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Efficient transformation of Medicago truncatula cv. Jemalong using the hypervirulent Agrobacterium tumefaciens strain AGL1

  • Genetic Transformation and Hybridization
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Abstract

The efficiency of Agrobacterium tumefaciens transformation of the model legume Medicago truncatula cv. Jemalong (genotype 2HA) was evaluated for strains LBA 4404, C58pMP90, C58pGV2260 and AGL1. Binary vectors carrying promoter-gus/gfp reporter gene fusions and the nptII gene as selectable marker were used for plant in vitro transformation/regeneration. The highest transformation efficiency was obtained with the disarmed hypervirulent strain AGL1 (Ti plasmid TiBo542), for which the percentage of explants forming kanamycin (Km)-resistant calli was double that obtained with each of the other three strains. In addition, we were able to reduce the time necessary for plant regeneration using AGL1, with 24% of the explants generating Km-resistant transgenic plantlets within only 4–5 months of culture. Transgene expression in planta was analysed and found to be conserved in the T1 descendents.

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Acknowledgements

We thank Dr. Ray Rose for kindly providing us with seeds of the highly regenerative genotype 2HA of M. truncatula cv. Jemalong, Dr. Lise Jouanin for the A. tumefaciens strains C58pGV2260, C58pMP90 and AGL1, Dr. Helena Carvalho for the plasmid pBin-prMtGSb-gus and Dr. Jim Haseloff for pBin-m-gfp5-ER. We also acknowledge Dr. Thierry Huguet, in whose group this work originally initiated. We are grateful to Beecham S.A. for providing free samples of the antibiotic Augmentin. Finally, we thank our colleagues for valuable discussions.

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  1. Laboratoire des Interactions Plantes-Microorganismes, INRA-CNRS, UMR215, BP 27 , 31326, Castanet Tolosan Cedex, France

    M. Chabaud, F. de Carvalho-Niebel & D. G. Barker

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  1. M. Chabaud
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  2. F. de Carvalho-Niebel
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  3. D. G. Barker
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Correspondence to M. Chabaud.

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Communicated by R.J. Rose

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Chabaud, M., de Carvalho-Niebel, F. & Barker, D.G. Efficient transformation of Medicago truncatula cv. Jemalong using the hypervirulent Agrobacterium tumefaciens strain AGL1. Plant Cell Rep 22, 46–51 (2003). https://doi.org/10.1007/s00299-003-0649-y

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