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Background <p><i>Gynostemma pentaphyllum</i> is an economically valuable medicinal and edible plant. The accumulation of secondary metabolites of <i>G</i>. <i>pentaphyllum</i> exhibits tissue-specificity and is influenced by abiotic factors. However, there is currently little known regarding the molecular mechanisms involved. RT-qPCR provides a clear understanding of the gene expression changes in plants under developmental conditions. This study aimed to identify and validate stable reference genes for use in RT-qPCR studies across different tissues and stress conditions in <i>G. pentaphyllum.</i></p> Methods and results <p>The 12 candidate reference genes (<i>CYP</i>, <i>DnaJ</i>, <i>EF-1α</i>, <i>EIF3</i>, <i>EIF5A2</i>, <i>GAPDH</i>, <i>H1</i>, <i>MUB</i>, <i>RPL</i>, <i>UBC</i>, <i>α-TUB</i> and <i>β-TUB</i>) were selected for measurement by RT-qPCR with different tissues (leaves, petioles, stems, roots, flowers, fruits and tendrils), and abiotic stresses (Cd, MeJA and Cd + MeJA treatment). The geNorm, NormFinder, BestKeeper and RefFinder analysis were used to evaluate stability value of reference gene. The comprehensive ranking of evaluation showed that <i>EF-1α</i> was the most suitable reference gene for different tissues and <i>UBC</i> were the most stable reference genes for Cd stress and MeJA treatments tested. However, <i>CYP</i> showed the lowest stability under different experimental conditions. Furthermore, <i>GpFPS</i> and <i>GpSE</i> were used to verify the applicability of the selected reference genes.</p> Conclusion <p>This study provides the groundwork for precise gene expression measurements in <i>G. pentaphyllum</i>, enabling accurate assessment of target gene expression under different experimental conditions.</p>

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Screening and validation of reference genes of Gynostemma pentaphyllum with different tissues and under different treatment conditions by RT-qPCR analysis

  • 期刊论文

  • 发表日期：

    2025年9月20日

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  • Yunyi Zhou,
  • Qiuyuan Meng,
  • Lixiang Yao,
  • Ying Li,
  • Liying Yu,
  • Chunliu Pan

摘要

Background

Gynostemma pentaphyllum is an economically valuable medicinal and edible plant. The accumulation of secondary metabolites of G. pentaphyllum exhibits tissue-specificity and is influenced by abiotic factors. However, there is currently little known regarding the molecular mechanisms involved. RT-qPCR provides a clear understanding of the gene expression changes in plants under developmental conditions. This study aimed to identify and validate stable reference genes for use in RT-qPCR studies across different tissues and stress conditions in G. pentaphyllum.

Methods and results

The 12 candidate reference genes (CYP, DnaJ, EF-1α, EIF3, EIF5A2, GAPDH, H1, MUB, RPL, UBC, α-TUB and β-TUB) were selected for measurement by RT-qPCR with different tissues (leaves, petioles, stems, roots, flowers, fruits and tendrils), and abiotic stresses (Cd, MeJA and Cd + MeJA treatment). The geNorm, NormFinder, BestKeeper and RefFinder analysis were used to evaluate stability value of reference gene. The comprehensive ranking of evaluation showed that EF-1α was the most suitable reference gene for different tissues and UBC were the most stable reference genes for Cd stress and MeJA treatments tested. However, CYP showed the lowest stability under different experimental conditions. Furthermore, GpFPS and GpSE were used to verify the applicability of the selected reference genes.

Conclusion

This study provides the groundwork for precise gene expression measurements in G. pentaphyllum, enabling accurate assessment of target gene expression under different experimental conditions.