Department of Applied Chemistry, Chaoyang University of Technology, Taichung, 41349, Taiwan.
Biotechnol Appl Biochem. 2020 Jan;67(1):95-104. doi: 10.1002/bab.1810. Epub 2019 Sep 24.
Dysosma pleiantha (Hance) Woodson is one of the endangered traditional Chinese medicinal herbs, highly valued for its medicinal properties by Taiwan's mountain tribes. The present study aims to develop an efficient protocol for callus biomass by optimizing suitable culture medium, carbon source culture condition, and enhanced production of pharmaceutically important podophyllotoxin, kaempferol, and quercetin from callus culture of D. pleiantha under the influence of different additives. Best callus induction was achieved in Gamborg's medium (B5) with 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) along with 0.2 mg/L kinetin under dark condition. Tender leaves of D. pleiantha showed the maximum of 86% callus induction among the different explants tested. Highest leaf callus proliferation was noted in B5 medium with 1 mg/L 2,4-D incubated under complete darkness. In addition, it was found that B5 medium with 1 mg/L 2,4-D along with 2 g/L peptone produced more leaf callus biomass and enhanced production of podophyllotoxin (16.3-fold), kaempferol (12.39-fold), and quercetin (5.03-fold) compared to control. Therefore, D. pleiantha callogenesis can provide an alternative source for enhanced production of secondary compounds regardless of the exploitation of its natural plant population.
三叶崖爬藤是一种濒危的传统中药材，因其药用价值而受到台湾山地部落的高度重视。本研究旨在通过优化合适的培养基、碳源培养条件以及在不同添加剂的影响下，从三叶崖爬藤愈伤组织培养中提高药用重要的鬼臼毒素、山奈酚和槲皮素的产量，来开发一种有效的愈伤组织生物量生产方法。最佳愈伤组织诱导是在黑暗条件下，B5 培养基中添加 1 mg/L 2,4-二氯苯氧乙酸（2,4-D）和 0.2 mg/L 激动素。在不同的外植体中，三叶崖爬藤的嫩叶表现出最高的 86%愈伤组织诱导率。在完全黑暗的条件下，B5 培养基中添加 1 mg/L 2,4-D 时，叶片愈伤组织的增殖率最高。此外，研究发现，B5 培养基中添加 1 mg/L 2,4-D 和 2 g/L 蛋白胨比对照可产生更多的叶片愈伤组织生物量，并可显著提高鬼臼毒素（16.3 倍）、山奈酚（12.39 倍）和槲皮素（5.03 倍）的产量。因此，三叶崖爬藤的愈伤组织发生可以为增强次生化合物的生产提供一种替代来源，而无需开发其天然植物种群。
