Evidence, Mechanism and Alternative Chemical Seedbank-Level Control of Glyphosate Resistance of a Rigid Ryegrass (Lolium rigidum) Biotype from Southern Spain

Rigid ryegrass (Lolium rigidum) is one of the most troublesome weeds in different crops in the Mediterranean region. A rigid ryegrass biotype from an olive grove in Jaén province (Andalusía, southern Spain), potentially resistant to glyphosate (RG), was tested for its resistance level through dose-response assays using a susceptible biotype (SG). To test the hypothesis of a non-target-site-based resistance, as point mutations are far less common mechanisms of glyphosate resistance, studies were also conducted to elucidate whether resistance was associated with biochemical, metabolism, molecular and/or physiological mechanisms. Alternative herbicide-based control options, including single-herbicide or herbicide mixtures with glyphosate, applied at seedling, tillering or full heading stages, were tested in field experiments for 2 years for their efficacy against rigid ryegrass plants and their effects on the soil seed bank. Resistance levels of the RG biotype were 23- (LD(50)) and 7-fold (GR(50)) higher compared to the SG biotype. The SG biotype exhibited a significantly greater shikimic acid accumulation than the RG one. At 96 HAT, 58 and 89% of applied (14)C-glyphosate was up taken by leaves of RG and SG biotype plants, respectively, and, at this time, a significantly higher proportion of the glyphosate taken up by the treated leaf remained in its tissue in RG plants compared to the SG ones. The RG biotype did not reveal any point mutation in the glyphosate target site EPSP synthase. Overall, results confirmed reduced glyphosate uptake and translocation as being the mechanism involved in glyphosate resistance in the RG biotype. RG biotype responses to the alternative treatments tested in situ indicated that herbicide applications at the later growth stage tended to be less effective in terms of immediate effects on population size than earlier applications, and that only in some cases, the removal of at least 85% of the RG biotype was achieved. However, with few exceptions, the alternative treatments tested appeared to be highly effective in reducing the seed bank irrespective of the growth stage. The frequency of the resistant phenotype in the progeny of surviving plants of the RG biotype was dependent on treatment. Results suggest that a potential exists for effective management of glyphosate-resistant rigid ryegrass in olive groves in southern Spain.