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Stimulatory effect of copper and zinc sulphate on plant regeneration, glutathione-S-transferase analysis and assessment of antioxidant activities in Mucuna pruriens L. (DC)

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Abstract

The effect of heavy metals, CuSO4 and ZnSO4 on morphogenic response of cotyledonary node explants of Mucuna pruriens (L.), a fast growing tropical medicinal legume was evaluated. The climber holds economic importance in traditional as well as modern pharmaceutical areas. Murashige and Skoog (Physiol Plant 15:473–497, 1962) medium supplemented with various strength (0.5, 2.5, 4.5, 6.5 or 8.5 µM) of BA (6-benzyladenine), Kn (Kinetin) and 2-iP (2-isopentenyl adenine) were screened to establish optimum culture conditions for in vitro regeneration using cotyledonary node explant. Optimized medium (MS + BA 2.5 µM) was augmented with different concentration of CuSO4 (0.5–25.0 µM) and ZnSO4 (5–150 µM) to evaluate the effect of heavy metals on regeneration potential of the explants. Among various concentrations tested, MS medium with optimized BA (2.5 µM) along with 5.0 µM CuSO4 or 80.0 µM ZnSO4 proved beneficial, inducing 11.20 and 13.40 shoots per explant, respectively. The results demonstrated that glutathione-S-transferase (GSTs) was involved in fighting heavy metal stress, and might also have a role in in vitro regeneration. Elongated healthy microshoots on transfer to half strength MS medium produced 3.40 roots per shoot after 28 days of incubation without the necessity of exogenous auxin. The in vitro grown regenerants were hardened in Soilrite, and then established in garden and sandy soil (1:1) under natural light where they showed normal growth and development.

Key message

The work highlights the enhanced multiplication and better growth of the regenerants using elevated concentration of CuSO4 and ZnSO4. GSTs overexpression activity not only confers to overcome the stress but enhanced the in vitro regeneration of M. pruriens also.

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Abbreviations

BA:

6-Benzyladenine

CDNB:

1-Chloro-2, 4-dinitrobenzene

CuSO4 :

Copper sulphate

EDTA:

Ethylenediamine tetra acitic acid

PVP:

Polyvinylpyrrolidone

ZnSO4 :

Zinc sulphate

2-iP:

2-Isopentenyl adenine

d:

Days

SOD:

Superoxide dismutase

GSH:

Glutathione

CA:

Carbonic anhydrase

MS:

Murashige and Skoog

DDW:

Double distilled water

ROS:

Reactive oxygen species

GST:

Glutathione-S-transferase

CAT:

Catalase

APX:

Ascorbate per oxidase

NaOH:

Sodium hydroxide

GR:

Glutathione reductase

HCL:

Hydrochloric acid

HgCl2 :

Mercuric chloride

NBT:

Nitroblue tetrazolium

DMRT:

Duncan’s multiple range tests

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Acknowledgement

The authors extend their appreciation to the International Scientific Partnership program (ISPP) at King Saud University for funding this research work through ISPP#0082. M. Anis gratefully acknowledges the University Grants Commission, Govt. of India, New Delhi for the award of UGC-BSR-Faculty Fellowship (2017).

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Authors and Affiliations

  1. Department of Botany, Aligarh Muslim University, Aligarh, 202 002, India

    Naushad Alam, Mohammad Anis & Saad Bin Javed

  2. Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh, 11451, Saudi Arabia

    Mohammad Anis & Abdulrahman A. Alatar

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  1. Naushad Alam
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  2. Mohammad Anis
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Alam, N., Anis, M., Javed, S.B. et al. Stimulatory effect of copper and zinc sulphate on plant regeneration, glutathione-S-transferase analysis and assessment of antioxidant activities in Mucuna pruriens L. (DC). Plant Cell Tiss Organ Cult 141, 155–166 (2020). https://doi.org/10.1007/s11240-020-01776-8

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