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Background  Western clover ( Trifolium occidentale ) is a perennial herb with characteristics compatible for its development as an attractive model species for genomics studies relating to the forage legume, white clover ( Trifolium repens ). Its characteristics such as a small diploid genome, self-fertility and ancestral contribution of one of the genomes of  T .  repens , facilitates its use as a model for genetic analysis of plants transformed with legume or novel genes.    Results  In this study, a reproducible transformation protocol was established following screening of  T .  occidentale  accessions originating from England, Ireland, France, Spain and Portugal. The protocol is based upon infection of cotyledonary explants dissected from mature seed with the  Agrobacterium tumefaciens  strain GV3101 carrying vectors which contain the  bar  selection marker gene. Transformation frequencies of up to 7.5% were achieved in 9 of the 17 accessions tested. Transformed plants were verified by PCR and expression of the  gusA  reporter gene, while integration of the T-DNA was confirmed by Southern blot hybridisation and segregation of progeny in the T 1  generation.    Conclusions  Development of this protocol provides a valuable contribution toward establishing  T .  occidentale  as a model species for white clover. This presents opportunities for further improvement in white clover through the application of biotechnology.

plant methods

Genetic transformation of western clover (Trifolium occidentale D. E. Coombe.) as a model for functional genomics and transgene introgression in clonal pasture legume species