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Changes in morphological phenotypes and withanolide composition of Ri-transformed roots of Withania somnifera

  • Genetic Transformation and Hybridization
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Abstract

Developmental variability was introduced into Withania somnifera using genetic transformation by Agrobacterium rhizogenes, with the aim of changing withasteroid production. Inoculation of W. somnifera with A. rhizogenes strains LBA 9402 and A4 produced typical transformed root lines, transformed callus lines, and rooty callus lines with simultaneous root dedifferentiation and redifferentiation. These morphologically distinct transformed lines varied in T-DNA content, growth rates, and withasteroid accumulation. All of the lines with the typical transformed root morphology contained the TL T-DNA, and 90% of them carried the TR T-DNA, irrespective of the strain used for infection. Accumulation of withaferin A was maximum (0.44% dry weight) in the transformed root line WSKHRL-1. This is the first detection of withaferin A in the roots of W. somnifera. All of the rooty callus lines induced by strain A4 contained both the TL and the TR-DNAs. In contrast, 50% of the rooty-callus lines obtained with strain LBA 9402 contained only the TR T-DNA. All the rooty callus lines accumulated both withaferin A and withanolide D. The callusing lines induced by LBA 9402 lacked the TL T-DNA genes, while all the callusing lines induced by strain A4 contained the TL DNA. Four of these callus lines produced both withaferin A (0.15–0.21% dry weight) and withanolide D (0.08–0.11% dry weight), and they grew faster than the transformed root lines. This is the first report of the presence of withasteroids in undifferentiated callus cultures of W. somnifera.

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Acknowledgements

The authors are grateful to the Programme Coordinator, Centre of Advanced Study, Department of Botany for the facilities provided 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 Avance) is gratefully acknowledged.

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

    Maumita Bandyopadhyay & Sumita Jha

  2. Laboratoire de Biologie de la Rhizosphére (Phytopharmacie), Institut National de la Recherche Agronomique, 78026, Versailles cédex, France

    David Tepfer

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  1. Maumita Bandyopadhyay
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  2. Sumita Jha
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  3. David Tepfer
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Correspondence to Sumita Jha.

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Communicated by P. Lakshmanan

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Bandyopadhyay, M., Jha, S. & Tepfer, D. Changes in morphological phenotypes and withanolide composition of Ri-transformed roots of Withania somnifera . Plant Cell Rep 26, 599–609 (2007). https://doi.org/10.1007/s00299-006-0260-0

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