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Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the over-expression of chalcone isomerase gene with the elicitation treatment

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Abstract

Economically important compounds, such as licorice flavonoids, are present in insufficient amounts in the hairy roots. To overcome this problem, we took the transgenic approach combined with the elicitation technique to increase the flavonoid production. The Glycyrrhiza uralensis Fisch cDNA encoding chalcone isomerase gene (chi) was over-expressed in hairy roots of G. uralensis Fisch mediated by the disarmed Agrobacterium rhizogenes A4. Stable genetic transformation was confirmed by Southern blot analysis. The transgenic and wild cultures were subsequently elicited with PEG8000 (2%) alone, yeast extract (YE) (0.1%) alone, or both of them, and then the total flavonoids were extracted and measured. The results showed that over a culture period of 3 weeks, the wild-type hairy roots, the untreated transgenic hairy roots, and the double-treated transgenic hairy roots accumulated 0.842, 1.394, and 2.838 (g/100 g DW) of total flavonoids, respectively. Moreover, the enhanced accumulation of flavonoids were correlated with the elevated level of chi transcripts and CHI activity, confirming the key role of chi in the flavonoids synthesis. This research demonstrated that the combination of the metabolic engineering and PEG8000-YE elicitation treatment was an effective strategy to increase the flavonoids production in hairy roots of G. uralensis Fisch.

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Abbreviations

CHI:

Chalcone isomerase (enzyme)

chi :

Chalcone isomerase (cDNA)

CTAB:

Cetyldimethylethylammonium bromide

DW:

Dry weight

FW:

Fresh weight

PCR:

Polymerase chain reaction

PEG8000:

Polyethylene glycol (molecular weight 8000)

RT-PCR:

Reverse transcriptase-polymerase chain reaction

YE:

Yeast extract

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Acknowledgments

This research was supported by grants from National Centre for Molecular Crop Design and from the National ‘863’ Progra (No.2007AA021504) in Peking University, China. The authors are very grateful to Professor Xi-Ping Wang and assistants Ying Xie and Hai-Min Chen of National Centre for Molecular Crop Design for their advices and help.

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Authors and Affiliations

  1. Department of Breeding and Genetics, China Pharmaceutical University, 210038, Nanjing, Jiangsu Province, China

    Hai-Chao Zhang & Shan-Lin Gao

  2. National Centre for Molecular Crop Design, 100085, Beijing, China

    Jing-Mei Liu

  3. Food Science and Technology College, Guangdong Ocean University, 524025, Zhanjiang, Guangdong Province, China

    Hong-Yu Lu

Authors
  1. Hai-Chao Zhang
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  2. Jing-Mei Liu
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  3. Hong-Yu Lu
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  4. Shan-Lin Gao
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Correspondence to Jing-Mei Liu or Shan-Lin Gao.

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Communicated by H. Judelson.

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Zhang, HC., Liu, JM., Lu, HY. et al. Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the over-expression of chalcone isomerase gene with the elicitation treatment. Plant Cell Rep 28, 1205–1213 (2009). https://doi.org/10.1007/s00299-009-0721-3

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  • DOI: https://doi.org/10.1007/s00299-009-0721-3

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