Department of Biotechnology, Quaid-i-Azam University, Islamabad 45320, Pakistan.
Department of Pharmaceutical Botany, Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand.
Molecules. 2020 Oct 21;25(20):4859. doi: 10.3390/molecules25204859.
Thai basil is a renowned medicinal plant and a rich source of bioactive antioxidant compounds with several health benefits, with actions to prevent of cancer, diabetes and cardiovascular disease. Plant cell and tissue culture technologies can be routinely established as an important, sustainable and low-cost biomass source to produce high-value phytochemicals. The current study aimed at developing an effective protocol to produce Thai basil leaf-derived callus cultures with sustainable and high production of biomass and antioxidants as an alternative of leaves production. MS basal medium with various concentrations of plant growth regulators (PGRs) compatible with nutraceutical applications (i.e., gibberellic acid (GA) and 6-benzylaminopurine (BAP) either alone or in combination with naphthalene acetic acid (NAA)) were evaluated. Among all tested PGRs, the combination BAP:NAA (5 mg/L:1 mg/L) yields the maximum biomass accumulation (fresh weight (FW): 190 g/L and dry weight (DW): 13.05 g/L) as well as enhanced phenolic (346.08 mg/L) production. HPLC quantification analysis indicated high productions of chicoric acid (35.77 mg/g DW) and rosmarinic acid (7.35 mg/g DW) under optimized callus culture conditions. Antioxidant potential was assessed using both in vitro cell free and in vivo cellular antioxidant assays. Maximum in vitro antioxidant activity DPPH (93.2% of radical scavenging activity) and ABTS (1322 µM Trolox equivalent antioxidant capacity) was also observed for the extracts from callus cultures grown in optimal conditions. In vivo cellular antioxidant activity assay confirmed the effective protection against oxidative stress of the corresponding extract by the maximum inhibition of ROS and RNS production. Compared to commercial leaves, callus extracts showed higher production of chicoric acid and rosmarinic acid associated with higher antioxidant capacity. In addition, this biological system also has a large capacity for continuous biomass production, thus demonstrating its high potential for possible nutraceutical applications.
泰国罗勒是一种著名的药用植物，是生物活性抗氧化化合物的丰富来源，具有多种健康益处，可以预防癌症、糖尿病和心血管疾病。植物细胞和组织培养技术可以常规建立，作为生产高价值植物化学物质的重要、可持续和低成本生物量来源。本研究旨在开发一种有效的方法，以生产泰国罗勒叶衍生的愈伤组织培养物，具有可持续和高生物量和抗氧化剂的生产，作为叶片生产的替代物。评估了适合营养应用的各种植物生长调节剂 (PGR) 的 MS 基本培养基（即赤霉素 (GA) 和 6-苄基氨基嘌呤 (BAP) 单独或与萘乙酸 (NAA) 组合）。在所有测试的 PGR 中，BAP:NAA（5 mg/L:1 mg/L）组合产生最大的生物量积累（鲜重 (FW)：190 g/L 和干重 (DW)：13.05 g/L）以及增强的酚类物质（346.08 mg/L）产量。HPLC 定量分析表明，在优化的愈伤组织培养条件下， chicoric 酸（35.77 mg/g DW）和迷迭香酸（7.35 mg/g DW）的产量较高。使用体外无细胞和体内细胞抗氧化测定法评估了抗氧化潜力。在优化条件下生长的愈伤组织提取物的 DPPH（93.2%自由基清除活性）和 ABTS（1322 µM Trolox 当量抗氧化能力）的体外抗氧化活性也达到最大值。体内细胞抗氧化活性测定证实了相应提取物对氧化应激的有效保护，最大程度地抑制了 ROS 和 RNS 的产生。与商业叶片相比，愈伤组织提取物显示出更高的 chicoric 酸和迷迭香酸产量，以及更高的抗氧化能力。此外，该生物系统还具有大容量的连续生物量生产能力，因此展示了其在可能的营养应用方面的巨大潜力。
