Reference genes for normalization of qPCR assays in sugarcane plants under water deficit

Autor(es)

de Andrade, Larissa Mara ; Brito, Michael dos Santos ; Peixoto Junior, Rafael Favero ; Ribeiro Marchiori, Paulo Eduardo ; Nobile, Paula Macedo ; Boer Martins, Alexandre Palma ; Ribeiro, Rafael Vasconcelos ; Creste, Silvana

• Tipo de documento: Artigo Científico
• Fonte: PLANT METHODS; v. 13, APR 17 2017.
• Citações Web of Science: 14

Resumo

Background: Sugarcane (Saccharum spp.) is the main raw material for sugar and ethanol production. Among the abiotic stress, drought is the main one that negatively impact sugarcane yield. Although gene expression analysis through quantitative PCR (qPCR) has increased our knowledge about biological processes related to drought, gene network that mediates sugarcane responses to water deficit remains elusive. In such scenario, validation of reference gene is a major requirement for successful analyzes involving qPCR. Results: In this study, candidate genes were tested for their suitable as reference genes for qPCR analyses in two sugarcane cultivars with varying drought tolerance. Eight candidate reference genes were evaluated in leaves sampled in plants subjected to water deficit in both field and greenhouse conditions. In addition, five genes were evaluated in shoot roots of plants subjected to water deficit by adding PEG8000 to the nutrient solution. NormFinder and RefFinder algorithms were used to identify the most stable gene(s) among genotypes and under different experimental conditions. Both algorithms revealed that in leaf samples, UBQ1 and GAPDH genes were more suitable as reference genes, whereas GAPDH was the best reference one in shoot roots. Conclusion: Reference genes suitable for sugarcane under water deficit were identified, which would lead to a more accurate and reliable analysis of qPCR. Thus, results obtained in this study may guide future research on gene expression in sugarcane under varying water conditions. (AU)

• Processo FAPESP: 08/57495-3 - Integrando características fisiológicas, morfológicas e anatômicas para entender o rendimento diferencial de sacarose em genótipos de cana-de-açúcar
• Beneficiário: Eduardo Caruso Machado
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Temático
• Processo FAPESP: 11/50661-8 - Caracterização de transcritos de cana-de-açúcar que respondem ao estresse hídrico visando à obtenção de transgênicos tolerantes à seca
• Beneficiário: Silvana Aparecida Creste Dias de Souza
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Regular