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通过高效薄层色谱法(HPTLC)检测发现,氯化镉(CdCl)诱导提高了萝芙木碱和阿吗碱的产量。

Cadmium chloride (CdCl) elicitation improves reserpine and ajmalicine yield in as revealed by high-performance thin-layer chromatography (HPTLC).

作者信息

Zafar Nadia, Mujib A, Ali Muzamil, Tonk Dipti, Gulzar Basit, Malik Moien Qadir, Mamgain Jyoti, Sayeed Rukaya

机构信息

Department of Botany, Cellular Differentiation and Molecular Genetics Section, Jamia Hamdard, New Delhi, 110062 India.

出版信息

3 Biotech. 2020 Aug;10(8):344. doi: 10.1007/s13205-020-02339-6. Epub 2020 Jul 20.

Abstract

In vitro cultures play a promising role for production of pharmaceutically important plant secondary metabolites and the use of elicitation can mitigate the low productivity of active compounds. In the present study, the influence of cadmium chloride (CdCl) elicitation on alkaloid yield was investigated in . This heavy metal was employed to enhance the yield of reserpine and ajmalicine in leaf derived callus, leaves, stems and roots of in vitro grown cultures. Different concentrations [0.05 mM (C1), 0.10 mM (C2), 0.15 mM (C3) and 0.20 mM (C4)] of CdCl were added to the MS medium. The elicitor's influence on callus biomass, biochemical attributes and the yield of alkaloids was monitored at regular intervals. The amendment of CdCl improved growth and maximum callus biomass (1.29 g fresh weight and 0.16 g dry weight) was noted at 0.15 mM (C3) after 6 days of elicitation. The addition of elicitor in medium caused cellular stress and to analyse the role of CdCl in plant defence responses various antioxidant enzymes, i.e., superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activities were measured in treated and non-treated cultures. The antioxidant enzyme activity increased linearly with elevated levels of CdCl in medium; highest APX (0.88 EU min mgprotein), SOD (5.40 EU min mgprotein) and CAT (4.21 EU min mgprotein) activity were observed in leaves of in vitro regenerated plants at C4. The quantitative analyses of reserpine and ajmalicine were conducted in different elicitated tissues using high-performance thin-layer chromatography (HPTLC) method. The study reveals enriched level of reserpine and ajmalicine in cultivated tissues and the enhancement was noted up to C3 (0.15 mM) elicitor level. Reserpine yield was maximum (0.191 mg g DW) in roots of in vitro regenerated plants. The accumulation of ajmalicine was, however, better in leaf derived callus at C3 (0.131 mg g DW). Higher elicitor dose (0.20 mM) inhibited callus biomass growth and subsequent alkaloid accumulation. The present study indicates the use of CdCl as a propitious method in enhancing reserpine and ajmalicine yield in .

摘要

体外培养在生产具有药学重要性的植物次生代谢产物方面发挥着有前景的作用,而使用诱导剂可以缓解活性化合物低产量的问题。在本研究中,研究了氯化镉(CdCl)诱导对萝芙木生物碱产量的影响。这种重金属被用于提高体外培养的萝芙木愈伤组织、叶片、茎和根中利血平和阿吗灵的产量。将不同浓度[0.05 mM(C1)、0.10 mM(C2)、0.15 mM(C3)和0.20 mM(C4)]的CdCl添加到MS培养基中。定期监测诱导剂对愈伤组织生物量、生化特性和生物碱产量的影响。CdCl的添加促进了生长,诱导6天后,在0.15 mM(C3)时记录到最大愈伤组织生物量(鲜重1.29 g,干重0.16 g)。培养基中添加诱导剂会引起细胞应激,为了分析CdCl在植物防御反应中的作用,在处理和未处理的培养物中测量了各种抗氧化酶,即超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)的活性。抗氧化酶活性随着培养基中CdCl水平的升高而呈线性增加;在C4处理的体外再生植物叶片中观察到最高的APX(0.88 EU min mg蛋白)、SOD(5.40 EU min mg蛋白)和CAT(4.21 EU min mg蛋白)活性。使用高效薄层色谱(HPTLC)方法对不同诱导组织中的利血平和阿吗灵进行了定量分析。研究表明,培养组织中利血平和阿吗灵的含量丰富,并且在诱导剂水平达到C3(0.15 mM)时含量增加。体外再生植物根中的利血平产量最高(0.191 mg g干重)。然而,在C3(0.131 mg g干重)时,阿吗灵在叶片来源的愈伤组织中积累得更好。较高的诱导剂剂量(0.20 mM)抑制了愈伤组织生物量的生长以及随后的生物碱积累。本研究表明,使用CdCl是提高萝芙木中利血平和阿吗灵产量的一种有利方法。

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