Genome size analysis of field grown and tissue culture regenerated Rauvolfia serpentina (L) by flow cytometry: Histology and scanning electron microscopic study for in vitro morphogenesis

Zafar, Nadia; Mujib, A.; Ali, Muhammad Asghar; Tonk, Dipti; Gulzar, Basit; Malik, Moien Qadir; Sayeed, Rukaya; Mamgain, Jyoti

doi:10.1016/j.indcrop.2018.11.049

Cited by 37 publications

(16 citation statements)

References 56 publications

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“…This is in accordance with the results that were reported by Singh et al (2009) and Rohela et al (2013) on R. serpentina. Contrary to our results, whitish calli from BA + IAA were obtained by Rani et al (2014) and friable calli from BA alone were reported by Zafar et al (2019). In comparison to control, wherein minute callus was observed to have formed from the petiolar end, it could be clearly stated that in PGR-supplemented media, especially auxins like 2,4-D were utilized for inducing friable calli, and cytokinins (BA and KIN) employed for inducing green embryogenic calli, proved to be much more efficient.…”

Section: Discussioncontrasting

confidence: 99%

“…Irrespective of PGR type, lower concentrations exhibited better performance than the higher ones, with the exception of BA. Similar results were published in the reports of Mallick et al (2012) and Zafar et al (2019) wherein it was claimed that shoots were more responsive to higher concentrations of BA during shoot induction. Most likely, a higher dose of BA was effective in shoot initiation and lower concentrations of the same induced better shoot growth and proliferation.…”

Section: Discussionsupporting

confidence: 89%

“…However, Rashmi and Trivedi (2016) further reported the use of BA + 2,4-D combination for callus induction. In the reports of Pandey et al (2007), Bahuguna et al (2011), Rohela et al (2013), and Zafar et al (2019, it was stated that 1-2 mg/l 2,4-D along NAA or BA or KIN was effective in higher callus inductions. However, reported the use of a higher dose of 2,4-D (5 mg/l) along with 2 mg/l NAA for callus induction in R. serpentina.…”

Section: Discussionmentioning

confidence: 97%

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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: Discussioncontrasting

confidence: 99%

Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 97%

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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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“…etc [1][2][3]. Among the different genus of Apocynaceae; the Rauwolfia genus is popularly known as the major source of pharmaceutical alkaloids [4,5]. Among the Rauwolfia species; initially Rauwolfia serpentina L. Benth.…”

Section: Introductionmentioning

confidence: 99%

High performance liquid chromatography based quantification of reserpine in Rauwolfia tetraphylla L. and enhanced production through precursor feeding

Rohela

Bylla

Pendli

et al. 2021

AChrom

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Rauwolfia tetraphylla L., is an important medicinal plant in Apocynaceae family and is recognized as an alternative source to Rauwolfia serpentina L., in terms of anti-hypertensive alkaloid production i.e. reserpine. In view of this, the present study is conducted to estimate the reserpine content in different parts (leaf, stem and root) of field grown plants (2 years old), tissue cultured plantlets (R1) (two months old) and cell suspensions cultures (two months old with and without precursor feeding) of R. tetraphylla by using high performance liquid chromatography (HPLC) technique. Overall maximum content of reserpine (in %) was estimated from the root samples. Roots of field grown plants has recorded high percent of reserpine (0.39%) followed by roots of tissue cultured plantlets (0.35%) and root callus based cell suspension cultures (0.38 %) which was fed with precursor amino acid (100 mg/L of tryptophan). In control type of root callus based cell suspension cultures, reserpine content was quantified as 0.14%; by precursor feeding (100 mg/L of tryptophan) it was enhanced to 0.38%. In conclusion, the reserpine content (0.35 and 0.38%) produced by the roots of tissue cultured plantlets (R1) and 100 mg/L tryptophan fed root callus based cell suspensions was comparable to that of the reserpine content (0.39%) of root parts of field grown plants. The present study demonstrates the reserpine production by in vitro cell suspension cultures throughout the year without sacrificing the medicinal plants.

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“…Also, the low alkaloidal content in the plants demands use of natural sources in larger quantities. Conjointly, the significant depletion of the natural sources has probed into the matter of environmental concern (Zafar et al, 2019). Therefore, sustainable and enhanced production through an elaborative study of the metabolic interactions involved in the synthesis of the alkaloid is required.…”

Section: Introductionmentioning

confidence: 99%

Conversion of Cytochrome P450 2D6 of Human Into a FRET-Based Tool for Real-Time Monitoring of Ajmalicine in Living Cells

Ambrin

Ahmad

Alqarawi

et al. 2019

Front. Bioeng. Biotechnol.

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Ajmalicine is naturally present in the root bark of Catharanthus roseus L. and Rauvolfia serpentina (L.) Benth ex.Kurz. It has been extensively utilized in the treatment of hypertension across the world. The increased demand, overconsumption, and low content of the alkaloid in the plants have raised the issue of the depletion of natural sources. The metabolic engineering approach has not been successful in improving the content of the ajmalicine because the metabolic regulation of this metabolite is not known. The regulation of a metabolite in the metabolic pathway requires a tool that can carry out real-time measurement of the flux of the metabolite in living system. Given this, the present study was conducted to develop a genetically encoded FRET-based nanosensor by engineering human Cytochrome P450-2D6, an ajmalicine binding protein. The Cytochrome P450-2D6 was sandwiched between two FRET fluorophores. The design of the nanosensor brings two fluorescent proteins in conjunction with the ajmalicine binding protein, such that it undergoes FRET (Fluorescence Resonance Energy Transfer) upon binding of the ligand. The nanosensor, named as FLIP-Ajn (Fluorescence Indicator Protein for Ajmalicine), was pH stable and ajmalicine specific. The affinity of the FLIP-Ajn was 582 μM. The FLIP-Ajn successfully performed real-time measurement of ajmalicine in prokaryotic (bacteria) and eukaryotic systems (yeast, animal cell line, and plant suspension culture), thereby, establishing its biocompatibility in monitoring of ajmalicine in living cells. Besides, several affinity mutants of the nanosensor were generated through mutations in the ajmalicine binding protein to increase the detection range of the nanosensor at varying physiological scales. The non-invasiveness and high spatial and temporal resolution of the tool holds a great significance in the bio-imaging of a highly compartmentalized metabolic pathway. The flux study of ajmalicine will help in identifying the regulatory steps involved in the synthesis of the alkaloids and, hence, will improve the production rate of ajmalicine from its natural sources.

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Genome size analysis of field grown and tissue culture regenerated Rauvolfia serpentina (L) by flow cytometry: Histology and scanning electron microscopic study for in vitro morphogenesis

Cited by 37 publications

References 56 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.

High performance liquid chromatography based quantification of reserpine in Rauwolfia tetraphylla L. and enhanced production through precursor feeding

Conversion of Cytochrome P450 2D6 of Human Into a FRET-Based Tool for Real-Time Monitoring of Ajmalicine in Living Cells

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