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Biochemical, molecular, and elemental profiling of Withania somnifera L. with response to zinc stress

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Abstract

Zn stress seriously induces various toxic responses in Withania somnifera L., when accumulated above the threshold level which was confirmed by investigating the responses of protein, expression of antioxidant enzymes, and elemental profiling on accumulation of Zn. Zn was supplemented in the form of ZnSO4 (0, 25, 50, 100, and 200 μM) through MS liquid medium and allowed to grow the in vitro germinated plants for 7 and 14 days. The study revealed that when the application of Zn increased, a significant reduction of growth characteristics was noticed with alterations of proteins (both disappearance and de novo synthesis). The activity of CAT, SOD, and GPX were increased up to certain concentrations and then declined, which confirmed through in-gel activity under different treatments. RT-PCR was conducted by taking three sets of genes from CAT (RsCat, Catalase1, Cat1) and SOD (SodCp, TaSOD1.2, MnSOD) and found that gene RsCat from CAT and MnSOD from SOD have shown maximum expression of desired genes under Zn stress, which indicate plant’s stress tolerance mechanisms. The proton-induced X-ray emission study confirmed an increasing order of uptake of Zn in plants by suppressing and expressing other elemental constituents which cause metal homeostasis. This study provides insights into molecular mechanisms associated with Zn causing toxicity to plants; however, cellular and subcellular studies are essential to explore molecule-molecule interaction during Zn stress in plants.

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Abbreviations

Zn:

Zinc

MS:

Murashige and Skoog

RT-PCR:

Reverse transcriptase polymerase chain reaction

CAT:

Catalase

SOD:

Superoxide dismutase

PIXE:

Proton-induced X-ray emission

ROS:

Reactive oxygen species

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

EDTA:

Ethylenediamine-tetraacetic acid

GPX:

Guaiacol peroxidase

POD:

Peroxidase

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Acknowledgements

The authors are grateful to the Director, Institute of Physics, Bhubaneswar, for providing PIXE facility to study the metal analysis.

Funding

This study received financial support from UGC-DAE Consortium for Scientific Research, Kolkata, India (Grant No. UGC-DAE-CSR-KC/CRS/2009/TE-01/1539).

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Authors and Affiliations

  1. School of Biological Sciences, AIPH University, Bhubaneswar, Odisha, 752101, India

    Jyoti Ranjan Rout

  2. Post Graduate Department of Biotechnology, Utkal University, VaniVihar, Bhubaneswar, Odisha, 751004, India

    Rout George Kerry

  3. Department of Life Science, National Institute of Technology Rourkela, Rourkela, Odisha, 769008, India

    Debasna Panigrahi

  4. Biochemistry and Molecular Biology Laboratory, Post Graduate Department of Botany, Utkal University, VaniVihar, Bhubaneswar, Odisha, 751004, India

    Jyoti Ranjan Rout, Santi Lata Sahoo & Chinmay Pradhan

  5. UGC-DAE Consortium of Scientific Research, Kolkata Centre, Bidhan Nagar, Salt Lake, Kolkata, 700098, India

    Shidharth Sankar Ram, Anindita Chakraborty & Mathummal Sudarshan

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  1. Jyoti Ranjan Rout
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  2. Rout George Kerry
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  3. Debasna Panigrahi
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Correspondence to Jyoti Ranjan Rout.

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Rout, J.R., Kerry, R.G., Panigrahi, D. et al. Biochemical, molecular, and elemental profiling of Withania somnifera L. with response to zinc stress. Environ Sci Pollut Res 26, 4116–4129 (2019). https://doi.org/10.1007/s11356-018-3926-6

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