ABSTRACT

Gardenia jasminoides J. Ellis. is widely used in horticulture and its fruit is heavily used in the natural pigment and traditional Chinese medicinal industries. Functional discovery and characterisation of genes in fruits generally require accurate transcript quantification. Therefore, we first identified twenty candidate genes and then set out to determine their stability in qRT-PCR assays using three software programs, i.e. geNorm (version 3.5), NormFinder (version 20), and RefFinder. The optimal number of ICGs required for qRT-PCR data normalisation was also determined. Our results indicated that two ICGs were sufficient for accurate qRT-PCR data normalisation in the organs tested. From the consensus genes of the three software programs, EIF3G and GAPDH were the top-ranked ICGs in pulp and seeds at three developmental stages, i.e. the green fruit stage, the orange fruit stage, and the red fruit stage. DEAD and 40S-S15a were the top-ranked ICGs in pulp, leaves, seeds, and stems at the red fruit stage. The reliability of the selected ICGs was confirmed through expression profile comparison of two genes, i.e. phytoene synthase and β-ring hydroxylase involved in carotenoid biosynthesis. This is the first report of ICG screening in G. jasminoides. Our results provide a basis for normalising gene expression levels and understanding of gene function in G. jasminoides.

摘要

子茉莉J.埃利斯。被广泛用于园艺，其果实被大量用于天然色素和传统中药工业。水果中基因的功能发现和表征通常需要准确的转录本定量。因此，我们首先鉴定了二十个候选基因，然后开始使用三种软件程序（即geNorm（3.5版），NormFinder（20版）和RefFinder）在qRT-PCR分析中确定其稳定性。还确定了qRT-PCR数据标准化所需的最佳ICG数。我们的结果表明，两个ICG足以在被测器官中进行准确的qRT-PCR数据标准化。从EIF3G和GAPDH这三个软件程序的共有基因在三个发育阶段，即绿色水果阶段，橙色水果阶段和红色水果阶段，果肉和种子中的ICG排名最高。DEAD和40S-S15a在红色果实阶段的果肉，叶片，种子和茎中是排名最高的ICG。通过比较两个基因（即类胡萝卜素生物合成中的八氢番茄红素合酶和β-环羟化酶）的表达谱比较，可以确定所选ICG的可靠性。这是关于茉莉G. ICG筛选的第一份报告。我们的研究结果为规范茉莉香菇的基因表达水平和了解基因功能提供了基础。