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

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Abstract

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.

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Abbreviations

APX:

Ascorbate peroxidase

BAP:

α 6-Benzyl aminopurine

CdCl2 :

Cadmium chloride

CAT:

Catalase

2,4-D:

2,4-Dichloro-phenoxy acetic acid

DW:

Dry weight

FW:

Fresh weight

H2O2 :

Hydrogen peroxide

MDA:

Malondialdehyde

MS:

Murashige and Skoog

NAA:

Naphthalene acetic acid

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

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Acknowledgments

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.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Banasthali University, Banasthali, 304022, Rajasthan, India

    Gyan Singh Shekhawat, Kusum Verma, Sonali Jana, Kusum Singh, Preeti Teotia & Archana Prasad

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  1. Gyan Singh Shekhawat
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  2. Kusum Verma
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  3. Sonali Jana
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  4. Kusum Singh
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Correspondence to Gyan Singh Shekhawat.

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Shekhawat, G.S., Verma, K., Jana, S. et al. In vitro biochemical evaluation of cadmium tolerance mechanism in callus and seedlings of Brassica juncea . Protoplasma 239, 31–38 (2010). https://doi.org/10.1007/s00709-009-0079-y

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