Germ-line transformation of Arabidopsis lasiocarpa.
In planta transformation methods have opened up the possibility of transforming plant species for which no regeneration protocols currently exist. In this study, the suitability of the germ-line transformation method developed for Arabidopsis thaliana was examined for four taxa in the Brassicaceae that have not been previously transformed: Arabidopsis griffithiana, Arabidopsis lasiocarpa, Arabidopsis petraea and Capsella bursa-pastoris. Numerous transformants were obtained for A. lasiocarpa. Transformation of A. lasiocarpa was confirmed at the phenotypic and molecular levels for stably transformed lines and for backcrossed lines segregating the T-DNA insert. Parameters affecting transformation efficiency of A. lasiocarpa were also explored. As with A. thaliana, sucrose and surfactant in the inoculation medium are required for high levels of transformation, although the suitable concentrations of these are different for A. lasiocarpa. Other components present in earlier versions of the inoculation medium had little effect on transformation efficiency. Vacuum infiltration (rather than simple floral dipping) led to higher rates of transformation and did not seriously affect seed production in A. lasiocarpa. Identification of species susceptible to germ-line transformation will aid in determining the factors important for applying this technology to more recalcitrant species.
Tague
B W
BW
Department of Biology, Wake Forest University, Winston-Salem, NC 27109, USA. taguebw@wfu.edu
D017360:Arabidopsis / Q000166:cytology* / Q000187:drug effects / Q000235:genetics* / Q000382:microbiology; D015139:Blotting, Southern; D003433:Crosses, Genetic; D005456:Fluorescent Dyes; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006824:Hybridization, Genetic; D007612:Kanamycin / Q000494:pharmacology; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008856:Microscopy, Fluorescence; D010641:Phenotype; D030821:Plants, Genetically Modified; D012231:Rhizobium / Q000235:genetics; D013395:Sucrose / Q000494:pharmacology; D014170:Transformation, Genetic
D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D013395:Sucrose; D007612:Kanamycin