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

## TL;DR

This study identifies the most stable reference genes for gene expression analysis in Lyophyllum decastes under various environmental conditions and growth stages.

## Contribution

The study provides a comprehensive evaluation of reference gene stability in Lyophyllum decastes under multiple environmental and developmental conditions.

## Key findings

- UBCE is the most stable gene across all developmental stages and environmental conditions.
- EF1b, β-ACT, HSD17B3, and Cyb are the most stable under cold, heat, acidic, and heavy metal conditions, respectively.
- UBCE and EF1b are the top-ranked genes for use across all tested conditions.

## 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.