Floral dip: a simplified method forAgrobacterium‐mediated transformation ofArabidopsis thaliana

Abstract

SummaryThe Agrobacterium vacuum infiltration method has made it possible to transform Arabidopsis thaliana without plant tissue culture or regeneration. In the present study, this method was evaluated and a substantially modified transformation method was developed. The labor-intensive vacuum infiltration process was eliminated in favor of simple dipping of developing floral tissues into a solution containing Agrobacterium tumefaciens, 5% sucrose and 500 microliters per litre of surfactant Silwet L-77. Sucrose and surfactant were critical to the success of the floral dip method. Plants inoculated when numerous immature floral buds and few siliques were present produced transformed progeny at the highest rate. Plant tissue culture media, the hormone benzylamino purine and pH adjustment were unnecessary, and Agrobacterium could be applied to plants at a range of cell densities. Repeated application of Agrobacterium improved transformation rates and overall yield of transformants approximately twofold. Covering plants for 1 day to retain humidity after inoculation also raised transformation rates twofold. Multiple ecotypes were transformable by this method. The modified method should facilitate high-throughput transformation of Arabidopsis for efforts such as T-DNA gene tagging, positional cloning, or attempts at targeted gene replacement.

Global translational reprogramming is a fundamental layer of immune regulation in plants

“…efr7 (SALK_205018; ERF7, a homologue of the ethylene responsive TF gene ERF1 ) and gcn2 (GABI_862B02) were from the Arabidopsis Biological Resource Center (ABRC). Transgenic plants were generated using the floral dip method 30 .…”

uORF-mediated translation allows engineered plant disease resistance without fitness costs

“…Xoo strains PXO347 and PXO99 were grown on nutrient agar medium (0.1% yeast extract, 0.3% beef extract, 0.5% polypeptone, and 1% sucrose) at 28 °C for 2 d before resuspension in sterile water and dilution to OD 600nm = 0.5 for inoculation. 5 to 10 leaves of each plant were inoculated by the leaf-clipping method at the booting (panicle development) stage 27, 28 . Disease was scored by measuring the lesion length at 14 d post inoculation (dpi).…”

Selective gene dosage by CRISPR‐Cas9 genome editing in hexaploid Camelina sativa

“…Camelina Celine (cultivar) was transformed following an improved method of the traditional Arabidopsis floral‐dip method (Clough and Bent, 1998). Flower of Camelina were dipped in a solution of agrobacterium supplemented with sucrose (73 m m ), MgCl 2 (12.6 m m ) and acetosyringone (100 μ m ) in a vacuum under low pressure (50 mbar).…”