Accounting for 53% of United States peanuts (Arachis hypogaea L.), Georgia is the top peanut-producing state with approximately 1.42 billion kg produced in 2023. Peanut producers often use the ALS imidazolinone herbicide imazapic but reduced yellow nutsedge (Cyperus esculentus L.) control was reported in Georgia peanuts after four-years of continuous imazapic use. This study aimed to determine the level of resistance (LD50, I50, and GR50), potential cross-resistance for the suspected resistant population, and identify the associated genetic mutations conferring resistance. A susceptible biotype was treated with 0, 0.0088, 0.0175, 0.035, 0.07, 0.14, 0.28, and 0.56 kg ai ha-1, and a resistant biotype was sprayed with 0, 0.07, 0.14, 0.28, 0.56, 1.13, 2.26, and 4.5 kg ai ha-1 of imazapic. To determine if the suspected resistant biotype was cross-resistant to halosulfuron-methyl, an ALS herbicide used to control nutsedge spp., both biotypes were treated with 0, 0.0117, 0.0233, 0.0466, 0.0933, 0.187, 0.373, and 0.746 g ai ha-1 of halosulfuron-methyl. Plants were rated for injury 7, 14, and 28 days after treatment (DAT), and above-ground biomass harvested at 28 DAT. For imazapic, LD50 was 0.041 and 1.503 kg ai ha-1 and the GR50 was estimated to be 0.0128 and 1.853 kg ai ha-1 for Sus and Res biotypes, respectively, indicating 36 and 145-fold increase in resistance of the Res biotype for I50 and GR50, respectively. Both biotypes responded similarly to applications of halosulfuron-methyl, with biomass reductionat rates greater than 0.023 kg ai ha-1. Transcriptome profiles revealed a mutation in the target-site gene of the resistant biotype, causing an amino acid substitution from Alanine to Valine at position 205 (Ala205Val). Growers should continue to rotate chemistries and implement integrated weed management approaches for control of C. esculentus as the use of imazapic over consecutive years has led to resistance in C. esculentus.

Palmer amaranth (Amaranthus palmeri S. Watson) was first reported in Iowa in 2013 and has continued to spread across the state over the last decade. Amaranthus palmeri is widely recognized as one of the more economically important weeds in production agriculture. The presence of A. palmeri in Iowa is concerning as the species has evolved resistance to ten herbicide sites of action, however, no formal characterization has been conducted on Iowa populations. Therefore, herbicide assays were conducted on an A. palmeri population collected in Harrison County, IA, in 2023 (Southwest Palmer Amaranth [SWPA]) and a known herbicide-susceptible population collected from Nebraska in 2001 (Palmer Amaranth Susceptible [PAS]). The two populations were treated with preemergence and postemergence herbicides commonly used in Iowa. The treatments included preemergence applications of atrazine, metribuzin, and mesotrione and postemergence applications of atrazine, imazethapyr, glyphosate, lactofen, mesotrione, glufosinate, 2,4-D, and dicamba at 1× and 4× the labeled rates. Survival frequency of SWPA was >90% when treated postemergence with 1× rates of imazethapyr, atrazine, glyphosate, and mesotrione compared with ≤6% for PAS. Both SWPA and PAS had 0% survival when treated with lactofen, glufosinate, 2,4-D, and dicamba at the 1× or 4× rates. Plant population density reduction for SWPA was 53% and 40% in response to 1× rates of preemergence-applied mesotrione and atrazine, respectively. Metribuzin applied preemergence reduced SWPA plant population density by >90% at both rates. Dose–response experiments revealed the 50% effective doses (ED50) of mesotrione, glyphosate, imazethapyr, and atrazine for SWPA were 9.5-,8.5-, 71-, and 40-fold greater than for PAS, respectively. The results confirm that SWPA is four-way multiple-herbicide resistant. Amaranthus palmeri infestations are likely to continue to spread within Iowa; therefore, diversified weed management programs that include early detection, rapid response, and effective multi-tactic management strategies will be required for control.

Experiments were initiated to characterize a waterhemp population (CHR) discovered in a central Illinois corn field after it was not controlled by the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor topramezone. Field experiments conducted during 2014–2015 indicated that acetolactate synthase (ALS)-, protoporphyrinogen oxidase (PPO)-, photosystem II (PSII)-, and HPPD-inhibiting herbicides and the synthetic auxin 2,4-D did not control the CHR population. Laboratory experiments confirmed target site–based resistance mechanisms to ALS- and PPO-inhibiting herbicides. Herbicide doses required to reduce dry biomass 50% (GR50) were determined in greenhouse dose–response experiments, and indicated 16-fold resistance to the HPPD inhibitor mesotrione, 9.5-fold resistance to the synthetic auxin 2,4-D, and 252-fold resistance to the PSII inhibitor atrazine. Complementary results from field, laboratory, and greenhouse investigations indicate that the CHR population has evolved resistance to herbicides from five sites of action (SOAs): ALS-, PPO-, PSII-, and HPPD-inhibiting herbicides and 2,4-D. Herbicide use history for the field in which CHR was discovered indicates no previous use of 2,4-D.

Giant ragweed is a highly competitive weed that continually threatens crop production systems due to evolved resistance to acetolactate synthase–inhibiting herbicides (ALS-R) and glyphosate (GR). Two biotypes of GR giant ragweed exist and are differentiated by their response to glyphosate, termed here as rapid response (RR) and non–rapid response (NRR). A comparison of data from surveys of Indiana crop fields done in 2006 and 2014 showed that GR giant ragweed has spread from 15% to 39% of Indiana counties and the NRR biotype is the most prevalent. A TaqMan® single-nucleotide polymorphism genotyping assay was developed to identify ALS-R populations and revealed 47% of GR populations to be ALS-R as well. The magnitude of glyphosate resistance for NRR populations was 4.6 and 5.9 based on GR50 and LD50 estimates, respectively. For RR populations, these values were 7.8 to 9.2 for GR50 estimates and 19.3 to 22.3 for LD50 estimates. A novel use of the Imaging-PAM fluorometer was developed to discriminate RR plants by assessing photosystem II quantum yield across the entire leaf surface. H2O2 generation in leaves of glyphosate-treated plants was also measured by 3,3′-diaminobenzidine staining and quantified using imagery analysis software. Results show photo-oxidative stress of mature leaves is far greater and occurs more rapidly following glyphosate treatment in RR plants compared with NRR and glyphosate-susceptible plants and is positively associated with glyphosate dose. These results suggest that under continued glyphosate selection pressure, the RR biotype may surpass the NRR biotype as the predominant form of GR giant ragweed in Indiana due to a higher level of glyphosate resistance. Moreover, the differential photo-oxidative stress patterns in response to glyphosate provide evidence of different mechanisms of resistance present in RR and NRR biotypes.

The widespread occurrence of Palmer amaranth resistant to acetolactate synthase inhibitors and/or glyphosate led to the increased use of protoporphyrinogen oxidase (PPO)-inhibiting herbicides. This research aimed to: (1) evaluate the efficacy of foliar-applied fomesafen to Palmer amaranth, (2) evaluate cross-resistance to foliar PPO inhibitors and efficacy of foliar herbicides with different mechanisms of action, (3) survey the occurrence of the PPO Gly-210 deletion mutation among PPO inhibitor–resistant Palmer amaranth, (4) identify other PPO target-site mutations in resistant individuals, and (5) determine the resistance level in resistant accessions with or without the PPO Gly-210 deletion. Seedlings were sprayed with fomesafen (263 gaiha−1), dicamba (280 gaiha−1), glyphosate (870 gaiha−1), glufosinate (549 g ai ha−1), and trifloxysulfuron (7.84 gaiha−1). Selected fomesafen-resistant accessions were sprayed with other foliar-applied PPO herbicides. Mortality and injury were evaluated 21 d after treatment (DAT). The PPX2L gene of resistant and susceptible plants from a selected accession was sequenced. The majority (70%) of samples from putative PPO-resistant populations in 2015 were confirmed resistant to foliar-applied fomesafen. The efficacy of other foliar PPO herbicides on fomesafen-resistant accessions was saflufenacil>acifluorfen=flumioxazin>carfentrazone=lactofen>pyraflufen-ethyl>fomesafen>fluthiacet-methyl. With small seedlings, cross-resistance occurred with all foliar-applied PPO herbicides except saflufenacil (i.e., 25% with acifluorfen, 42% with flumioxazin). Thirty-two percent of PPO-resistant accessions were multiple resistant to glyphosate and trifloxysulfuron. Resistance to PPO herbicides in Palmer amaranth occurred in at least 13 counties in Arkansas. Of 316 fomesafen survivors tested, 55% carried the PPO Gly-210 deletion reported previously in common waterhemp. The PPO gene (PPX2L) in one accession (15CRI-B), which did not encode the Gly-210 deletion, encoded an Arg-128-Gly substitution. The 50% growth reduction values for fomesafen in accessions with Gly-210 deletion were 8- to 15-fold higher than that of a susceptible population, and 3- to 10-fold higher in accessions without the Gly-210 deletion.

A fixed-plot management study for control of acetolactate synthase (ALS)–resistant common cocklebur in soybean was initiated in 1994 at Fayetteville, AR. Three susceptible and three imazaquin-resistant common cocklebur plants were transplanted into the field, and seed (burs) were distributed throughout the plots in the fall of 1994. Herbicide treatments included imazaquin, chlorimuron, and chlorimuron plus metribuzin applied each year from 1995 through 1999 and herbicide rotations containing ALS inhibitors and herbicides with alternative modes of action. Effectiveness of management systems and the dynamics of the development of common cocklebur resistance, including development of resistance to imazaquin and chlorimuron, were evaluated. Imazaquin controlled susceptible common cocklebur populations in 1995 but not the resistant population, resulting in significant soybean yield reduction. By the end of the 1996 season, the resistant biotype dominated imazaquin plots, and a high level of cross-resistance to chlorimuron was observed in the population. Resistant populations were reduced by non-ALS herbicide programs of sulfentrazone plus clomazone applied preemergence (PRE), metribuzin plus clomazone applied PRE followed by bentazon applied postemergence (POST), and transgenic herbicide programs of glyphosate and glufosinate applied POST. Rotating ALS inhibitors with non–ALS-inhibiting heribicides may slow the development of resistance, but resistant individuals may eventually dominate the population.

In 2006 and 2007, farmers from two counties in Illinois reported failure to control waterhemp with glyphosate. Subsequent onsite field experiments revealed that the populations might be resistant to multiple herbicides. Greenhouse experiments therefore were conducted to confirm glyphosate resistance, and to test for multiple resistance to other herbicides, including atrazine, acifluorfen, lactofen, and imazamox. In glyphosate dose-response experiments, both populations responded similarly to a previously characterized glyphosate-resistant population (MO1). Both Illinois populations also demonstrated high frequencies of resistance to the acetolactate synthase (ALS) inhibitor, imazamox. Additionally, one of the populations demonstrated high frequencies of resistance to both atrazine and the protoporphyrinogen oxidase (PPO) inhibitor, lactofen. Furthermore, using combinations of sequential and tank-mix herbicide applications, individual plants resistant to herbicides spanning all four site-of-action groups were identified from one population. Molecular experiments were performed to provide an initial characterization of the resistance mechanisms and to provide confirmation of the presence of multiple resistance traits within the two populations. Both populations contained the W574L ALS mutation and the ΔG210 PPO mutation, previously shown to confer resistance to ALS and PPO inhibitors, respectively. Atrazine resistance in both populations is suspected to be metabolism-based, because a triazine target-site mutation was not identified. A P106S EPSPS mutation, previously reported to confer glyphosate resistance, was identified in one population. This mutation was identified in both resistant and sensitive plants from the population; however, and so more research is needed to determine the glyphosate-resistance mechanism(s). This is the first known case of a weed population in the United States possessing multiple resistance to herbicides from four site-of-action groups.

Imazamox-resistant wheat varieties carry the Imi1 allele, which confers resistance to the imidazolinone (IMI) herbicide imazamox. This resistance trait allows the selective control of jointed goatgrass, a difficult-to-control winter annual grass weed. Allele movement between IMI-resistant wheat and jointed goatgrass may occur via hybridization and backcross events. Hybrids (F1) of IMI-resistant wheat and jointed goatgrass were identified in 2008 in a commercial wheat field in Eastern Oregon. In 2009 and 2010, surveys were conducted in Eastern Oregon to determine the prevalence of the Imi1 allele in wheat × jointed goatgrass hybrids. Using polymerase chain reaction assays we detected the presence of the Imi1 allele. A total of 128 sites were surveyed over the 2 yr. Of 1,548 plants sampled, 1,100 were positive for the Imi1 alelle and of those, 1,087 were heterozygous and 13 were homozygous for the allele. We assessed hybrid yield components and how these components varied across the sampled sites. The association between the proportion of IMI-resistant hybrids and the area or management practice in the commercial fields was determined. Nonagricultural sites or production of IMI-resistant wheat in consecutive years were two factors associated with a greater proportion of IMI-resistant hybrids. Our results demonstrate that the Imi1 allele is moving from IMI-resistant wheat to jointed goatgrass, producing resistant hybrids and backcross plants. This is the first report of natural occurrence of IMI-resistant backcross plants in commercial wheat fields. Therefore, it is important to implement field management practices that reduce IMI-resistant hybrid production and to effectively manage nonagricultural areas with jointed goatgrass infestations to prevent introgression of the IMI-resistance allele.

A set of universal and degenerate primers has been designed to clone (by polymerase chain reaction [PCR]) the conserved domains of the acetolactate synthase (ALS) gene where mutations confer resistance to ALS herbicides in plants. These primers were successful in cloning conserved domains of ALS in all monocotyledonous and dicotyledonous plants tested to date, as well as that of bacteria. Total genomic DNA was used as the source of target DNA because no introns were found in the sequences to be amplified. The design of the universal primers was performed after subtle modifications of the consensus degenerate hybrid oligonucleotide primers approach, which implies the synthesis of hybrid oligonucleotide primers containing fixed clamp 5′ and degenerate core 3′ sequences. Optimizations of PCR reactions were done according to a Taguchy approach described for the first time with degenerate oligonucleotides. This method optimizes a PCR reaction using four variables (deoxynucleoside triphosphate, DNA, primers, and Mg2+) under three different concentrations per variable using just nine reactions. The ALS herbicide-binding domains from many susceptible and resistant plants can be cloned and sequenced in a few hours by using only 100 mg of starting plant material, like one leaf or several small seedlings or seeds.

In 1996 a common sunflower population near Howard, SD, was suspected to be cross-resistant to imazethapyr and chlorimuron. Whole-plant acetolactate synthase (ALS) assays confirmed ALS-inhibitor resistance in the Howard biotype. The I 50 values (inhibition of 50% of the enzyme activity) indicated that the resistant population required 39 and 9 times more imazethapyr and chlorimuron, respectively, to obtain the same level of enzyme inhibition compared with the sensitive biotype. Herbicide dose response data supported the whole-plant enzyme assay data; control (> 90%) was not achieved with less than a four-times application rate of chlorimuron. Control with imazethapyr was not achieved even with a 16-times rate. Chlorimuron and imazethapyr controlled 70 and 95% of the population, respectively, when a four-times rate of each herbicide was applied separately. Differences in 14C-herbicide absorption were observed, suggesting that there may be physical or chemical differences in leaf surface composition between the resistant and sensitive biotypes. Although translocation of 14C-herbicide was less in the resistant biotype than in the sensitive biotype, the differences were not enough to explain chlorimuron and imazethapyr selectivity between the two biotypes. Overall results suggested that the differences in the common sunflower populations were attributed to an altered site of action on the ALS enzyme.