Single-Cell Complementation of Barley mlo Mutants Using a PDS-1000/He Hepta System
نویسنده
چکیده
Transient expression via particle bombardment is widely used as a means of gene transfer to bacteria, yeast, animals, and plants. In this study we describe the use of the Hepta adaptor for the PDS-1000/He biolistic system for transformation and transient expression of Mlo in single epidermal cells of detached barley leaves. Barley Mlo is known to dampen plant defense and its expression in single epidermal cells of mlo resistant mutants restores susceptibility to attack from the powdery mildew fungal pathogen, Blumeria graminis. Consistently high transformation efficiencies were obtained upon delivery of a plasmid carrying both Green Fluorescent Protein (GFP) and Mlo. After bombardment and fungal spore inoculation, we found leaf sectors with numerous green fluorescent epidermal cells supporting growth of the pathogen. Likewise, we observed a high transformation efficiency of Arabidopsis leaf cells upon delivery of a GUS (β-glucuronidase) reporter gene construct. Application of the Hepta adaptor reduces the number of biolistic transfers necessary to obtain sufficient numbers of transformed plant cells for quantitative scoring of single-cell traits.
منابع مشابه
Molecular characterization of mlo mutants in North American two- and six-rowed malting barley cultivars.
SUMMARY Barley lines PRU1, URS1 and URS2 represent three candidate mlo mutants induced in either the two-rowed cultivar Prudentia or the six-rowed cultivar Ursula. Both Prudentia and Ursula are North American malting barley varieties with specific malting properties. Here, we analysed the three candidate mutants at the molecular level. We identified lesions in the Mlo gene of all three lines, c...
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