During the 2017 to 2019 growing seasons, samples of waterhemp and Palmer amaranth that had reportedly survived field-rate applications of protoporphyrinogen oxidase (PPO)–inhibiting herbicides were collected from the American Midwest and tested for target-site mutations known at the time to confer resistance. Target-site resistance was identified in nearly all (135 of 145) tested common waterhemp populations but in only 8 of 13 Palmer amaranth populations. Follow-up research on one population of Palmer amaranth (W-8), which tested negative for all such mutations, confirmed it was resistant to lactofen, with a magnitude of resistance comparable to that conferred by the ΔG210 PPO2 mutation. Gene sequences from both isoforms of PPO (PPO1 and PPO2) were compared between W-8 and known PPO inhibitor–sensitive sequence. A glycine-to-alanine substitution at the 399th amino acid position (G399A) of PPO2, recently identified to reduce target-site herbicide sensitivity, was observed in a subset of resistant W-8 plants. Because no missense mutation completely delimited resistant and sensitive sequences, we initially suspected the presence of a secondary, non-target-site resistance mechanism in this population. To isolate G399A, a segregating F2 population was produced and screened with a delimiting rate of lactofen. χ2 goodness-of-fit analysis of dead/alive ratings indicated single-locus inheritance of resistance in the F2 population, and molecular markers for the W-8 parental PPO2 coding region co-segregated tightly, but not perfectly, with resistance. More research is needed to fully characterize Palmer amaranth PPO inhibitor–resistance mechanisms, which appear to be more diverse than those found in common waterhemp.

Herbicide applications performed with pulse width modulation (PWM) sprayers to deliver specific spray droplet sizes could maintain product efficacy, minimize potential off-target movement, and increase flexibility in field operations. Given the continuous expansion of herbicide-resistant Palmer amaranth populations across the southern and midwestern United States, efficacious and cost-effective means of application are needed to maximize Palmer amaranth control. Experiments were conducted in two locations in Mississippi (2016, 2017, and 2018) and one location in Nebraska (2016 and 2017) for a total of 7 site-years. The objective of this study was to evaluate the influence of a range of spray droplet sizes [150 (Fine) to 900 μm (Ultra Coarse)] on lactofen and acifluorfen efficacy for Palmer amaranth control. The results of this research indicated that spray droplet size did not influence lactofen efficacy on Palmer amaranth. Palmer amaranth control and percent dry-biomass reduction remained consistent with lactofen applied within the aforementioned droplet size range. Therefore, larger spray droplets should be used as part of a drift mitigation approach. In contrast, acifluorfen application with 300-μm (Medium) spray droplets provided the greatest Palmer amaranth control. Although percent biomass reduction was numerically greater with 300-μm (Medium) droplets, results did not differ with respect to spray droplet size, possibly as a result of initial plant injury, causing weight loss, followed by regrowth. Overall, 900-μm (Ultra Coarse) droplets could be used effectively without compromising lactofen efficacy on Palmer amaranth, and 300-μm (Medium) droplets should be used to achieve maximum Palmer amaranth control with acifluorfen.

Glyphosate-resistant (GR) Palmer amaranth is a widespread problem in southeastern cotton production areas. Herbicide programs to control this weed in no-till cotton commonly include flumioxazin applied with preplant burndown herbicides approximately 3 wk before planting followed by fomesafen applied PRE and then glufosinate or glyphosate applied POST. Flumioxazin and fomesafen are both protoporphyrinogen oxidase (PPO) inhibitors. Multiple yearly applications of PPO inhibitors in cotton, along with widespread use of PPO inhibitors in rotational crops, raise concerns over possible selection for PPO resistance in Palmer amaranth. An experiment was conducted to determine the potential to substitute diuron for one of the PPO inhibitors in no-till cotton. Palmer amaranth control by diuron and fomesafen applied PRE varied by location, but fomesafen was generally more effective. Control by both herbicides was inadequate when timely rainfall was not received for activation. Palmer amaranth control was more consistent when programs included a preplant residual herbicide. Applied preplant, flumioxazin was more effective than diuron. Programs with diuron preplant followed by fomesafen PRE were as effective as flumioxazin preplant followed by fomesafen only if fomesafen was activated in a timely manner. Programs with flumioxazin preplant followed by diuron PRE were as effective as flumioxazin preplant followed by fomesafen PRE at all locations, regardless of timely activation of the PRE herbicide. As opposed to flumioxazin preplant followed by fomesafen PRE, which exposes Palmer amaranth to two PPO-inhibiting herbicides, one could reduce selection pressure by using flumioxazin preplant followed by diuron PRE without sacrificing Palmer amaranth control or cotton yield.

Molecular assays are often implemented by weed scientists for detection of herbicide-resistant individuals; however, the utility of these assays can be limited if multiple mechanisms of evolved resistance exist. Waterhemp resistant to protoporphyrinogen oxidase (PPO)– inhibiting herbicides is conferred by a target-site mutation in PPX2L (a gene coding for PPO), resulting in the loss of a glycine at position 210 (ΔG210). This ΔG210 mutation of PPX2L is the only known mechanism responsible for PPO-inhibitor resistance (PPO-R) in waterhemp from five states (Illinois, Indiana, Iowa, Kansas, and Missouri); however, a limited number of populations have been tested, especially in Illinois. To verify the ubiquity of the ΔG210 in PPO-R waterhemp populations in Illinois, a previously published allele-specific PCR (asPCR) was used for the detection of the ΔG210 mutation to associate this mutation with phenotypic resistance in 94 Illinois waterhemp populations. The ΔG210 mutation was detected in all populations displaying phenotypic resistance to lactofen (220 g ai ha−1), indicating the deletion is likely the only mechanism of resistance. With evidence that the ΔG210 mutation dominates PPO-R waterhemp biotypes, molecular detection techniques have considerable utility. Unfortunately, the previously published asPCR is time consuming, very sensitive to PCR conditions, and requires additional steps to eliminate the possibility of false negatives. To overcome these limitations, a streamlined molecular method using the TaqMan® technique was developed, utilizing allele-specific, fluorescent probes for high-throughput, robust discrimination of each allele (resistant and susceptible) at the 210th amino acid position of PPX2L.

A field study was conducted in 2007 and 2008 near Murphysboro, IL to determine the effect of plant height and addition of glyphosate on control of glyphosate-resistant horseweed with saflufenacil. Saflufenacil was applied at rates ranging from 25 to 125 g ai ha−1 alone and in combination with glyphosate at 840 g ae ha−1, and the efficacy compared to paraquat at 840 g ai ha−1. Control of horseweed with glyphosate applied alone was less than 30%, confirming the presence of glyphosate-resistant plants. At 14 d after application, all treatments with saflufenacil or paraquat provided at least 90% control. Saflufenacil applied alone at the lowest rate of 25 g ha−1 provided less control (92%) than all other treatments that included saflufenacil, and efficacy was reduced as horseweed height at application increased. Horseweed control from saflufenacil at 50 g ha−1 was reduced as plant height increased in 2007 but not in 2008. However, saflufenacil applied at 50 g ha−1 or greater resulted in at least 98% control, regardless of horseweed height at application or tank mixture with glyphosate. Combining glyphosate with saflufenacil at 25 g ha−1 increased horseweed control compared with saflufenacil applied alone and resulted in control similar to saflufenacil applied at 50 g ha−1. Control of horseweed from paraquat declined over time as the growth continued from the apical meristem. The extent of horseweed regrowth from applications of saflufenacil alone was less than that observed from paraquat. The addition of glyphosate to saflufenacil further reduced the frequency of horseweed regrowth compared with saflufenacil applied alone.

Greenhouse studies were conducted to determine the influence of spray-solution pH, adjuvant, light intensity, temperature, and glyphosate on the efficacy of saflufenacil on horseweed. Control of glyphosate-resistant horseweed from saflufenacil alone was greatest with a spray-solution pH of 5, compared with pH 7 or 9. However, when glyphosate was added to saflufenacil, similar GR50 values were measured with spray solutions adjusted to pH 5 and 9, and horseweed control at pH 9 was 38% greater than at pH 7. The efficacy of saflufenacil on horseweed was 36% greater when crop oil concentrate was used as an adjuvant compared with nonionic surfactant, regardless of the addition of glyphosate or the sensitivity of the horseweed population to glyphosate (resistant vs. susceptible). The addition of glyphosate to low rates of saflufenacil increased control over saflufenacil applied alone on glyphosate-susceptible and -resistant horseweed. Saflufenacil activity was greater under low light intensity (300 μmol m−2 s−1) than high light intensity (1,000 μmol m−2 s−1). Although initial horseweed control was greater under high temperature (27 C) compared with low temperature (10 C), by 21 d after treatment horseweed dry weight was similar from saflufenacil applied under high and low temperatures.