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Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of bacopa saponins in transformed calli and plants

Plant Cell Reports volume 30, pages 941–954 (2011)Cite this article

Abstract

We have developed an efficient transformation system for Bacopa monnieri, an important Indian medicinal plant, using Agrobacterium rhizogenes strains LBA 9402 and A4. Transformed roots induced by strain LBA 9402 spontaneously dedifferentiated to callus while excised roots induced by strain A4 spontaneously showed induction of shoot buds within 10 days. PCR and RT-PCR analysis revealed the presence and expression of the rolAB and rolC genes at the transcription level in pRi A4 transformed cultures indicating that the TL-DNA was integrated retained and expressed in the A4-Ri transformed shoots. Transformed calli showed the presence of rolAB or rol A, TR and ags genes. Transformed plants showed morphological features typically seen in transgenic plants produced by A. rhizogenes. Growth and biomass accumulation was significantly higher in the transformed shoots (twofold) and roots (fourfold) than in the non-transformed (WT) plants. In pRi A4-transformed plants, the content of bacopasaponin D, bacopasaponin F, bacopaside II and bacopaside V was enhanced significantly as compared to WT plants of similar age while bacoside A3 and bacopasaponin C content was comparable with that of WT plants. Significant increase in content of five bacopa saponins could be detected in pRi 9402-transformed callus cultures. There is an overall stimulatory effect on accumulation of bacopa saponins in transformed plants and cells of B. monnieri establishing the role of endogenous elicitation by Ri T-DNA of A. rhizogenes.

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Acknowledgments

Sukanya Majumdar is indebted to the University Grants Commission for award of Junior and Senior Research Fellowship, to Dr. B. Chaubey, Reader Department of Botany for help in molecular analysis and to the Programme Coordinator, CAS, Dept of Botany, CU & Coordinator, BC Guha Centre for Biotechnology, CU, for facilities provided.

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  1. Centre of Advanced Study, Department of Botany, University of Calcutta, 35 Ballygunge Circular Road, Kolkata, 700019, India

    Sukanya Majumdar & Sumita Jha

  2. Indian Institute of Chemical Biology, 4 Raja SC Mullick Road, Jadavpur, Kolkata, 700032, India

    Saraswati Garai

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  1. Sukanya Majumdar
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  2. Saraswati Garai
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  3. Sumita Jha
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Correspondence to Sumita Jha.

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A contribution to the Special Issue: Plant Biotechnology in Support of the Millennium Development Goals.

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Majumdar, S., Garai, S. & Jha, S. Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of bacopa saponins in transformed calli and plants. Plant Cell Rep 30, 941–954 (2011). https://doi.org/10.1007/s00299-011-1035-9

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Keywords

  • Bacopa monnieri
  • Agrobacterium rhizogenes
  • Medicinal plants
  • Plant regeneration
  • Agropine strains
  • Bacopa saponins
  • Jujubogenin
  • Glycosides
  • Transformed cultures