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Generation of transgenic Lolium temulentum plants by Agrobacterium tumefaciens-mediated transformation

  • Genetic Transformation and Hybridization
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Abstract

Lolium temulentum L. (Darnel ryegrass) has been proposed to be used as a model species for functional genomics studies in forage and turf grasses, because it is a self-fertile, diploid species with a short life cycle and is closely related to other grasses. Embryogenic calluses were induced from mature embryos of a double haploid line developed through anther culture. The calluses were broken up into small pieces and used for Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harboring pCAMBIA1301 and pCAMBIA1305.2 vectors were used to infect embryogenic callus pieces. Hygromycin was used as a selection agent in stable transformation experiments. Hygromycin resistant calluses were obtained after 4–6 weeks of selection and transgenic plants were produced in 10–13 weeks after Agrobacterium-mediated transformation. Fertile plants were readily obtained after transferring the transgenics to the greenhouse. Transgenic nature of the regenerated plants was demonstrated by Polymerase chain reaction (PCR), Southern hybridization analysis, and GUS staining. Progeny analysis showed Mendelian inheritance of the transgenes. The transformation system provides a valuable tool for functionality tests of candidate genes in forage and turf grasses.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxy acetic acid

ESTs:

Expressed sequence tags

PCR:

Polymerase chain reaction

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Acknowledgements

The authors would like to thank Elane Wright and Tina Norton for critical reading of the manuscript. This work was supported by the Samuel Roberts Noble Foundation.

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Authors and Affiliations

  1. Forage Improvement Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA

    Yaxin Ge, Xiaofei Cheng, Andrew Hopkins & Zeng-Yu Wang

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  1. Yaxin Ge
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  2. Xiaofei Cheng
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  3. Andrew Hopkins
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  4. Zeng-Yu Wang
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Correspondence to Zeng-Yu Wang.

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Communicated by G. D. May

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Ge, Y., Cheng, X., Hopkins, A. et al. Generation of transgenic Lolium temulentum plants by Agrobacterium tumefaciens-mediated transformation. Plant Cell Rep 26, 783–789 (2007). https://doi.org/10.1007/s00299-006-0288-1

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