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.

Original languageEnglish
JournalPlant Cell, Tissue and Organ Culture
DOIs
Publication statusAccepted/In press - 6 Dec 2014

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sesquiterpenoids
acetosyringone
Rhizobium rhizogenes
Persicaria minor
vitamins
culture media
genetically modified organisms
metabolites
leaves
methodology

Keywords

  • Hairy root culture
  • Persicaria minor
  • Sesquiterpene
  • β-Caryophyllene

ASJC Scopus subject areas

  • Horticulture

Cite this

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title = "Establishment of Persicaria minor hairy roots and analysis of secreted β-caryophyllene in medium broth",
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.",
keywords = "Hairy root culture, Persicaria minor, Sesquiterpene, β-Caryophyllene",
author = "Ashraf, {Mehdi Farshad} and {Che Mohd Zain}, {Che Radziah} and Zamri Zainal and {Mohd Noor}, Normah and Nurina Anuar and Masturah Markom and Ismanizan Ismail",
year = "2014",
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AU - Ashraf, Mehdi Farshad

AU - Che Mohd Zain, Che Radziah

AU - Zainal, Zamri

AU - Mohd Noor, Normah

AU - Anuar, Nurina

AU - Markom, Masturah

AU - Ismail, Ismanizan

PY - 2014/12/6

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N2 - 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.

AB - 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.

KW - Hairy root culture

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