Identification of glyphosate-resistant Italian ryegrass (Lolium multiflorum) in Oregon

A suspected glyphosate-resistant Italian ryegrass biotype was collected from a filbert orchard near Portland, OR, where glyphosate was applied multiple times per year for about 15 yr. Greenhouse studies were conducted to determine if this biotype was glyphosate resistant. The plants were sprayed with glyphosate (0.01 to 3.37 kg ae ha(-1)) 14 d after planting and shoot biomass was determined 3 wk after herbicide treatment. Based on the dose-response experiments conducted in the greenhouse, the suspected Italian ryegrass biotype was approximately fivefold more resistant to glyphosate than the susceptible biotype. Plants from both susceptible and resistant biotypes were treated with glyphosate (0.42 and 0.84 kg ha-1) and shikimic acid was extracted 12, 24, 48, and 96 h after treatment. The susceptible biotype accumulated between three and five times more shikimic acid than did the resistant biotype. Leaf segments from both susceptible and resistant biotypes were incubated with different glyphosate concentrations (0-5 to 3000 mu M) for 14 h under continuous light. Shikimic acid was extracted from each leaf segment and quantified. At a concentration up to 100 mu M, leaf segments from the susceptible biotype accumulated more shikimic acid than leaf segments from the resistant biotype. The epsps gene was amplified and sequenced in both susceptible and resistant biotypes; however, no amino acid change was found in the resistant biotype. The level of resistance in this biotype is similar to that reported for a glyphosate-resistant Italian ryegrass biorype from Chile.