Selection and Validation of Endogenous Reference Genes for RT-qPCR Normalization in Different Stresses and Tissues of the Tiger Milk Mushroom, Pleurotus tuber-regium

Author: mycolabadmin
2021-09-20
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Summary

This research identified the most reliable genes to use as internal controls when studying gene expression in the tiger milk mushroom. This is important for ensuring accurate scientific results when studying this nutritionally and medicinally valuable mushroom species. Impact on everyday life: – Helps improve the accuracy of research on medicinal mushrooms – Contributes to better understanding of how mushrooms respond to environmental stress – Supports development of improved mushroom cultivation methods – Aids in quality control for mushroom-based products – Advances our knowledge of fungal biology and genetics

Background

Pleurotus tuber-regium is a sclerotium-producing mushroom with pharmacological and nutritional value that is popular in China, Africa, and the United States. Gene expression analysis using RT-qPCR requires appropriate reference genes for accurate results, but reference genes for P. tuber-regium have not yet been validated. Proper reference genes must have stable expression across different experimental conditions to minimize bias in RT-qPCR data normalization.

Objective

To identify and validate suitable reference genes for RT-qPCR data normalization in P. tuber-regium under different stress conditions and tissue types. The study aimed to screen and evaluate 14 candidate reference genes for their expression stability across different temperatures, drought levels, salinity shifts, fruitbody, and sclerotium samples.

Results

The analysis identified E3upl and Tif-5a as the most stably expressed gene combination across all experimental conditions. Different genes showed optimal stability under specific conditions: β-tub was most stable across developmental stages, Wlp under different temperatures, E3upl under water deficit, and U2ribo under salinity shifts. GAPDH consistently showed the least stability across conditions.

Conclusion

E3upl and Tif-5a were validated as the optimal reference gene combination for RT-qPCR data normalization in P. tuber-regium across various stress conditions and tissue types. This study provides fundamental information for reference gene selection in P. tuber-regium molecular studies and will contribute to future gene function analyses.

Published in:Mycoscience,
Study Type:Laboratory Research,
Source: 10.47371/mycosci.2021.05.001