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In vitro Micropropagation of Acorus calamus L. Using Rhizome Nodal Segments: Optimization of Surface Sterilization, Hormonal Regimes, and Acclimatization
Subject area: Biological & Medical Sciences · Area of research: Biotechnology
DOI: https://doi.org/10.64388/IREV9I11-1717903
Abstract
Acorus calamus L. (family Acoraceae), commonly known as Sweet Flag or 'Vacha', is a pharmaceutically important semi-aquatic perennial herb whose bioactive rhizomes are extensively exploited in Ayurveda, Unani, and Traditional Chinese Medicine. Conventional propagation through rhizome segments is severely limited by low multiplication rates and susceptibility to soilborne pathogens, necessitating the development of reliable micropropagation protocols. The present study, conducted at the State Forest Research Institute (SFRI), Jabalpur, Madhya Pradesh, established a reproducible and efficient in vitro propagation protocol for A. calamus using rhizome nodal segments as explants. An optimized two-step surface sterilization regimen 1.0% Bavistin (carbendazim) for 10 minutes followed by 0.1% HgCl₂ for 3 minutes effectively eliminated microbial contamination while preserving explant viability. Shoot induction was most effectively achieved on Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzylaminopurine (BAP), yielding a maximum of 4.8 ± 0.6 shoots per explant within approximately 9 days. Shoot multiplication was optimized on MS medium containing 3.0 mg/L BAP combined with 3.0 mg/L naphthaleneacetic acid (NAA), producing 9.4 ± 1.2 shoots per explant with a mean shoot length of 5.1 ± 0.9 cm. In vitro rooting was best accomplished on half-strength MS medium supplemented with 1.0 mg/L NAA, generating fibrous, well-differentiated roots within 8 days. Ex vitro acclimatization of in vitro-raised plantlets in a soil : sand : organic manure (1:1:1) substrate under stepwise humidity reduction achieved an 85% survival rate after four weeks under greenhouse conditions. The protocol developed provides a scalable tool for the clonal propagation of A. calamus, with applications in conservation of threatened natural populations, commercial cultivation for the essential oil and pharmaceutical industries, and production of certified disease-free planting material.
Keywords
Acorus Calamus, Micropropagation, Rhizome Nodal Explant, BAP, NAA, Shoot Multiplication, Acclimatization, Ayurvedic Medicinal Plant
How to cite this paper
@article{1717903,
author = {Shailendra Singh Yadav, Ananya Shrivastava, Rajni Nigam, Sandeep Fellows, Pradeep Vasudeva},
title = {In vitro Micropropagation of Acorus calamus L. Using Rhizome Nodal Segments: Optimization of Surface Sterilization, Hormonal Regimes, and Acclimatization},
journal = {Iconic Research And Engineering Journals},
year = {2026},
volume = {9},
number = {11},
pages = {1940-1950},
issn = {2456-8880},
url = {https://www.irejournals.com/formatedpaper/1717903.pdf},
abstract = {Acorus calamus L. (family Acoraceae), commonly known as Sweet Flag or 'Vacha', is a pharmaceutically important semi-aquatic perennial herb whose bioactive rhizomes are extensively exploited in Ayurveda, Unani, and Traditional Chinese Medicine. Conventional propagation through rhizome segments is severely limited by low multiplication rates and susceptibility to soilborne pathogens, necessitating the development of reliable micropropagation protocols. The present study, conducted at the State Forest Research Institute (SFRI), Jabalpur, Madhya Pradesh, established a reproducible and efficient in vitro propagation protocol for A. calamus using rhizome nodal segments as explants. An optimized two-step surface sterilization regimen 1.0% Bavistin (carbendazim) for 10 minutes followed by 0.1% HgCl₂ for 3 minutes effectively eliminated microbial contamination while preserving explant viability. Shoot induction was most effectively achieved on Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzylaminopurine (BAP), yielding a maximum of 4.8 ± 0.6 shoots per explant within approximately 9 days. Shoot multiplication was optimized on MS medium containing 3.0 mg/L BAP combined with 3.0 mg/L naphthaleneacetic acid (NAA), producing 9.4 ± 1.2 shoots per explant with a mean shoot length of 5.1 ± 0.9 cm. In vitro rooting was best accomplished on half-strength MS medium supplemented with 1.0 mg/L NAA, generating fibrous, well-differentiated roots within 8 days. Ex vitro acclimatization of in vitro-raised plantlets in a soil : sand : organic manure (1:1:1) substrate under stepwise humidity reduction achieved an 85% survival rate after four weeks under greenhouse conditions. The protocol developed provides a scalable tool for the clonal propagation of A. calamus, with applications in conservation of threatened natural populations, commercial cultivation for the essential oil and pharmaceutical industries, and production of certified disease-free planting material.},
keywords = {Acorus Calamus, Micropropagation, Rhizome Nodal Explant, BAP, NAA, Shoot Multiplication, Acclimatization, Ayurvedic Medicinal Plant},
month = {May},
doi = {https://doi.org/10.64388/IREV9I11-1717903}
}