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

Autor: de Andrade LM; Centro de Cana, Instituto Agronômico (IAC), P.O. Box 206, Ribeirão Preto, SP 14001-970 Brazil.; Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP 14049-900 Brazil., Dos Santos Brito M; Centro de Cana, Instituto Agronômico (IAC), P.O. Box 206, Ribeirão Preto, SP 14001-970 Brazil., Fávero Peixoto Junior R; Centro de Cana, Instituto Agronômico (IAC), P.O. Box 206, Ribeirão Preto, SP 14001-970 Brazil.; Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP 14049-900 Brazil., Marchiori PER; Centro de Ecofisilogia e Biofísica, IAC, P.O. Box 1481, Campinas, SP 13012-970 Brazil., Nóbile PM; Centro de Cana, Instituto Agronômico (IAC), P.O. Box 206, Ribeirão Preto, SP 14001-970 Brazil., Martins APB; Centro de Cana, Instituto Agronômico (IAC), P.O. Box 206, Ribeirão Preto, SP 14001-970 Brazil.; Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP 14049-900 Brazil., Ribeiro RV; Departamento de Biologia Vegetal, Instituto de Biologia, Universidade Estadual de Campinas, P.O. Box 6109, Campinas, SP 13083-970 Brazil., Creste S; Centro de Cana, Instituto Agronômico (IAC), P.O. Box 206, Ribeirão Preto, SP 14001-970 Brazil.
Jazyk: angličtina
Zdroj: Plant methods [Plant Methods] 2017 Apr 17; Vol. 13, pp. 28. Date of Electronic Publication: 2017 Apr 17 (Print Publication: 2017).
DOI: 10.1186/s13007-017-0178-2
Abstrakt: 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.
Databáze: MEDLINE
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