Abstract

The present investigation aimed to obtain salt-tolerant Artemisia vulgaris L. to develop a constant form through in vitro mutagenesis with ethyl methane sulphonate (EMS) as the chemical mutagen. NaCl tolerance was evaluated by the ability of the callus to maintain its growth under different concentrations, ranges from (0 mM to 500 mM). However, NaCl salinity concentration at (500 mM) did not show any development of callus, slight shrinking, and brown discoloration taking place over a week. Thus, all the biochemical and antioxidant assays were limited to (0–400 mM) NaCl. On the other hand, selected calluses were treated with 0.5% EMS for 30, 60, and 90 min and further subcultured on basal media fortified with different concentrations of 0–400 mM NaCl separately. Thus, the callus was treated for 60 min and was found to induce the mutation on the callus. The maximum salt-tolerant callus from 400 mM NaCl was regenerated in MS medium fortified with suitable hormones. Biochemical parameters such as chlorophyll, carotenoids, starch, amino acids, and phenol contents decreased under NaCl stress, whereas sugar and proline increased. Peroxidase (POD) and superoxide dismutase (SOD) activities peaked at 200 mM NaCl, whereas catalase (CAT) was maximum at 100 mM NaCl. Enhanced tolerance of 0.5% the EMS-treated callus, attributed to the increased biochemical and antioxidant activity over the control and NaCl stress. As a result, the mutants were more tolerant of salinity than the control plants.

Highlights

  • A. vulgaris L. is a medicinally useful traditional plant and is widely being used for the healing of diabetes

  • After 3 weeks of incubation, explants cultured on Murashige and Skoog (MS) medium regenerated the callus BioTech 2021, 10, x FOR PEER REVIEWand regeneration of the whole plant of A. vulgaris from contro6 olf c14allus (0 mM NaCl stress) (Figure 2a,b)

  • We found a further increase in total amino acid when NaCl stressed the callus of A. vulgaris treated with ethyl methane sulphonate (EMS)

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Summary

Introduction

A. vulgaris L. (mugwort) is a medicinally useful traditional plant and is widely being used for the healing of diabetes. The in vitro regeneration of the high artemisinin-producing somaclonal variation against (NaCl) salt-tolerance and the improvement of sequence-characterized amplified region (SCAR) markers in ‘Artemisia annua L.’ were reported by [3]. Salt stress is a major environmental limitation that has a global impact on plant yield and distribution. Salinity (NaCl) inhibits plant growth by altering homeostasis in water status and ionic distribution and causing oxidative stress. Osmotic pressure, mineral shortages, and a mix of physiological and biochemical abnormalities are all common symptoms of salt stress [6]. The plant cell adaptation and variation towards high level salinity involve osmotic change and the compartmentalization of toxic ions, whereas an increasing body of evidence suggests that high salinity stimulates the formation of oxidative stress and reactive oxygen species (ROS) [7]

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