title: Improvements in the transformation of Arabidopsis thaliana C24 leaf-discs by Agrobacterium tumefaciens.
abstract: We report here an efficient Arabidopsis leafdisc transformation protocol yielding an average transformation frequency of 1.6 transgenic shoots per leaf explant 4 weeks after the bacterial infection period. Subsequent cultivation in vitro is such that a high percentage (85-90%) of the primary transformants produces seeds with an average seed yield of 100-300 seeds per plant. This improved transformation protocol yields mainly (70%) transformants segregating for a single T-DNA locus of which 68% actually contain one T-DNA insert. The objective is to generate a pool of independent transformants harboring an activator T-DNA construct in a gene tagging approach to isolate genes involved in morphogenesis and auxin signal transduction.
authors.affiliation: Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333, AL Leiden, The Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: van der Graaff
authors.forename: P J
authors.initials: PJ
authors.lastname: Hooykaas