Arbuscular mycorrhizal fungi enhance antioxidant capacity of in vitro propagated garden thyme (Thymus vulgaris L.)

Abstract

Garden thyme (Thymus vulgaris L., Lamiaceae) is an important aromatic herb used for its medicinal values including antioxidant and antimicrobial properties. The present study was performed to analyze the changes in natural antioxidants after inoculation of in vitro propagated garden thyme plants with arbuscular mycorrhizal fungi (AMF). An efficient and low-cost protocol for large-scale multiplication of this aromatic plant was developed. The explants were cultured on full and half strength Murashige and Skoog (MS) medium containing indole-3-butyric acid (IBA). The maximum number of shoots and roots was obtained on ½ MS medium supplemented with 0.1 mg L−1 IBA after 4 weeks of culture. The successfully adapted in vitro plants (survival rate 95%) were inoculated with arbuscular mycorrhizal fungi (Claroideoglomus claroideum, ref. EEZ 54). Plants were then transferred into field conditions. Mycorrhizal fungi enhanced the activity of some soil enzymes, acid and alkaline phosphatase, urease as well as the levels of extractable glomalin-related proteins in plant rhizosphere. Arbuscular mycorrhizal associations with higher plants promote the accumulation of antioxidant metabolites such as phenols and flavonoids and increase the activity of antioxidant enzymes. The results from the present study suggest enhanced antioxidant capacity of the inoculated T. vulgaris plants which was due mainly to increased accumulation of phenolic compounds (total phenols and flavonoids) together with stimulation of the activity of superoxide dismutase (SOD) and guaiacol peroxidase (GPO).

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Abbreviations

AMF:

Arbuscular mycorrhizal fungi

APX:

Ascorbate peroxidase

CAT:

Catalase

DPPH :

2,2-diphenylpycril-hydrazyl

FRAP:

Ferric ion reducing antioxidant power

GPO:

Guaiacol peroxidase

MS:

Murashige and Skoog

NM:

Non-mycorrhizal plants

SOD:

Superoxide dismutase

BA:

6-benzyladenine

NAA:

ɑ-naphtalene acetic acid

IBA:

Indole-3-butyric acid

IAA:

Indole-3-acetic acid

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Acknowledgements

This study was conducted with financial support from Slovak Research and Development Agency and Bulgarian Science Fund at the Ministry of Education and Science, Bilateral project SK_BG 01/3-30.09.2016.

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Correspondence to Ira Stancheva.

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Zayova, E., Stancheva, I., Geneva, M. et al. Arbuscular mycorrhizal fungi enhance antioxidant capacity of in vitro propagated garden thyme (Thymus vulgaris L.). Symbiosis 74, 177–187 (2018). https://doi.org/10.1007/s13199-017-0502-7

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Keywords

  • Antioxidants
  • Arbuscular mycorrhizal fungi
  • Garden thyme
  • In vitro culture
  • Micro propagation
  • Soil enzymes