Pineapple (Ananas comosus L.), a popular tropical fruit, is a good model for evolutionary analysis and genetic research on adaptation to drought habitats, multiple fruits, and crassulacean acid metabolism (CAM) photosynthesis. Reliable reference genes for the normalization of the levels of development-related and/or stress-responsive genes is important for elucidating the mechanisms of plant development as well as their adaptation to various environments. In this study, ten candidate reference genes were selected, and the expression stability of each gene was assessed across 105 pineapple samples, consisting of different reproductive organs (stigma, petal, sepal, ovule, anther, flower, and fruitlet), abiotic stress (salinity, drought, and cold), and hormones (abscisic acid, ethylene, jasmonic acid, and salicylic acid). Our results revealed that PP2A and UBQ were stably expressed during pineapple reproductive development. PP2A and CYC were the stable reference genes across abiotic stress, whereas RAN and EF1α/PP2A was the best candidates for various hormones. To validate the feasibility of using these stably expressed genes for further experiments, we evaluated the expression profile of AcMYB30 across different samples. Our results provide important insights on the growth and development of the pineapple plant as well as information on stress-tolerance genes and stress-signaling pathways in this important fruit.

中文翻译：

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菠萝（Ananas comosus L.）生殖发育过程中参考基因的差异表达分析及对非生物胁迫，激素刺激的响应

菠萝（菠萝L.）是一种流行的热带水果，是进行进化分析和遗传研究以适应干旱生境，多种水果和克拉苏酸代谢（CAM）光合作用的良好模型。标准化与发育相关和/或胁迫响应基因水平的可靠参考基因，对于阐明植物发育的机制及其对各种环境的适应性至关重要。在这项研究中，选择了十个候选参考基因，并评估了105个菠萝样品中每个基因的表达稳定性，这些样品由不同的生殖器官（柱头，花瓣，萼片，胚珠，花药，花和小果），非生物胁迫（盐度，干旱和寒冷）和激素（脱落酸，乙烯，茉莉酸和水杨酸）。我们的结果表明PP2A和UBQ在菠萝生殖发育过程中稳定表达。PP2A和CYC是克服非生物胁迫的稳定参考基因，而RAN和EF1α / PP2A是各种激素的最佳候选基因。为了验证使用这些稳定表达的基因进行进一步实验的可行性，我们评估了AcMYB30在不同样品中的表达谱。我们的结果提供了有关菠萝植物生长和发育的重要见解，以及有关该重要果实中的耐胁迫基因和胁迫信号通路的信息。