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

Plant Methods

Greg Bryan¹  Chris S Jones²  Dorothy A Maher²  Kim A Richardson¹ 

[1] AgResearch Ltd, Grasslands Research Centre, Private Bag 11008, Palmerston North 4442, New Zealand;Pastoral Genomics, c/o Grasslands Research Centre, Private Bag 11008, Palmerston North 4442, New Zealand

关键词: Accession;    Organogenic regeneration;    Genetic transformation;    Trifolium occidentale;    Agrobacterium-mediated transformation;   

Others  :  820338
DOI  :  10.1186/1746-4811-9-25

 received in 2013-05-02, accepted in 2013-07-01,  发布年份 2013

【 摘 要 】

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 T₁ 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.

【 授权许可】

   
2013 Richardson et al.; licensee BioMed Central Ltd.

【 图 表 】

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