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Agrobacterium-mediated genetic transformation of the desiccation tolerant resurrection plant Ramonda myconi (L.) Rchb.

  • Genetic Transformation and Hybridization
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Abstract

In this paper we describe the first procedure for Agrobacterium tumefaciens-mediated genetic transformation of the desiccation tolerant plant Ramonda myconi (L.) Rchb. Previously, we reported the establishment of a reliable and effective tissue culture system based on the integrated optimisation of antioxidant and growth regulator composition and the stabilisation of the pH of the culture media by means of a potassium phosphate buffer. This efficient plant regeneration via callus phase provided a basis for the optimisation of the genetic transformation in R. myconi. For gene delivery, both a standard (method A) and a modified protocol (method B) have been applied. Since the latter has previously resulted in successful transformation of another resurrection plant, Craterostigma plantagineum, an identical protocol was utilized in transformation of R. myconi, as this method may prove general for dicotyledonous resurrection plants. On this basis, physical and biochemical key variables in transformation were evaluated such as mechanical microwounding of plant explants and in vitro preinduction of vir genes. While the physical enhancement of bacterial penetration was proved to be essential for successful genetic transformation of R. myconi, an additional two-fold increase in the transformation frequency was obtained when the above physical and biochemical treatments were applied in combination. All R 0 and R 1 transgenic plants were fertile, and no morphological abnormalities were observed on the whole-plant level.

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Abbreviations

GM medium:

Germination medium (Toldi et al. 1994)

RA medium:

Ramonda medium (Tóth et al. 2004)

MSI medium:

Modified MS medium for bacterial infection (Borsics et al. 2002)

CPY medium:

Culture medium for Agrobacterium

BAP:

6-Benzyladenine

NAA:

1-Naphtaleneacetic acid

sgm explants:

Leaf segment explants

int explants:

Intact, but microwounded explants

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Acknowledgements

This project was supported by an OECD Co-operative Research Programme entitled as “Biological Resource Management for Sustainable Agriculture Systems.” The authors would like to thank Gábor Takács for the excellent photographic work and Dr. Rainer Höfgen (MPI of Molecular Plant Physiology, Golm, Germany) for kindly providing p35SGUSINT.

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Authors and Affiliations

  1. Agricultural Biotechnology Center, H-2101, Gödöllő, P.O. Box 411, Hungary

    Sándor Tóth, Csaba Kiss, Gabriella Kovács & Ottó Toldi

  2. School of Life Sciences, University of Sussex, Falmer, Brighton, BN1 9QG, UK

    Peter Scott

  3. MTT Agrifood Research Finland, Institute of Horticulture, Toivonlinnantie 518, FIN-21500, Piikkiö, Finland

    Seppo Sorvari

Authors
  1. Sándor Tóth
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  2. Csaba Kiss
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  3. Peter Scott
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  4. Gabriella Kovács
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  5. Seppo Sorvari
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  6. Ottó Toldi
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Corresponding author

Correspondence to Ottó Toldi.

Additional information

Communicated by D. Dudits

Collaborator via a fellowship under the OECD Co-operative Research Programme: Biological Resource Management for Sustainable Agriculture Systems

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Tóth, S., Kiss, C., Scott, P. et al. Agrobacterium-mediated genetic transformation of the desiccation tolerant resurrection plant Ramonda myconi (L.) Rchb.. Plant Cell Rep 25, 442–449 (2006). https://doi.org/10.1007/s00299-005-0083-4

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  • DOI: https://doi.org/10.1007/s00299-005-0083-4

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