biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 64:253-257, 2020 | DOI: 10.32615/bp.2020.016

Transcriptome-based validation of proper reference genes for reverse trascription quantitative PCR analysis of Sinocalycanthus chinensis

C. ZHANG, Q.-N. JIANG, Y.-G. WANG, J.-X. FU, B. DONG, L.-H. ZHOU*, H.-B. ZHAO*
Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants,
Key Laboratory of National Forestry and Grassland Administration on Germplasm Innovation and Utilization for Southern Garden Plants, School of Landscape Architecture, Zhejiang Agriculture and Forestry University,
Hangzhou 311300, Zhejiang, P.R. China

Reverse transcription quantitative PCR is a widely used method to detect gene expressions. To obtain accurate expression results, the selection of proper reference genes is important and necessary. However, related works concerning reference gene selection have not been carried on many plant species, especially endangered ones. The aim of the present study was to select dependable reference genes for expression normalization of an endangered plant species with medicinal and ornamental value: Sinocalycanthus chinensis (Calycanthaceae). Nine reference genes with stable expressions were chosen for further analysis according to transcriptomic sequencing data from S. chinensis. The expression stability of these candidate genes in inner and outer petals at different floral developmental stages and in many different tissues was then analyzed with the geNorm and NormFinder software. The reference genes were evaluated by normalizing the expression of the anthocyanidin synthase gene in the outer and inner petals at different floral developmental stages to further verify the expression stability of these genes. Elongation factor 1-alpha (ScEF) and 50S ribosomal protein L27 were found to be the two most stable genes in the overall ranking of all the samples and different tissues. Furthermore, ScEF and protein phosphatase 2A were stably expressed in all petal samples. Moreover, among the nine candidate reference genes, phosphoglycerate kinase performed poorly in all sample sets. Our results will help to obtain reliable expression data in molecular studies of S. chinensis.

Keywords: anthocyanidin synthase, floral developmental stages, geNorm, NormFinder, petals.

Received: September 5, 2019; Revised: January 17, 2020; Accepted: January 28, 2020; Published online: March 19, 2020  Show citation

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ZHANG, C., JIANG, Q.-N., WANG, Y.-G., FU, J.-X., DONG, B., ZHOU, L.-H., & ZHAO, H.-B. (2020). Transcriptome-based validation of proper reference genes for reverse trascription quantitative PCR analysis of Sinocalycanthus chinensis. Biologia plantarum, 64, Article 253-257. https://doi.org/10.32615/bp.2020.016
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