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Enhanced Production of Phenylethanoids Mediated Through Synergistic Approach of Precursor Feeding and Light Regime in Cell Suspension Culture of Rhodiola imbricata (Edgew.)

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Abstract

Precursor feeding is a potential strategy for increasing specialized metabolite production in plant cell culture systems. In the present study, cell suspension cultures were developed and subsequently evaluated for precursor feeding investigations. Cell suspension cultures were established in Murashige and Skoog (MS) medium containing 0.5 mg/L thidiazuron (TDZ) + 1 mg/L α-naphthalene acetic acid (NAA). The growth biomass and metabolite pattern were analyzed to identify specific culture days required for prolific biomass production. The maximum cell dry weight (DW) was observed in leaf cell suspension (1.22 g/100 mL) and root cell suspension culture (1.12 g/100 mL) on day 21. Afterward, the effect of precursor concentrations (tyrosol; 0.5, 1, 2, and 3 mM) along with two light regimes, photoperiod (16L/8D h, 70 µmol/m2/s) and dark (24 h), was evaluated for cell growth and metabolite accumulation. The results revealed that leaf cell suspension treated with 3 mM tyrosol concentration detected maximum salidroside content (26.05 mg/g DW) on day 15, incubated under photoperiod (16L/8D h) condition. Similarly, under photoperiod (16L/8D h), root cell suspension treated with 3 mM tyrosol produced maximum salidroside content (26.62 mg/g DW) on day 12. Moreover, the total phenolics content increased significantly (44.21 mg/g DW) on day 12 in 3 mM tyrosol treatment under photoperiod (16L/8D h). However, precursor concentrations did not influence the total flavonoids content. The present investigation suggests that the immediate pathway precursor, tyrosol, has a strong effect on enhanced production of salidroside, irrespective of explant type and light regimes.

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Abbreviations

MS:

Murashige and Skoog

16L/8D h:

16-H light/8-h dark

TDZ:

Thidiazuron

NAA:

α-Naphthalene acetic acid

RPM:

Rotation per minutes

PAL:

Phenylalanine ammonia-lyase

CHS:

Chalcone synthase

LCS:

Leaf cell suspension

RCS:

Root cell suspension

FW:

Fresh weight

DW:

Dry weight

PTFE:

Polytetrafluoroethylene

UPLC–PDA:

Ultra-performance liquid chromatography-photodiode array

SE:

Standard error

TPC:

Total phenolics content

TFC:

Total flavonoids content

mM:

Mili molar

SPSS:

Statistical Package for the Social Sciences

MS:

Microsoft Office

ANOVA:

Analysis of variance

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Acknowledgements

SR acknowledges UGC, New Delhi, for providing research fellowship and Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, India, for Ph. D. enrolment. The authors are thankful to Director, CSIR-IHBT, Palampur, for providing the necessary facilities. CSIR-IHBT publication number for this manuscript is 4892.

Funding

The authors acknowledge the Council of Scientific and Industrial Research (CSIR), Government of India, under the project “Biotechnological interventions for sustainable bio-economy generation through characterization, conservation, prospection, and utilization of Himalayan bioresources” (MLP-0201) and National Medicinal Plants Board (NMPB), “Development of Probiotics for Plant Tissue Culture Boosting the performance of micro propagated plant materials by supplementing plant associated useful endophytes” (GAP-0274) for providing financial support.

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Authors and Affiliations

  1. Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, 176061, Himachal Pradesh, India

    Shiv Rattan & Ashish R. Warghat

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India

    Shiv Rattan, Anil Kumar, Dinesh Kumar & Ashish R. Warghat

  3. Chemical Technology Division, CSIR-Institute of Himalayan Bioresource Technology, Palampur, 176061, Himachal Pradesh, India

    Anil Kumar & Dinesh Kumar

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  1. Shiv Rattan
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  2. Anil Kumar
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Contributions

Conceptualization: Shiv Rattan and Ashish R. Warghat; methodology: Shiv Rattan and Ashish R. Warghat; data analysis and investigation: Shiv Rattan, Anil Kumar, Dinesh Kumar, and Ashish R. Warghat; original manuscript preparation: Shiv Rattan and Ashish R. Warghat; manuscript-review and editing: Shiv Rattan, and Ashish R. Warghat.

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Correspondence to Ashish R. Warghat.

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Rattan, S., Kumar, A., Kumar, D. et al. Enhanced Production of Phenylethanoids Mediated Through Synergistic Approach of Precursor Feeding and Light Regime in Cell Suspension Culture of Rhodiola imbricata (Edgew.). Appl Biochem Biotechnol 194, 3242–3260 (2022). https://doi.org/10.1007/s12010-022-03914-8

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