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Identification and validation of qRT-PCR reference genes for analyzing grape infection with gray mold
Lina Tan1, Lijuan Lu2, Wen Sun1
1Laboratory of Modern Biotechnology, School of Forestry and Landscape Architecture, Anhui Agricultural University, Hefei, China.
BMC Genomics
|October 25, 2024
Summary
This study identifies stable reference genes for accurate gene expression analysis in grapes using quantitative real-time PCR (qRT-PCR). VIT-17s0000g02750 and VIT-06s0004g04280 are validated as reliable internal controls for studying grape disease resistance.
Area of Science:
- Plant Science
- Molecular Biology
- Genomics
Background:
- Grapes possess significant nutritional and economic value, with ongoing research into their development, quality, and stress resistance.
- Gray mold, caused by Botrytis cinerea, poses a major threat to grape crops, affecting various plant parts.
- Quantitative real-time PCR (qRT-PCR) is vital for understanding gene expression in plant-pathogen interactions.
Purpose of the Study:
- To identify stable internal reference genes for precise gene expression analysis in grapes.
- To ensure accurate qRT-PCR results in studies involving grape disease resistance.
- To provide reliable molecular tools for grape research.
Main Methods:
- Transcriptome data from grapes under disease stress were analyzed.
- Twelve candidate reference genes with high expression were selected.
- Gene stability was evaluated using multiple algorithms (delta-CT, geNorm, NormFinder, BestKeeper, RefFinder) on Botrytis cinerea-treated grape varieties.
Main Results:
- Twelve candidate reference genes were identified from transcriptome data.
- Stability analysis revealed two highly reliable reference genes.
- VIT-17s0000g02750 and VIT-06s0004g04280 demonstrated consistent expression across different grape varieties and treatments.
Conclusions:
- VIT-17s0000g02750 and VIT-06s0004g04280 are validated as suitable reference genes for qRT-PCR in grapes.
- This finding supports accurate molecular studies on grape biological processes and disease resistance.
- Establishes a foundation for future research into grape molecular mechanisms.
