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Establishment of Persicaria minor hairy roots and analysis of secreted β-caryophyllene in medium broth

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Abstract

A method for hairy root induction of Persicaria minor, was developed and investigated, and a qualitative evaluation of the sesquiterpenes in the culture medium was performed. The transgenic status was confirmed by PCR analysis using rolB and virD specific primers. The efficiency with which four different strains of Agrobacterium rhizogenes (A4, ATCC43056, ATCC15834 and ATCC13333) and different concentrations of acetosyringone induced hairy roots from in vitro leaf segments of P. minor was evaluated. Strains A4 and ATCC13333 were able to induce hairy root in an average of 6 days of co-cultivation and exhibited significantly higher transformation efficiencies, 20 and 21.3 %, respectively, compared to the other strains in the presence of 100 µM acetosyringone. Half-strength Murashige–Skoog basal medium supplemented with Gamborg vitamins supported the greatest increase (35-fold) of growth index of hairy roots relative to the initial weight after 8 weeks of culture. GC–MS results showed that β-caryophyllene was the main sesquiterpenes detected and was quantified with standards, with total peak areas of 7.934. Results suggested that hairy roots secreted sesquiterpenes into the medium while preserving viable hairy roots as a source of specialized metabolites.

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Abbreviations

AS:

Acetosyringone

GI:

Growth index

HR (s):

Hairy root (s)

MS:

Murashige–Skoog

TF:

Transformation frequency

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Acknowledgments

The authors gratefully thank the Ministry of Science, Technology and Innovation (MOSTI) of Malaysia through the Science Fund research grant (02-01-02-SF0909) and UKM Research University grant (DLP-2013-008) for funding this research.

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Authors and Affiliations

  1. Plant Biotechnology Centre, Institute of Systems Biology (INBIOSIS), Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor, Malaysia

    Mehdi Farshad Ashraf, Zamri Zainal, Normah Mohd Noor & Ismanizan Ismail

  2. Faculty of Science and Technology, School of Biosciences and Biotechnology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor, Malaysia

    Mehdi Farshad Ashraf, Che Radziah Che Mohd Zain, Zamri Zainal & Ismanizan Ismail

  3. Department of Chemical and Process Engineering, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor, Malaysia

    Nurina Anuar & Masturah Markom

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  1. Mehdi Farshad Ashraf
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  2. Che Radziah Che Mohd Zain
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  3. Zamri Zainal
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Correspondence to Mehdi Farshad Ashraf or Ismanizan Ismail.

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Ashraf, M.F., Che Mohd Zain, C.R., Zainal, Z. et al. Establishment of Persicaria minor hairy roots and analysis of secreted β-caryophyllene in medium broth. Plant Cell Tiss Organ Cult 121, 11–20 (2015). https://doi.org/10.1007/s11240-014-0674-2

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