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Alkanna orientalis (L.) Boiss. plant isolated cultures and antimicrobial activity of their extracts: phenomenon, dependence on different factors and effects on some membrane-associated properties of bacteria

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Abstract

Herbal medicine requires searching for new sources with antimicrobial activity. Alkanna sp. (Boraginaceae) is widely used in medicine due to detoxification and antibacterial effects. The aim of this study was to obtain Alkanna orientalis (L.) Boiss. plant callus extracts, to investigate antimicrobial activity of extracts against bacteria and yeast and to reveal responsible mechanisms. Callus tissue cultures have been obtained using different nutrient media. Antimicrobial activity and minimal inhibitory concentrations (MIC) were determined by the methods of extract diffusion in agar and dilution using different test-microorganisms. Quantity and quality of naphthoquinones were determined using spectrophotometric and high performance liquid chromatographic analyses. H+/K+ exchange by whole cells was assayed using selective electrodes, ATPase activity and SH-groups number of membrane vesicles—by spectrophotometric methods. It was revealed that callus extracts containing naphthoquinones (0.19 ± 0.01 %) possessed bacteriostatic activity against gram-positive bacteria including pathogenic ones (MIC were 125–750 μg/mL−1), and bactericidal activity against lactic acid bacteria (Lactobacillus acidophilus, L. rhamnosus) (MIC was 250 μg/mL). MIC against Enterococcus hirae was 250 μg/mL of callus extract or 31.25 μg/mL of shikonin. It depended on the medium content, cultivation duration and the activity remained for 10–11 years. Moreover, intact root extracts inhibited H+/K+ exchange of E. hirae but callus extracts had a stronger effect. The inhibitor, N,N′-dicyclohexylcarbodiimide (DCCD)-sensitive H+/K+ exchange was changed; ATPase activity and SH-groups number were lowered by two and more fold under the influence of the extracts. Alkanet root and callus tissues extracts were concluded to have a high antimicrobial activity. So, they might directly affect the FOF1-ATPase which in turn regulates the bacterial growth. These results are useful for further investigation of alkanet extracts composition and their application as an alternative antimicrobial agent in pharmaceutical and food industry as well as in medicine.

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Abbreviations

MS:

Murashige and Skoog

BAP:

6-Benzylaminopurine

DCCD:

N,N′-dicyclohexylcarbodiimide

FOF1 :

Proton-translocating ATPase

IAA:

Indole-3-acetic acid

∆µ +H :

Proton-motive force

MEP:

Multidrug efflux pump

MIC:

Minimal inhibitory concentration

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Acknowledgments

This study was done in the frame of Basic research support by State Committee on Science, Ministry of Education and Science of Armenia (#10-3/9).

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Authors and Affiliations

  1. Department of Microbiology and Plants and Microbes Biotechnology, Biology Faculty, Yerevan State University, 1 A. Manoogian Str., 0025, Yerevan, Armenia

    Margarit Petrosyan, Naira Sahakyan, Anna Poladyan, Yuri Popov & Armen Trchounian

  2. Institute of Botany of National Academy of Sciences, 1 Acharyan Str., 0063, Yerevan, Armenia

    Yekaterina Shcherbakova

  3. Department of Biophysics, Biology Faculty, Yerevan State University, 1 A. Manoogian Str., 0025, Yerevan, Armenia

    Zaruhi Vardanyan

Authors
  1. Margarit Petrosyan
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  2. Yekaterina Shcherbakova
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  3. Naira Sahakyan
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  4. Zaruhi Vardanyan
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  5. Anna Poladyan
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  6. Yuri Popov
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  7. Armen Trchounian
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Correspondence to Armen Trchounian.

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Petrosyan, M., Shcherbakova, Y., Sahakyan, N. et al. Alkanna orientalis (L.) Boiss. plant isolated cultures and antimicrobial activity of their extracts: phenomenon, dependence on different factors and effects on some membrane-associated properties of bacteria. Plant Cell Tiss Organ Cult 122, 727–738 (2015). https://doi.org/10.1007/s11240-015-0806-3

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  • DOI: https://doi.org/10.1007/s11240-015-0806-3

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