中药川贝母组织培养材料中小泡体外培养及等甾体生物碱的LC-MS/MS分析。

IF 4.1 3区 生物学 Q1 PLANT SCIENCES
Hung-Chi Chang, Hui-Min Xie, Maw-Rong Lee, Chiu-Ying Lin, Mei-Kuen Yip, Dinesh Chandra Agrawal, Hsin-Sheng Tsay
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引用次数: 11

摘要

背景:川贝母是一种重要的中草药,是医药工业高度开发利用的三级保护物种。野生物种数量的减少无法满足市场需求。因此,本研究旨在建立一种体外培养球茎的方法。此外,该研究旨在对组织培养衍生的小球和愈伤组织进行LC-MS/MS分析,以确定其存在的异甾体生物碱(佩米辛、垂替辛和垂替辛酮),并将其数量与市购的原料药样品进行比较。结果:在添加6-苄基氨基嘌呤(1 mg L-1)和α-萘乙酸(0.4 mg L-1)的Murashige和Skoog基础培养基(MSBM)上,肝硬化F.肝硬化体外种子萌发率为91%。将发芽的种子从培养皿转移到含有无激素MSBM的玻璃瓶中,3个月后,37.5%的幼苗发育成球。在1/2 × MSBM上培养小球的横切面,实现小球的再生和增殖。每隔2个月进行重复传代培养,第5次传代末可产生3072个,重1270 g。LC-MS/MS分析表明,离体小球中存在大量的培米辛,而黑暗培养的愈伤组织中也存在培米辛和垂向培米辛。结论:该研究报告了一种有效的体外培养方法,可以产生肝硬化肝硬化小细胞,并在组织培养的小细胞和愈伤组织中存在一些等甾体生物碱。这项研究可能对全年在实验室条件下的肝硬化小细胞和愈伤组织的产生有巨大的帮助。此外,这些结果可以进一步用于研究在商业规模的生物反应器中使用液体和细胞悬浮培养生产异甾体生物碱。因此,我们不仅可以减少对自然栖息地的依赖,而且还可以帮助这一重要物种的就地保护。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

In vitro propagation of bulblets and LC-MS/MS analysis of isosteroidal alkaloids in tissue culture derived materials of Chinese medicinal herb Fritillaria cirrhosa D. Don.

In vitro propagation of bulblets and LC-MS/MS analysis of isosteroidal alkaloids in tissue culture derived materials of Chinese medicinal herb Fritillaria cirrhosa D. Don.

In vitro propagation of bulblets and LC-MS/MS analysis of isosteroidal alkaloids in tissue culture derived materials of Chinese medicinal herb Fritillaria cirrhosa D. Don.

In vitro propagation of bulblets and LC-MS/MS analysis of isosteroidal alkaloids in tissue culture derived materials of Chinese medicinal herb Fritillaria cirrhosa D. Don.

Background: Fritillaria cirrhosa, an important Chinese medicinal herb, is a Class-III protected and highly exploited species by pharmaceutical industry. Dwindling wild populations of species are unable to meet market demand. Therefore, this study was carried out to develop an in vitro propagation method for bulblet production. Also, the study aimed to carry out LC-MS/MS analysis of tissue culture-derived bulblets and callus for the presence of isosteroidal alkaloids (peimissine, verticine, and verticinone), and compare its quantities with commercially available crude drug samples.

Results: In vitro seed germination (91%) of F. cirrhosa was achieved on Murashige and Skoog's basal medium (MSBM) supplemented with 6-benzylaminopurine (1 mg L-1) and α-naphthalene-acetic-acid (0.4 mg L-1). On transfer of germinated seeds from Petri-dishes to glass bottles containing hormone-free MSBM, 37.5% of seedlings developed bulblets after 3 months of incubation. Regeneration and multiplication of bulblets were achieved by culture of transverse sections of bulblets on 1/2 X MSBM. By repeated subcultures at an interval of 2 months, 3072 bulblets weighing 1270 g could be produced at the end of 5th subculture. LC-MS/MS analysis showed a significant presence of peimissine in in vitro bulblets while callus incubated in the dark showed presence of peimissine and verticine.

Conclusion: The study reports an efficient in vitro propagation method of bulblets production of F. cirrhosa and presence of some isosteroidal alkaloids in tissue culture-derived bulblets and callus. The study could be of immense help in production of F. cirrhosa bulblets and callus under laboratory conditions round the year. Also, these results can be used further to investigate production of isosteroidal alkaloids in bioreactors at commercial scale using liquid and cell suspension cultures. Thus, we not only can reduce our dependence on collections from natural habitats, but also can help in in situ conservation of this important species.

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来源期刊
Botanical Studies
Botanical Studies PLANT SCIENCES-
CiteScore
4.80
自引率
2.90%
发文量
32
审稿时长
13 weeks
期刊介绍: Botanical Studies is an open access journal that encompasses all aspects of botany, including but not limited to taxonomy, morphology, development, genetics, evolution, reproduction, systematics, and biodiversity of all plant groups, algae, and fungi. The journal is affiliated with the Institute of Plant and Microbial Biology, Academia Sinica, Taiwan.
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