Aluminum chloride elicitation (amendment) improves callus biomass growth and reserpine yield in Rauvolfia serpentina leaf callus

Zafar, Nadia; Mujib, A.; Ali, Muhammad Asghar; Tonk, Dipti; Gulzar, Basit

doi:10.1007/s11240-017-1230-7

Cited by 37 publications

(21 citation statements)

References 37 publications

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“…However, such enhancement was obtained only through salicylic acid and tryptamine elicitations. Similar usage of elicitors for reserpine enhancement was reported by Nurcahyani et al (2008), Harisaranraj et al (2009), Zafar et al (2017); thus, differing from the results of the present experiment, wherein enhancement of the secondary metabolite was affected without the use of any elicitor. There are multiple reports related to reserpine estimation from in vitro shoot or calli or cell suspension culture or synthetic seeds or hairy roots individually (reviewed by Mukherjee et al 2019).…”

Section: Discussionsupporting

confidence: 52%

Ameliorated reserpine production via in vitro direct and indirect regeneration system in Rauvolfia serpentina (L.) Benth. ex Kurz.

et al. 2020

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Rauvolfia serpentina (L.) Benth. ex Kurz., popularly known as Indian Snakeroot plant, belonging to Apocynaceae family, holds immense medicinal importance, owing to its rich source of multiple secondary metabolites such as ajmaline, ajmalicine, reserpine, and serpentine. To meet the constant demands for the key secondary metabolite (reserpine) by majority of the pharmaceutical industries, the present study assessed the effects of direct and indirect regeneration system on amelioration of reserpine accumulation in shoots of R. serpentina. In vitro multiple shoot cultures were established using shoot tip explants. Best results for shoot initiation, multiplication, and biomass production were obtained in case of Murashige and Skoog medium, supplemented with 1 mg/l N 6 -benzyladenine. The multiple shoots were then sub-cultured on cytokinin-auxin combination media for further proliferation. Highest shoot and leaf multiplication rates and the most enhanced biomass were obtained in case of 1-1.5 mg/l Kinetin + 0.2 mg/l α-naphthalene acetic acid (NAA). Callus induction and its subsequent proliferation was obtained using 1.5 mg/l 2,4-dichlorophenoxyacetic acid. The best indirect shoot regeneration with highest shoot and leaf proliferation from calli was observed in case of 1 mg/l thidiazuron + 0.2 mg/l NAA. Reserpine content estimation via HPTLC from in vitro shoots (direct regeneration) and calli (indirect regeneration) were recorded to undergo an almost three-fold and two-fold increment (respectively) in comparison to that of the mother plant. Thus, in vitro direct regeneration system proved to be more effective and efficient in ameliorating the reserpine content.

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Section: Discussionsupporting

confidence: 52%

Ameliorated reserpine production via in vitro direct and indirect regeneration system in Rauvolfia serpentina (L.) Benth. ex Kurz.

et al. 2020

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“…and chemical stresses . The practice of adding these biotic and abiotic elicitors in culture gradually became a booming strategy in stimulating target product synthesis by slashing time required to get enhanced levels of products (Zafar et al 2017). The growing use of chemical elicitors especially metals in pharmaceutical sectors and several agro-industries made these compounds most important, predominant abiotic stress agents (Cai et al 2013).…”

Section: Introductionmentioning

confidence: 99%

Cadmium chloride (CdCl2) elicitation improves reserpine and ajmalicine yield in Rauvolfia serpentina as revealed by high-performance thin-layer chromatography (HPTLC)

Zafar¹,

Mujib²,

Ali³

et al. 2020

3 Biotech

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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 2 ) elicitation on alkaloid yield was investigated in Rauvolfia serpentina. 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 2 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 2 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 2 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 2 in medium; highest APX (0.88 EU min −1 mg −1 protein), SOD (5.40 EU min −1 mg −1 protein) and CAT (4.21 EU min −1 mg −1 protein) 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 −1 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 −1 DW). Higher elicitor dose (0.20 mM) inhibited callus biomass growth and subsequent alkaloid accumulation. The present study indicates the use of CdCl 2 as a propitious method in enhancing reserpine and ajmalicine yield in R. serpentina.

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“…In the present study, an increased level of proline was noted in tissues amended with 2,4-D, which suggests the generation of stress; furthermore, in the callus grown in an elevated stress level, a higher level of taxol was recorded when compared with that grown in less stressful conditions. A higher level of proline accumulation in response to stresses was also observed in several plants such as Arabidopsis and Rauvolfia (Szabados and Savoure, 2010;Liang et al, 2013;Nadia et al, 2017). A higher accumulation of proline in stressful conditions was earlier reported to be caused by an enhanced expression of the P5C6 gene, which is associated with proline synthesis (Chen et al, 2009).…”

Section: Antioxidant Enzymes and Proline As Stress Markers In Responsmentioning

confidence: 58%

“…When added to the medium, these PGRs induce cellular stress and are used as signalling molecules to trigger a cascade of gene expression in biosynthetic pathways of alkaloid synthesis (Binder et al, 2009;Zahid and Mujib, 2012;Mujib et al, 2016). The biotic/abiotic stress induced in cultured tissues is assayed by investigating the responses of antioxidant enzymes in cultured cells (Cai et al, 2012;Nadia et al, 2017). Stress marker enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) are studied to estimate stress levels in cultivated plant cells (Samar et al, 2015).…”

mentioning

confidence: 99%

Quantification of taxol by high-performance thin layer chromatography in Taxus wallichiana callus cultivated in vitro

Mujib¹,

Tonk²,

Gulzar³

et al. 2020

bta

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Aluminum chloride elicitation (amendment) improves callus biomass growth and reserpine yield in Rauvolfia serpentina leaf callus

Cited by 37 publications

References 37 publications

Ameliorated reserpine production via in vitro direct and indirect regeneration system in Rauvolfia serpentina (L.) Benth. ex Kurz.

Ameliorated reserpine production via in vitro direct and indirect regeneration system in Rauvolfia serpentina (L.) Benth. ex Kurz.

Cadmium chloride (CdCl2) elicitation improves reserpine and ajmalicine yield in Rauvolfia serpentina as revealed by high-performance thin-layer chromatography (HPTLC)

Quantification of taxol by high-performance thin layer chromatography in Taxus wallichiana callus cultivated in vitro

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