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

Autor(es)

Takamori, Luciana Midori ; Carrion Pereira, Alyne Valeria ; Souza, Gustavo Maia ; Esteves Vieira, Luiz Gonzaga ; Ribas, Alessandra Ferreira

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 7, AUG 17 2017.
• Citações Web of Science: 3

Resumo

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 degrees 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. (AU)

• Processo FAPESP: 14/07830-1 - Estabelecimento dos parâmetros de transformação para U. brizantha via bombardeamento de partículas e obtenção de plantas transgênicas
• Beneficiário: Alyne Valéria Carrion Pereira
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico
• Processo FAPESP: 13/04819-4 - Transformação genética de Urochloa brizantha visando aumentar a tolerância a estresses abióticos
• Beneficiário: Alessandra Ferreira Ribas
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores