Zhang Chen, Li Tong, Hou Cheng-Lin, Shen Xiao-Ye
College of Life Science, Capital Normal University, Beijing, 100048, People's Republic of China.
AMB Express. 2017 Dec;7(1):14. doi: 10.1186/s13568-016-0314-9. Epub 2017 Jan 3.
Stable reference genes are necessary to analyse quantitative real-time reverse transcription PCR (qRT-PCR) data and determine the reliability of the final results. For further studies of the valuable fungus Shiraia bambusicola, the identification of suitable reference genes has become increasingly urgent. In this study, three conventional reference genes and nine novel candidates were evaluated under different light conditions (all-dark, all-light and 12-h light/dark) and in different media (rice medium, PD medium, and Czapek-Dox medium). Three popular software programs (geNorm, NormFinder and BestKeeper) were used to analyse these genes, and the final ranking was determined using RefFinder. SbLAlv9, SbJsn1, SbSAS1 and SbVAC55 displayed the best stability among the genes, while SbFYVE and SbPKI showed the worst. These emerging genes exhibited significantly better properties than the three existing genes under almost all conditions. Furthermore, the most reliable reference genes were identified separately under different nutrient and light conditions, which would help accessible to make the most of the existing data. In summary, a group of novel housekeeping genes from S. bambusicola with more stable properties than before was explored, and these results could also provide a practical approach for other filamentous fungi.
稳定的内参基因对于分析定量实时逆转录PCR（qRT-PCR）数据以及确定最终结果的可靠性至关重要。对于珍贵真菌竹黄的进一步研究而言，鉴定合适的内参基因变得愈发迫切。在本研究中，对三个传统内参基因和九个新候选基因在不同光照条件（全暗、全光和12小时光照/黑暗）以及不同培养基（大米培养基、PD培养基和察氏培养基）下进行了评估。使用三个常用软件程序（geNorm、NormFinder和BestKeeper）对这些基因进行分析，并使用RefFinder确定最终排名。SbLAlv9、SbJsn1、SbSAS1和SbVAC55在这些基因中表现出最佳稳定性，而SbFYVE和SbPKI表现最差。在几乎所有条件下，这些新出现的基因都比三个现有基因表现出明显更好的特性。此外，在不同营养和光照条件下分别鉴定出了最可靠的内参基因，这将有助于充分利用现有数据。总之，探索了一组来自竹黄的比以前具有更稳定特性的新管家基因，这些结果也可为其他丝状真菌提供一种实用方法。
