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Background: Early detection of herbicide resistance in weeds is crucial for successful implementation of integrated weed management. We conducted a herbicide resistance survey of the winter annual grasses feral rye (Secale cereale), downy brome (Bromus tectorum), and jointed goatgrass (Aegilops cylindrica) from Colorado winter wheat production areas for resistance to imazamox and quizalofop. Results: All samples were susceptible to quizalofop. All downy brome and jointed goatgrass samples were susceptible to imazamox. Out of 314 field collected samples, we identified three feral rye populations (named A, B, and C) that were imazamox resistant. Populations B and C had a target‐site mechanism with mutations in the Ser653 residue of the acetolactate synthase (ALS) gene to Asn in B and to Thr in C. Both populations B and C had greatly reduced ALS in vitro enzyme inhibition by imazamox. ALS feral rye protein modeling showed that steric interactions induced by the amino acid substitutions at Ser653 impaired imazamox binding. Individuals from population A had no mutations in the ALS gene. The ALS enzyme from population A was equally sensitive to imazamox as to known susceptible feral rye populations. Imazamox was degraded two times faster in population A compared with a susceptible control. An oxidized imazamox metabolite formed faster in population A and this detoxification reaction was inhibited by malathion. Conclusion: Population A has a nontarget‐site mechanism of enhanced imazamox metabolism that may be conferred by cytochrome P450 enzymes. This is the first report of both target‐site and metabolism‐based imazamox resistance in feral rye. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Winter annual grass species such as jointed goatgrass (Aegilops cylindrica Host), downy brome (Bromus tectorum L.), and feral rye (Secale cereale L.) negatively affect winter wheat (Triticum aestivum L.) yields. To generate a novel in‐crop selective herbicide trait for winter annual grass control, mutagenesis was used to generate multiple winter wheat lines resistant to the acetyl co‐A carboxylase (ACCase) inhibitor quizalofop p‐ethyl (QPE). Field trials were performed to assess crop safety for wheat lines with one or two homoeologous mutations and to assess winter annual grass control with QPE treatments. Crop safety parameters including visual injury rating, plant height, grain yield, and kernel weight were evaluated following QPE treatment at multiple growth stages. The susceptible winter wheat cultivar Hatcher displayed 100% crop injury when treated at tillering in early spring or at jointing in late spring. Single‐gene lines generally had crop injury and yield loss, especially at the jointing application timing. The two‐mutation cultivars Incline AX and LCS Fusion AX had low to no injury or yield loss from QPE applications at all timings and no injury from high rate QPE applications (93 and 185 g ha−1) in either the fall or the spring. Control of downy brome and feral rye was highest (92–99%), while jointed goatgrass control ranged from a low of 73% at 93 g a.i. ha−1 to a high of 98% at 109 a.i. ha−1 QPE. Our data indicate that the targeted winter annual grass weeds are controlled by QPE at rates that have acceptable crop safety for two‐mutation QPE‐resistant wheat cultivars. Core Ideas: Winter wheat cultivars with two acetyl co‐A carboxylase mutations had no yield loss from quizalofop treatment.Feral rye, downy brome, and jointed goatgrass were controlled by quizalofop.Quizalofop is a second postemergence selective mode of action for winter annual grasses. [ABSTRACT FROM AUTHOR]

Genomic‐based epidemiology can provide insight into the origins and spread of herbicide resistance mechanisms in weeds. We used kochia (Bassia scoparia) populations resistant to the herbicide glyphosate from across western North America to test the alternative hypotheses that (i) a single EPSPS gene duplication event occurred initially in the Central Great Plains and then subsequently spread to all other geographical areas now exhibiting glyphosate‐resistant kochia populations or that (ii) gene duplication occurred multiple times in independent events in a case of parallel evolution. We used qPCR markers previously developed for measuring the structure of the EPSPS tandem duplication to investigate whether all glyphosate‐resistant individuals had the same EPSPS repeat structure. We also investigated population structure using simple sequence repeat markers to determine the relatedness of kochia populations from across the Central Great Plains, Northern Plains and the Pacific Northwest. We found that the original EPSPS duplication genotype was predominant in the Central Great Plains where glyphosate resistance was first reported. We identified two additional EPSPS duplication genotypes, one having geographical associations with the Northern Plains and the other with the Pacific Northwest. The EPSPS duplication genotype from the Pacific Northwest seems likely to represent a second, independent evolutionary origin of a resistance allele. We found evidence of gene flow across populations and a general lack of population structure. The results support at least two independent evolutionary origins of glyphosate resistance in kochia, followed by substantial and mostly geographically localized gene flow to spread the resistance alleles into diverse genetic backgrounds. [ABSTRACT FROM AUTHOR]

BACKGROUND: Amaranthus palmeri is an aggressive and prolific weed species with major impact on agricultural yield and is a prohibited noxious weed across the Midwest. Morphological identification of A. palmeri from other Amaranthus species is extremely difficult in seeds, which has led to genetic testing for seed identification in commercial seed lots. RESULTS: We created an inexpensive and reliable genetic test based on novel, species‐specific, single nucleotide polymorphisms (SNPs) from GBS (Genotyping by Sequencing) data. We report three SNP‐based genetic tests for identifying A. palmeri alone or in a mixed pool of Amaranthus spp. Sensitivity ranged from 99.8 to 100%, specificity from 99.59 to 100%. Accuracy for all three tests is > 99.7%. All three are capable of reliably detecting one A. palmeri seed in a pool of 200 Amaranthus spp. seeds. The test was validated across 20 populations of A. palmeri, along with eight other Amaranthus species, the largest and most genetically diverse panel of Amaranthus samples to date. CONCLUSION: Our work represents a marked improvement over existing commercial assays resulting in an identification assay that is (i) accurate, (ii) robust, (iii) easy to interpret and (iv) applicable to both leaf tissue and pools of up to 200 seeds. Included is a data transformation method for calling of closely grouped competitive fluorescence assays. We also present a comprehensive GBS dataset from the largest geographic panel of Amaranthus populations sequenced. Our approach serves as a model for developing markers for other difficult to identify species. © 2021 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND: Bidens subalternans (greater beggarticks) is a tetraploid and troublesome weed infesting annual crops in most tropical regions of the world. A glyphosate‐resistant (GR) B. subalternans biotype was detected in a soybean field from Paraguay. A series of physiological and molecular analyses were conducted to elucidate its resistance mechanisms. RESULTS: The GR biotype had a high level of resistance (> 15‐fold LD50), relative to a glyphosate‐susceptible (GS) biotype. Shikimate accumulation was up to ten‐fold greater for GS compared with GR. We found no differences in sensitivity when plants were treated and kept under lower (10/4 °C) or higher temperatures (25/20 °C). GS and GR had the same relative EPSPS gene copy number, and similar glyphosate absorption and translocation rates. Neither biotype metabolized glyphosate. A double amino acid substitution (TIPT – Thr102Ile and Pro106Thr) was found in only one EPSPS allele from one of the two EPSPS homoeologs present in tetraploid GR B. subalternans. CONCLUSION: This is the first report of a TIPT double mutation conferring high levels of glyphosate resistance in a weed species. The presence of both wild‐type and TIPT mutant EPSPS on the polyploid genome of GR B. subalternans may offset a potential fitness cost, requiring additional research to confirm the absence of deleterious effects. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND: We previously identified a glyphosate‐resistant A. trifida phenotype from Wisconsin USA that showed a non‐rapid response to glyphosate. The mechanism of glyphosate resistance in this phenotype has yet to be elucidated. We conducted experiments to investigate non‐target‐site resistance and target‐site resistance mechanisms. The roles of glyphosate absorption, translocation, and metabolism in resistance of this phenotype have not been reported previously, nor have EPSPS protein abundance or mutations to the full‐length sequence of EPSPS. RESULTS: Whole‐plant dose–response results confirmed a 6.5‐level of glyphosate resistance for the resistant (R) phenotype compared to a susceptible (S) phenotype. Absorption and translocation of 14C‐glyphosate were similar between R and S phenotypes over 72 h. Glyphosate and AMPA concentrations in leaf tissue did not differ between R and S phenotypes over 96 h. In vivo shikimate leaf disc assays confirmed that glyphosate EC50 values were 4.6‐ to 5.4‐fold greater for the R than S phenotype. Shikimate accumulation was similar between phenotypes at high glyphosate concentrations (>1000 μM), suggesting that glyphosate entered chloroplasts and inhibited EPSPS. This finding was supported by results showing that EPSPS copy number and EPSPS protein abundance did not differ between R and S phenotypes, nor did EPSPS sequence at Gly101, Thr102, and Pro106 positions. Comparison of full‐length EPSPS sequences found five nonsynonymous polymorphisms that differed between R and S phenotypes. However, their locations were distant from the glyphosate target site and, therefore, not likely to affect enzyme‐glyphosate interaction. CONCLUSION: The results suggest that a novel mechanism confers glyphosate resistance in this A. trifida phenotype. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Increased copy number of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene confers resistance to glyphosate, the world's most-used herbicide. There are typically three to eight EPSPS copies arranged in tandem in glyphosate-resistant populations of the weed kochia (Kochia scoparia). Here, we report a draft genome assembly from a glyphosate-susceptible kochia individual. Additionally, we assembled the EPSPS locus from a glyphosate-resistant kochia plant by sequencing select bacterial artificial chromosomes from a kochia bacterial artificial chromosome library. Comparing the resistant and susceptible EPSPS locus allowed us to reconstruct the history of duplication in the structurally complex EPSPS locus and uncover the genes that are coduplicated with EPSPS , several of which have a corresponding change in transcription. The comparison between the susceptible and resistant assemblies revealed two dominant repeat types. Additionally, we discovered a mobile genetic element with a FHY3/FAR1-like gene predicted in its sequence that is associated with the duplicated EPSPS gene copies in the resistant line. We present a hypothetical model based on unequal crossing over that implicates this mobile element as responsible for the origin of the EPSPS gene duplication event and the evolution of herbicide resistance in this system. These findings add to our understanding of stress resistance evolution and provide an example of rapid resistance evolution to high levels of environmental stress. [ABSTRACT FROM AUTHOR]

Abstract: There have been previous calls for, and efforts focused on, realizing the power and potential of weed genomics for better understanding of weeds. Sustained advances in genome sequencing and assembly technologies now make it possible for individual research groups to generate reference genomes for multiple weed species at reasonable costs. Here, we present the outcomes from several meetings, discussions, and workshops focused on establishing an International Weed Genomics Consortium (IWGC) for a coordinated international effort in weed genomics. We review the ‘state of the art’ in genomics and weed genomics, including technologies, applications, and on‐going weed genome projects. We also report the outcomes from a workshop and a global survey of the weed science community to identify priority species, key biological questions, and weed management applications that can be addressed through greater availability of, and access to, genomic resources. Major focus areas include the evolution of herbicide resistance and weedy traits, the development of molecular diagnostics, and the identification of novel targets and approaches for weed management. There is increasing interest in, and need for, weed genomics, and the establishment of the IWGC will provide the necessary global platform for communication and coordination of weed genomics research. © 2018 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: Synthetic auxins such as 2,4‐dichlorophenoxyacetic acid (2,4‐D) have been widely used for selective control of broadleaf weeds since the mid‐1940s. In 2009, an Amaranthus tuberculatus (common waterhemp) population with 10‐fold resistance to 2,4‐D was found in Nebraska, USA. The 2,4‐D resistance mechanism was examined by conducting [14C] 2,4‐D absorption, translocation and metabolism experiments. RESULTS: No differences were found in 2,4‐D absorption or translocation between resistant and susceptible A. tuberculatus plants. Resistant plants metabolized [14C] 2,4‐D more rapidly than did susceptible plants. The half‐life of [14C] 2,4‐D in susceptible plants was 105 h, compared with 22 h in resistant plants. Pretreatment with the cytochrome P450 inhibitor malathion inhibited [14C] 2,4‐D metabolism in resistant plants and reduced the 2,4‐D dose required for 50% growth inhibition (GR50) of resistant plants by 7‐fold to 27 g ha‐1, similar to the GR50 for susceptible plants in the absence of malathion. CONCLUSION: Our results demonstrate that rapid 2,4‐D metabolism is a contributing factor to resistance in A. tuberculatus, potentially mediated by cytochrome P450. Metabolism‐based resistance to 2,4‐D could pose a serious challenge for A. tuberculatus control because of the potential for cross‐resistance to other herbicides. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: Resistance to the synthetic auxin herbicide dicamba is increasingly problematic in Kochia scoparia. The resistance mechanism in an inbred dicamba‐resistant K. scoparia line (9425R) was investigated using physiological and transcriptomics (RNA‐Seq) approaches. RESULTS: No differences were found in dicamba absorption or metabolism between 9425R and a dicamba‐susceptible line, but 9425R was found to have significantly reduced dicamba translocation. Known auxin‐responsive genes ACC synthase (ACS) and indole‐3‐acetic acid amino synthetase (GH3) were transcriptionally induced following dicamba treatment in dicamba‐susceptible K. scoparia but not in 9425R. Chalcone synthase (CHS), the gene regulating synthesis of the flavonols quertecin and kaemperfol, was found to have twofold higher transcription in 9425R both without and 12 h after dicamba treatment. Increased CHS transcription co‐segregated with dicamba resistance in a forward genetics screen using an F2 population. CONCLUSION: Prior work has shown that the flavonols quertecin and kaemperfol compete with auxin for intercellular movement and vascular loading via ATP‐binding cassette subfamily B (ABCB) membrane transporters. The results of this study support a model in which constitutively increased CHS expression in the meristem produces more flavonols that would compete with dicamba for intercellular transport by ABCB transporters, resulting in reduced dicamba translocation. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: The glyphosate‐resistant rapid response (GR RR) resistance mechanism in Ambrosia trifida is not due to target‐site resistance (TSR) mechanisms. This study explores the physiology of the rapid response and the possibility of reduced translocation and vacuolar sequestration as non‐target‐site resistance (NTSR) mechanisms. RESULTS: GR RR leaf discs accumulated hydrogen peroxide within minutes of glyphosate exposure, but only in mature leaf tissue. The rapid response required energy either as light or exogenous sucrose. The combination of phenylalanine and tyrosine inhibited the rapid response in a dose‐dependent manner. Reduced glyphosate translocation was observed in GR RR, but only when associated with tissue death caused by the rapid response. Nuclear magnetic resonance studies indicated that glyphosate enters the cytoplasm and reaches chloroplasts, and it is not moved into the vacuole of GR RR, GR non‐rapid response or glyphosate‐susceptible A. trifida. CONCLUSION: 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 [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: Glyphosate‐resistant (GR) Ambrosia trifida is now present in the midwestern United States and in southwestern Ontario, Canada. Two distinct GR phenotypes are known, including a rapid response (GR RR) phenotype, which exhibits cell death within hours after treatment, and a non‐rapid response (GR NRR) phenotype. The mechanisms of resistance in both GR RR and GR NRR remain unknown. Here, we present a description of the RR phenotype and an investigation of target‐site mechanisms on multiple A. trifida accessions. RESULTS: Glyphosate resistance was confirmed in several accessions, and whole‐plant levels of resistance ranged from 2.3‐ to 7.5‐fold compared with glyphosate‐susceptible (GS) accessions. The two GR phenotypes displayed similar levels of resistance, despite having dramatically different phenotypic responses to glyphosate. Glyphosate resistance was not associated with mutations in EPSPS sequence, increased EPSPS copy number, EPSPS quantity, or EPSPS activity. CONCLUSION: 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 [ABSTRACT FROM AUTHOR]

Glyphosate-resistant (GR) Kochia scoparia has evolved in dryland chemical fallow systems throughout North America and the mechanism of resistance involves 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene duplication. Agricultural fields in four states were surveyed for K. scoparia in 2013 and tested for glyphosate-resistance level and EPSPS gene copy number. Glyphosate resistance was confirmed in K. scoparia populations collected from sugarbeet fields in Colorado, Wyoming, and Nebraska, and Montana. Glyphosate resistance was also confirmed in K. scoparia accessions collected from wheat-fallow fields in Montana. All GR samples had increased EPSPS gene copy number, with median population values up to 11 from sugarbeet fields and up to 13 in Montana wheat-fallow fields. The results indicate that glyphosate susceptibility can be accurately diagnosed using EPSPS gene copy number. [ABSTRACT FROM AUTHOR]

Weed populations can have high genetic plasticity and rapid responses to environmental selection pressures. For example, 100-fold amplification of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene evolved in the weed species Amaranthus palmeri to confer resistance to glyphosate, the world’s most important herbicide. However, the gene amplification mechanism is unknown. We sequenced the EPSPS gene and genomic regions flanking EPSPS loci in A. palmeri, and searched for mobile genetic elements or repetitive sequences. The EPSPS gene was 10,229 bp, containing 8 exons and 7 introns. The gene amplification likely proceeded through a DNA-mediated mechanism, as introns exist in the amplified gene copies and the entire amplified sequence is at least 30 kb in length. Our data support the presence of two EPSPS loci in susceptible (S) A. palmeri, and that only one of these was amplified in glyphosate-resistant (R) A. palmeri. The EPSPS gene amplification event likely occurred recently, as no sequence polymorphisms were found within introns of amplified EPSPS copies from R individuals. Sequences with homology to miniature inverted-repeat transposable elements (MITEs) were identified next to EPSPS gene copies only in R individuals. Additionally, a putative Activator (Ac) transposase and a repetitive sequence region were associated with amplified EPSPS genes. The mechanism controlling this DNA-mediated amplification remains unknown. Further investigation is necessary to determine if the gene amplification may have proceeded via DNA transposon-mediated replication, and/or unequal recombination between different genomic regions resulting in replication of the EPSPS gene. [ABSTRACT FROM AUTHOR]

A previously unknown glyphosate resistance mechanism, amplification of the 5-enolpyruvyl shikimate-3-phosphate synthase gene, was recently reported in Amaranthus palmeri. This evolved mechanism could introgress to other weedy Amaranthus species through interspecific hybridization, representing an avenue for acquisition of a novel adaptive trait. The objective of this study was to evaluate the potential for this glyphosate resistance trait to transfer via pollen from A. palmeri to five other weedy Amaranthus species ( Amaranthus hybridus, Amaranthus powellii, Amaranthus retroflexus, Amaranthus spinosus, and Amaranthus tuberculatus). Field and greenhouse crosses were conducted using glyphosate-resistant male A. palmeri as pollen donors and the other Amaranthus species as pollen recipients. Hybridization between A. palmeri and A. spinosus occurred with frequencies in the field studies ranging from <0.01% to 0.4%, and 1.4% in greenhouse crosses. A majority of the A. spinosus × A. palmeri hybrids grown to flowering were monoecious and produced viable seed. Hybridization occurred in the field study between A. palmeri and A. tuberculatus (<0.2%), and between A. palmeri and A. hybridus (<0.01%). This is the first documentation of hybridization between A. palmeri and both A. spinosus and A. hybridus. [ABSTRACT FROM AUTHOR]

Inheritance of glyphosate resistance in a Palmer amaranth biotype from North Carolina was studied. Glyphosate rates for 50% survival of glyphosate-resistant (GR) and glyphosate-susceptible (GS) biotypes were 1288 and 58 g ha-1, respectively. These values for F1 progenies obtained from reciprocal crosses (GR × GS and GS × GR) were 794 and 501 g ha-1, respectively. Dose response of F1 progenies indicated that resistance was not fully dominant over susceptibility. Lack of significant differences between dose responses for reciprocal F1 families suggested that genetic control of glyphosate resistance was governed by nuclear genome. Analysis of F1 backcross (BC1F1) families showed that 10 and 8 BC1F1 families out of 15 fitted monogenic inheritance at 2000 and 3000 g ha-1 glyphosate, respectively. These results indicate that inheritance of glyphosate resistance in this biotype is incompletely dominant, nuclear inherited, and might not be consistent with a single gene mechanism of inheritance. Relative 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) copy number varied from 22 to 63 across 10 individuals from resistant biotype. This suggested that variable EPSPS copy number in the parents might be influential in determining if inheritance of glyphosate resistance is monogenic or polygenic in this biotype. [ABSTRACT FROM AUTHOR]

The herbicide glyphosate became widely used in the United States and other parts of the world after the commercialization of glyphosate-resistant crops. These crops have constitutive overexpression of a glyphosate-insensitive form of the herbicide target site gene, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Increased use of glyphosate over multiple years imposes selective genetic pressure on weed populations. We investigated recently discovered glyphosate-resistantAmaranthuspalmeri populations from Georgia, in comparison with normally sensitive populations. EPSPS enzyme activity from resistant and susceptible plants was equally inhibited by glyphosate, which led us to use quantitative PCR to measure relative copy numbers of the EPSPS gene. Genomes of resistant plants contained from 5-fold to more than 160-fold more copies of the EPSPS gene than did genomes of susceptible plants. Quantitative RT-PCR on cDNA revealed that EPSPS expression was positively correlated with genomic EPSPS relative copy number. Immunoblot analyses showed that increased EPSPS protein level also correlated with EPSPSgenomic copy number. EPSPS gene amplification was heritable, correlated with resistance in pseudo-F2 populations, and is proposed to be the molecular basis of glyphosate resistance. FISH revealed that EPSPS genes were present on every chromosome and, therefore, gene amplification was likely not caused by unequal chromosome crossing over. This occurrence of gene amplification as an herbicide resistance mechanism in a naturally occurring weed population is particularly significant because it could threaten the sustainable use of glyphosate-resistant crop technology. [ABSTRACT FROM AUTHOR]

Field studies using glyphosate-resistant corn (Zea mays L.), sugarbeet (Beta vulgaris L.), and spring wheat (Triticum aestivum L.) were conducted at Scottsbluff, NE, Torrington, WY, and Fort Collins, CO, over 6 yr to evaluate weed population dynamics under glyphosate- and conventional herbicide-based weed control strategies. We report the response of common lambsquarters (Chenopodium album L.), wild buckwheat (Polygonum convolvulus L.), and all grasses combined to four management strategies imbedded in two crop rotations. There were no consistent or highly significant benefits from a diverse crop rotation versus continuous corn when weed control was considered. Rotating herbicide mode of action for general weed control provided no benefit above that provided by glyphosate at 0.8 kg acid equivalent (ae) ha-1 applied twice each year. There was no evidence that any weed species developed resistance to glyphosate. The most striking finding of these studies was the buildup of common lambs- quarters and wild buckwheat that occurs when glyphosate is applied at 0.4 kg ae ha-1 twice each year. Such a low-use-rate approach is to be completely discouraged since it appears to enrich the gene pool for individuals that survive low rates of glyphosate. The continuing decline in the real cost of glyphosate should eliminate any legitimate reason for growers to consider using less than the full labeled rate of glyphosate. All treatments that included glyphosate at all three locations over 6 yr provided significantly better grass control than the nonglyphosate conventional treatments. [ABSTRACT FROM AUTHOR]

The potential introduction of wheat (Triticum aestivum L.) cultivars with transgenic traits has generated increased interest in pollen-mediated gene flow (PMGF). The objectives of this study were to estimate wheat PMGF between commercial fields across multiple years and locations, and to compare estimates from large fields to those from smaller experimental plots. The study was conducted in a total of 56 commercial field locations in eastern Colorado in 2003, 2004, and 2005. We measured PMGF by tracking the movement of an imidazolinone herbicide resistance gene from resistant to susceptible cultivars, sampled at distances of 0.23 to 61 m. At least one sample from all 56 fields and from all 18 evaluated cultivars had detectable PMGF. The highest observed PMGF was 5.3% at 0.23 m. The farthest distance at which PMGF was detected was 61 m and the highest PMGF at that distance was 0.25%. Higher levels and greater distances of PMGF were detected in commercial fields than in experimental plots. Based on estimates from a generalized linear mixed model with a random location effect, the distance required to ensure 95% confidence that 95% of locations have PMGF less than 0.9% is 41.1 m for cultivars heading earlier than the pollen source and 0.7 m for cultivars heading later than the pollen source. These confidence limits should represent the highest levels of PMGF expected to occur in winter wheat in the west-central Great Plains and will be useful for wheat biotechnology regulation. [ABSTRACT FROM AUTHOR]