais identificou GDE, com 3150, seguido pelo de seis (2494 GDE) e 12h (525 GDE). A análise de enriquecimento funcional foi realizada com o teste exato de Fisher, com os GDE associados aos seus respectivos termos Gene Ontology (GO), por meio do aplicativo Blast2GO. Como total de 90 termos GO, enriquecidos não redundantes, foi verificado que nossos resultados apontam para uma rápida resposta inicial das raízes do Eucalyptus induzidas pelo PEG 8000 em hidroponia. As características mais marcantes dessa resposta, em condições de hidroponia, são: o aumento do consumo energético via catabolismo de carboidratos, ativação da via glicolítica, repressão da cadeia respiratória nas mitocôndrias, aumento da via de fermentação, execução parcial do ciclo do acido cítrico e repressão do metabolismo primário (replicação do DNA e biossíntese de RNA, desenvolvimento e crescimento). Os termos enriquecidos apontam para uma predominância do estresse de hipóxia, induzido pela alta viscosidade do PEG em hidroponia, em comparação ao estresse hídrico.Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2018-10-04T13:32:15Z No. of bitstreams: 2 Dissertação - Lucas França Fagundes - 2017.pdf: 2859645 bytes, checksum: a3a69c2ef467d243d394e4ee270a7680 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-10-05T14:14:11Z (GMT) No. of bitstreams: 2 Dissertação - Lucas França Fagundes - 2017.pdf: 2859645 bytes, checksum: a3a69c2ef467d243d394e4ee270a7680 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2018-10-05T14:14:11Z (GMT). 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dc.title.eng.fl_str_mv RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
dc.title.alternative.eng.fl_str_mv RNA-Seq indicates hypoxia stress induced by polyethylene glycol on Eucalyptus roots in hydroponics
title RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
spellingShingle RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
Fagundes, Lucas França
Hidroponia
Raíz
RNA-Seq
Eucalyptus
Hipóxia
Hydropony
Root
Eucalyptus
Hypoxia
CIENCIAS AGRARIAS::AGRONOMIA
title_short RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
title_full RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
title_fullStr RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
title_full_unstemmed RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
title_sort RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia
author Fagundes, Lucas França
author_facet Fagundes, Lucas França
author_role author
dc.contributor.advisor1.fl_str_mv Novaes, Evandro
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/0568272239145336
dc.contributor.referee1.fl_str_mv Novaes, Evandro
dc.contributor.referee2.fl_str_mv Antunes, Adriana Maria
dc.contributor.referee3.fl_str_mv Moraes, Moemy Gomes de
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/1960687146765217
dc.contributor.author.fl_str_mv Fagundes, Lucas França
contributor_str_mv Novaes, Evandro
Novaes, Evandro
Antunes, Adriana Maria
Moraes, Moemy Gomes de
dc.subject.por.fl_str_mv Hidroponia
Raíz
RNA-Seq
Eucalyptus
Hipóxia
topic Hidroponia
Raíz
RNA-Seq
Eucalyptus
Hipóxia
Hydropony
Root
Eucalyptus
Hypoxia
CIENCIAS AGRARIAS::AGRONOMIA
dc.subject.eng.fl_str_mv Hydropony
Root
Eucalyptus
Hypoxia
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::AGRONOMIA
description Due in most part to the rapid growth and adaptability of the Eucalyptus in Brazil, the country's forestry sector is one of the most competitive of the world. Drought is one of the main obstacles of the culture, which has been expanding to the country’s interior regions such as the Cerrado biome. The objective of this work was to identify genes and biological processes involved in the responses of the Eucalyptus roots to drought. The experiment was conducted in a completely randomized design with 2 treatments (control and deficit) x 3 root RNA gathering time (3, 6 and 12h after start of treatment) and three biological replicates (commercial clones 1528, I144, VM1). The plants grew in hydroponics for 50 days, before the experiment start by using PEG8000 to induce water deficit (osmotic potential of -0.9 MPa). The root RNA of the seedlings was extracted using the RNeasy Plant Mini® kit (Quiagen). RNA samples were sequenced on the HiSeq 2000 platform (Illumina), yielding approximately 626 million 50 bp sequences in single end mode. After quality control, the sequences were aligned in the reference genome of Eucalyptus grandis. After counting reads aligned on each gene, differential expression analyses were performed with edgeR package of R/Bioconductor. A total of 6169 differentially expressed genes (DEG) (FDR ≤ 5% and Log2 FoldChange > |2|) were identified. Of these, 3350 were up-regulated and 2819 were repressed under osmotic stress, considering all the time-series evaluations. At three hours after treatment the highest number of DEG was obtained, with 3150, followed by six hours (2494 DEG) and 12h (525 DEG). Functional enrichment analysis was performed with the Fisher exact test, by software Blast2GO, with the DEG associated with their respective Gene Ontology (GO) terms. As a total of 90 GO terms, enriched and non-redundant, was verified that our results indicate a rapid initial response of Eucalyptus roots induced by PEG 8000 in hydroponics. The most remarkable characteristics of this response under hydroponic conditions are: increased energy consumption via carbohydrate catabolism, activation of the glycolytic pathway, repression of the respiratory chain in the mitochondria, increase of the fermentation pathway, partial execution of the citric acid cycle and repression of the primary metabolism (DNA replication and RNA biosynthesis, development and growth). These enriched functional terms point to the predominance of hypoxia stress, induced by the high viscosity of PEG in hydroponics, as compared to the osmotic stress.
publishDate 2017
dc.date.issued.fl_str_mv 2017-12-18
dc.date.accessioned.fl_str_mv 2018-10-05T14:14:11Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv FAGUNDES, L. F. RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia. 2017. 85 f. Dissertação (Mestrado em Genética e Melhoramento de Plantas) - Universidade Federal de Goiás, Goiânia, 2017.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/8948
dc.identifier.dark.fl_str_mv ark:/38995/001300000dshf
identifier_str_mv FAGUNDES, L. F. RNA-Seq indica que Polietilenoglicol induz estresse de hipóxia nas raízes de Eucalyptus em hidroponia. 2017. 85 f. Dissertação (Mestrado em Genética e Melhoramento de Plantas) - Universidade Federal de Goiás, Goiânia, 2017.
ark:/38995/001300000dshf
url http://repositorio.bc.ufg.br/tede/handle/tede/8948
dc.language.iso.fl_str_mv por
language por
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600
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dc.relation.cnpq.fl_str_mv -3091138714907603907
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dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
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dc.publisher.none.fl_str_mv Universidade Federal de Goiás
dc.publisher.program.fl_str_mv Programa de Pós-graduação em Genética e Melhoramento de Plantas (EA)
dc.publisher.initials.fl_str_mv UFG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Escola de Agronomia - EA (RG)
publisher.none.fl_str_mv Universidade Federal de Goiás
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