The production of crocin, an uncommon and valuable apocarotenoid with strong biological activity, was obtained in a cell suspension culture of saffron (Crocus sativus L.) established from style-derived calli to obtain an in-vitro system for metabolite production. Salycilic acid (SA) was used at different concentrations to elicit metabolite production, and its effect was analyzed after a 4 days of treatment. HPLC-DAD analysis was used for total crocin quantification while the Folin-Ciocâlteu method was applied for phenolic compounds (PC) content. Interestingly, despite cell growth inhibition, a considerable exudation was observed when the highest SA concentration was applied, leading to a 7-fold enhanced production of crocin and a 4-fold increase of phenolics compared to mock cells. The maximum antioxidant activity of cell extracts was evidenced after SA 0.1 mM elicitation. Water-soluble extracts of saffron cells at concentrations of 1, 0.5, and 0.1 µg mL⁻¹ showed significant inhibitory effects on MDA-MB-231 cancer cell viability. The heterologous vacuolar markers RFP-SYP51, GFPgl133Chi, and AleuRFP, were transiently expressed in protoplasts derived from the saffron cell suspensions, revealing that SA application caused a rapid stress effect, leading to cell death. Cell suspension elicitation with SA on the 7th day of the cell growth cycle and 24 h harvest time was optimized to exploit these cells for the highest increase of metabolite production in saffron cells.

中文翻译：

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水杨酸在藏红花悬浮培养的细胞中引起crocin和酚类物质的渗出。

藏红花（Crocus sativusL.）从样式衍生的愈伤组织建立，以获得用于代谢产物生产的体外系统。使用不同浓度的水杨酸（SA）引发代谢产物的产生，并在处理4天后对其效果进行了分析。HPLC-DAD分析用于总crocro定量，而Folin-Ciocâlteu方法用于酚类化合物（PC）含量。有趣的是，尽管细胞生长受到抑制，但当施用最高的SA浓度时仍观察到相当大的渗出，与模拟细胞相比，可导致番红花的产量增加7倍，酚类增加4倍。SA 0.1 mM激发后，证明了细胞提取物的最大抗氧化活性。藏红花细胞的水溶性提取物的浓度分别为1、0.5和0.1 µg mL ^-1对MDA-MB-231癌细胞的存活率显示出显着的抑制作用。异源液泡标记RFP-SYP51，GFPgl133Chi和AleuRFP在藏红花细胞悬液衍生的原生质体中瞬时表达，表明SA的应用引起了快速的应激效应，导致细胞死亡。在细胞生长周期的第7天和24小时的收获时间用SA诱导细胞悬浮液进行了优化，以利用这些细胞在藏红花细胞中最大程度地提高代谢产物的产量。