Abstract
Quantitative real-time PCR (qPCR) is a popular and powerful tool used to understand the molecular mechanisms of flower development. However, the accuracy of this approach depends on the stability of reference genes. The capitulum of chrysanthemums is very special, which is consisting of ray florets and disc florets. There are obvious differences between the two types of florets in symmetry, gender, histological structure, and function. Furthermore, the ray florets have various shapes. The objective of present study was to identify the stable reference genes in Chrysanthemum morifolium and Chrysanthemum lavandulifolium during the process of flower development. In this study, nine candidate reference genes were selected and evaluated for their expression stability acrosssamples during the process of flower development, and their stability was validated by four different algorithms (Bestkeeper, NormFinder, GeNorm, and Ref-finder). SAND (SAND family protein) was found to be the most stably expressed gene in all samples or different tissues during the process of C. lavandulifolium development. Both SAND and PGK (phosphoglycerate kinase) performed most stable in Chinese large-flowered chrysanthemum cultivars, and PGK was the best in potted chrysanthemums. There were differences in best reference genes among varieties as the genetic background of them were complex. These studies provide guidance for selecting reference genes for analyzing the expression pattern of floral development genes in chrysanthemums.
Highlights
Asteraceae is one of the largest families of flowering plants (Tähtiharju et al, 2012)
QRT-PCR has become an important technique for the analysis of gene expression because of its sensitivity, accuracy, and high throughput nature (Bustin, 2000)
The ideal reference genes should be stably expressed under all experimental conditions and independent of organs, tissues, developmental stages, and different treatments (Huggett et al, 2005)
Summary
Asteraceae is one of the largest families of flowering plants (Tähtiharju et al, 2012). Members of this family have typical capitulum, which is consists of ray florets and disc florets. Ray florets are highly specialized in pollinator attraction and disc florets assume the reproductive function. The combination of two types florets improve the chances of reproductive success and are more evolved than a single flower (Lane, 1994; Funk et al, 2009). There are obvious differences between the two types of florets in symmetry, gender, histological structure, and function.
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