These encompassing results suggest that resistance to glyphosate in these GR RR A. trifida accessions is not conferred by a target-site resistance mechanism. © 2017 Society of Chemical Industry.

The GR RR mechanism of resistance is not associated with vacuole sequestration of glyphosate, and the observed reduced translocation is likely a consequence of rapid tissue death. Rapid cell death was inhibited by exogenous application of aromatic amino acids phenylalanine and tyrosine. The mechanism by which these amino acids inhibit rapid cell death in the GR RR phenotype remains unknown, and it could involve glyphosate phytotoxicity or other agents generating reactive oxygen species. Implications of these findings are discussed. The GR RR mechanism is distinct from the currently described glyphosate TSR or NTSR mechanisms in other species. © 2017 Society of Chemical Industry.

Resistance to glyphosate and paraquat has evolved in some populations of Conyza spp. from California, USA. This study evaluated whether herbicide absorption and translocation were involved in the mechanism of resistance to both herbicides. Three lines of each species were used: glyphosate-paraquat-susceptible (GPS), glyphosate-resistant (GR) and glyphosate-paraquatresistant (GPR). Radiolabelled herbicide was applied to a fully expanded leaf, and absorption and movement out of the treated leaf were monitored for up to 24 h for paraquat and 72 h for glyphosate. Plants treated with paraquat were incubated in darkness for the first 16 h and then subjected to light conditions. More glyphosate was absorbed in C. bonariensis (52.9-58.3%) compared with C. canadensis (28.5-37.6%), but no differences in absorption were observed among lines within a species. However, in both species, the GR and GPR lines translocated less glyphosate out of the treated leaf when compared with their respective GPS lines. Paraquat absorption was similar among lines and across species (71.3-77.6%). Only a fraction of paraquat was translocated in the GPR lines (3% or less) when compared with their respective GPS or GR lines (20% or more) in both species. Taken together, these results indicate that reduced translocation is involved in the mechanism of resistance to glyphosate and paraquat in C. bonariensis and C. canadensis.

Herbicide resistance is a challenge for modern agriculture further complicated by cases of resistance to multiple herbicides. Conyza bonariensis and Conyza canadensis are invasive weeds of field crops, orchards, and non-cropped areas in many parts of the world. In California, USA, Conyza populations resistant to the herbicides glyphosate and paraquat have recently been described. Although the mechanism conferring resistance to glyphosate and paraquat in these species was not elucidated, reduced translocation of these herbicides was observed under experimental conditions in both species. Glyphosate and paraquat resistance associated with reduced translocation are hypothesized to be a result of sequestration of herbicides into the vacuole, with the possible involvement of over-expression of genes encoding tonoplast transporters of ABC-transporter families in cases of glyphosate resistance or cationic amino acid transporters (CAT) in cases of paraquat resistance. However, gene expression in response to herbicide treatment has not been studied in glyphosate and paraquat resistant populations. In the current study, we evaluated the transcript levels of genes possibly involved in resistance using real-time PCR. First, we evaluated eight candidate reference genes following herbicide treatment and selected three genes that exhibited stable expression profiles; ACTIN, HEAT-SHOCK-PROTEIN-70, and CYCLOPHILIN. The reference genes identified here can be used for further studies related to plant-herbicide interactions. We used these reference genes to assay the transcript levels of EPSPS, ABC transporters, and CAT in response to herbicide treatment in susceptible and resistant Conyza spp. lines. No transcription changes were observed in EPSPS or CAT genes after glyphosate or paraquat treatment, suggesting that these genes are not involved in the resistance mechanism. Transcription of the two ABC transporter genes increased following glyphosate treatment in all Conyza spp. lines. Transcription of ABC transporters also increased after paraquat treatment in all three lines of C. bonariensis. However, in C. canadensis, paraquat treatment increased transcription of only one ABC transporter gene in the susceptible line. The increase in transcription of ABC transporters after herbicide treatment is likely a stress response based on similar response observed across all Conyza lines regardless of resistance or sensitivity to glyphosate or paraquat, thus these genes do not appear to be directly involved in the mechanism of resistance in Conyza spp.

Italian ryegrass [Lolium perenne L. spp. multiflorum (Lam.) Husnot] is one of the most troublesome weeds worldwide. L. multiflorum is also a grass seed crop cultivated on 50,000 ha in Oregon, where both diploid and tetraploid cultivars are grown. A survey was conducted to understand the distribution, frequency, and susceptibility of L. multiflorum to selected herbicides used to control L. multiflorum. The herbicides selected were clethodim, glufosinate, glyphosate, mesosulfuron-methyl (mesosulfuron), paraquat, pinoxaden, pyroxsulam, quizalofop-P-ethyl (quizolafop), pronamide, flufenacet + metribuzin, and pyroxasulfone. The ploidy levels of the populations were also tested. A total of 150 fields were surveyed between 2017 and 2018, of which 75 (50%) had L. multiflorum present. Herbicide-resistant populations were documented in 88% of the 75 populations collected. The most frequent mechanisms of action were resistance to Acetyl-CoA carboxylase (ACCase), Acetolactate Synthase (ALS), 5-enolpyruvylshikimate-3-phosphate (EPSPs) inhibitors, and combinations thereof. Multiple and cross-resistance, found in 75% of the populations, were the most frequent patterns of resistance. Paraquat-resistant biotypes were confirmed in six orchard crop populations for the first time in Oregon. Herbicide resistance was spatially clustered, with most cases of resistance in the northern part of the surveyed area. ALS and ACCase resistant populations were prevalent in wheat (Triticum aestivum L.) fields. Multiple-resistance was positively correlated with plant density. Tetraploid feral populations were identified, but no cases of herbicide resistance were documented. This is the first survey of herbicide resistance and ploidy diversity in L. multiflorum in western Oregon. Resistant populations were present across the surveyed area, indicating that the problem is widespread.

Reduced control of some glyphosate-resistant hairy fleabane populations with paraquat has raised concerns about evolved multiple resistance to both glyphosate and paraquat. The objective of this study was to confirm the presence of multiple-resistant (glyphosate and paraquat) hairy fleabane populations in California. A series of dose-response experiments was conducted to evaluate the effect of glyphosate and paraquat in a known susceptible (S) and putative multiple-resistant (R) population of hairy fleabane. The greenhouse experiments were conducted during summer, fall, and winter under controlled temperature and natural light conditions. Multiple-resistant hairy fleabane was identified; however, the level of resistance to glyphosate varied substantially among seasons. During the summer, the glyphosate rate required to reduce growth by 50% (GR50) for the R population was 0.94 kg ae ha−1, 5.2-fold more than for the S population. In the fall and winter experiments, however, the R population response to glyphosate was similar to the S population with a GR50of 0.22 kg ae ha−1or less. Multiple-resistant plants were controlled in the fall and winter at rates that did not control the same population during summer. GR50of paraquat varied among seasons (0.94, 0.24, and 0.07 kg ai ha−1during summer, fall, and winter, respectively); however, plant mortality was more consistent. This is the first reported case of glyphosate–paraquat resistance in hairy fleabane and the multiple-resistant population could pose a significant challenge to annual no-till and perennial cropping systems in California. Further research on the mechanisms of resistance and the physiological factors underlying the seasonally variable response to glyphosate is needed.

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BACKGROUND Poa annua is a widespread winter annual weed species in California. Recently, poor control of this species with glyphosate was reported by growers in an almond orchard in California with a history of repetitive glyphosate use. The objectives of this research were to evaluate the level of glyphosate resistance in a developed S4 P. annua line (R) and identify the mechanisms of resistance involved. RESULTS Whole‐plant dose–response experiments confirmed glyphosate resistance in R, which required 18‐fold more glyphosate to achieve a 50% growth reduction compared with a susceptible line (S), results that were supported by the lower shikimate accumulation observed in R compared with S. No differences in glyphosate absorption, translocation, or metabolism were observed, suggesting that non‐target‐site mechanisms of resistance are not involved in the resistance phenotype. A missense single nucleotide polymorphism was observed in EPSPS coding position 106 in R, resulting in a leucine to proline substitution. This polymorphism was observed exclusively in P. supina EPSPS homeologs. A seven‐fold increase in the number of copies of EPSPS alleles was observed in R compared with S. CONCLUSIONS We report the first case of glyphosate resistance associated with both EPSPS duplication and target‐site mutation at position 106, leading to high levels of glyphosate resistance in the allotetraploid weed species Poa annua L. Data obtained in this research will be useful for the development of diagnostic tools for rapid glyphosate resistance identification, monitoring and containment. © 2018 Society of Chemical Industry