, Volume 239, Issue 1–4, pp 31–38 | Cite as

In vitro biochemical evaluation of cadmium tolerance mechanism in callus and seedlings of Brassica juncea

  • Gyan Singh ShekhawatEmail author
  • Kusum Verma
  • Sonali Jana
  • Kusum Singh
  • Preeti Teotia
  • Archana Prasad
Original Article


In vitro grown callus and seedlings of Brassica juncea were treated with equimolar concentrations of cadmium and compared for their respective tolerance to cadmium. Calli cultures were grown on Murashige and Skoog medium supplemented with α 6-benzyl aminopurine (200 µg L−1, naphthalene acetic acid 200 µg L−1) and 2,4-dichloro-phenoxy acetic acid (65 µg L−1) while the seedlings grown on Hoagland's nutrient solution have been carried out. Cellular homeostasis and detoxification to cadmium in B. juncea were studied by analyzing the growth in terms of fresh weight and dry weight, lipid peroxidation, proline accumulation, and antioxidative enzymes (superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT)). At 200 µM cadmium, callus and seedlings showed 73.61% and 74.76% reduction in tolerance, respectively. A significant increase in malondialdehyde (MDA) content was found in both calli and seedlings; however, the amount of MDA content was more in seedlings. Proline content increased on lower concentration of cadmium (up to 50 µM), and it further decreased (up to 200 µM). But the accumulation of proline was higher in callus cultures. The overall activity of antioxidative enzymes (SOD, CAT, and APX) was found to be higher in callus in comparison to seedlings of B. juncea. Callus and seedlings showed a significant (P ≤ 0.5) increase in SOD activity in a concentration-dependent manner up to 50 µM cadmium concentration but decreased further. APX activity increased significantly at low cadmium levels but CAT activity decreased significantly throughout on increasing cadmium concentrations from 5 to 200 µM, respectively. Hence, it was observed that callus of B. juncea was more tolerant in comparison to seedlings exposed to equimolar concentrations of cadmium. Thus, from the present studies, it is concluded that calli were more tolerant toward cadmium-induced oxidative stress. Hence, it is suitable material for the study of cadmium tolerance mechanisms and for the manipulations within them for better understanding of cadmium detoxification strategies in B. juncea.


Antioxidant Brassica juncea Callus Cadmium Metal Tolerance 



Ascorbate peroxidase


α 6-Benzyl aminopurine


Cadmium chloride




2,4-Dichloro-phenoxy acetic acid


Dry weight


Fresh weight


Hydrogen peroxide




Murashige and Skoog


Naphthalene acetic acid


Reactive oxygen species


Superoxide dismutase



G.S. Shekhawat is thankful to Prof. Aditya Shastri, Vice Chancellor, Banasthali University, for providing all the necessary requirements to successfully carry out this research work.

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Gyan Singh Shekhawat
    • 1
    Email author
  • Kusum Verma
    • 1
  • Sonali Jana
    • 1
  • Kusum Singh
    • 1
  • Preeti Teotia
    • 1
  • Archana Prasad
    • 1
  1. 1.Department of Bioscience and BiotechnologyBanasthali UniversityBanasthaliIndia

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