Phenolic compounds, antioxidant capacity and antimutagenic activity in different growth stages of in vitro raised plants of Origanum vulgare L.
Abstract
Efficient micropropagation procedure was developed for Origanum vulgare, a high-value culinary herb, and the phytochemicals, phenolic content, antioxidant and antimutagenic activity of leaf and stem, derived from different growing stages were analyzed. The agar solidified Murashige and Skoog (MS) medium supplemented with a combination of 6-benzylaminopurine and α-naphthaleneacetic acid was optimized as best shoot-multiplication-medium. Shoots were rooted best on 1/2 strength MS medium supplemented with 50 µM indole-3-butyric acid (IBA). The plantlets were successfully acclimatized ex vitro in a soil, sand and farmyard manure mixture (2:1:1 v/v/v) with 100% survival rate in greenhouse. The total anthocyanin and total phenolic content were observed significantly higher in leaves of in vitro-raised plants. However, total tannin, flavonoid and antioxidant activity remained higher in leaves of mother plant maintained under ployhouse condition. All the plant extracts have shown significant antimutagenic activity except in vitro-growing plants. A total of 13 polyphenolic compounds were detected in different extracts using high performance liquid chromatography. Among these, catechin was detected maximum in in vitro-growing cultures and chlorogenic acid in leaves of mother plant. These findings will help the farmers, medicinal plant growers, and industries for mass multiplication and effective extraction of phytochemicals from O. vulgare.
Keywords
Anthocyanin Shoot elongation Micropropagation Polyphenolics HPLCAbbreviations
- AAE
Ascorbic acid equivalent
- ABTS
2,2-Azinobis (3-ethylbenzothiazoline-6-sulphonic acid)
- asl
Above mean sea level
- BAP
6-Benzylaminopurine
- CN
Cyanidin 3-glucoside
- DPPH
2,2-Diphenyl-1-picryhydrazyl
- DAD
Diode-array detection
- GA3
Gibberellic acid
- GAE
Gallic acid equivalent
- HPLC
High performance liquid chromatography
- IBA
Indole-3-butyric acid
- IL
In vitro-raised plant leaf
- IS
In vitro-raised plant stem
- IVG
In vitro-growing cultures
- MPL
Mother plant leaf
- MPS
Mother plant stem
- MS
Murashige and Skoog
- NAA
α-Naphthaleneacetic acid
- PBS
Phosphate-buffered saline
- PGRs
Plant growth regulators
- TAE
Tannic acid equivalent
- TBE
Tris borate ethylenediaminetetraacetic acid
- QE
Quercetin equivalent
- µM
Micro mole
Notes
Acknowledgements
Authors thank Director G. B. Pant National Institute of Himalayan Environment and Sustainable Development, for his encouragement and facilities. Authors also thank Head, Department of Biotechnology, Bhimtal Campus, Kumaun University Nainital for facilities and encouragement during the initial stage of experimentation. Colleagues of Biodiversity Conservation and Management theme are thanked for cooperation and help during the study. Anonymous reviewers are gratefully acknowledged for providing useful inputs to improve the manuscript draft.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
Supplementary material
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