The resistance to fenoxaprop-P-ethyl, a herbicide that inhibits acetyl-CoA carboxylase (ACCase), has emerged in shortawn foxtail (Alopecurus aequalis Sobol.) since the 1990s, presenting a considerable challenge to wheat (Triticum aestivum L.) production in China. One of the primary mechanisms responsible for this high-level resistance is the presence of mutations at codons 1781, 2041, and 2078 in the ACCase gene. However, the conventional methods used to detect these mutations, such as polymerase chain reaction (PCR) and gene sequencing, are time-consuming and labor-intensive. To address this issue and enable the prompt and effective detection of these common ACCase mutations in A. aequalis, a loop-mediated isothermal amplification (LAMP) strategy was developed. The LAMP assay specifically targets the Ile-1781-Leu and Asp-2078-Gly mutations within the ACCase gene of A. aequalis. Through the optimization of primers, systems, and conditions, the LAMP assay enables rapid differentiation between wild-type individuals and mutants of A. aequalis carrying either of these two mutations. Including SYBR Green I dye in the final reaction mixtures enables detection of the target mutation through a noticeable color change that can be observed with the naked eye. It is noteworthy that the sensitivity of the LAMP assay was approximately 104-fold greater than that of conventional PCR methods. Additionally, a derived cleaved amplified polymorphic sequence (dCAPS) assay was established for each mutation to distinguish between homozygous and heterozygous mutants. Overall, the developed LAMP assay could efficiently detect the Ile-1781-Leu and Asp-2078-Gly mutations in the ACCase gene of A. aequalis, offering significant advantages for the monitoring and management of fenoxaprop-P-ethyl resistance.

Quizalofop-resistant rice allows for over-the-top applications of quizalofop, a herbicide that inhibits acetyl-coenzyme A carboxylase. However, previous reports have indicated that quizalofop applied postemergence may cause significant injury to quizalofop-resistant rice. Therefore, field experiments were conducted to evaluate the response of quizalofop-resistant rice cultivars to quizalofop applications across different planting dates. Under controlled conditions, the effects of soil moisture content, air temperature, and light intensity on quizalofop-resistant rice sensitivity to quizalofop were investigated. In the planting date experiment, injury of more than 11 percentage points was observed on early-planted rice compared with late-planted rice at the 5-leaf stage, with higher injury observed under saturated soil conditions. However, quizalofop applications at the labeled rate caused ≤16% reduction in yield regardless of planting environment. Quizalofop-resistant cultivars exhibited more injury by at least 25 percentage points when soil was maintained at 90% or 100% of field capacity because rice cultivars ‘PVL01’, ‘PVL02’, and ‘RTv7231 MA’ exhibited ≥42%, 30%, and ≥54% injury, respectively, compared with ≤10%, ≤5%, and ≤22% injury, respectively, at 40% or 50% of field capacity, pooled over rating timing. Greater injury ranging from 18% to 31% was observed on quizalofop-resistant rice grown under low light intensity (600 µmol m−2s−1) compared with 5% to 14% injury under high light intensity (1,150 µmol m−2s−1). The injury persisted from 7 to 28 d after 5-leaf stage application (DAFT), averaged over quizalofop-resistant cultivars and air temperatures (20/15 C and 30/25 C day/night, respectively). At 7 DAFT, greater injury (by 5 to 21 percentage points) was observed on quizalofop-resistant cultivars; PVL01, PVL02, and RTv7231 MA exhibited 33%, 9%, and 58% injury, respectively, under 20/15 C temperature conditions compared with 13%, 4%, and 37% injury, respectively, under 30/25 C day/night conditions averaged over light intensities. Overall, quizalofop is likely to cause a greater risk for injury to quizalofop-resistant rice if it is applied under cool, cloudy, and moist soil conditions.

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.

Chinese sprangletop [Leptochloa chinensis (L.) Nees] is one grass weed severely affecting rice (Oryza sativa L.) growth in paddies in China. Cyhalofop-butyl is the main herbicide used to control grass weeds in Chinese paddy fields, especially for controlling L. chinensis; however, L. chinensis has evolved resistance to cyhalofop-butyl due to continuous and extensive application. To investigate cyhalofop-butyl resistance levels and mechanisms in L. chinensis in some of the Chinese rice areas, 66 field populations were collected and treated with cyhalofop-butyl. Of these tested populations, 10 showed a high level of resistance to cyhalofop-butyl; the 50% effective dose ranged within 108.4 to 1,443.5 g ai ha−1 with resistance index values of 9.1 to 121.8 when compared with the susceptible population. Acetyl-coenzyme A carboxylase genes (ACCase) of susceptible and all 10 resistant populations were amplified and sequenced. Among them, Ile-1781-Leu, Trp-2027-Cys, Trp-2027-Ser, and Ile-2041-Asn mutations were found in five resistant populations. No known resistance-related mutations were found in the other five resistant populations, indicating that resistance to cyhalofop-butyl in these populations was likely to be endowed by non–target site resistance mechanisms. Notably, the Ile-1781-Leu and Trp-2027-Cys substitutions have previously been reported, but this is the first report of Trp-2027-Ser and Ile-2041-Asn mutations in L. chinensis. Furthermore, three derived cleaved amplified polymorphic sequence methods were developed to rapidly detect these mutations in L. chinensis.

Zoysia germplasm exhibit different levels of sensitivity to fluazifop-P-butyl, but the genetic factors responsible for such differences are unknown. Segregation patterns of the fluazifop-P-butyl tolerance trait were studied under greenhouse conditions. In total, 244 F1 lines were generated from multiple crosses between the tolerant line 5337-2 (non–target site tolerance) and three more-sensitive lines (123, 252, and 5330-23). Progeny segregation showed that fluazifop-P-butyl tolerance within zoysiagrass (Zoysia spp.) is expressed as a quantitative trait with a wide range of intermediate phenotypes between parental phenotypes. Transgressive segregation was extensive and largely favored susceptibility in most families, but was especially evident for 5337-2 × 123 and 5337-2 × 5330-23. The segregation patterns for biomass reduction and percent injury were different within reciprocal crosses and among three different family crosses. Reciprocal effects were observed in growth reduction for 5337-2 × 5330-23, in percent injury at 3 wk after the treatment (WAT), and for 5337-2 × 252 at 6 WAT. This indicated that fluazifop-P-butyl tolerance was not completely controlled by nuclear genetic factors in 5337-2 and maternal/cytoplasmic inheritance was also partially responsible. These results suggested that fluazifop-P-butyl tolerance may be attributed to multiple genetic mechanisms, which could present a challenge for future breeding efforts because of the difficulty of fixing multiple traits within a breeding population.

Breeding herbicide tolerance into new cultivars can improve safety and weed control in turfgrass systems. The sensitivity to fluazifop-P-butyl of 27 zoysiagrass (Zoysia spp.) lines was screened under greenhouse conditions to identify potential tolerant germplasm for breeding programs. The herbicide rate that caused 50% biomass reduction (GR50) and the rate that caused 50% injury (ID50) were calculated to select the three most-tolerant and the five most-susceptible lines for studying the physiological mechanisms responsible for fluazifop-P-butyl tolerance. The differences in GR50 and ID50 between susceptible and tolerant lines ranged from 4-fold to more than 10-fold. Cytochrome P450–mediated metabolism was not detected in fluazifop-P-butyl–tolerant lines. Sequencing of the ACCase gene confirmed that none of the seven previously reported mutations conferring resistance to acetyl-CoA carboxylase (ACCase)-inhibiting herbicides in other species were present in any of the tolerant or susceptible zoysiagrass lines studied. An Ala-2073-Thr substitution was identified in two tolerant lines, but this mutation did not completely explain the tolerant phenotype. No clear differences in absorption and translocation rates of 14C-radiolabeled fluazifop-P-butyl were observed among most lines, with the exception of a susceptible line that exhibited greater translocation than two of the tolerant lines. Metabolite profiles did not differ between tolerant and susceptible lines. Our results suggest that the diversity in tolerance to fluazifop-P-butyl in zoysiagrass germplasm is most likely the result of a combination of different, minor, additive non–target site mechanisms such as translocation rate and compartmentation after absorption.

With the widespread occurrence of herbicide-resistant weeds in midsouthern U.S. rice, new technologies are needed to achieve adequate weed control. A new non–genetically modified rice trait has been commercialized that is resistant to quizalofop, an acetyl coenzyme A carboxylase (ACCase)-inhibiting herbicide. The addition of quizalofop-resistant rice to production systems will increase the use of quizalofop, possibly increasing the risk for injury to other grass crops. Experiments were conducted in 2014 and 2015 to determine the sensitivity of corn, grain sorghum, and conventional rice to low rates of quizalofop (1/10× to 1/200× of 160 g ai ha–1). Conventional rice was not affected by quizalofop rate or application timing. Corn displayed the greatest response to the 1/10× quizalofop rate at the two- to three-leaf stage, with 50% to 65% injury and 35% to 37% relative yield compared to the nontreated check. Grain sorghum was injured 31% to 34% by the 1/10× quizalofop rate applied at the two- to three-leaf stage, and there was 20% to 26% injury at the panicle exertion growth stage. The highest rate of quizalofop at the panicle exertion stage reduced yields 28% to 46%. Overall, risk for injury to any of the three evaluated crops from quizalofop appears low, with greatest injury observed at the highest quizalofop drift rate, with minimal injury at lower rates.

Italian ryegrass has invaded wheat fields in China and is becoming a predominant, troublesome weed. Fenoxaprop-P-ethyl has been widely used for weed control on Chinese farms since the 1990s. However, overuse has led to fenoxaprop-P-ethyl resistance in Italian ryegrass in Chinese wheat fields. In this study, we identified a putative fenoxaprop-P-ethyl–resistant population of Italian ryegrass, HZYC-6, from Henan province, China. Mutations involving Asp-2078-Gly and Ile-1781-Leu substitutions were identified in the carboxyl-transferase domain of acetyl-coenzyme A carboxylase in this population, and these mutations are the likely cause of the target site–based resistance to fenoxaprop-P-ethyl. In addition, we identified cytochrome P450–mediated metabolism of herbicides (non–target site based resistance) in the HZYC-6 population, indicating that multiple mechanisms of resistance may be segregating in this population. Furthermore, HZYC-6 was also highly resistant to haloxyfop-R-methyl and quizalofop-P-ethyl, moderately resistant to clodinafop-propargyl and sethoxydim, and had low resistance to clethodim and pinoxaden.

Herbicide-resistant weeds pose a considerable threat to agriculture, but their resistance mechanisms are poorly understood. Differential gene expression analysis of a weed subjected to herbicide treatment is a key step toward more mechanistic studies. Such an analysis, often involving quantitative real-time PCR (qPCR), requires suitable reference genes as internal controls. In this study, we identified optimal reference genes in the noxious weed, Japanese foxtail. This weed has evolved resistance to acetyl-coenzyme A carboxylase (ACCase) inhibitors. We analyzed the stability of eight commonly used candidate reference genes (glyceraldehyde-3-phosphate dehydrogenase [GAPDH]; ubiquitin [UBQ]; capsine phosphatase [CAP]; beta-tubulin [TUB]; eukaryotic initiation factor 4a [EIF4A]; elongation factor-1 alpha [EF1]; 18S ribosomal RNA [18S]; 25S ribosomal RNA [25S]) from root, stem, and leaf tissue of plants that were either resistant or sensitive to ACCase inhibitors, with or without herbicide stress, using qPCR. The results were further ranked and analyzed using geNorm, NormFinder, and BestKeeper software. These analyses identified EF1 and UBQ in roots, EF1, TUB, CAP, and 18S in stems, and EF1, GAPDH, and 18S in leaves as suitable references for qPCR normalization. We have identified a set of reference genes that can be used to study herbicide resistance mechanisms in Japanese foxtail.

Inheritance of resistance to diclofop was studied in three wild oat biotypes (designated B, C., and H) from the Willamette Valley of Oregon. Cultivated oat (cultivar ‘Monida’) was crossed, including reciprocals, to three wild oat biotypes. Leaves of each F1 plant were spotted with diclofop as a nondestructive test for resistance or susceptibility. All F1 hybrids were resistant, indicating that resistance is dominant and is under nuclear control. The F2 plants where Monida was the maternal parent were screened with diclofop, and F2 plants of the Monida/C cross were screened with fenoxaprop because the parent C biotype was resistant to fenoxaprop. At lower doses, a 3:1 (R:S) segregation ratio in F2 was observed and at higher doses a 1:3 (R:S) segregation ratio was often observed. The F2:3 families segregated in a 1:2:1 (all resistant : segregating resistant and susceptible all susceptible) ratio when treated with a 1.1-kg ae ha−1 dose of diclofop. This confirms that resistance to diclofop in the B, C, and H biotypes is primarily under monogenic control, with resistance being dominant to susceptibility at lower herbicide doses. At increased doses, susceptibility becomes dominant. Knowledge of the inheritance of resistance may help in the development of containment measures to prevent the spread of herbicide-resistance genes.

A diclofop-methyl-resistant biotype of Italian ryegrass was characterized to determine the expression and inheritance of herbicide resistance and whether this trait was due to the presence of a diclofop-insensitive form of acetyl-coenzyme A carboxylase (ACCase). At the whole plant level, the resistant biotype was > 93-fold more resistant to diclofop-methyl than the susceptible biotype. Crosses of diclofop-resistant and –susceptible plants were performed to produce F1 plants. No maternal effects were evident in responses of reciprocal F1 plants to diclofop. GR50 diclofop rates determined for resistant, F1, and susceptible plants were 15, 6.3, and 0.16 kg ha−1, respectively. F2 populations treated with a 7.5 kg ha−1 rate of diclofop exhibited three injury response phenotypes 3 wk after treatment: a susceptible (S) phenotype which was killed, an intermediate resistance (I) phenotype with severe injury, and a resistant (R) phenotype with little or no injury. Testcross progeny exhibited only I and S phenotypes. Observed segregation of phenotypes in F2 and testcross populations conformed to segregation ratios predicted for a trait with inheritance controlled by a single partially dominant nuclear gene. ACCase activity determined in crude cell-free extracts of resistant, F1, and susceptible biotypes exhibited I50 values of 50, 20, and 0.7 μM diclofop, respectively. A positive relationship between the injury response phenotype and site of action (ACCase) response to diclofop was evident in both F1 and F2 populations. In extracts from R, I, and S phenotype F2 plants, 20 μM diclofop acid inhibited ACCase-mediated incorporation of 14C by 27.1, 45.1, and 78.9%, respectively. The ACCase data are consistent with the hypothesis that diclofop resistance in Italian ryegrass is conferred by a diclofop-insensitive form of ACCase.

Studies were conducted to test if dicamba reduces haloxyfop herbicidal activity and to determine if this interaction by auxin-type herbicides or bentazon was caused by preventing the haloxyfop-induced inhibition of acetyl-CoA carboxylase (ACCase). Addition of dicamba at concentrations of 2.17 or 4.34 mM reduced haloxyfop activity on tall fescue shoots. Haloxyfop at 14 to 20 μM inhibited the activity of ACCase in cell-free extracts from tall fescue by nearly 50%. Dicamba did not prevent the inhibitory effect of haloxyfop on ACCase activity in these cell-free enzyme preparations. Dicamba alone at concentrations as high as 3.2 mM had no significant effect on the in vitro activity of the enzyme. Bentazon concentrations up to 10 mM did not prevent haloxyfop inhibition of ACCase activity. Bentazon alone or mixed with haloxyfop significantly inhibited ACCase activity at bentazon concentrations starting at 1 mM with near 40% inhibition at 10 mM. of the numerous possible explanations for the interference of dicamba or bentazon with haloxyfop action, interaction directly at the enzyme level appears to have been eliminated.

Resistance to fenoxaprop-P and other aryloxyphenoxypropionate and cyclohexanedione herbicides in the wild oat population, UM1, is controlled by a single, partially dominant, nuclear gene. In arriving at this conclusion, parents, F1 hybrids, and F2 plants derived from reciprocal crosses between UM1 and a susceptible wild oat line, UM5, were treated with fenoxaprop-P over a wide range of dosages. Based on these experiments, a dosage of 400 g ai ha−1 fenoxaprop-P was selected to discriminate between three response types. At this dosage, susceptible plants were killed and resistant plants were unaffected, whereas plants characterized as intermediate in response were injured but recovered. Treated F2 plants segregated in a 1:2:1 (R, I, S) ratio, indicative of single nuclear gene inheritance. This was confirmed by selfing F2 plants and screening several F3 families. Families derived from intermediate F2 plants segregated for the three characteristic response types, whereas those derived from resistant F2 plants were uniformly resistant. Chisquare analysis indicated the F2 segregation ratios fit those expected for a single partially dominant nuclear gene system. In addition, F2 populations from both crosses were screened with a mixture of fenoxaprop-Pand sethoxydim. The dosages of both herbicides (150 g ai ha−1 fenoxaprop-P and 100 g ha−1 sethoxydim) were sufficient to control only susceptible plants. Treated F2 populations segregated in a 3:1 (R:S) pattern, thereby confirming that resistance to the two chemically unrelated herbicides results from the same gene alteration.

Silky windgrass is a serious weed in central and northern Europe. Its importance has escalated in recent years because of its growing resistance to acetolactate synthase (ALS)-inhibiting herbicides. This study investigated the resistance level for three herbicide sites of action in eight silky windgrass populations, collected in fields neighboring a field where iodosulfuron sodium salt–resistant silky windgrass had previously been found. Target site resistance (TSR) and non–target site resistance (NTSR) mechanisms were identified, and a spatial gradient distribution hypothesis of ALS resistance was tested. Populations showed large variations in ED50 values to iodosulfuron, with resistance indices (RIs) ranging from 0.1 to 372. No cross-resistance was found to other herbicide groups with the same site of action as iodosulfuron. In contrast, resistance was observed to the acetyl-CoA carboxylase inhibitor, fenoxaprop ethyl ester (RI from 0.7 to 776), while the activity of prosulfocarb, an inhibitor of long-chain fatty-acid synthesis, was unaffected. Iodosulfuron-resistant phenotypes were associated with NTSR, while fenoxaprop ethyl ester resistance was caused by both NTSR and TSR (Ile-1781-Leu mutation). A large-scale trend in the spatial distribution of resistance to ALS indicated a decreasing resistance with increased distance from an epicenter. After finer-scale analysis, less than 0.05% of the residual variation could be attributed to spatial autocorrelation. The spatial resistance pattern was not correlated with the dominant wind direction, while there was a correlation between the resistant phenotype and type of crop. This study underlines that NTSR mechanisms do not always confer broad resistance to different herbicide subclasses and site of action, hence the complex relationship to resistant phenotype. NTSR mechanisms, in particular detoxification, were present at different levels for the herbicides tested in the silky windgrass populations of this study. The factors contributing to the spatial distribution of resistance remain elusive.

The herbicide pinoxaden is a phenylpyrazoline inhibitor of acetyl coenzyme A carboxylase. Research was conducted to determine the effects of pinoxaden (90 g ai ha−1) alone and in combination with herbicide safeners on creeping bentgrass injury as well as perennial ryegrass and roughstalk bluegrass control. Greenhouse experiments determined that herbicide safeners cloquintocet-mexyl, fenchlorazole-ethyl, and mefenpyr-diethyl were more effective in reducing creeping bentgrass injury from pinoxaden than benoxacor, isoxadifen-ethyl, and naphthalic-anhydride. Other experiments determined that creeping bentgrass injury from pinoxaden decreased as rates (0, 23, 45, 68, 90, 225, or 450 g ha−1) of cloquintocet-mexyl, fenchlorazole-ethyl, and mefenpyr-diethyl increased. On the basis of creeping bentgrass responses to various safener rates, safeners were applied at 68 and 450 g ha−1 in additional experiments to evaluate their effects on pinoxaden (90 g ha−1) injury to creeping bentgrass and efficacy against perennial ryegrass and roughstalk bluegrass. In these experiments, safeners mefenpyr-diethyl and cloquintocet-mexyl reduced pinoxaden-induced creeping bentgrass injury (from 25 to ≤ 5%) more than fenchlorazole-ethyl at 2 wk after treatment. Safeners reduced pinoxaden efficacy against roughstalk bluegrass. Perennial ryegrass was controlled > 80% by pinoxaden and herbicide safeners did not reduce control. Field experiments should evaluate pinoxaden in combination with cloquintocet-mexyl and mefenpyr-diethyl to optimize safener : herbicide ratios and rates for creeping bentgrass safety as well as perennial ryegrass and roughtstalk bluegrass control in different climates and seasons.

Understanding the mechanism of herbicide resistance is fundamental for designing sustainable weed control strategies and exploiting herbicides rationally. Shortawn foxtail is a problem grass weed infesting several important crops in China. The repeated use of acetyl-CoA carboxylase (ACCase)-inhibiting herbicides has resulted in herbicide resistance in this weed. The ACCase gene of resistant individuals of a shortawn foxtail population (JSLS-1) has an Ile-2041-Thr mutation. F2 generation seeds, originated from the same heterozygous plant, were harvested, and two homozygous mutant (JSLS-1RR) and wild (JSLS-1SS) populations for the Ile-2041-Thr mutation were obtained. In whole plants, the JSLS-1RR population conferred high resistance to fenoxaprop and clodinafop, moderate resistance to haloxyfop, low resistance to pinoxaden, and no obvious resistance to clethodim and sethoxydim, compared with JSLS-1SS and a proven susceptible population (HNXY-1). A derived cleaved amplified polymorphic sequence (dCAPS) marker was developed to rapidly detect the rare Ile-2041-Thr mutation in the shortawn foxtail population. This is the first report of the cross-resistance pattern of Ile-2041-Thr mutation, and the robust dCAPS marker could quickly detect this mutation in shortawn foxtail.

Japanese foxtail is a predominant tetraploid grass weed in wheat and oilseed rape fields in eastern China. In China, pyroxsulam is mainly used to manage annual grass weeds, especially those resistant to acetyl coenzyme A carboxylase (ACCase)-inhibiting herbicides. Using dose–response studies, a pyroxsulam-resistant population, ACTC-1, was identified with a resistance index value of 58. Additionally, ACTC-1 was cross-resistant to sulfonylureas, imidazolinones, triazolopyrimidines, pyrimidinyl-benzoates, and sulfonylaminocarbonyl-triazolinones and multiresistant to ACCase and photosystem II inhibitors. Sequence analysis revealed four gene fragments encoding acetolactate synthase (ALS) from ACTC-1, and three from JNXW-1, a pyroxsulam-sensitive population. An Asp-376-Glu substitution was found in ALS1;2 and an Ile-2041-Asn in Acc1;1, which may be responsible for its resistance to pyroxsulam and ACCase inhibitors, respectively. In vitro assays of ALS activity revealed that in ACTC-1, the sensitivity of ALS to pyroxsulam was lower, and the basal ALS activity was twofold higher than that of sensitive population JNXW-1. Additionally, the combined application of pyroxsulam with malathion or piperonyl butoxide increased the sensitivity of ACTC-1 to pyroxsulam, although it could not completely overcome the resistance. It was inferred that both target-site-based resistance and nontarget-site-based resistance may be involved in the resistance to pyroxsulam.

Acetyl-coenzyme A carboxylase (ACCase) assays and absorption, translocation, and metabolism experiments were conducted to investigate the mechanism(s) responsible for resistance in a johnsongrass biotype that exhibited low levels of resistance to the cyclohexanedione (CHD) herbicide sethoxydim and the aryloxyphenoxypropionate (APP) herbicides quizalofop-P and fluazifop-P. The rate of [14C]quizalofop-ethyl absorption was significantly higher in the resistant compared to the susceptible biotype 8, 24, and 48 h after treatment (HAT), but by 72 HAT, there was no significant difference in the amount of [14C]quizalofop-ethyl detected in either biotype. Additionally, little or no differences in the translocation of [14C]quizalofop-ethyl were observed in the resistant and susceptible biotypes at any time interval after application. In [14C]quizalofop-ethyl metabolism experiments, similar levels of quizalofop-ethyl and quizalofop metabolites were observed in the resistant and susceptible biotypes 8, 24, 48, and 72 HAT, but slightly higher levels of quizalofop acid were detected in the resistant biotype 48 and 72 HAT. In ACCase assays, the concentrations of quizalofop-P, clethodim, and sethoxydim that inhibited ACCase activity by 50% (I50) were statistically similar in the two biotypes, indicating that the resistant johnsongrass biotype contains an ACCase that is sensitive to the APP and CHD herbicides. In the absence of APP or CHD herbicides, however, the specific activity of ACCase in the resistant biotype was two to three times greater than that of the susceptible biotype. The specific activity of ACCase in the resistant biotype was also significantly greater than that of the susceptible biotype in the presence of all concentrations of quizalofop-P and sethoxydim and in the presence of 0.1, 1, and 10 µM clethodim. These results suggest that resistance to quizalofop-P and sethoxydim is conferred by an overproduction of ACCase in the resistant johnsongrass biotype.

Herbicides targeting grass plastidic acetyl coenzyme A carboxylase (ACC) are effective selective graminicides. Their intensive use worldwide has selected for resistance genes in a number of grass weed species. Biochemistry and molecular biology have been the means of determining the herbicidal activity and selectivity toward crop plants of ACC-inhibiting herbicides. In recent years, elucidation of the tridimensional structure of ACC and identification of five amino acid residues within the ACC carboxyl transferase domain that are critical determinants for herbicide sensitivity shed light on the basis of ACC-based resistance to herbicides. However, metabolism-based resistance to ACC-inhibiting herbicides is much less well known, although this type of resistance seems to be widespread. A number of genes thus endow resistance to ACC-inhibiting herbicides, with the possibility for various resistance genes that confer dominant resistance at the herbicide field rate to accumulate within a single weed population or plant. This, together with a poor knowledge of the genetic parameters driving resistance, renders the evolution of resistance to ACC-inhibiting herbicides unpredictable. Future research should consider developing tactics to slow the spread of resistance. For this purpose, it is crucial that our understanding of metabolism-based resistance improves rapidly because this mechanism is complex and can confer resistance to herbicides with different target sites.

Several seed production fields of the Italian ryegrass cultivar ‘Tetrone’ were destroyed in 1988 by 280 to 350 g ai ha−1 racemic fenoxaprop applied for wild oat control. Because similar rates of fenoxaprop had possessed adequate safety when applied to ‘Oregon common’ Italian ryegrass, suspicion arose that the cultivars differed in tolerance. Seedlings of 21 commonly grown cultivars were screened in the greenhouse at the three-leaf growth stage to determine their fresh weight GR50 for fenoxaprop. The GR50 values for the two most tolerant cultivars, ‘Marshall’ and ‘Torero’, were more than threefold greater than the two most sensitive cultivars, ‘Futaharu’ and ‘Ace’. Cultivars could be separated into sensitive, intermediate, and tolerant groups, but the distribution of the GR50 values appeared to be continuous rather than discrete. Tolerance increased with growth stage, and the average GR50 for tillered plants was 80% higher than that for the two-leaf stage and 41% higher than that for the four-leaf stage seedlings. Cultivars differed slightly in the specific activity of acetyl–coenzyme A carboxylase (ACCase) (EC 6.4.1.2) and in the I50 values for the inhibition by fenoxaprop, but the only clear relationship between these biochemical factors and whole-plant tolerance was a threefold increase in ACCase activity at the tillered stage over that present in the younger seedlings.