Evolved resistance to the herbicide glyphosate has been reported in eleven weed species, including Lolium multiflorum. Two glyphosate-resistant L. multiflorum populations were collected, one from Chile (SF) and one from Oregon, USA (OR), and the mechanisms conferring glyphosate resistance were studied. Based on a Petri dish dose-response bioassay, the OR and the SF populations were two and fivefold more resistant to glyphosate when compared to the susceptible (S) population, respectively; however, based on a whole-plant dose-response bioassay, both OR and SF populations were fivefold more resistant to glyphosate than the S population, implying that different resistance mechanisms might be involved. The S population accumulated two and three times more shikimic acid in leaf tissue 96 h after glyphosate application than the resistant OR and SF populations, respectively. There were no differences between the S and the glyphosate-resistant OR and SF populations in 14C-glyphosate leaf uptake; however, the patterns of 14C-glyphosate translocation were significantly different. In the OR population, a greater percentage of 14C-glyphosate absorbed by the plant moved distal to the treated section and accumulated in the tip of the treated leaf. In contrast, in the S and in the SF populations, a greater percentage of 14C-glyphosate moved to non-treated leaves and the stem. cDNA sequence analysis of the EPSP synthase gene indicated that the glyphosate-resistant SF population has a proline 106 to serine amino acid substitution. Here, we report that glyphosate resistance in L. multiflorum is conferred by two different mechanisms, limited translocation (nontarget site-based) and mutation of the EPSP synthase gene (target site-based).
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 Ϫ1 ) 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 Ϫ1 ) 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 M) for 14 h under continuous light. Shikimic acid was extracted from each leaf segment and quantified. At a concentration up to 100 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 biotype from Chile.Nomenclature: Glyphosate; Italian ryegrass, Lolium multiflorum Lam. LOLMU; filbert, Corylus avellana L.
Summary Primisulfuron‐resistant (AR and MR) and ‐susceptible (AS and MS) Bromus tectorum biotypes were collected from a Poa pratensis field at Athena, Oregon, and in research plots at Madras, Oregon. Studies were conducted to characterize the resistance of the B. tectorum biotypes. Whole plant bioassay and acetolactate synthase (ALS) enzyme assay revealed that the AR biotype was highly resistant to the sulfonylurea (SU) herbicides, primisulfuron and sulfosulfuron and to a sulfonylaminocarbonyltriazolinone (SCT) herbicide, propoxycarbazone‐sodium. However, the AR biotype was not resistant to imazamox, an imidazolinone (IMI) herbicide. Results of the whole plant bioassay studies showed that the MR biotype was moderately resistant to all ALS inhibitors tested. However, there were no differences in ALS sensitivities between the MR and MS biotypes. The nucleotide and amino acid sequence analysis of the als gene demonstrated a single‐point mutation from C to T, conferring the exchange of the amino acid proline to serine at position 197 in the AR biotype. However, this mutation was not found in the MR biotype. Results of this research indicate that: the resistance of the AR biotype to SU and SCT herbicides is based on an altered target site due to a single‐point mutation; resistance in the MR biotype is not due to a target site mutation.
Studies were conducted to determine the relative fitness and competitive ability of an acetolactate synthase (ALS) inhibitor–resistant (R) downy brome biotype compared with a susceptible (S) biotype. In previous research, the mechanism of resistance was determined to be an altered ALS enzyme. Seed germination of the R biotype was compared with that of the S biotype at 5, 15, and 25 C. There were no different germination characteristics between R and S biotypes at 15 and 25 C. However, the R biotype germinated 27 h earlier than the S biotype and had reached over 60% germination when the S biotype initially germinated at 5 C. Under noncompetitive greenhouse conditions, growth of the R biotype was similar to that of the S biotype on the basis of shoot dry weight, leaf area, and plant height. Seed production of the R biotype was 83%, when compared with the S biotype, but seeds of the R biotype were larger than those of the S biotype. Replacement series experiments were conducted in the greenhouse to determine the relative competitive ability of R and S biotypes. No difference in competitive ability was observed between R and S biotypes on the basis of shoot dry weight, leaf area, or plant height. Thus, it appears that ALS-resistance trait is not associated with growth penalty in either noncompetitive or competitive conditions. In the absence of ALS inhibitors, these results suggest that the R biotype would remain at a similar frequency in a population of R and S biotypes.
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