Characterization of glyphosate resistance in cloned Amaranthus palmeri plants

Glyphosate-resistant Palmer amaranth from Georgia (GA), USA, possesses multiple copies of the gene that encodes 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), the enzyme target site of this herbicide. The cloned plants of glyphosate-resistant and glyphosate-susceptible Palmer amaranth biotypes from Mississippi (MS), USA, and GA were evaluated for glyphosate injury (digital imaging) in leaf disc bioassays. Four groups (three resistant groups: two from MS [G and R] and one from GA [C7]; one susceptible group from GA [C3]) were chosen for cloning to facilitate long-term studies. After exposure to glyphosate (1.0 mmol L-1, 144 h), the level of injury (mean value) was low in the resistant groups, while a higher level of injury was found in the susceptible group. However, the individual injury values within all groups varied widely. The mean EPSPS gene copy number of these groups was G >= R > C7 >>> C3. However, a higher copy number did not always convey increased resistance in these bioassays. When the copy number was high (>20), 81.5% of the bioassayed plants exhibited little or no injury and only similar to 20% were significantly injured, while 50% of the plants with a low copy number (<20) remained healthy. Overall, no strong statistical correlation of the copy number versus injury occurred in these cloned plants and no statistical relationship of resistance and copy number with the sex of the MS plants was observed. The results suggest that although an elevated copy number of the EPSPS gene can instill resistance, other mechanisms might contribute to the overall glyphosate resistance of Palmer amaranth in these plants.