Insertional mutagenesis in arabidopsis thaliana: presonication of germinating seeds increases the efficiency of their transformation

An efficient procedure of transforming Arabidopsis thaliana germinating seeds was elaborated on the basis of the method of Feldmann and Marks. The procedure involves microdamaging Tl seeds by sonication before culturing with a vector Agrobacterium strain and yields more than 1% T2 transformants. Germinating seeds were transformed with Agrobacterium timefaciens hypervirulent strain A281 carrying plasmid pLD3 derived from pBI 121. A collection of 54 A. thaliana T2 transformants was obtained; some of them showed marked morphological alterations. The transgenic nature of plants that acquired resistance to a marker antibiotic was confirmed histochemically and by PCR amplification of a T-DNA region.