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Selection of reliable reference genes in Colletotrichum scovillei during different growth stages, host interactions, and plant extract treatment for qRT-PCR

Summary

Researchers identified the best internal control genes for measuring fungal gene expression in Colletotrichum scovillei, a fungus that causes serious disease in chili peppers. Different genes worked best under different conditions: one for normal growth, another during infection, and a third when treated with plant extracts. This discovery will help scientists study how this fungus develops and causes disease, potentially leading to better ways to control it using natural plant-based treatments.

Background

Colletotrichum scovillei, belonging to the C. acutatum species complex, causes aggressive anthracnose in chili peppers worldwide. Quantitative real-time PCR (qRT-PCR) is a highly sensitive method for evaluating gene expression, but its accuracy depends on reliable reference genes. Stable reference genes have not yet been identified for C. scovillei under various conditions.

Objective

To select and validate reliable internal control genes for qRT-PCR analysis in C. scovillei during different growth stages (conidia, conidial germination, mycelium growth), host interactions with chili pepper, and treatment with common solvents and plant extracts. The study aimed to identify the most suitable reference genes for accurate gene expression normalization under these diverse conditions.

Results

CsPP2A showed the best stability during C. scovillei growth stages, CsTUB demonstrated superior stability during host interactions, and CsUCE was more stable after DMSO and Cestrum glanduliferum extract treatment. When combining all conditions, no single gene exhibited consistent stability across all algorithms, indicating condition-specific reference gene requirements.

Conclusion

The study identified condition-specific reference genes for C. scovillei qRT-PCR analysis: CsPP2A for growth stages, CsTUB for host interactions, and CsUCE for plant extract treatment. The research provides the first validated reference genes for C. scovillei and demonstrates the necessity of selecting appropriate reference genes tailored to specific experimental conditions rather than using universal housekeeping genes.
  • Published in:BMC Plant Biology,
  • Study Type:Experimental Research Study,
  • Source: PMID: 40615801, DOI: 10.1186/s12870-025-06733-1
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