Agrobacterium‐Mediated Genetic Transformation of Switchgrass

摘要: Although Agrobacterium tumefaciens has been successfully used to transfer genes a wide range of plant species, it received little attention for transformation forage grasses. Therefore, the objective present study was demonstrate Agrobacterium-mediated switchgrass (Panicum virgatum L.). The A. strain AGL 1 carrying binary vector pDM805, coding phosphinothricin acetyltransferase (bar) and β-glucuronidase (gus) genes, utilized in these experiments. Somatic embryos, embryogenic calluses, mature caryopses, plantlet segments served as target tissues infection. Treated cultures were selected presence 10 mg L -1 bialaphos resultant plantlets treated with herbicide Basta [monoammonium 2-amino-4(hydroxymethylphosphinyl)butanoate]. T-DNA delivery efficiency affected by genotype, explant or absence acetosyringone (3',5'-dimethoxy-4'-hydroxyacetophenone) during inoculation cocultivation. Approximately 600 transgenic plants produced, efficiencies ranged from 0 nearly 100%. Stable integration, expression, inheritance both transgenes confirmed molecular genetic analyses. 90% tested appeared have only one two copies inserts. Controlled crosses between To nontransgenic 'Alamo' indicated expected ratio 1:1 (transgenic:nontransgenic) T according X 2 test at P = 0.05. These results indicate that method is effective transferring foreign into switchgrass.