Acta Physiologiae Plantarum

, Volume 36, Issue 10, pp 2683–2693 | Cite as

Stimulation of in vitro morphogenesis, antioxidant activity and over expression of kaurenoic acid 13-hydroxylase gene in Stevia rebaudiana Bertoni by chlorocholine chloride

  • Sayanti Kundu
  • Avishek Dey
  • Abhijit BandyopadhyayEmail author
Original Paper


Phytoconstituents from medicinal plants are considered as important source of raw materials of drugs for pharmaceutical industries. Biotechnology has become an inevitable approach in the area of research and development of medicinal plants for many decades. The present work has been carried out to ascertain the role of chlorocholine chloride (CCC) on in vitro morphogenesis, antioxidant activity and expression level of kaurenoic acid 13-hydroxylase (KA13H) gene in Stevia rebaudiana. To fulfill these purposes chlorocholine chloride was applied in the Murashige and Skoog (Physiol Plant 15(3):473–497, 1962) medium in combination with other plant growth regulators such as 1-naphthalene acetic acid, kinetin and thidiazuron. Chlorocholine chloride was found to contribute significant role on in vitro morphogenesis of S. rebaudiana as evidenced by the formation of embryogenic calli and increase in callusing and microshooting efficiency of explant, i.e., cotyledonary leaf. Moreover, antioxidant enzyme activity as well as ascorbic acid content of the calli and leaves was also stimulated after application of chlorocholine chloride. Q-PCR amplification using gene-specific primers revealed that CCC also promoted the expression level of KA13H gene in S. rebaudiana leaves. The overall study highlighted the promising role of chlorocholine chloride on regeneration efficiency of cotyledonary leaf, significant promotion in antioxidant potential and expression of KA13H gene in S. rebaudiana.


Stevia rebaudiana Chlorocholine chloride In vitro morphogenesis Antioxidant activity Kaurenoic acid 13-hydroxylase 



Analysis of variance


Chlorocholine chloride






Murashige and Skoog


Normality of solution


1 Naphthalene acetic acid


Nitroblue tetrazolium chloride




Quantitative polymerase chain reaction


Polyvinyl pyrrolidone


Relative humidity


Rotation per minute


Trichloro acetic acid






Weight by volume


Volume by volume



The authors are grateful to their supervisor for valuable and constructive suggestions. Authors also express their thankfulness to the Head, Department of Botany, University of Burdwan for providing laboratory and infrastructure facilities. Authors are also grateful to Dr. Anupam Basu, Associate Professor and Head of the Department, Department of Zoology, University of Burdwan for his assistance in Q-PCR analysis. Financial support in form of fellowship from Government of West Bengal, India is duly acknowledged.


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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2014

Authors and Affiliations

  • Sayanti Kundu
    • 1
  • Avishek Dey
    • 1
  • Abhijit Bandyopadhyay
    • 1
    Email author
  1. 1.Department of Botany, UGC Centre for Advanced StudyThe University of BurdwanBurdwanIndia

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