Key Takeaways & Executive Findings
- •• Seven candidate reference genes were evaluated for qRT-PCR normalization in Lycoris aurea under seven exogenous hormone treatments. • Expression stability was assessed using geNorm, BestKeeper, NormFinder, and RefFinder algorithms, providing a comprehensive ranking. • The study fills a gap in reference gene validation for Lycoris species under hormone stress conditions, crucial for accurate gene expression analysis. • Identified stable reference genes will facilitate molecular studies on bulb growth and development, aiding in optimizing artificial reproduction.
Abstract
The bulbs of Lycoris aurea Herb can produce unique alkaloids, which have been shown to have antiviral, anti-tumor, anti-malarial, and immunostimulatory properties and are effective in the symptomatic treatment of Alzheimer’s disease. As a consequence, the demand for Lycoris bulbs has dramatically increased, especially for L. aurea, which is native to southern China and is highly important for increasing bulb yield. Previous research in this area reported that the use of exogenous hormones is a way to optimize the artificial reproduction of their bulbs. Therefore, studying the molecular mechanism of bulb growth and development is highly important. Quantitative real-time PCR (qRT-PCR) is widely used in gene expression analysis to explore gene functions that regulate plant growth and development because of its high specificity, accuracy, sensitivity and efficiency, and selecting reference genes correctly is crucial for obtaining proper results and interpretations via qRT-PCR analysis. The selection and validation of reference genes of some Lycoris species in different floral development stages and tissues have been carried out, but under some abnormal conditions, including abiotic stress and hormone treatments, have not been assessed. In this study, we aimed to identify suitable reference genes for L. aurea under seven hormone treatments. A total of seven candidate reference genes were selected for investigation. The expressions of these candidate genes were measured by qRT-PCR. The expression stability of candidate genes was comprehensively evaluated by four different statistical algorithms, including geNorm, BestKeeper, NormFinder and RefFinder.
1. Introduction
The bulbs of Lycoris aurea Herb can produce unique alkaloids, which have been shown to have antiviral, anti-tumor, anti-malarial, and immunostimulatory properties and are effective in the symptomatic treatment of Alzheimer’s disease [1]. As a consequence, the demand for Lycoris bulbs has dramatically increased, especially for L. aurea, which is native to southern China and is highly important for increasing bulb yield [2]. Previous research in this area reported that the use of exogenous hormones is a way to optimize the artificial reproduction of their bulbs [3]. Therefore, studying the molecular mechanism of bulb growth and development is highly important.
Quantitative real-time PCR (qRT-PCR) is widely used in gene expression analysis to explore gene functions that regulate plant growth and development because of its high specificity, accuracy, sensitivity and efficiency, and selecting reference genes correctly is crucial for obtaining proper results and interpretations via qRT-PCR analysis [4]. The selection and validation of reference genes of some Lycoris species in different floral development stages and tissues have been carried out, but under some abnormal conditions, including abiotic stress and hormone treatments, have not been assessed [5,6]. In this study, we aimed to identify suitable reference genes for L. aurea under seven hormone treatments. A total of seven candidate reference genes were selected for investigation. The expressions of these candidate genes were measured by qRT-PCR. The expression stability of candidate genes was comprehensively evaluated by four different statistical algorithms, including geNorm, BestKeeper, NormFinder and RefFinder [7–10].
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Wei Zhao, Ying Tian, Yi Wang, Yun Wu, Ziming Ren, Kehu Li (2026). Selection of reference genes for quantitative real-time PCR analysis in exogenous hormone-treated Lycoris aurea. Acta Biochimica et Biophysica Sinica. https://doi.org/10.3724/abbs.2024197
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Frequently Asked Questions
What is the purpose of this study?
The study aims to identify suitable reference genes for quantitative real-time PCR (qRT-PCR) analysis in Lycoris aurea under seven exogenous hormone treatments, as stable reference genes are crucial for accurate gene expression analysis.
Which candidate reference genes were evaluated?
Seven candidate reference genes were evaluated: AP4, UBQ4, TUB2, EIF, UBCE, TBP, and UBC24.
What statistical algorithms were used to assess gene stability?
The expression stability of candidate genes was evaluated using four statistical algorithms: geNorm, BestKeeper, NormFinder, and RefFinder.
Why is it important to validate reference genes under hormone treatments?
Reference gene expression can vary under different conditions, including hormone treatments. Validating stable reference genes under these conditions ensures accurate normalization in qRT-PCR, which is essential for reliable gene expression studies.
What are the potential applications of this research?
The identified stable reference genes will facilitate molecular studies on bulb growth and development in Lycoris aurea, aiding in optimizing artificial reproduction and understanding the effects of exogenous hormones.
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