Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics and Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing, 210037, China.
Institute of Mountain Resources, Guizhou Academy of Science, Guiyang, 550001, China.
Mol Biol Rep. 2022 Jul;49(7):5985-5995. doi: 10.1007/s11033-022-07382-5. Epub 2022 Mar 31.
Passiflora edulis is a tropical fruit with high nutrient and medicinal values that is widely planted in southern China. However, the molecular biology of P. edulis has not been well studied. There are few reports regarding the choice of reference genes for gene expression studies of passion fruit.
By using three algorithms, implemented in geNorm, NormFinder and BestKeeper, we have selected ten candidate reference genes to explore their transcriptional expression stability in various tissues and under cold stress conditions. EF1 and HIS were stably expressed in five tissues. Ts and OTU were stably in vegetative organs. 50 S and Liom were stably in reproductive organs. The transcriptional abundance of EF1 and UBQ was stable in cold-treated and recovery treated leaf samples of P. edulis. In all samples, EF1 and Ts exhibited the highest expression stability. Evaluation of selected genes using simple statistical methods (ANOVA and post hoc analysis). Overall, EF1 emerged as the optimum reference gene for qRT-PCR normalize in P. edulis. In addition, the qRT-PCR analysis revealed that expression of ICE1 increases with the duration of cold treatment.
In this study, we successfully screened stable reference genes from 10 candidates in P. edulis and verified the results by analyzing the expression level of ICE1. The results provide reliable and effective reference genes for future research on gene expression analysis in P. edulis, and lay a foundation for follow-up research on functional genes in P. edulis.
西番莲是一种营养和药用价值都很高的热带水果，在中国南方广泛种植。然而，西番莲的分子生物学尚未得到很好的研究。关于西番果基因表达研究中参考基因的选择，报道较少。
我们使用 geNorm、NormFinder 和 BestKeeper 中的三种算法，从十个候选参考基因中选择了十个候选参考基因，以研究它们在不同组织和冷胁迫条件下的转录表达稳定性。EF1 和 HIS 在五种组织中表达稳定。Ts 和 OTU 在营养器官中表达稳定。50S 和 Liom 在生殖器官中表达稳定。EF1 和 UBQ 在冷处理和恢复处理的西番莲叶片样本中的转录丰度稳定。在所有样本中，EF1 和 Ts 表现出最高的表达稳定性。使用简单的统计方法（ANOVA 和事后分析）对选定基因进行评估。总体而言，EF1 是 P. edulis 中 qRT-PCR 归一化的最佳参考基因。此外，qRT-PCR 分析表明 ICE1 的表达随着冷处理时间的延长而增加。
本研究成功筛选了 10 个候选基因在西番莲中的稳定参考基因，并通过分析 ICE1 的表达水平验证了结果。这些结果为西番莲基因表达分析的后续研究提供了可靠和有效的参考基因，为西番莲功能基因的后续研究奠定了基础。
