Key Laboratory of Forest Plant Ecology, Ministry of Education, Heilongjiang Provincial Key Laboratory of Ecological Utilization of Forestry-Based Active Substances, College of Chemistry, Chemical Engineering and Resource Utilization, Northeast Forestry University, Harbin, 150040, China.
College of Forest Science, Kim Il Sung University, Pyongyang, 999093, Democratic People's Republic of Korea.
Appl Microbiol Biotechnol. 2022 Nov;106(21):7027-7037. doi: 10.1007/s00253-022-12194-5. Epub 2022 Sep 29.
Atractylodes chinensis is a medicinal plant widely used for the treatment of gastric disorders, and its main bioactive compounds are atractylon and β-eudesmol. This study was purposed to establish the adventitious root culture system of A. chinensis for in vitro production of atractylon and β-eudesmol. The main parameters in the adventitious root induction and suspension cultures were optimized to maximize the culture efficiency. Adventitious roots were induced most efficiently from leaf explants on Murashige and Skoog (MS) solid medium containing 1.5 mg/L naphthaleneacetic acid (NAA) and 30 g/L sucrose with the highest root induction rate of approximately 92% and 12.9 roots per explant. During the adventitious root suspension culture, the root biomass and the accumulated content of the target compounds simultaneously increased to reach the maximum values after 8 weeks of culture. The maximum yield of the target compounds (total concentration 3.38 mg/g DW, total yield 2.66 mg) was achieved in the roots cultured in ½ MS liquid medium supplemented with 2.0 mg/L IBA, 3.2 mg/L NAA, and 40 g/L sucrose with the inoculum density of 8 g/L. Through the central composite design experiment, it was found that the combined use of different types of auxins in the suspension culture could further improve root growth and metabolite accumulation than the application of only one type of auxin. This work provides a new possibility to have a promising candidate for the industrial production of A. chinensis pharmaceuticals without relying on wild resources or field cultivation. KEY POINTS: • The induction culture was optimized for efficient root induction. • Suspension culture was optimized for the atractylon and β-eudesmol production. • Combined use of different auxins improves root growth and metabolite accumulation.
白术是一种广泛用于治疗胃病的药用植物,其主要生物活性化合物为苍术酮和β-桉叶醇。本研究旨在建立白术的不定根培养体系,用于体外生产苍术酮和β-桉叶醇。优化了不定根诱导和悬浮培养的主要参数,以最大限度地提高培养效率。叶片外植体在含有 1.5mg/L 萘乙酸(NAA)和 30g/L 蔗糖的 Murashige 和 Skoog(MS)固体培养基上诱导不定根的效率最高,不定根诱导率约为 92%,每个外植体产生 12.9 条不定根。在不定根悬浮培养过程中,不定根生物量和目标化合物的积累含量同时增加,培养 8 周后达到最大值。在补充了 2.0mg/L IBA、3.2mg/L NAA 和 40g/L 蔗糖的 ½ MS 液体培养基中,以 8g/L 的接种密度进行培养,可获得目标化合物(总浓度 3.38mg/g DW,总产率 2.66mg)的最大产量。通过中心组合设计实验发现,与单独使用一种生长素相比,在悬浮培养中组合使用不同类型的生长素可以进一步促进根的生长和代谢产物的积累。这项工作为工业生产白术药物提供了一种新的可能性,而无需依赖野生资源或田间种植。关键点:• 优化了诱导培养以实现高效生根。• 优化了悬浮培养以生产苍术酮和β-桉叶醇。• 组合使用不同的生长素可促进根的生长和代谢产物的积累。