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Clovers (Trifolium spp.)

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Agrobacterium Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1223))

Abstract

Clovers (Trifolium spp.) constitute one of the major forage legumes widely grown for its rich protein content and its major role in maintaining environmental sustainability by improving the soil fertility. Gene technology can assist plant improvement efforts in clovers (Trifolium spp.), aiming to improve forage quality, yield, and adaptation to biotic and abiotic stresses. An efficient and reproducible protocol for Agrobacterium-mediated transformation of a range of Trifolium species, using cotyledonary explants and different selectable marker genes, is described. The protocol is robust and allows for genotype and Agrobacterium strain-independent transformation of clovers. Stable meiotic transmission of transgenes has been demonstrated for selected transgenic clovers carrying single T-DNA inserts recovered from Agrobacterium-mediated transformation. This methodology can also be successfully used for “isogenic transformation” in clovers: the generation of otherwise identical plants with and without the transgene from the two cotyledons of a single seed. Stable transgenes may be used in further functional genomics, develop new traits and profile gene expression using reporters, and facilitate purification of tissue or single cells.

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Authors and Affiliations

  1. School of Applied Systems Biology, AgriBio, Centre for AgriBioscience, La Trobe University, Bundoora, VIC, 3083, Australia

    Samira Rahimi-Ashtiani, John Mason & German Spangenberg

  2. Biosciences Research Division, Department of Environment and Primary Industries, AgriBio, Centre for AgriBioscience, La Trobe University, 5 Ring Road, Bundoora, VIC, 3083, Australia

    Sareena Sahab, Stephen Panter, John Mason & German Spangenberg

Authors
  1. Samira Rahimi-Ashtiani
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  2. Sareena Sahab
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  3. Stephen Panter
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  4. John Mason
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  5. German Spangenberg
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Corresponding author

Correspondence to German Spangenberg .

Editor information

Editors and Affiliations

  1. Plant Sciences Institute, and Department of Agronomy, Iowa State University, Center for Plant Transformation, Ames, Iowa, USA

    Kan Wang

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Rahimi-Ashtiani, S., Sahab, S., Panter, S., Mason, J., Spangenberg, G. (2015). Clovers (Trifolium spp.). In: Wang, K. (eds) Agrobacterium Protocols. Methods in Molecular Biology, vol 1223. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-1695-5_18

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  • DOI: https://doi.org/10.1007/978-1-4939-1695-5_18

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-1694-8

  • Online ISBN: 978-1-4939-1695-5

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