Three types of calli were induced from Origanum vulgare (O. vulgare) aseptic seedlings, and the friable calli with white appearance and high growth rate were further screened and used to develop cell suspension culture to produce polyphenols. Murashige and Skoog (MS) medium with 3.0 mg/L Kinetin (KT) and 0.5 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) was suitable for both O. vulgare cells growth and polyphenols accumulation. To further enhance the polyphenols accumulation, O. vulgare cells were treated by phenylalanine (Phe) feeding and salicylic acid (SA) elicitation. Compared with the individual Phe feeding and SA elicitation, SA elicitation combined with Phe feeding showed a much better promotion effect on the polyphenols synthesis in O. vulgare cells, especially rosmarinic acid (RosA) accumulation. With the combined treatment of 200 µM SA and 100 µM Phe, total polyphenols content and yield were 41.36 mg/g and 752.93 mg/L, respectively. RosA content and yield reached 31.25 mg/g and 570.37 mg/L, which were 5.44 and 5.47 times that of the control. Furthermore, the total polyphenols extracted from the cultured cells treated by SA elicitation combined with Phe feeding displayed a much higher antioxidant capacity than that of untreated cells, meanwhile its 1,1-diphenyl-2-trinitrophenyl hydrazine (DPPH) and superoxide anion radical-scavenging activity were much stronger than that of vitamin C. What’s more, our results also showed that RosA was the principal contributor to the fine antioxidant capacity of the total polyphenols extracted from the SA and Phe treated cells. Our research indicated that SA elicitation combined with Phe feeding significantly improved the polyphenols yield and antioxidant capacity of the cultured O. vulgare cells, and therefore has a promising application prospect in natural polyphenols production.

从Origanum vulgare ( O. vulgare )无菌苗中诱导出3种愈伤组织，进一步筛选出外观白色、生长速度快的脆性愈伤组织，用于细胞悬浮培养，生产多酚类物质。含有 3.0 mg/L 激动素 (KT) 和 0.5 mg/L 2,4-二氯苯氧基乙酸 (2,4-D) 的 Murashige 和 Skoog (MS) 培养基适用于O. vulgare细胞生长和多酚积累。为了进一步增强多酚的积累，O. vulgare通过苯丙氨酸（Phe）喂养和水杨酸（SA）诱导处理细胞。与单独饲喂 Phe 和 SA 诱导相比，SA 诱导联合 Phe 饲喂对O. vulgare多酚合成有更好的促进作用细胞，尤其是迷迭香酸 (RosA) 积累。在 200 µM SA 和 100 µM Phe 的联合处理下，总多酚含量和产量分别为 41.36 mg/g 和 752.93 mg/L。RosA 含量和产量分别达到 31.25 mg/g 和 570.37 mg/L，分别是对照的 5.44 和 5.47 倍。此外，从SA诱导联合Phe喂养处理的培养细胞中提取的总多酚比未处理细胞显示出更高的抗氧化能力，同时其1,1-二苯基-2-三硝基苯肼（DPPH）和超氧阴离子自由基-清除活性比维生素 C 强得多。此外，我们的结果还表明 RosA 是从 SA 和 Phe 处理的细胞中提取的总多酚的优良抗氧化能力的主要贡献者。O. vulgare细胞，因此在天然多酚生产中具有广阔的应用前景。