Plant Cell Reports

, Volume 21, Issue 11, pp 1103–1107

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

Genetic Transformation and Hybridization

DOI: 10.1007/s00299-003-0633-6

Cite this article as:
Shi, H. & Kintzios, S. Plant Cell Rep (2003) 21: 1103. doi:10.1007/s00299-003-0633-6

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.

Keywords

Agrobacterium rhizogenesHairy rootsPueraria phaseoloides Puerarin

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

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.College of Life ScienceSouth China Normal UniversityGuangzhouChina
  2. 2.Laboratory of Plant Physiology, Department of Agricultural BiotechnologyAgricultural University of AthensAthensGreece
  3. 3.BERA Biosensors CorporationNew YorkUSA