Selection of reliable reference genes for RT-qPCR analysis during developmental stages and abiotic stress in Setaria viridis

Autor(es)

Martins, Polyana Kelly ^[1] ; Mafra, Valeria ^[2] ; de Souza, Wagner Rodrigo ^[1] ; Ribeiro, Ana Paula ^[1] ; Vinecky, Felipe ^[1] ; Basso, Marcos Fernando ^[1] ; Dias Brito da Cunha, Barbara Andrade ^[1] ; Kobayashi, Adilson Kenji ^[1] ; Correa Molinari, Hugo Bruno ^[1]

Afiliação do(s) autor(es)

^[1] Embrapa Agroenergy CNPAE, Genet & Biotechnol Lab, BR-70770901 Brasilia, DF - Brazil

^[2] Brazilian Ctr Res Energy & Mat, Brazilian Bioethanol Sci & Technol Lab, BR-13083100 Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 6, JUN 20 2016.
• Citações Web of Science: 32

Resumo

Real-time PCR (RT-qPCR) expression analysis is a powerful analytical technique, but reliable results depend on the use of stable reference genes for proper normalization. This study proposed to test the expression stability of 13 candidate reference genes in Setaria viridis, a monocot species recently proposed as a new C4 model plant. Gene expression stability of these genes was assayed across different tissues and developmental stages of Setaria and under drought or aluminum stress. In general, our results showed Protein Kinase, RNA Binding Protein and SDH as the most stable genes. Moreover, pairwise analysis showed that two reference genes were sufficient to normalize the gene expression data under each condition. By contrast, GAPDH and ACT were the least stably expressed genes tested. Validation of suitable reference genes was carried out to profile the expression of P5CS and GolS during abiotic stress. In addition, normalization of gene expression of SuSy, involved in sugar metabolism, was assayed in the developmental dataset. This study provides a list of reliable reference genes for transcript normalization in S. viridis in different tissues and stages of development and under abiotic stresses, which will facilitate genetic studies in this monocot model plant. (AU)

• Processo FAPESP: 14/07918-6 - Caracterização funcional do gene Target of Rapamycin using Setaria como planta modelo: dissecando o papel da via TOR regulando crescimento e metabolismo em plantas C4
• Beneficiário: Valéria Mafra Cota
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado