Journal of Plant Biochemistry and Biotechnology

, Volume 23, Issue 4, pp 366–378 | Cite as

Effect of potassium chloride and calcium chloride induced stress on in vitro cultures of Bacopa monnieri (L.) Pennell and accumulation of medicinally important bacoside A

  • M. L. Ahire
  • S. Laxmi
  • P. R. Walunj
  • P. B. Kavi Kishor
  • T. D. NikamEmail author
Original Article


Growth, osmotic adjustment, antioxidant enzyme defense and principle medicinal component bacoside A was studied in in vitro raised shoots of Bacopa monnieri under different concentrations of KCl and CaCl2 (0, 50, 100, 150 or 200 mM). Significant reduction was observed in shoot number per culture; shoot length, fresh weight, dry weight and tissue water content (TWC) when shoots were exposed to increasing KCl and CaCl2 concentrations (50–200 mM) as compared to control. Minimum damage to the membrane as assessed by malondialdehyde (MDA) content was noticed in control in contrast to sharp increase in KCl and CaCl2 stressed shoots. Higher amounts of free proline, glycine betaine and total soluble sugars (TSS) accumulated in KCl and CaCl2 exposed shoots compared to the controls. Among different concentrations of KCl and CaCl2, increasing concentration of CaCl2 showed more increase in osmolyte accumulation. Na+ content decreased with increasing concentrations of KCl and CaCl2. Accumulation of K+ increased significantly in KCl (50–100 mM) stressed shoots as compared to control, while it decreased in CaCl2 treated shoots indicating that it prevents the uptake of K+ ions. Ca2+ accumulation significantly increased with increasing concentrations of CaCl2 up to 150 mM but decreased at higher concentrations. Shoots treated with KCl and CaCl2 (0–100 mM) showed higher antioxidant enzyme (SOD, CAT, APX and GPX) activities but KCl suppressed the activities at higher concentrations. Accumulation of bacoside A was enhanced with an increase in KCl and CaCl2 concentration up to 100 mM. It appears from the data that accumulation of osmolytes, and elevated activities of antioxidant enzymes play an important role in osmotic adjustment in shoot cultures of Bacopa and the two salts tested have a positive effect on bacoside accumulation.


Antioxidant enzymes Bacopa monnieri Bacoside A In vitro growth responses Osmolyte accumulation Salinity Shoot cultures 



Ascorbate peroxidase






Guaiacol peroxidase




Murashige and Skoog


Nitroblue tetrazolium chloride


Superoxide dismutase


Total soluble sugars


Tissue water content



The authors wish to acknowledge the financial support from the Department of Botany, University of Pune, under UGC, SAP-DRS III program sanctioned by UGC, New Delhi.


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Copyright information

© Society for Plant Biochemistry and Biotechnology 2013

Authors and Affiliations

  • M. L. Ahire
    • 1
    • 2
  • S. Laxmi
    • 1
  • P. R. Walunj
    • 1
  • P. B. Kavi Kishor
    • 3
  • T. D. Nikam
    • 1
    Email author
  1. 1.Department of BotanyUniversity of PunePuneIndia
  2. 2.Department of BotanyYashavantrao Chavan Institute of ScienceSataraIndia
  3. 3.Department of GeneticsOsmania UniversityHyderabadIndia

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