Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato
Autor(a) principal: | |
---|---|
Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFLA |
Texto Completo: | http://repositorio.ufla.br/jspui/handle/1/40634 |
Resumo: | The anthocyanins are compounds from the secondary metabolism of plants and belong to the flavonoids group. They are responsible for the blue, red and purple pigmentation in many plant tissues, as leaves, flowers, seeds and fruits. For plants they are mainly responsible for the growth and development, defense against pathogens, drought tolerance and protection against UV radiation. In addition, the anthocyanin are extremely beneficial to human health, they fight and prevent a range of chronic diseases. Among these benefits, antioxidant effects, prevention against cardiovascular diseases and cancer stand out. The main vegetables and fruits present in our diet accumulate anthocyanin just in small quantities, and in most cases being restricted to the epidermal layers of the edible parts. Since the peel usually represents less than 5% of the total mass of edible parts of the plant, a high anthocyanin accumulation is desirable, and the tomato is a great candidate for this improvement, since it is the principal vegetable in worldwide. microRNAs (miRNAs) are a class of small non-coding RNAs that contain 21-24 nucleotides and regulate gene expression post transcriptionally, by mRNA cleavage or suppressing translation. In tomato, the miRNAs role in the anthocyanin biosynthesis pathway is still not well characterized. Aiming for a better understanding of the action of the miRNAs in this pathway, we performed the prediction, validation and knockout of some miRNAs as well as the expression analyses for some of the anthocyanin pathway related genes. The miRNA target prediction identified the genes Anthocyanidin synthase (ANS), SlAN1 (bHLH) and Chalcone Synthase (CHS) being possible regulated by the miR5368, miR6024-3p and miR9471b-3p, respectively. We analyzed the expression of these miRNAs and anthocyanin related genes in Micro-Tom triple mutant Aft/atv/hp2 and Micro-Tom Wild-Type by RT-qPCR. Afterwards, we performed the knockout of each miRNA precursor in Micro-Tom Wild-Type by CRISPR-Cas9. As a result, we observed transformed explants with a purple phenotype in leaf veins and stem, indicating that the miR5368, miR6024 and miR9471 promote negative regulation in the anthocyanin biosynthesis pathway acting in the genes ANS, SlAN1 and CHS respectively. |
id |
UFLA_23e8590292c30972233278cee5a422d8 |
---|---|
oai_identifier_str |
oai:localhost:1/40634 |
network_acronym_str |
UFLA |
network_name_str |
Repositório Institucional da UFLA |
repository_id_str |
|
spelling |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomatoNovos microRNAs controlando a rota de biossíntese de antocianina em tomateAntocianinas - BiossínteseMicroRNAsCRISPR-Cas9Anthocyanin - BiosynthesisGenética vegetalThe anthocyanins are compounds from the secondary metabolism of plants and belong to the flavonoids group. They are responsible for the blue, red and purple pigmentation in many plant tissues, as leaves, flowers, seeds and fruits. For plants they are mainly responsible for the growth and development, defense against pathogens, drought tolerance and protection against UV radiation. In addition, the anthocyanin are extremely beneficial to human health, they fight and prevent a range of chronic diseases. Among these benefits, antioxidant effects, prevention against cardiovascular diseases and cancer stand out. The main vegetables and fruits present in our diet accumulate anthocyanin just in small quantities, and in most cases being restricted to the epidermal layers of the edible parts. Since the peel usually represents less than 5% of the total mass of edible parts of the plant, a high anthocyanin accumulation is desirable, and the tomato is a great candidate for this improvement, since it is the principal vegetable in worldwide. microRNAs (miRNAs) are a class of small non-coding RNAs that contain 21-24 nucleotides and regulate gene expression post transcriptionally, by mRNA cleavage or suppressing translation. In tomato, the miRNAs role in the anthocyanin biosynthesis pathway is still not well characterized. Aiming for a better understanding of the action of the miRNAs in this pathway, we performed the prediction, validation and knockout of some miRNAs as well as the expression analyses for some of the anthocyanin pathway related genes. The miRNA target prediction identified the genes Anthocyanidin synthase (ANS), SlAN1 (bHLH) and Chalcone Synthase (CHS) being possible regulated by the miR5368, miR6024-3p and miR9471b-3p, respectively. We analyzed the expression of these miRNAs and anthocyanin related genes in Micro-Tom triple mutant Aft/atv/hp2 and Micro-Tom Wild-Type by RT-qPCR. Afterwards, we performed the knockout of each miRNA precursor in Micro-Tom Wild-Type by CRISPR-Cas9. As a result, we observed transformed explants with a purple phenotype in leaf veins and stem, indicating that the miR5368, miR6024 and miR9471 promote negative regulation in the anthocyanin biosynthesis pathway acting in the genes ANS, SlAN1 and CHS respectively.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)As antocianinas são compostos pertencentes ao metabolismo secundário das plantas derivadas do grupo dos flavonoides. Elas são responsáveis pela coloração azul, vermelha ou roxa em diferentes tecidos da planta, como folhas, flores, sementes e frutos. Nas plantas, elas são principalmente responsáveis pelo crescimento e desenvolvimento, defesa contra patógenos, tolerância a seca e proteção contra radiação UV. Além disso, as antocianinas são extremamente benéficas para a saúde humana, elas combatem e previnem uma série de doenças crônicas. Dentre estes benefícios, se destacam os efeitos antioxidantes, prevenção contra doenças cardiovasculares e câncer. Os principais vegetais e frutas presentes em nossa dieta acumulam antocianina apenas em pequenas quantidades, sendo a maioria restrita as camadas epidérmicas. Pela casca representar menos de 5% da massa total das partes comestíveis da planta, um aumento nos teores de antocianina é desejável, e o tomate se mostra um ótimo candidato para este aumento, visto que ele representa o principal vegetal consumido em todo o mundo. Os microRNAs (miRNA) são uma classe de pequenos RNAs não codificadores de proteínas, de tamanho variável entre 20-24 nucleotídeos, atuando no controle da expressão de genes codificadores de proteínas, por meio da clivagem do mRNA ou bloqueando a sua tradução. No tomate, a atuação dos miRNAs na rota de biossíntese de antocianina ainda é pouco estudada. Visando um melhor entendimento da atuação dos miRNAs nesta rota, realizamos a predição, validação e knockout de alguns miRNAs bem como análises de expressão para alguns genes relacionados à rota de biossíntese da antocianina. A predição de alvos para os miRNAs identificou os genes Anthocyanidin synthase (ANS), SlAN1 (bHLH) e Chalcone Synthase (CHS) sendo possivelmente regulados por miR5368, miR6024-3p e miR9471b-3p respectivamente. Analisamos a expressão destes miRNAs e dos genes relacionados a rota da antocianina no Micro-Tom Aft/atv/hp2 e Micro-Tom Selvagem por RT-qPCR. Posteriormente, realizamos o knockout de cada precursor destes miRNAs no Micro-Tom Selvagem pela técnica CRISPR-Cas9. Como resultados, observamos um fenótipo roxo nas nervuras e na haste dos explantes transformados, indicando que o miR5368, miR6024 e miR9471 promovem uma regulação negativa na rota de biossíntese de antocianina atuando nos genes ANS, SlAN1 e CHS respectivamente.Universidade Federal de LavrasPrograma de Pós-Graduação em Biotecnologia VegetalUFLAbrasilNão especifica vinculação com nenhum departamentoChalfun Junior, AntonioBrum, Christiane Noronha FernandesBenedito, Vagner AugustoChalfun Junior, AntonioSilva, Éder Marques daBenedito, Vagner AugustoReis, Gabriel Lasmar dos2020-05-06T17:03:10Z2020-05-06T17:03:10Z2020-05-062020-02-20info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfREIS, G. L. dos. Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato. 2020. 52 p. Dissertação (Mestrado em Biotecnologia Vegetal)-Universidade Federal de Lavras, Lavras, 2020.http://repositorio.ufla.br/jspui/handle/1/40634enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFLAinstname:Universidade Federal de Lavras (UFLA)instacron:UFLA2023-04-12T14:11:55Zoai:localhost:1/40634Repositório InstitucionalPUBhttp://repositorio.ufla.br/oai/requestnivaldo@ufla.br || repositorio.biblioteca@ufla.bropendoar:2023-04-12T14:11:55Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA)false |
dc.title.none.fl_str_mv |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato Novos microRNAs controlando a rota de biossíntese de antocianina em tomate |
title |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato |
spellingShingle |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato Reis, Gabriel Lasmar dos Antocianinas - Biossíntese MicroRNAs CRISPR-Cas9 Anthocyanin - Biosynthesis Genética vegetal |
title_short |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato |
title_full |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato |
title_fullStr |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato |
title_full_unstemmed |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato |
title_sort |
Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato |
author |
Reis, Gabriel Lasmar dos |
author_facet |
Reis, Gabriel Lasmar dos |
author_role |
author |
dc.contributor.none.fl_str_mv |
Chalfun Junior, Antonio Brum, Christiane Noronha Fernandes Benedito, Vagner Augusto Chalfun Junior, Antonio Silva, Éder Marques da Benedito, Vagner Augusto |
dc.contributor.author.fl_str_mv |
Reis, Gabriel Lasmar dos |
dc.subject.por.fl_str_mv |
Antocianinas - Biossíntese MicroRNAs CRISPR-Cas9 Anthocyanin - Biosynthesis Genética vegetal |
topic |
Antocianinas - Biossíntese MicroRNAs CRISPR-Cas9 Anthocyanin - Biosynthesis Genética vegetal |
description |
The anthocyanins are compounds from the secondary metabolism of plants and belong to the flavonoids group. They are responsible for the blue, red and purple pigmentation in many plant tissues, as leaves, flowers, seeds and fruits. For plants they are mainly responsible for the growth and development, defense against pathogens, drought tolerance and protection against UV radiation. In addition, the anthocyanin are extremely beneficial to human health, they fight and prevent a range of chronic diseases. Among these benefits, antioxidant effects, prevention against cardiovascular diseases and cancer stand out. The main vegetables and fruits present in our diet accumulate anthocyanin just in small quantities, and in most cases being restricted to the epidermal layers of the edible parts. Since the peel usually represents less than 5% of the total mass of edible parts of the plant, a high anthocyanin accumulation is desirable, and the tomato is a great candidate for this improvement, since it is the principal vegetable in worldwide. microRNAs (miRNAs) are a class of small non-coding RNAs that contain 21-24 nucleotides and regulate gene expression post transcriptionally, by mRNA cleavage or suppressing translation. In tomato, the miRNAs role in the anthocyanin biosynthesis pathway is still not well characterized. Aiming for a better understanding of the action of the miRNAs in this pathway, we performed the prediction, validation and knockout of some miRNAs as well as the expression analyses for some of the anthocyanin pathway related genes. The miRNA target prediction identified the genes Anthocyanidin synthase (ANS), SlAN1 (bHLH) and Chalcone Synthase (CHS) being possible regulated by the miR5368, miR6024-3p and miR9471b-3p, respectively. We analyzed the expression of these miRNAs and anthocyanin related genes in Micro-Tom triple mutant Aft/atv/hp2 and Micro-Tom Wild-Type by RT-qPCR. Afterwards, we performed the knockout of each miRNA precursor in Micro-Tom Wild-Type by CRISPR-Cas9. As a result, we observed transformed explants with a purple phenotype in leaf veins and stem, indicating that the miR5368, miR6024 and miR9471 promote negative regulation in the anthocyanin biosynthesis pathway acting in the genes ANS, SlAN1 and CHS respectively. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-05-06T17:03:10Z 2020-05-06T17:03:10Z 2020-05-06 2020-02-20 |
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.uri.fl_str_mv |
REIS, G. L. dos. Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato. 2020. 52 p. Dissertação (Mestrado em Biotecnologia Vegetal)-Universidade Federal de Lavras, Lavras, 2020. http://repositorio.ufla.br/jspui/handle/1/40634 |
identifier_str_mv |
REIS, G. L. dos. Novel microRNAs controlling the anthocyanin biosynthesis pathway in tomato. 2020. 52 p. Dissertação (Mestrado em Biotecnologia Vegetal)-Universidade Federal de Lavras, Lavras, 2020. |
url |
http://repositorio.ufla.br/jspui/handle/1/40634 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Lavras Programa de Pós-Graduação em Biotecnologia Vegetal UFLA brasil Não especifica vinculação com nenhum departamento |
publisher.none.fl_str_mv |
Universidade Federal de Lavras Programa de Pós-Graduação em Biotecnologia Vegetal UFLA brasil Não especifica vinculação com nenhum departamento |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFLA instname:Universidade Federal de Lavras (UFLA) instacron:UFLA |
instname_str |
Universidade Federal de Lavras (UFLA) |
instacron_str |
UFLA |
institution |
UFLA |
reponame_str |
Repositório Institucional da UFLA |
collection |
Repositório Institucional da UFLA |
repository.name.fl_str_mv |
Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA) |
repository.mail.fl_str_mv |
nivaldo@ufla.br || repositorio.biblioteca@ufla.br |
_version_ |
1807835144501854208 |