VEGETOS: An International Journal of Plant Research & Biotechnology
(Society For Plant Research)

Research Articles


Volume: 33, Issue: 2, June 2020

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Page Visits: 145

Doi: 10.1007/s42535-020-00107-8
Doi Link:
First Page: 296
Last Page: 303
Published: 19 March, 2020

Micropropagation, polyphenol content and biological properties of Sweet Flag (Acorus calamus): a potent medicinal and aromatic herb


Acorus calamus L. (family-Acoraceae) is commonly known as ‘Sweet Flag’ which grows naturally in marshy land, shallow water and the edges of the pond. In recent years, there is a growing demand of this plant due to its multidimensional implications including food, cosmetics and pharmaceuticals. In the present study, an efficient and rapid in vitro regeneration system along with higher accumulation of secondary metabolites has been developed for A. calamus. The rhizome bud explant cultured on Murashige and Skoog’s medium supplemented with a combination of BAP (3.0 mg/l) and IAA (1.0 mg/l) resulted in the production of maximum numbers of shoots (11.3 ± 0.4). Microshoots were transferred to MS medium supplemented with various concentrations and combinations of auxins for in vitro rooting. The maximum contents of total phenolics, flavonoids and higher antioxidant properties were reported in the rhizomes of tissue culture raised plants as compared to field grown ones. Methanolic and acetone extract of rhizomes of micropropagated plants showed slightly elevated antibacterial activity against five bacterial pathogens. The current findings indicated that, the phytochemicals together with the bioactivity of Sweet Flag is significantly high in the tissue culture derived plantlets.



Antibacterial activity, Antioxidant properties, Flavonoids, Micropropagation, Phenolics, Sweet Flag


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

  2. Altaf A, Shashidhara S, Rajasekharan PE, Hareesh Kumar V, Honnesh NH (2010) In vitro regeneration of Acorus calamus—an important medicinal plant. J Curr Pharm Res 2(1):36–39

  3. Amoo SO, Van Staden J (2012) Influence of plant growth regulators on shoot proliferation and secondary metabolite production in micropropagated Huernia hystrix. Plant Cell Tiss Org Cult 112(2):249–256

  4. Asha Devi S, Ganjewala D (2011) Antioxidant activities of methanolic extracts of Sweet-flag (Acorus calamus) leaves and rhizomes. J Herbs Spices Med Plants 17:1–11

  5. 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(1):93–99

  6. Balyan U, Verma SP, Sakrar B (2019) Phenolic compounds from Syzygium cumini (L.) Skeels leaves: extraction and membrane purification. J Appl Res Med Aromat Plants 12:43–58

  7. Benzie IF, Strain JJ (1996) Ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: the FRAP assay. Ann Biochem 239:70–76

  8. Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free radical method to evaluate antioxidant activity. LWT Food Sci Technol 28:25–30

  9. Chandra D, Prasad K (2017) Phytochemicals of Acorus calamus (Sweet Flag). J Med Plants Stud 5(5):277–281

  10. Chang C, Yang M, Wen H, Chern J (2002) Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 10:178–182

  11. Chaturvedi M, Chaturvedi AK (2017) Acorus calamus: a divine drug for the human being. Imp J Interdiscip Res 3(1):1499–1503

  12. Chavan JJ, Gaikwad NB, Kshirsagar PR, Dixit GB (2013b) Total phenolics, flavonoids and antioxidant properties of three Ceropegia species from Western Ghats of India. S Afr J Bot 88:273–277

  13. Chavan JJ, Gaikwad NB, Umdale SD, Kshirsagar PR, Bhat KV, Yadav SR (2014b) Efficiency of direct and indirect shoot organogenesis, molecular profiling, secondary metabolite production and antioxidant activity of micropropagated Ceropegia santapaui. Plant Growth Regulat 72:1–15

  14. Chavan JJ, Gaikwad NB, Yadav SR (2013a) High multiplication frequency and genetic stability analysis of Ceropegia panchganiensis, a threatened ornamental plant of Western Ghats: Conservation implications. Sci Hortic 161:134–142

  15. Chavan JJ, Nalawade AS, Gaikwad NB, Gurav RV, Dixit GB, Yadav SR (2014a) An efficient in vitro regeneration of Ceropegia noorjahaniae: an endemic and critically endangered medicinal herbof the Western Ghats. Physiol Mol Biol Plants 20(3):405–410

  16. Devaki M, Nirupama R, Nirupama M, Yajurvedi HN (2016) Protective effect of rhizome extracts of the herb, vacha (Acorus calamus) against oxidative damage: an in vivo and in vitro study. Food Sci Hum Wellness 5(2):76–84

  17. Hettiarachchi A, Fernando KKS, Jayasuriya AHM (1997) In vitro propagation of Wadakha (Acorus calamus L). J Natl Sci Counc Sri Lanka 25(3):151–157

  18. Howes MR, Houghton PJ (2003) Plants used in Chinese and Indian traditional medicine for improvement of memory and cognitive function. Pharmacol Biochem Behav 75:513–527

  19. Irshad S, Mahmood M, Perveen F (2012) In vitro antibacterial activities of three medicinal plants using agar well diffusion method. Res J Biol 2:1–8

  20. Kim WJ, Hwang KH, Park DG, Kim TJ, Kim DW, Choi DK, Moon WK, Lee KH (2011) Major constituents and antimicrobial activity of Korean herb Acorus calamus. Nat Prod Res. 25(13):1278–1281

  21. Kumar V, Singh R, Joshi V (2014) Antimicrobial activity of rhizome extract of Acorus calamus against different micro-organisms. Octa J Biosci 2(1):59–63

  22. Marongiu B, Piras A, Porcedda S, Scorpciapino A (2005) Chemical composition of the essential oil and supercritical CO2 extract of Commiphora myrrha (Nees) Engl. and of Acorus calamus L. J Agric Food Chem 53:7939–7943

  23. McGaw LJ, Jager AK, Van Staden J (2002) Isolation of β-asarone, an antibacterial and anthelmintic compound from Acorus calamus in South Africa. S Afr J Bot 68:31–35

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

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

  26. Ncube B, Ngunge VNP, Finnie JF, Van Staden J (2011) A comparative study of the antimicrobial and phytochemical properties between outdoor grown and micropropagated Tulbaghia violacea Harv. plants. J Ethnopharmacol 134(3):775–780

  27. Palani S, Raja S, Praveen Kumar R, Venkadesan D, Devi K, Sivaraj A, Senthil Kumar B (2009) Therapeutic efficacy of antihepatotoxic and antioxidant activities of Acorus calamus on acetaminophen-induced toxicity in rat. Int J Integr Biol 7(1):39–43

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

  29. Si MM, Lou JS, Zhou CX, Shen JN, Wu HH (2010) Insulin releasing and alpha-glucosidase inhibitory activity of ethyl acetate fraction of Acorus calamus in vitro and in vivo. J Ethnopharmacol 28:154–159

  30. Singh KK, Prakash A (1994) Indigenous phytotherapy among the Gond tribes of Uttar Pradesh, India. Ethnobot 6:37–41

  31. Singh R, Sharma PK, Malviya R (2011) Pharmacological properties and Ayurvedic value of Indian Buch plant (Acorus calamus): a short review. Adv Biol Res 5(3):145–154

  32. Singleton VL, Rossi JA (1965) Colorimetry of total phenolics with phosphor-molybdic phosphotungstic acid reagents. Am J Enol Vitic 16:144–158

  33. Verma S, Singh N (2012) In vitro mass multiplication of Acorus calamus L.—an endangered medicinal plant. Am-Eurasian J Agric Environ Sci 12(11):1514–1521





Acknowledgements :

Rashtriya Uchchtar Shiksha Abhiyan (RUSA), Ministry of Human Resource Development (MHRD), Govt. of India is gratefully acknowledged for instrumentation facilities created under Component 8. Authors are indebted to the Principal, Yashavantrao Chavan Institute of Science, Satara (Autonomous) for providing necessary laboratory facilities.

Author Information:

Prajakta S. Babar
Department of Biotechnology, Yashavantrao Chavan Institute of Science, Satara, India

Anuradha V. Deshmukh
Department of Biotechnology, Yashavantrao Chavan Institute of Science, Satara, India

Supriya S. Salunkhe
Department of Biotechnology, Yashavantrao Chavan Institute of Science, Satara, India

Jaykumar J. Chavan
Department of Biotechnology, Yashavantrao Chavan Institute of Science, Satara, India

Pdf Download