Selection of the Reference Gene for Expression Normalization in Papaver somniferum L. under Abiotic Stress and Hormone Treatment.

Zhaoping Zhang,Yucheng Zhao,Junqing Zhang,Yujie Wei,Yanrong Li,Fang Chen,Yujie Su,Yongfu Gong,Changjian Li

doi:10.3390/genes11020124

Zhaoping Zhang, Yucheng Zhao + Show 7 more

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https://doi.org/10.3390/genes11020124

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Abstract

Papaver somniferum L. is an important medical plant that produces analgesic drugs used for the pain caused by cancers and surgeries. Recent studies have focused on the expression genes involved in analgesic drugs biosynthesis, and the real-time quantitative polymerase chain reaction (RT-qPCR) technique is the main strategy. However, no reference genes have been reported for gene expression normalization in P. somniferum. Herein, nine reference genes (actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cyclophilin 2 (CYP2), elongation factor 1-alpha (EF-1α), glyceraldehyde-3-phosphate dehydrogenase 2, cytosolic (GAPC2), nuclear cap-binding protein subunit 2 (NCBP2), protein phosphatase 2A (PP2A), TIP41-like protein (TIP41), and tubulin beta chain (TUB)) of P. somniferum were selected and analyzed under five different treatments (cold, drought, salt, heavy metal, and hormone stress). Then, BestKeeper, NormFinder, geNorm, and RefFinder were employed to analyze their gene expression stability. The results reveal that NCBP2 is the most stable reference gene under various experimental conditions. The work described here is the first report regarding on reference gene selection in P. somniferum, which could be used for the accurate normalization of the gene expression involved in analgesic drug biosynthesis.

Highlights

Papaver somniferum L., known as the opium poppy, originated from Europe and now is grown around the world
P. somniferum is attractive for researchers, and research efforts have been undertaken in many aspects, including the pharmacological activity and metabolism of alkaloids in the plant [5,6]
The most stably expressed genes were able to normalize the data of real-time quantitative polymerase chain reaction (RT-qPCR), which is a commonly used tool for gene expression analysis

Summary

Introduction

Papaver somniferum L., known as the opium poppy, originated from Europe and now is grown around the world. It is famous as an analgesic, as well as a narcotic and alkaloid, and morphine, codeine, papaverine, and thebaine are its main compounds [1,2]. Due to their analgesic activities, pharmaceutical opiates are universally applied for relieving the pain caused by cancer, surgery, and wounds [3]. P. somniferum is attractive for researchers, and research efforts have been undertaken in many aspects, including the pharmacological activity and metabolism of alkaloids in the plant [5,6]

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