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In vitro propagation and establishment of adventitious root cultures of Rheum emodi Wall. ex Meisn for quantification of emodin production

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Abstract

Rheum emodi Wall. ex Meisn is an important medicinal herb used in treating type II diabetes mellitus, arthritis, inflammation, and fever. Roots and rhizomes of this plant possess anthraquinones showing anticancer, antiviral, anti-inflammatory, anti-ulcerogenic, immunosuppressive, and pro-apoptotic activities. In the present study, in vitro propagation and adventitious root establishment were achieved for the detection and quantification of important secondary metabolites. In micropropagation experiments, high rate of callus induction was obtained from mid rib (100%) as well as leaf explants (97.67 ± 1.20%). The highest percent shoot induction was observed from leaf explants (76.25 ± 2.39%) as compared to callus (66.67 ± 2.20%). Regenerated shoots showed 70% rooting response with healthy long roots (5.73 ± 0.15 cm). Adventitious root culture response from nodal and leaf explants was better in liquid medium (93.33 ± 1.67% and 73.13 ± 2.87%) compared to semi-solid medium (85.0 ± 2.89% and 76.67 ± 3.33%). The highest DPPH (2–2 diphenyl, 1-picrylhydrazyl) free radical scavenging activity (92.73 ± 0.07%) and total antioxidant activity (6.24 ± 0.07 µg ascorbic acid equivalent (AAE) mg−1) was recorded in in vitro roots. Maximum phenol content (14.70 ± 0.29 µg gallic acid equivalent (GAE) mg−1) was present in adventitious root culture whereas flavonoid content (82.37 ± 0.12 µg quercetin equivalent (QE) mg−1) in in vitro shoots. The identification and quantification of secondary metabolites in in vitro shoots, in vitro roots, and adventitious root cultures were evaluated via high-performance liquid chromatography-mass spectroscopy (HPLC–MS) analysis which revealed the presence of three anthraquinones, emodin, physcion, and chrysophanol, in adventitious root cultures. HPLC–UV detector analysis showed the highest emodin concentration (2.24%) in in vitro shoots. The present study gives a highly reproducible micropropagation protocol for R. emodi as well as provides an alternative source of anticancer compound, emodin, from in vitro shoots and root cultures.

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Acknowledgements

The authors are thankful to the Department of Biological Sciences, C.B.S.H., G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, India, for providing the necessary funding and facilities. The authors are also thankful to Dr. D.S. Rawat from the Department of Biological Sciences, G.B. Pant University of Agriculture and Technology for kind help rendered in collection and identification of the plant material. They are also thankful to SAIF (Sophisticated Analytical Instrument Facility), IIT Bombay for HPLC-MS analysis, and Prof. Anjana Srivastava from the Department of Chemistry, G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, for HPLC analysis. The corresponding author is thankful to University Grant Commission (UGC) for providing her fellowship.

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  1. Department of Biological Sciences, CBSH, G. B. Pant University of Agriculture & Technology, Pantnagar 263145, Udham Singh Nagar, Uttarakhand, India

    Ruchi Singh & Preeti Chaturvedi

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  1. Ruchi Singh
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  2. Preeti Chaturvedi
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RS and PC conceived the research idea; RS performed the experiment, analyzed the data, and wrote the paper; and PC supervised the research and designed the experiment. All authors have read and approved the final manuscript.

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Correspondence to Preeti Chaturvedi.

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Singh, R., Chaturvedi, P. In vitro propagation and establishment of adventitious root cultures of Rheum emodi Wall. ex Meisn for quantification of emodin production. In Vitro Cell.Dev.Biol.-Plant 58, 80–92 (2022). https://doi.org/10.1007/s11627-021-10220-1

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