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|Title:||Selection and validation of castor bean (ricinus communis) reference genes for quantitative PCR (RT-qPCR) in developing and germinating seeds and expression pattern of four ricin-family genes|
|Author:||Rocha, Antonio Jose|
de Oliveira Barsottini, Mario Ramos
da Rocha, Soraya Lilia
|Abstract:||This study aims to expand the set of internal control genes used for RT-qPCR experiments with Castor bean (Ricinus communis) seeds by evaluating candidate genes across several seed tissues and developmental stages. Nine reference genes were selected, including actin-11 (ACT11), tubulin alpha-2 (T alpha 2), elongation factor 1-alpha (EF1-alpha), protein phosphatase 2A-2 (PP2A2), polyubiquitin-3 (PUB3) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Biological samples consisted of R. communis seeds in 15 stages of maturation and germination. We demonstrate that PP2A2, PUB3 and EF1-alpha are the most stably expressed genes across the tested conditions and therefore appropriate for RT-qPCR. Subsequently, those reference genes were used for the analysis of the expression of four R. communis ricin-family genes. In developing seeds, the highest ricin expression levels was seen in the nucellus and in the endosperm, whereas in germinating seeds a peak expression occurs 4-6 days after germination. The four tested ricin isoforms exhibited differential expression patterns across tissues and seed developmental stages, which may indicate distinct biological roles for each ricin gene|
|Appears in Collections:||IB - Artigos e Outros Documentos|
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