Selection of reference genes for expression Study in pulp and seeds of Theobroma grandiflorum (Willd. ex Spreng.) Schum

Ferraz dos Santos Lucas, Santana Silva Raner José, Do Amaral Daniel Oliveira Jordão, Barbosa de Paula Márcia Fabiana, Ludke Falcão Loeni, Legavre Thierry, Alves Rafael Moyses, Marcellino Lucilia Helena, Micheli Fabienne. 2016. Selection of reference genes for expression Study in pulp and seeds of Theobroma grandiflorum (Willd. ex Spreng.) Schum. PloS One, 11 (8):e0160646, 12 p.

https://doi.org/10.1371/journal.pone.0160646

Article de revue ; Article de revue à facteur d'impact Revue en libre accès total

Prévisualisation

Version publiée - Anglais
Sous licence CC0 1.0 Sans restriction de droits pour le monde entier.
2016 Santos et al PlosOne ref genes.pdf
Télécharger (524kB) | Prévisualisation

Quartile : Q1, Sujet : MULTIDISCIPLINARY SCIENCES

Liste HCERES des revues (en SHS) : oui

Thème(s) HCERES des revues (en SHS) : Psychologie-éthologie-ergonomie; Staps

Résumé : 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.

Mots-clés Agrovoc : Theobroma, génotype, gène, identification, expression des gènes, stabilité génétique, pulpe de fruits, graine, PCR, amélioration des plantes, Theobroma grandiflorum