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Journal of Experimental Botany, Vol. 52, No. 361, pp. 1697-1702, August 1, 2001
© 2001 Oxford University Press

Original Papers

Transformation of Lotus japonicus using the herbicide resistance bar gene as a selectable marker

Dasharath P. Lohar1, Kathy Schuller2, Diana M. Buzas3, Peter M. Gresshoff3 and Jiri Stiller3,4

1 Center for Legume Research, M409 Walters Life Sciences Building, University of Tennessee, Knoxville, TN 37996-0845, USA
2 Biological Sciences, Flinders University, Adelaide, SA 5001, Australia
3 Department of Botany, University of Queensland, St Lucia, QLD 4072, Australia

Transgenic plants of the model legume Lotus japonicus were regenerated by hypocotyl transformation using a bar gene as a selectable marker. The bar encodes for Phosphinothricin Acetyl Transferase that detoxifies phosphinothricin (PPT), the active ingredient of herbicides such as Ignite (AgrEvo) and Basta (Hoechst). Transgenic L. japonicus plants resistant to PPT were positive upon PCR by bar gene-specific primers. In 5 out of 7 independent lines tested, PPT resistance segregated as a single dominant allele indicating a single T-DNA insertion into the plant genome. All regenerated plants were fertile and void of visible somaclonal abnormalities contrary to 14% infertility when antibiotic selectable markers were used. The lack of somaclonal variation, ease of PPT application and low cost of PPT makes this protocol an attractive alternative for the regeneration of transgenic L. japonicus. The production of PPT herbicide-resistant L. japonicus plants may have significant commercial applications in crop production.

Key words: Lotus, transformation, somaclonal variation, phosphinothricin.


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