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Volume 26
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10.3390/ijms262211004
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Screening and Identification of Reference Genes Under Different Conditions and Growth Stages of Lyophyllum decastes

by
Yun-Qi Hui
1,2,†,
Huan-Ling Yang
2,†,
Yu-Qing Zhang
1,2,
Chen-Zhao Zhu
2,
Li-Ping Xi
2,
Chun-Yan Song
2,
Zheng-Peng Li
2,
E-Xian Li
3,
Shu-Hong Li
3,
Yong-Nan Liu
1,* and
Rui-Heng Yang
2,*
1
Hunan Provincial Key Laboratory of Forestry Biotechnology, Central South University of Forestry & Technology, Changsha 410004, China
2
Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
3
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory for Southwest Microbial Diversity of the Ministry of Education, Yunnan University, Kunming 650032, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Int. J. Mol. Sci. 2025, 26(22), 11004; https://doi.org/10.3390/ijms262211004 (registering DOI)
Submission received: 11 October 2025 / Revised: 4 November 2025 / Accepted: 5 November 2025 / Published: 13 November 2025
(This article belongs to the Section Molecular Plant Sciences)
Versions Notes

Abstract

Internal reference genes are a prerequisite for ensuring the accuracy of gene verification experiments, but few relevant studies on Lyophyllum decastes have investigated the growth cycle and different environmental conditions. In this study, the qPCR results of 22 house-keeping genes were analyzed using GeNorm, BestKeeper, NormFinder and RefFinder. The results revealed that the most stable gene differed under different conditions. Across all developmental stages and under hot, cold, acidic, alkaline, and salt conditions, UBCE gene displays the greatest expression stability. However, EF1b, β-ACT, HSD17B3, and Cyb presented the greatest stability under cold, heat, and acidic conditions, and heavy metal exposure, respectively. To screen for genes suitable for all conditions, RefFinder’s ranking results revealed that UBCE and EF1b ranked in the top 2, demonstrating the highest gene expression stability. In contrast, Cyb was positioned at the bottom of the comprehensive ranking table. This study not only revealed potential factors affecting the suitability of reference genes but also identified optimal reference genes from a set of candidate genes across diverse conditions.
Keywords: Lyophyllum decastes; real-time fluorescence quantitative PCR; internal reference gene; culture conditions Lyophyllum decastes; real-time fluorescence quantitative PCR; internal reference gene; culture conditions

Share and Cite

MDPI and ACS Style

Hui, Y.-Q.; Yang, H.-L.; Zhang, Y.-Q.; Zhu, C.-Z.; Xi, L.-P.; Song, C.-Y.; Li, Z.-P.; Li, E.-X.; Li, S.-H.; Liu, Y.-N.; et al. Screening and Identification of Reference Genes Under Different Conditions and Growth Stages of Lyophyllum decastes. Int. J. Mol. Sci. 2025, 26, 11004. https://doi.org/10.3390/ijms262211004

AMA Style

Hui Y-Q, Yang H-L, Zhang Y-Q, Zhu C-Z, Xi L-P, Song C-Y, Li Z-P, Li E-X, Li S-H, Liu Y-N, et al. Screening and Identification of Reference Genes Under Different Conditions and Growth Stages of Lyophyllum decastes. International Journal of Molecular Sciences. 2025; 26(22):11004. https://doi.org/10.3390/ijms262211004

Chicago/Turabian Style

Hui, Yun-Qi, Huan-Ling Yang, Yu-Qing Zhang, Chen-Zhao Zhu, Li-Ping Xi, Chun-Yan Song, Zheng-Peng Li, E-Xian Li, Shu-Hong Li, Yong-Nan Liu, and et al. 2025. "Screening and Identification of Reference Genes Under Different Conditions and Growth Stages of Lyophyllum decastes" International Journal of Molecular Sciences 26, no. 22: 11004. https://doi.org/10.3390/ijms262211004

APA Style

Hui, Y.-Q., Yang, H.-L., Zhang, Y.-Q., Zhu, C.-Z., Xi, L.-P., Song, C.-Y., Li, Z.-P., Li, E.-X., Li, S.-H., Liu, Y.-N., & Yang, R.-H. (2025). Screening and Identification of Reference Genes Under Different Conditions and Growth Stages of Lyophyllum decastes. International Journal of Molecular Sciences, 26(22), 11004. https://doi.org/10.3390/ijms262211004

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