Stable Reference Gene Selection for Ophiocordyceps sinensis Gene Expression Studies Under Different Developmental Stages and Light-Induced Conditions

Summary

This research identified the most reliable genes to use as reference points when studying how the medicinal fungus Ophiocordyceps sinensis develops and responds to light. This is important because having stable reference points allows scientists to accurately measure how other genes behave during fungal growth and development. Impact on everyday life: – Helps improve cultivation methods for this valuable medicinal fungus – Advances our understanding of how fungi develop and respond to environmental conditions – Contributes to more efficient production of traditional Chinese medicine – Enables better quality control in medicinal fungal products

Background

Chinese cordyceps (Ophiocordyceps sinensis) is a traditional Chinese medicine formed by an entomopathogenic fungus that parasitizes insect larvae. The formation process includes asexual proliferation in host hemolymph and sexual development through fruiting body formation. Understanding the molecular mechanisms requires reliable reference genes for RT-qPCR analysis, but no stable reference genes have been reported for O. sinensis fruiting body development stages.

Objective

To identify and validate stable reference genes for RT-qPCR analysis of O. sinensis under different developmental stages and light conditions. The study aimed to evaluate 10 candidate reference genes during asexual reproduction, fruiting body development, and light-induced conditions.

Results

The study found that Tef1 and Tub1 were the most stable reference genes during asexual reproduction, while Tyr and Cox5 were most stable during fruiting body development. Under light-induced conditions, Tyr and Tef1 showed the highest stability. The analysis revealed that no more than 2 reference genes are required for RT-qPCR normalization in all three experimental conditions.

Conclusion

This first comprehensive screening of reference genes in O. sinensis provides reliable tools for studying gene expression during morphological transformation, fruiting body development, and light stress response. The identified stable reference genes will enable more accurate gene expression studies in different experimental models of O. sinensis development.
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