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.
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Doi: 10.1007/s42535-021-00287-x
First Page: 122
Last Page: 132
Views: 467

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


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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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
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