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Genetic transformation of Tylophora indica with Agrobacterium rhizogenes A4: growth and tylophorine productivity in different transformed root clones

  • Genetic Transformation and Hybridization
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Abstract

We have developed an efficient transformation system for Tylophora indica, an important medicinal plant in India, using Agrobacterium rhizogenes strains LBA9402 and A4 to infect excised leaf and stem explants and intact shoots at different sites. The induction of callus and transformed roots was dependent on the bacterial strain, explant type and inoculation site used. Transformed roots were induced only in explants infected with A. rhizogenes strain A4, while an optimal transformation frequency of up to 60% was obtained with intact shoots inoculated at the nodes. The presence of the left-hand transferred DNA (TL-DNA) in the genome of T. indica roots induced by A. rhizogenes was confirmed by PCR amplification of the rooting locus genes of A. rhizogenes. Root growth and the production of tylophorine, the major alkaloid of the plant, varied substantially among the nine root clones studied. Both parameters increased over time in liquid cultures, with maximum biomass and tylophorine accumulation occurring within 4–6 weeks of growth in fresh medium. Interestingly, in liquid culture, the culture medium also accumulated tylophorine up to concentrations of 9.78±0.21 mg l−1.

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Abbreviations

BM:

Basal medium

%DW:

Percentage dry weight

HPLC:

High performance liquid chromatography

NT:

Non-transformed

Ri:

Root-inducing

YMB:

Yeast mannitol broth

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Acknowledgements

The authors are grateful to the Programme Coordinator, Centre of Advanced Study, Department of Botany for the facilities provided, Dr. F. Abe, Fukuoka University, Japan for authentic samples of tylophorine, to Dr. Albert Kollman, INRA, Versailles, France for his help in tylophorine analysis and to Mr. N. Roy for photography. Financial assistance from Indo-French Centre for the Promotion of Advanced Research (Centre Franco-Indien pour la Promotion de la Recherche Avancée) is gratefully acknowledged

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Authors and Affiliations

  1. Centre of Advanced Study in Cell and Chromosome Research, Department of Botany, University of Calcutta, 35 Ballygunge Circular Road, Calcutta, 700019, India

    Kuntal Narayan Chaudhuri & Sumita Jha

  2. Department of Botany, RKMVC College, 24 Parganas (North), Rahara, West Bengal, India

    Biswajit Ghosh

  3. Laboratoire de Biologie de la Rhizosphère (Phytopharmacie), Institut National de la Recherche Agronomique, 78026, Versailles cedex, France

    David Tepfer

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  1. Kuntal Narayan Chaudhuri
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  2. Biswajit Ghosh
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  3. David Tepfer
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  4. Sumita Jha
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Correspondence to Sumita Jha.

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Communicated by P. Lakshmanan

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Chaudhuri, K.N., Ghosh, B., Tepfer, D. et al. Genetic transformation of Tylophora indica with Agrobacterium rhizogenes A4: growth and tylophorine productivity in different transformed root clones. Plant Cell Rep 24, 25–35 (2005). https://doi.org/10.1007/s00299-004-0904-x

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  • DOI: https://doi.org/10.1007/s00299-004-0904-x

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