Mitotic-inhibiting herbicides, like prodiamine and dithiopyr, are used to control annual bluegrass (Poa annua L.) preemergence in managed turfgrass; however, resistance to mitotic-inhibiting herbicides has evolved due to repeated applications of herbicide from a single mechanism of action. Three suspected resistant populations (R1, R2, and R3) were collected in Alabama and Florida and screened for resistance to prodiamine. Part of the α-tubulin gene was sequenced for known target-site mutations. Target-site mutations were reported in all three R populations, with each containing an amino acid substitution at position 239 from threonine to isoleucine (Thr-239-Ile). Previous research has indicated that the Thr-239-Ile mutation confers resistance to dinitroaniline herbicides in other species. Dose–response screens using prodiamine and dithiopyr were conducted and I50 values were calculated for R1, R2, and R3 using regression models based on seedling emergence. For prodiamine, I50 values for R1, R2, and R3 were 35.3, 502.7, and 91.5 g ai ha−1, respectively, resulting in 2.9-, 41.9-, and 7.6-fold resistance, respectively, when compared with a susceptible (S) population. For dithiopyr, I50 values for R1, R2, and R3 were 154.0, 114.2, and 190.1 g ai ha−1, respectively, resulting in 3.6-, 2.7-, and 4.5-fold resistance, respectively, when compared with an S population. When comparing I90 values with the highest labeled use rates, R2 had a 2.9-fold level of resistance to prodiamine, and R1, R2, and R3 had a 2.4-, 2.0-, and 3.2-fold levels of resistance to dithiopyr, respectively. This is the first report of a variable response in P. annua to prodiamine despite each R population possessing the same mutation.

The mitotic-inhibiting herbicide pronamide controls susceptible annual bluegrass (Poa annua L.) pre- and postemergence, but in some resistant populations, postemergence activity is compromised, hypothetically due to a target-site mutation, lack of root uptake, or an unknown resistance mechanism. Three suspected pronamide-resistant (LH-R, SC-R, and SL-R) and two pronamide-susceptible (BS-S and HH-S) populations were collected from Mississippi golf courses. Dose–response experiments were conducted to confirm and quantify pronamide resistance, as well as resistance to flazasulfuron and simazine. Target sites known to confer resistance to mitotic-inhibiting herbicides were sequenced, as were target sites for herbicides inhibiting acetolactate synthase (ALS) and photosystem II (PSII). Pronamide absorption and translocation were investigated following foliar and soil applications. Dose–response experiments confirmed pronamide resistance of LH-R, SC-R, and SL-R populations, as well as instances of multiple resistance to ALS- and PSII-inhibiting herbicides. Sequencing of the α-tubulin gene confirmed the presence of a mutation that substituted isoleucine for threonine at position 239 (Thr-239-Ile) in LH-R, SC-R, SL-R, and BS-S populations. Foliar application experiments failed to identify differences in pronamide absorption and translocation between the five populations, regardless of harvest time. All populations had limited basipetal translocation—only 3% to 13% of the absorbed pronamide—across harvest times. Soil application experiments revealed that pronamide translocation was similar between SC-R, SL-R, and both susceptible populations across harvest times. The LH-R population translocated less soil-applied pronamide than susceptible populations at 24, 72, and 168 h after treatment, suggesting that reduced acropetal translocation may contribute to pronamide resistance. This study reports three new pronamide-resistant populations, two of which are resistant to two modes of action (MOAs), and one of which is resistant to three MOAs. Results suggest that both target site– and translocation-based mechanisms may be associated with pronamide resistance. Further research is needed to confirm the link between pronamide resistance and the Thr-239-Ile mutation of the α-tubulin gene.

Glufosinate resistance in Palmer amaranth (Amaranthus palmeri S. Watson) was recently detected in three accessions from Arkansas, USA. Amaranthus palmeri is the first and only broadleaf weed species resistant to this herbicide, and the resistance mechanism is still unclear. A previous study characterized the glufosinate resistance level in the accessions from Arkansas. A highly glufosinate-resistant accession was further used to investigate the mechanism conferring glufosinate resistance in A. palmeri. Experiments were designed to sequence the herbicide target enzyme cytosolic and chloroplastic glutamine synthetase isoforms (GS1 and GS2, respectively) and quantify copy number and expression. Absorption, translocation, and metabolism of glufosinate using the 14C-labeled herbicide were also evaluated in the resistant and susceptible accessions. The glufosinate-resistant accession had an increase in copy number and expression of GS2 compared with susceptible plants. All accessions showed only one GS1 copy and no differences in expression. No mutations were identified in GS1 or GS2. Absorption (54% to 60%) and metabolism (13% to 21%) were not different between the glufosinate-resistant and glufosinate-susceptible accessions. Most residues of glufosinate (94% to 98%) were present in the treated leaf. Glufosinate translocation to tissues above the treated leaf and in the roots was not different among accessions. However, glufosinate translocation to tissues below the treated leaf (not including roots) was greater in the resistant A. palmeri (2%) compared with the susceptible (less than 1%) accessions. The findings of this paper strongly indicate that gene amplification and increased expression of the chloroplastic glutamine synthetase enzyme are the mechanisms conferring glufosinate resistance in the A. palmeri accession investigated. Thus far, no additional resistance mechanism was observed, but further investigations are ongoing.

Acetolactate synthase (ALS) inhibitors provide postemergence control of green kyllinga (Kyllinga brevifolia Rottb.) in turfgrass and other cropping systems. A suspected resistant (R) biotype of K. brevifolia was collected from a golf course and evaluated for resistance to ALS inhibitors. In greenhouse experiments, the sulfosulfuron rates required to cause 50% shoot biomass reduction from the nontreated at 4 wk after treatment (WAT) were 10 and 792 g ai ha−1 for the susceptible (S) and R biotypes, respectively. The rates required to cause 50% injury at 4 WAT were 189 and >3,360 g ai ha−1, respectively. In other experiments, shoot mass of the R biotype was not reduced by imazaquin, trifloxysulfuron-sodium, pyrimisulfan, thiencarbazone + foramsulfuron + halosulfuron, florasulam + halauxifen-methyl, and bentazon compared with the nontreated, while sulfentrazone reduced biomass similarly for both R and S biotypes. Gene sequencing of the R biotype revealed a mutation at Asp-376-Glu that has previously conferred resistance to five families of ALS inhibitors. This is the first report of ALS-inhibitor resistance in K. brevifolia.

Dithiopyr and dinitroanilines are preemergence-applied, mitotic-inhibiting herbicides used to control goosegrass [Eleusine indica (L.) Gaertn.] in turfgrass. A suspected resistant E. indica population was collected from a golf course putting green and was evaluated for possible resistance to dithiopyr and prodiamine. After dose–response evaluation, the α-tubulin gene was sequenced for known target-site mutations that have been reported to confer resistance to mitotic-inhibiting herbicides. A mutation was discovered that resulted in an amino acid substitution at position 136 from leucine to phenylalanine (Leu-136-Phe). Previous research has indicated that Leu-136-Phe does confer resistance to dinitroaniline herbicides. The level of resistance indicated by regression models and I50 values indicates that there is 54.1-, 4.7-, >100-, and >100-fold resistance to dithiopyr, prodiamine, pendimethalin, and oryzalin, respectively, when compared with the susceptible population based on seedling emergence response and 88.4-, 7.8-, >100-, and >100-fold resistance to dithiopyr, prodiamine, pendimethalin, and oryzalin, respectively, when compared with the susceptible population based on biomass reduction response. This is the first report of less resistance to prodiamine compared with pendimethalin or oryzalin due to a target-site α-tubulin mutation and the first report of a target-site α-tubulin mutation associated with dithiopyr resistance.

Research was conducted using a functional malachite green colorimetric assay to evaluate acetyl-coenzyme A carboxylase (ACCase) activity previously identified as resistant to sethoxydim and select aryloxyphenoxypropionate (FOPs) herbicides, fenoxaprop, and fluazifop. Two resistant southern crabgrass [Digitaria ciliaris (Retz.) Koeler] biotypes, R1 and R2, containing an Ile-1781-Leu amino acid substitution and previously identified as resistant to sethoxydim, pinoxaden, and fluazifop but not clethodim was utilized as the resistant chloroplastic ACCase source compared with known susceptible (S) ACCase. Dose-response studies with sethoxydim, clethodim, fluazifop-p-butyl, and pinoxaden (0.6 to 40 µM) were conducted to compare the ACCase–herbicide interactions of R1, R2, and S using the malachite green functional assay. Assay results indicated that R biotypes required more ACCase-targeting herbicides to inhibit ACCase activity compared with S. IC50 values of all four herbicides for R biotypes were consistently an order of magnitude greater than those of S. No sequencing differences in the carboxyltransferase domain was observed for R1 and R2; however, R2 IC50 values were greater across all herbicides. These results indicate the malachite green functional assay is effective in evaluating ACCase activity of R and S biotypes in the presence of ACCase-targeting herbicides, which can be used as a replacement for the 14C-based radiometric functional assays.

A goosegrass [Eleusine indica (L.) Gaertn.] population uncontrolled by paraquat (R) in a vegetable production field in St. Clair County, AL, was collected in summer 2019. Research was conducted to assess the level of resistance of the suspected resistant population compared with three populations with no suspected paraquat resistance (S1, S2, and S3). Visual injury at all rating dates and biomass reduction at 28 d after treatment (DAT) of S populations occurred exponentially to increasing paraquat rates. S biotypes were injured more than R at 3 DAT, with biomass recovery at 28 DAT only occurring at rates <0.28 kg ha−1. Plant death or biomass reduction did not occur for any rate at any date for R. Paraquat rates that induced 50% or 90% injury or reduced biomass 50% or 90% compared with the non-treated (I50 or I90, respectively) ranged from 10 to 124 times higher I50 for R compared with S and 54 to 116 times higher I90 for R compared with S biotypes. These data confirm a paraquat-resistant E. indica biotype in Alabama, providing additional germplasm for study of resistance to photosystem I electron-diverting (PSI-ED) resistance mechanisms.

Few options are available for controlling bermudagrass invasion of seashore paspalum. Bermudagrass and seashore paspalum tolerance to topramezone, triclopyr, or the combination of these two herbicides were evaluated in both greenhouse and field conditions. Field treatments included two sequential applications of topramezone (15.6 g ai ha−1) alone and five rates of topramezone + triclopyr (15.6 + 43.2, 15.6 + 86.3, 15.6 + 172.6, 15.6 + 345.2, or 15.6 g ai ha−1 + 690.4 g ae ha−1). Secondary greenhouse treatments included a single application of topramezone (20.8 g ha−1) or triclopyr (258.9 g ha−1) alone, or in combination at 20.8 + 258.9 or 20.8 + 517.8 g ha−1, respectively. Greenhouse and field results showed that topramezone applications in combination with triclopyr present opposite responses between bermudagrass and seashore paspalum. Topramezone increased bermudagrass injury and decreased seashore paspalum bleaching injury compared to topramezone alone. In field evaluations, topramezone + triclopyr at 15.6 + 690.4 g ha−1 used in sequential applications resulted in >90% injury to bermudagrass, however, injury decreased over time. Furthermore, sequential applications of topramezone + triclopyr at 15.6 + 690.4 g ha−1 resulted in >50% injury to seashore paspalum. Application programs including topramezone plus triclopyr should increase bermudagrass suppression and reduce seashore paspalum injury compared to topramezone alone. However, additional studies are needed because such practices will likely require manipulation of topramezone rate, application timing, application interval, and number of applications in order to maximize bermudagrass control and minimize seashore paspalum injury.

Goosegrass control options in bermudagrass are limited. Topramezone is one option that offers excellent control of mature goosegrass, but application to bermudagrass results in unacceptable symptoms of bleaching and necrosis typical of hydroxyphenylpyruvate dioxygenase inhibitors. Previous research has shown that adding chelated iron reduced the phytotoxicity of topramezone without reducing the efficacy of the herbicide, resulting in safening when applied to bermudagrass. Our objective was to examine additional iron sources to determine whether similar safening effects occur with other sources. Field trials were conducted in the summers of 2016 to 2018 (Auburn University). Mixtures of topramezone and methylated seed oil were combined with six different commercial iron sources, including sodium ferric ethylenediamine di-o-hydroxyphenyl-acetate (FeEDDHA), ferrous diethylenetriamine pentaacetic acid (FeDTPA), iron citrate, FeSO4, and a combination of iron oxide/sucrate/sulfate, some of which contained nitrogen. Bermudagrass necrosis and bleaching symptoms were visually rated on a 0% to 100% scale. Reflectance (normalized difference vegetation index) and clipping yield measurements were also collected. Application of FeDTPA and FeSO4 reduced symptoms of bleaching and necrosis when applied with topramezone. Other treatments that contained nitrogen did not reduce injury but did reduce bermudagrass recovery time following the appearance of necrosis. Inclusion of small amounts of nitrogen often negated the safening effects of FeSO4. The iron oxide/sucrate/sulfate product had no effect on bleaching or necrosis. Data suggest that the iron source had a differential effect on bleaching and necrosis reduction when applied in combination with topramezone to bermudagrass. Overall, FeSO4 and FeDTPA safened topramezone the most on bermudagrass.

POST goosegrass and other grassy weed control in bermudagrass is problematic. Fewer herbicides that can control goosegrass are available due to regulatory pressure and herbicide resistance. Alternative herbicide options that offer effective control are needed. Previous research demonstrates that topramezone controls goosegrass, crabgrass, and other weed species; however, injury to bermudagrass may be unacceptable. The objective of this research was to evaluate the safening potential of topramezone combinations with different additives on bermudagrass. Field trials were conducted at Auburn University during summer and fall from 2015 to 2018 and 2017 to 2018, respectively. Treatments included topramezone mixtures and methylated seed oil applied in combination with five different additives: triclopyr, green turf pigment, green turf paint, ammonium sulfate, and chelated iron. Bermudagrass bleaching and necrosis symptoms were visually rated. Normalized-difference vegetative index measurements and clipping yield data were also collected. Topramezone plus chelated iron, as well as topramezone plus triclopyr, reduced bleaching potential the best; however, the combination of topramezone plus triclopyr resulted in necrosis that outweighed reductions in bleaching. Masking agents such as green turf paint and green turf pigment were ineffective in reducing injury when applied with topramezone. The combination of topramezone plus ammonium sulfate should be avoided because of the high level of necrosis. Topramezone-associated bleaching symptoms were transient and lasted 7 to 14 d on average. Findings from this research suggest that chelated iron added to topramezone and methylated seed oil mixtures acted as a safener on bermudagrass.

Annual sedge (Cyperus compressus L.) populations with resistance to halosulfuron were identified in turfgrass at two new locations in Georgia. Research was conducted to evaluate (1) resistance levels to two acetolactate synthase (ALS) inhibitors, (2) ALS enzyme susceptibility, (3) genetic differences associated with resistance, and (4) differential levels of ALS gene expression in these biotypes. In dose–response experiments, the biotypes were >160 times resistant to halosulfuron but only 12 times more resistant to imazaquin compared with a susceptible biotype. In vitro enzyme assays indicated that resistant (R) biotypes required 6.1-fold greater concentrations of imazaquin to reduce ALS activity 50% compared with the susceptible (S) biotype. Both R biotypes had a similar Pro-197-Ser amino acid substitution in the ALS gene that confers resistance to sulfonylureas. Compared with the S biotype, R biotypes had 4.4 times higher ALS gene expression than the S biotype. No differences in gene copy number were found between any biotypes for the ALS gene. Overall, ALS-resistant C. compressus selected by halosulfuron use in turfgrass may be exhibiting partial susceptibility to imazaquin but complete resistance to sulfonylureas. Differential levels of susceptibility to ALS inhibitors for these biotypes are associated with the Pro-197-Ser substitution and enhanced expression of the ALS gene.

Pinoxaden is a POST acetyl coenzyme A carboxylase (ACCase) inhibitor in the phenylpyrazolin chemical family and is labelled for turfgrass use at broadcast rates of 35.5 to 71 g ai ha−1 and spot spray rates of 156 to 310 g ai ha−1. A greenhouse rate-response study was conducted to characterize the efficacy of pinoxaden against common grassy weeds. Weed species examined in this study were yellow foxtail, southern sandbur, annual bluegrass, roughstalk bluegrass, large crabgrass, dallisgrass, bahiagrass, goosegrass, and perennial ryegrass. Nonlinear regressions were modelled to determine visible injury rates (the application rate at which 50% of the weed species were injured and the 90% [I90] rate) and weight reduction rates (the application rate at which there was a 50% reduction in fresh weight and 90% reduction [WR90]) for each weed species. Only annual bluegrass, bahiagrass, and goosegrass had visible injury I90 values greater than the maximum labelled spot spray rate of 310 g ai ha−1. Annual bluegrass, bahiagrass, southern sandbur, and goosegrass all had WR90 values greater than the maximum labelled spot spray rate of 310 g ai ha−1. Results from this study indicate that the evaluated weed species can be ranked, according to visible injury I90 values, from most to least susceptible: perennial ryegrass > yellow foxtail > dallisgrass > large crabgrass > southern sandbur > roughstalk bluegrass > bahiagrass > goosegrass > annual bluegrass.

Southern crabgrass [Digitaria ciliaris (Retz.) Koeler] is an annual grass weed that commonly infests turfgrass, roadsides, wastelands, and cropping systems throughout the southeastern United States. Two biotypes of D. ciliaris (R1 and R2) with known resistance to cyclohexanediones (DIMs) and aryloxyphenoxypropionates (FOPs) previously collected from sod production fields in Georgia were compared with a separate susceptible biotype (S) collected from Alabama for the responses to pinoxaden and to explore the possible mechanisms of resistance. Increasing rates of pinoxaden (0.1 to 23.5 kg ha−1) were evaluated for control of R1, R2, and S. The resistant biotypes, R1 and R2, were resistant to pinoxaden relative to S. The S biotype was completely controlled at rates of 11.8 and 23.5 kg ha−1, resulting in no aboveground biomass at 14 d after treatment. Pinoxaden rates at which tiller length and aboveground biomass would be reduced 50% (I50) and 90% (I90) for R1, R2, and S ranged from 7.2 to 13.2 kg ha−1, 6.9 to 8.6 kg ha−1, and 0.7 to 2.1 kg ha−1, respectively, for tiller length, and 7.7 to 10.2 kg ha−1, 7.2 to 7.9 kg ha−1, and 1.6 to 2.3 kg ha−1, respectively, for aboveground biomass. Prior selection pressure from DIM and FOP herbicides could result in the evolution of D. ciliaris cross-resistance to pinoxaden herbicides. Amplification of the carboxyl-transferase domain of the plastidic ACCase by standard PCR identified a point mutation resulting in an Ile-1781-Leu amino acid substitution only for the resistant biotype, R1. Further cloning of PCR product surrounding the 1781 region yielded two distinct ACCase gene sequences, Ile-1781 and Leu-1781. The amino acid substitution, Ile-1781-Leu in both resistant biotypes (R1 and R2), however, was revealed by next-generation sequencing of RNA using Illumina platform. A point mutation in the Ile-1781 codon leading to herbicide insensitivity in the ACCase enzyme has been previously reported in other grass species. Our research confirms that the Ile-1781-Leu substitution is present in pinoxaden-resistant D. ciliaris.

A 3-yr watermelon experiment was established in fall 2013 to evaluate cover crop, polyethylene mulch, tillage, and herbicide application components for weed control, yield, and profitability. Conservation tillage, either with a cereal rye cover crop alone or integrated with polyethylene mulch, was compared to the standard industry practice of conventional tillage with bedded polyethylene mulch. The study also used a non-bedded conventional tillage system without polyethylene to determine polyethylene and cover crop residue effects. Within each of the four systems, herbicide treatments comprised halosulfuron applied (1) at 26.3 g ai ha–1 PRE, (2) at 26.3 g ai ha–1 POST, or (3) sequentially at 26.3 g ai ha–1 PRE and POST. Each system also had a nontreated control. In addition, clethodim was applied in all plots twice POST at 140 g ai ha–1, except for nontreated in each system. In 2014, polyethylene or cereal rye cover crop effectively controlled tall morningglory, coffee senna, and carpetweed early season in nontreated plots, whereas the integration of the two was effective at controlling common purslane. Tall morningglory and purslane control was insufficient late season regardless of production system and herbicide application. In 2015, polyethylene effectively controlled cutleaf eveningprimrose, sicklepod, and arrowleaf sida early season in nontreated plots. Yellow nutsedge control was insufficient late season regardless of production system and herbicide application. Utilizing sequential halosulfuron applications did not increase weed control over PRE or POST alone in all years. Polyethylene use resulted in yields higher than systems without in all years. Across all 3 yr, net returns were highest for polyethylene mulch systems. The results of this experiment underscore the need for more progress in developing integrated conservation systems for watermelon production. Effective herbicides, low-disturbance cultivation, and/or hand weeding are most likely the key to success in conservation specialty crop systems.

Goosegrass biotypes from golf courses in Richmond, VA (CCV) and New Bern, NC (RB) historically treated with oxadiazon were identified as resistant compared to susceptible standard (PBU) based on comparisons of oxadiazon applied preemergence at increasing rates (0.03 to 2.24 kg ha-1). Oxadiazon at rates ≤ 2.24 kg ha-1 rate did not prevent emergence of suspected resistant CCV and RB seedlings. PBU emergence was completely prevented at 0.14 kg ha-1. Based on percent seedling emergence relative to non-treated and percent above-ground biomass reduction relative to non-treated, the oxadiazon rate at which emergence would be reduced 50% (I50) or 90% (I90) ranged from 0.12 to 0.18 kg ha-1 or 10.83 to 85.57 kg ha-1, respectively for suspected resistant CCV and RB, compared to 0.03 to 0.4 kg ha-1 or 0.12 to 0.19 kg ha-1, respectively for susceptible standard PBU. Seedling emergence data predicted 7.9 and 3.0 times greater I90 values for CCV and RB, respectively compared to biomass data. All three biotypes were completely controlled by preemergence applied labeled rates of prodiamine and indaziflam. This is the first peer-reviewed report of evolved weed resistance to oxadiazon.

Methiozolin is a selective herbicide that has been reported to control annual bluegrass in creeping bentgrass putting greens. Golf course managers frequently tank-mix fertilizers with herbicides to reduce time and labor, but no information is available regarding such mixtures with methiozolin. Research was conducted to evaluate methiozolin for annual bluegrass control and creeping bentgrass safety when tank-mixed with ammonium sulfate or iron sulfate. Mixtures with ammonium sulfate did not influence annual bluegrass control while they did reduce creeping bentgrass injury in some instances. Mixtures with iron sulfate varied by experimental run but annual bluegrass control was either similar or increased while creeping bentgrass injury did not vary. Paclobutrazol was included as an alternative agrochemical comparison for annual bluegrass management; its application resulted in similar control and injury with and without iron sulfate addition, and injury and control were similar to methiozolin at appropriate rates. While some differences were observed, overall annual bluegrass and creeping bentgrass response to methiozolin was not affected by tank-mix nutrient partner relative to methiozolin applied alone.

Experiments were conducted in environmental chambers to the evaluate effects of photoperiod and temperature on Florida betony growth and development. Plants were exposed to two photoperiods, short day (9 h) and long day (9 + 3 h night interruption), and three day/night temperature regimes, 18/14, 22/18, and 26/22 C. After 10 wk of growth, shoot length and weight were 3.4 and 3.5 times greater, respectively, in the long-day photoperiod and with the 26 and 22 than with the 22 and 18 C day and night temperature regime, respectively. Shoot number, however, was greatest in the short-day photoperiod and at a lower temperature of 22/18 C. Shoot number in long day 22/18 C and 26/22 C environments increased asymptotically. No difference in root weight was observed between long- and short-day environments, but root weight increased with increasing temperature. Flowering and tuber production only occurred in long-day environments, with greater production of both at higher temperatures. Results provide a general framework for understanding Florida betony growth and development characteristics in the field and provide insights that should be considered in developing control strategies.