Institute for Biological Research "Siniša Stanković"-National Institute of Republic of Serbia, University of Belgrade, Bulevar despota Stefana 142, Belgrade, 11060, Serbia.
Faculty of Biology, University of Belgrade, Studentski trg 16, Belgrade, Serbia.
Protoplasma. 2021 May;258(3):587-599. doi: 10.1007/s00709-020-01588-9. Epub 2020 Nov 27.
In this study, tansy (Tanacetum vulgare L.) in vitro culture was established from seeds collected from natural populations. The multiplication of plantlets was conducted through shoot tips that exhibited potent apical growth and regeneration capacities on basal medium (BM), without the addition of any plant growth regulators (PGRs). PGRs were also omitted for the establishment and cultivation of tansy root cultures. Both abaxial and adaxial leaf surfaces of in vitro micropropagated plantlets were covered with glandular biseriate trichomes. Histochemical staining showed that glandular secretions were rich in lipid and terpene compounds, confirmed by GC-MS analysis of essential oil (EO). In the total EO, similar portions of oxygenated monoterpenes (38.5% m/m) and oxygenated sesquiterpenes (22.6% m/m) were detected. Chemical profiles of methanol extracts of in vitro cultured tansy shoots and roots varied in quantity and quality from those obtained from wild-growingtansy. HPLC analysis indicated that the methanol extracts of in vitro cultured roots were the richest in 3,5-O-dicaffeoylquinic acid (3,5-O-DCQA), in which the concentration was 6 times higher (10.220 mg/g DW) than that in the extract obtained from roots of wild-growing tansy (1.684 mg/g DW). This result is noticeable in the manner of industrial production of biologically active 3,5-O-DCQA that has been shown to have antioxidant, hepatoprotective, antiviral, antimutagenic, and immunomodulatory activity. Biotechnological interventions on secondary metabolite production taking place in trichomes could further enhance the production of some important tansy metabolites and further investigation will be directed toward the elucidation of the pharmaceutical potential of tansy in vitro obtained metabolites, as mixtures or single moieties.
在这项研究中,从天然种群中采集的种子建立了蓬蒿(Tanacetum vulgare L.)的体外培养。通过在基础培养基(BM)上表现出强大的顶端生长和再生能力的茎尖进行植物的繁殖,无需添加任何植物生长调节剂(PGRs)。在建立和培养蓬蒿根培养物时也省略了 PGRs。体外微繁殖植物的下表皮和上表皮都覆盖着腺二列二叉毛。组织化学染色表明,腺分泌物富含脂质和萜烯化合物,这通过对精油(EO)的 GC-MS 分析得到了证实。在总 EO 中,检测到氧化单萜(38.5%m/m)和氧化倍半萜(22.6%m/m)的含量相似。从野生蓬蒿中获得的化学特征与从体外培养的蓬蒿茎和根的甲醇提取物的化学特征在数量和质量上有所不同。HPLC 分析表明,体外培养的根甲醇提取物中 3,5-O-二咖啡酰奎宁酸(3,5-O-DCQA)的含量最丰富,浓度为野生生长的蓬蒿根提取物的 6 倍(10.220mg/g DW)(1.684mg/g DW)。这一结果在工业生产具有抗氧化、保肝、抗病毒、抗突变和免疫调节活性的生物活性 3,5-O-DCQA 方面是显而易见的。在毛状体中进行的次生代谢产物生产的生物技术干预可以进一步提高一些重要的蓬蒿代谢产物的产量,进一步的研究将针对阐明体外获得的蓬蒿代谢产物的药用潜力,无论是混合物还是单一成分。