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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
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]
Total Authors: 9
Affiliation:
[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
Total Affiliations: 2
Document type: Journal article
Source: SCIENTIFIC REPORTS; v. 6, JUN 20 2016.
Web of Science Citations: 32
Abstract

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)

FAPESP's process: 14/07918-6 - Functional characterization of the Target of Rapamycin (TOR) gene using Setaria as the model plant: dissecting the role of TOR pathway regulating growth and metabolism in C4 plants
Grantee:Valéria Mafra Cota
Support Opportunities: Scholarships in Brazil - Post-Doctoral