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. 2021 May 13;12(5):732.
doi: 10.3390/genes12050732.

Identification and Validation of Reference Genes for Gene Expression Analysis in Schima superba

Zhongyi Yang  1   2   3 , Rui Zhang  1   3 , Zhichun Zhou  1   3
Affiliations

Identification and Validation of Reference Genes for Gene Expression Analysis in Schima superba

Zhongyi Yang et al. Genes (Basel). .

Abstract

Real-time quantitative PCR (RT-qPCR) is a reliable and high-throughput technique for gene expression studies, but its accuracy depends on the expression stability of reference genes. Schima superba is a fast-growing timber species with strong resistance. However, thus far, reliable reference gene identifications have not been reported in S. superba. In this study, 19 candidate reference genes were selected and evaluated for their expression stability in different tissues of S. superba. Three software programs (geNorm, NormFinder, and BestKeeper) were used to evaluate the reference gene transcript stabilities, and comprehensive stability ranking was generated by the geometric mean method. Our results show that SsuACT was the most stable reference gene and that SsuACT + SsuRIB was the best reference gene combination for different tissues. Finally, the stable and less stable reference genes were verified using SsuSND1 expression in different tissues. To our knowledge, this is the first report to verify appropriate reference genes for normalizing gene expression in S. superba for different tissues, which will facilitate the future elucidation of gene regulations in this species and useful references for relative species.

Keywords: Schima superba; real-time quantitative PCR; reference gene; stability evaluation; tissues.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Expression stability values (M) of candidate reference genes calculated by geNorm.
Figure 2
Figure 2
Pairwise variation (V) of candidate reference genes calculated by geNorm.
Figure 3
Figure 3
Expression stability of candidate reference genes analyzed by NormFinder.
Figure 4
Figure 4
Relative expression of the SsuSND1 gene using various different reference genes for normalization in different tissues of S. superba.
See this image and copyright information in PMC

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