Laboratory of Modern Biotechnology, School of Forestry and Landscape Architecture, Anhui Agricultural University, Hefei, China.
Horticulture Institute, Anhui Academy of Agricultural Sciences, Hefei, China.
BMC Genomics. 2024 Oct 24;25(1):997. doi: 10.1186/s12864-024-10889-9.
Grapes are highly valued for their nutritional and economic benefits, and have been widely studied for their biological attributes such as fruit development, quality formation, and stress resistance. One significant threat to grape quality is gray mold, caused by Botrytis cinerea, which can infect the flowers, fruits, leaves, and stems. The quantitative real-time PCR (qRT-PCR), known for its high sensitivity and quantitative accuracy, is an essential tool for analyzing gene expression related to the pathogenesis of gray mold, thereby providing deeper insights into the disease.
In this study, we aim to identify stable internal reference genes crucial for accurate gene expression analysis via qRT-PCR. Utilizing transcriptome data from grapes under various disease stresses, we identified twelve candidate reference genes with consistently high expression levels. The stability of these genes was assessed through delta-CT, geNorm, NormFinder, BestKeeper, and RefFinder analyses after establishing the cycling thresholds (Ct) in different grape varieties treated with Botrytis cinerea.
Our findings reveal that VIT-17s0000g02750 and VIT-06s0004g04280 exhibit stable expression and are suitable as new reference genes. This foundational work supports further research into the molecular mechanisms of grape biological processes.
葡萄因其营养价值和经济价值而备受重视，其生物学特性如果实发育、品质形成和抗逆性等也得到了广泛研究。灰霉病是葡萄品质的一个重大威胁，由 Botrytis cinerea 引起，可感染花、果实、叶片和茎。定量实时 PCR（qRT-PCR）以其高灵敏度和定量准确性而著称，是分析与灰霉病发病机制相关的基因表达的重要工具，从而深入了解该疾病。
本研究旨在通过 qRT-PCR 鉴定对准确基因表达分析至关重要的稳定内参基因。利用不同疾病胁迫下葡萄的转录组数据，我们鉴定出了十二个候选内参基因，它们的表达水平始终较高。在建立了不同葡萄品种经 Botrytis cinerea 处理后的循环阈值（Ct）后，通过 delta-CT、geNorm、NormFinder、BestKeeper 和 RefFinder 分析评估了这些基因的稳定性。
我们的研究结果表明，VIT-17s0000g02750 和 VIT-06s0004g04280 表达稳定，适合作为新的内参基因。这项基础性工作为进一步研究葡萄生物过程的分子机制提供了支持。
