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Genetic transformation of Pueraria phaseoloides with Agrobacterium rhizogenes and puerarin production in hairy roots

  • Genetic Transformation and Hybridization
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Abstract

An efficient transformation system for the medicinal plant Pueraria phaseoloides was established by using agropine-type Agrobacterium rhizogenes ATCC15834. Hairy roots could be obtained directly from the cut edges of petioles of leaf explants or via callus 10 days after inoculation with the bacteria. The highest frequency of explant transformation by A. rhizogenes ATCC15834 was about 70% after infection for 30 days. Hairy roots could grow rapidly on solid, growth regulator-free Murashige and Skoog medium and had characteristics of transformed roots such as fast growth and high lateral branching. Paper electrophoresis revealed that bacteria-free hairy roots of P. phaseoloides could synthesize agropine and mannopine. The polymerase chain reaction amplification of rooting locus genes showed that left-hand transferred DNA of the root inducing plasmid of A. rhizogenes was inserted into the genome of transformed P. phaseoloides hairy roots. The content of puerarin in hairy roots reached a level of 1.190 mg/g dry weight and was 1.067 times the content in the roots of untransformed plants.

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Abbreviations

HPLC :

High performance liquid chromatography

MS :

Murashige and Skoog medium

PCR :

Polymerase chain reaction

Ri :

Root inducing

rolB:

Rooting locus B

rolC:

Rooting locus C

YEB :

Yeast extract broth

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Acknowledgements

The authors express their gratitude to Prof. Jacques Tempé, INRS, France, for kindly providing agropine and mannopine standards. This work was supported by the Ministry of Science and Technology of China and the Ministry of Development of Greece.

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  1. S. Kintzios

    Present address: BERA Biosensors Corporation, New York, NY, USA

Authors and Affiliations

  1. College of Life Science, South China Normal University, 510631, Guangzhou, China

    H.-P. Shi

  2. Laboratory of Plant Physiology, Department of Agricultural Biotechnology, Agricultural University of Athens, Athens, Greece

    S. Kintzios

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  1. H.-P. Shi
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  2. S. Kintzios
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Correspondence to H.-P. Shi.

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Communicated by D. Dudits

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Shi, HP., Kintzios, S. Genetic transformation of Pueraria phaseoloides with Agrobacterium rhizogenes and puerarin production in hairy roots. Plant Cell Rep 21, 1103–1107 (2003). https://doi.org/10.1007/s00299-003-0633-6

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  • DOI: https://doi.org/10.1007/s00299-003-0633-6

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