Selection and validation of reference genes for quantitative real-time PCR in Cymbidium sinense

Yunfang Tian; Zhigang Chu; Huiyu Wang; Guoxia Wang; Si Wu; Yuzhen Yang

doi:10.2144/btn-2021-0073

Selection and validation of reference genes for quantitative real-time PCR in Cymbidium sinense

Biotechniques. 2022 Feb;72(2):51-59. doi: 10.2144/btn-2021-0073. Epub 2022 Jan 17.

Authors

Yunfang Tian^{1

2}, Zhigang Chu^{1

2}, Huiyu Wang^{1

2}, Guoxia Wang^{1

2}, Si Wu^{1

2}, Yuzhen Yang^{1

2}

Affiliations

¹ College of Life Sciences, Zhengzhou Normal University, Zhengzhou, 450044, People's Republic of China.
² Zhengzhou Key Laboratory of Ornamental & Medicinal Resources Plants, Zhengzhou, 450044, People's Republic of China.

PMID: 35037484
DOI: 10.2144/btn-2021-0073

Abstract

Selection of reference genes (RGs) is important for the accurate analysis of real-time quantitative PCR (RT-qPCR) results. This study screened RGs of Cymbidium sinense for more accurate quantification of target genes. The two most stable RGs for all tissues were ACT and EF1α, those for vegetative organs were UBQ3 and ACT, while those for reproductive organs as well as organs in the full flowering stage were EF1α and ACT. The AGAMOUS (CsAG1) expression level was verified using EF1α, ACT, GAPDH, UBQ2 and UBQ3 as RG. The expression profile of CsAG1 was consistent when normalized with EF1α, ACT and UBQ3. The results have practical value for the expression of key genes during the development of C. sinense.

Keywords: Cymbidium sinense; real-time fluorescent quantitative PCR; reference genes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gene Expression Profiling*
Genes, Plant*
Real-Time Polymerase Chain Reaction / methods
Reference Standards