Selection and Validation of Reference Genes in the Seeds of Paeonia delavayi in Quantitative Real-time PCR Analysis

doi:10.13560/j.cnki.biotech.bull.1985.2019-1208

Abstract

Abstract: The objective of this work is to analyze the expression stability of internal reference genes in the seeds at different developmental stages of Paeonia delavayi by real-time quantitative PCR（RT-qPCR）,and to screen the most stable reference genes in the maturation of seeds. RT-qPCR was used to analyze their expression differences of 8 housekeeping internal reference genes such as GAPC,PEPC,CYC,ACT,EF2α,TUB,ACP1 and RPL1 in P. delavayi seeds at different developmental stages. Three software（GeNorm,Norm Finder and BestKeeper）were applied to evaluate the expression stabilities of above genes in the seeds at different developmental stages of P. delavayi. The results from RT-qPCR and software demonstrated that the expression stabilities of 8 reference genes varied,the most stable ones were CYC and EF2,while the least sable one was ACT. RT-qPCR analysis of some related genes in the lipid metabolism and synthesis pathway of P. delavayi presented the same expression trend,which verified that CYC and EF2α were suitable as reference genes. In conclusion,2 internal reference genes,CYC and EF2α,are obtained to be the suitable internal reference genes for the seeds of P. delavayi.

Key words: Paeonia delavayi, seed, RT-q PCR, reference gene

PAN Mo-han, LU Tian-quan, TIAN Bo. Selection and Validation of Reference Genes in the Seeds of Paeonia delavayi in Quantitative Real-time PCR Analysis[J]. Biotechnology Bulletin, 2020, 36(9): 218-226.

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