Large-scale production of in vitro plantlets of sweet flag (Acorus calamus L.) and quantitative analysis of α- and β-asarone contents using high pressure liquid chromatography

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Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Website:www.vegetosindia.org
Pub Email: contact@vegetosindia.org
Doi: 10.1007/s42535-021-00287-x
First Page: 122
Last Page: 132
Views: 456


Keywords: n Acorus calamusn , Rhizome, Micropropagation, Plantlets, HPLC analysis, Asarone contents


Abstract


Acorus calamus is a perennial; an endangered and potential medicinal plant that belongs to the Acoraceae family has a range of medicinal properties and fragrant oils, which are being used in various industries. In the current investigation, the highly proficient and reproducible method for comprehensive plantlet regeneration was established and compared the α- and β-asarone contents in between the in vivo as well as in vitro raised plantlets. Amongst the combinations of plant growth regulators (PGRs), maximum shoots per explant (7.23) were observed with KIN. Later, the shoots were subcultured on MS medium supplemented with BAP, IAA and GA3 showed a maximum percentage of responses in shoot multiplication and elongation. The elongated shoots were subcultured on MS medium fortified with IBA for root initiation and development, and successively the plantlets were acclimatized in plastic pots in the culture room and greenhouse. Quantitative analysis of α- and β-asarone was performed using HPLC from in vitro and in vivo raised plantlets. A significant difference was noted in α-asarone content of in vitro plantlets than in vivo plantlets whereas the least difference was noted in β-asarone content. The results of the present study revealed the potential reproducible protocol for largescale multiplication of A. calamus in 130 days. Further, the regeneration protocol will be a very useful platform for in vitro breeding with different PGRs to analyze the impact on improvement of bioactive molecules such as α- and β-asarone content and fragrant oils, which have huge demand in pharmaceutical and cosmetic trades and owing to their high industrial values.

n              Acorus calamusn            , Rhizome, Micropropagation, Plantlets, HPLC analysis, Asarone contents


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References


Ahmed A (2010) Tissue culture studies of Acorus calamus Linn. and evaluation of secondary metabolites (Doctoral dissertation), RGUHS, Bangalore


Ahmed MB, Ahmed S, Salahin M, Sultana R, Khatun M, Razvy MA, Hossain MM (2007) Standardization of a suitable protocol for in vitro clonal propagation of Acorus calamus L. an important medicinal plant in Bangladesh. Am Eurasian J Sci Res 2:136–140


Anu A, Babu KN, John CZ, Peter KV (2001) In vitro clonal Multiplication of Acorus calamus L. J Plant Biochem Biotechnol 10:53–55


Babar PS, Deshmukh AV, Salunkhe SS, Chavan JJ (2020) Micropropagation, polyphenol content and biological properties of Sweet Flag (Acorus calamus): a potent medicinal and aromatic herb. Vegetos 33:296–303


Bahukhandi A, Rawat S, Bhatt ID, Rawal RS (2013) Influence of solvent types and source of collection on total phenolic content and antioxidant activities of Acorus calamus L. Natl Acad Sci Lett 36:93–99


Balakumbahan R, Rajamani K, Kumanan K (2010) Acorus calamus: an overview. J Med Plant Res 4:2740–2745


Bhat KG (2003) Flora of Udupi. Indian Naturalist (Regd.), Inchara, Chitpady, Udupi, pp 675–676


Chopra IC, Handa KL, Abrol BK (1958) Indian calamus: its scope in modern therapeutics. J Sci Ind Res 17A:8–11


Das BK, Swamy AV, Koti BC, Gadad PC (2019) Experimental evidence for use of Acorus calamus (asarone) for cancer chemoprevention. Heliyon 5:e01585


Deepalakshmi PD, Odgerel K, Thirugnanasambantham P, Yungeree O, Khorolragchaa A, Senthil K (2016) Metabolite profiling of in vitro cultured and field grown rhizomes of Acorus calamus from Mongolia Using GC–MS. Chromatographia 79:1359–1371


Devi NS, Kishor R, Sharma GJ (2012) Microrhizome induction in Acorus calamus Linn. An important medicinal and aromatic plant. Hortic Environ Biotechnol 53:410–414


Farheen M, Sonu S (2012) Protocol for in-vitro clonal propagation of Acorus calamus L. an important endangered medicinal plant. Indian Forester 138:63–69


Gomez KA, Gomez AA (1984) Statistical procedure for agricultural research, 2nd edn. Wiley, New York, p 704





Khare CP (2007) Indian medicinal plants: an illustrated dictionary. Springer Science and Business Media, Berlin


Kumar R, Sharma S, Sharma S, Kumar N (2016) Drying methods and distillation time affects essential oil content and chemical compositions of Acorus calamus L. in the Western Himalayas. J App Res Med Aromatic Plants 3:136–141





Lee MH, Chen YY, Tsai JW, Wang SC, Watanabe T, Tsai YC (2011) Inhibitory effect of β-asarone, a component of Acorus calamus essential oil, on inhibition of adipogenesis in 3T3-L1 cells. Food Chem 126:1–7





Mittal N, Ginwal HS, Varshney VK (2009) Pharmaceutical and biotechnological potential of Acorus calamus Linn.: an indigenous highly valued medicinal plant species. Phcog Rev 3:93–103


Mukherjee PK (2002) Quality control of herbal drugs: an approach to evaluation of botanicals. Business Horizons, New Delhi, pp 692–694


Mukherjee PK, Kumar V, Mal M, Houghton PJ (2007a) In vitro acetyl cholinesterase inhibitory activity of the essential oil from Acorus calamus and its main constituents. Planta Med 73:283–284


Mukherjee PK, Kumar V, Mal M, Houghton PJ (2007b) Acorus calamus: scientific validation of ayurvedic tradition from natural resources. Pharm Biol 45(8):651–666


Mukherjee PK, Mao AA (2021) Compendium of antiviral medicinal plants of North East India. Institute of Bioresources and Sustainable Development, Imphal and Botanical Survey of India, Kolkata, pp 5–10


Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497





Rajasekharan PE, Ravish BS, Vasantha Kumar T (2008) Optimization of protocols for the in vitro multiplication and conservation of Acorus calamus, an endangered medicinal plant. In: IV International symposium on acclimatization and establishment of micropropagated plants, vol 865, pp 37–41


Rani AS, Subhadra VV, Reddy VD (2000) In vitro propagation of Acorus calamus Linn. A medicinal plant. Indian J Exp Biol 38:730–732


Rasheed NMA, Nagaiah K, Waheed MA (2013) Recent analytical techniques in quality control of indigenous system of medicine. Ann Phytomed 2:44–58





Sandhyarani N, Kishor R, Sharma GJ (2011) Clonal propagation of triploid Acorus calamus Linn. using dual-phase culture system. J Crop Sci Biotechnol 14:213–217


Sharma V, Singh I, Chaudhary P (2014) Acorus calamus (The Healing Plant): a review on its medicinal potential, micropropagation and conservation. Nat Prod Res 28:1454–1466


Sharma V, Sharma R, Gautam DS, Kuca K, Nepovimova E, Martins N (2020) Role of vacha (Acorus calamus Linn.) in neurological and metabolic disorders: evidence from ethnopharmacology, phytochemistry, pharmacology and clinical study. J Clin Med 9:1176


Singh M, Kamal YT, Parveen R, Ahmad S (2011) Separation and simultaneous quantification of α-and β-Asarone in Acorus calamus Linn. from Indian sub-continent on caffeine modified silica. Asian J Chem 23:2046


Singh S, Srivastava R, Choudhary S (2010) Antifungal and HPLC analysis of the crude extracts of Acorus calamus, Tinospora cordifolia and Celestrus paniculatus. J Agric Technol 6:149–158


Subramoniam A (2014) Present scenario, challenges and future perspectives in plant-based medicine development. Ann Phytomed 3:31–36

 


Acknowledgements


We thank Manipal Academy of Higher Education (MAHE), Manipal, Karnataka, India, and TIFAC-CORE and FIST, DST New Delhi and K-FIST, VGST, Govt. of Karnataka for the facilities. We are indebted to Prof. K. Satyamoorthy, Director, Manipal School of Life Sciences, MAHE for support. The authors, Swathy PS, Sachin AT, Arya K, and Kiran KR are grateful to MAHE for Dr. T.M.A. Pai Ph.D. scholarship. Thanks are also due to Mrs. M. Ramya for the collection of A. calamus and Ms. P. Bhagyashree for experimental assistance. We are grateful to anonymous reviewers and editorial comments for the improvement of earlier version of the manuscript.


Author Information


Muthusamy Annamalai
Department of Plant Sciences, Manipal School of Life Sciences (MSLS), Manipal Academy of Higher Education (MAHE), Manipal, India
a.msamy@manipal.edu
Tantry Shashikala
Department of Plant Sciences, Manipal School of Life Sciences (MSLS), Manipal Academy of Higher Education (MAHE), Manipal, India


Thorat Sachin Ashok
Department of Plant Sciences, Manipal School of Life Sciences (MSLS), Manipal Academy of Higher Education (MAHE), Manipal, India


Kaniyassery Arya
Department of Plant Sciences, Manipal School of Life Sciences (MSLS), Manipal Academy of Higher Education (MAHE), Manipal, India

Kiran Kodsara Ramachandra
Department of Plant Sciences, Manipal School of Life Sciences (MSLS), Manipal Academy of Higher Education (MAHE), Manipal, India