Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses

Abstract Urochloa brizantha is one of the most important warm season forage grasses in tropical countries. Despite its importance, there are few studies on gene expression in this species under stressful conditions. Real-time (RT-qPCR) is an accurate technique for gene quantification analysis, but r...

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Autores principales:	Luciana Midori Takamori, Alyne Valéria Carrion Pereira, Gustavo Maia Souza, Luiz Gonzaga Esteves Vieira, Alessandra Ferreira Ribas
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b84d630e4c9d442eb8bf048fe202621a
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spelling	oai:doaj.org-article:b84d630e4c9d442eb8bf048fe202621a2021-12-02T12:32:56ZIdentification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses10.1038/s41598-017-09156-72045-2322https://doaj.org/article/b84d630e4c9d442eb8bf048fe202621a2017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-09156-7https://doaj.org/toc/2045-2322Abstract Urochloa brizantha is one of the most important warm season forage grasses in tropical countries. Despite its importance, there are few studies on gene expression in this species under stressful conditions. Real-time (RT-qPCR) is an accurate technique for gene quantification analysis, but reference genes must be validated under the same conditions used to assess the expression of the target genes. Here, we evaluated the stability of nine reference genes: Actin 12, Eukaryotic initiation factor 4 A, Elongation factor-1 alpha, FTSH protease 4, U2 auxiliary fator, Succinol Co-enzyme A, Tubulin alfa-5, Tubulin beta-6, Ubiquitin conjugating enzyme. Total RNA was extract from leaf tissues of U. brizantha subjected to 6, 12 and 24 h of cold and heat stresses (10 and 45 °C, respectively), and drought, including moderate (−0.5 to −0.7 MPa), severe (−1.1 to −1.8 MPa) and recovery after re-watering. The RefFinder web-based tool was used to rank the most stable reference genes for each stress. Elongation factor-1 alpha, Elongation factor-1 alpha or Ubiquitin conjugating enzyme, and Eukaryotic initiation factor 4 A were the most stable genes for heat, cold and drought stress, respectively. The expression of Rubisco large subunit gene was normalized against the most stable gene selected by ReFfinder for each stress.Luciana Midori TakamoriAlyne Valéria Carrion PereiraGustavo Maia SouzaLuiz Gonzaga Esteves VieiraAlessandra Ferreira RibasNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Luciana Midori Takamori Alyne Valéria Carrion Pereira Gustavo Maia Souza Luiz Gonzaga Esteves Vieira Alessandra Ferreira Ribas Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses
description	Abstract Urochloa brizantha is one of the most important warm season forage grasses in tropical countries. Despite its importance, there are few studies on gene expression in this species under stressful conditions. Real-time (RT-qPCR) is an accurate technique for gene quantification analysis, but reference genes must be validated under the same conditions used to assess the expression of the target genes. Here, we evaluated the stability of nine reference genes: Actin 12, Eukaryotic initiation factor 4 A, Elongation factor-1 alpha, FTSH protease 4, U2 auxiliary fator, Succinol Co-enzyme A, Tubulin alfa-5, Tubulin beta-6, Ubiquitin conjugating enzyme. Total RNA was extract from leaf tissues of U. brizantha subjected to 6, 12 and 24 h of cold and heat stresses (10 and 45 °C, respectively), and drought, including moderate (−0.5 to −0.7 MPa), severe (−1.1 to −1.8 MPa) and recovery after re-watering. The RefFinder web-based tool was used to rank the most stable reference genes for each stress. Elongation factor-1 alpha, Elongation factor-1 alpha or Ubiquitin conjugating enzyme, and Eukaryotic initiation factor 4 A were the most stable genes for heat, cold and drought stress, respectively. The expression of Rubisco large subunit gene was normalized against the most stable gene selected by ReFfinder for each stress.
format	article
author	Luciana Midori Takamori Alyne Valéria Carrion Pereira Gustavo Maia Souza Luiz Gonzaga Esteves Vieira Alessandra Ferreira Ribas
author_facet	Luciana Midori Takamori Alyne Valéria Carrion Pereira Gustavo Maia Souza Luiz Gonzaga Esteves Vieira Alessandra Ferreira Ribas
author_sort	Luciana Midori Takamori
title	Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses
title_short	Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses
title_full	Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses
title_fullStr	Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses
title_full_unstemmed	Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses
title_sort	identification of endogenous reference genes for rt-qpcr expression analysis in urochloa brizantha under abiotic stresses
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/b84d630e4c9d442eb8bf048fe202621a
work_keys_str_mv	AT lucianamidoritakamori identificationofendogenousreferencegenesforrtqpcrexpressionanalysisinurochloabrizanthaunderabioticstresses AT alynevaleriacarrionpereira identificationofendogenousreferencegenesforrtqpcrexpressionanalysisinurochloabrizanthaunderabioticstresses AT gustavomaiasouza identificationofendogenousreferencegenesforrtqpcrexpressionanalysisinurochloabrizanthaunderabioticstresses AT luizgonzagaestevesvieira identificationofendogenousreferencegenesforrtqpcrexpressionanalysisinurochloabrizanthaunderabioticstresses AT alessandraferreiraribas identificationofendogenousreferencegenesforrtqpcrexpressionanalysisinurochloabrizanthaunderabioticstresses
_version_	1718393943035478016

Identification of Endogenous Reference Genes for RT-qPCR Expression Analysis in Urochloa brizantha Under Abiotic Stresses

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