title: Selection of Reference Genes for Expression Study in Pulp and Seeds of Theobroma grandiflorum (Willd. ex Spreng.) Schum.
abstract: Cupuassu (Theobroma grandiflorum [Willd. ex Spreng.] Schum) is a species of high economic importance in Brazil with great potential at international level due to the multiple uses of both its seeds and pulp in the industry of sweets and cosmetics. For this reason, the cupuassu breeding program focused on the selection of genotypes with high pulp and seed quality-selection associated with the understanding of the mechanisms involved in fruit formation. Gene expression is one of the most used approaches related to such understanding. In this sense, quantitative real-time PCR (qPCR) is a powerful tool, since it rapidly and reliably quantifies gene expression levels across different experimental conditions. The analysis by qPCR and the correct interpretation of data depend on signal normalization using reference genes, i.e. genes presenting a uniform pattern of expression in the analyzed samples. Here, we selected and analyzed the expression of five genes from cupuassu (ACP, ACT, GAPDH, MDH, TUB) to be used as candidates for reference genes on pulp and seed of young, maturing and mature cupuassu fruits. The evaluation of the gene expression stability was obtained using the NormFinder, geNorm and BestKeeper programs. In general, our results indicated that the GAPDH and MDH genes constituted the best combination as reference genes to analyze the expression of cupuassu samples. To our knowledge, this is the first report of reference gene definition in cupuassu, and these results will support subsequent analysis related to gene expression studies in cupuassu plants subjected to different biotic or abiotic conditions as well as serve as a tool for diversity analysis based on pulp and seed quality.
authors: Universidade Estadual de Santa Cruz (UESC), Departamento de Ciências Biológicas (DCB), Centro de Biotecnologia e Genética (CBG), Rodovia Ilhéus-Itabuna, km 16, 45662-900 Ilhéus-BA, Brazil.
authors: Lucas
authors: L
authors: Ferraz Dos Santos
authors: Universidade Estadual de Santa Cruz (UESC), Departamento de Ciências Biológicas (DCB), Centro de Biotecnologia e Genética (CBG), Rodovia Ilhéus-Itabuna, km 16, 45662-900 Ilhéus-BA, Brazil.
authors: Raner José
authors: RJ
authors: Santana Silva
authors: Universidade Estadual de Santa Cruz (UESC), Departamento de Ciências Biológicas (DCB), Centro de Biotecnologia e Genética (CBG), Rodovia Ilhéus-Itabuna, km 16, 45662-900 Ilhéus-BA, Brazil.
authors: Daniel
authors: D
authors: Oliveira Jordão do Amaral
authors: Universidade Estadual de Santa Cruz (UESC), Departamento de Ciências Biológicas (DCB), Centro de Biotecnologia e Genética (CBG), Rodovia Ilhéus-Itabuna, km 16, 45662-900 Ilhéus-BA, Brazil.
authors: Márcia Fabiana
authors: MF
authors: Barbosa de Paula
authors: Embrapa Recursos Genéticos e Biotecnologia, Brasília-DF, 70770-917, Brazil.
authors: Loeni
authors: L
authors: Ludke Falcão
authors: CIRAD, UMR AGAP, F-34398 Montpellier, France.
authors: Thierry
authors: T
authors: Legavre
authors: Embrapa Amazônia Oriental, 66095-903 Belém-PA, Brazil.
authors: Rafael Moyses
authors: RM
authors: Alves
authors: Embrapa Recursos Genéticos e Biotecnologia, Brasília-DF, 70770-917, Brazil.
authors: Lucilia Helena
authors: LH
authors: Marcellino
authors: Universidade Estadual de Santa Cruz (UESC), Departamento de Ciências Biológicas (DCB), Centro de Biotecnologia e Genética (CBG), Rodovia Ilhéus-Itabuna, km 16, 45662-900 Ilhéus-BA, Brazil.|CIRAD, UMR AGAP, F-34398 Montpellier, France.
authors: Fabienne
authors: F
authors: Micheli
mesh_terms: D017931:DNA Primers; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D019660:Malvaceae / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012639:Seeds / Q000235:genetics
chemical_list: D017931:DNA Primers