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Apomictic Bahiagrass Expressing the bar Gene Is Highly Resistant to Glufosinate under Field Conditions

^a Agronomy Dep., Plant Molecular and Cellular Biology Program, Genetics Institute, Univ. of Florida-IFAS, Gainesville, FL 32611
^b NFREC, Agronomy Dep., Univ. of Florida-IFAS, Marianna, FL

View larger version (19K): [in this window] [in a new window]   Figure 1. Diagrammatic representation of the bar gene expression cassette used for genetic transformation of bahiagrass (Paspalum notatum Flugge).

View larger version (92K): [in this window] [in a new window]   Figure 2. Generation and evaluation of herbicide-resistant bahiagrass: (a) regeneration of plants from embryogenic bahiagrass callus following gene transfer; (b) transgenic bahiagrass plants established in soil in the greenhouse; (c) response of nontransgenic, wild-type bahiagrass (WT) and transgenic bahiagrass line 4 or (d) line 24 expressing the bar gene, 14 d after application of 0.2, 0.5, or 1.0% glufosinate ammonium under greenhouse conditions; (e) seed-derived progeny plants of transgenic bahiagrass; (f) transgenic bahiagrass lines expressing the bar gene (center) surrounded by nontransgenic bahiagrass plants before and (g) 14 d after application of 0.6% glufosinate ammonium under field conditions; (h) immunodetection of bar-encoded phosphinothricin acetyltransferase (PAT) in T0 transgenic lines or (i) seed-derived progeny of transgenic plants by lateral flow membrane assay (Libertylink Stix test). Protein extracts with detectable levels of PAT display a PAT-specific band indicated by the arrow, whereas extracts from nontransgenic, wild-type (WT) bahiagrass show a separate band, common to all lines, which indicates the functionality of the lateral flow membrane strip.

View larger version (89K): [in this window] [in a new window]   Figure 3. Southern blot analysis of transgenic bahiagrass. Genomic DNA (20 µg) from nontransgenic wild-type (WT) and transgenic bahiagrass lines (4–52) was digested with BglII, which cuts once in the expression cassette of the bar gene (Fig. 1). The full length coding region of the bar gene was used as a probe and labeled with 32P-deoxy cytidine 5'-triphosphate. Linearized pJFBar plasmid (20 pg) was used as a positive control (PC).