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

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 (Tα2), elongation factor 1-alpha (EF1-α), 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-α 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.

Source: Gene Expression Patterns - September 17, 2019 Category: Genetics & Stem Cells Source Type: research