Synthesis and thylakoid membrane binding of the radioactively labeled herbicide dinoseb

2,4-Dinitro-6-isobutylphenol (i-dinoseb), an isomer of the phenolic herbicide dinoseb and equally active as an inhibitor of photosynthetic electron transport and photophosphorylation, has been synthesized, ³H-labeled with a specific activity of 490 mCi/mmol. Its binding to broken chloroplasts is strongly pH dependent and biphasic representing a high- and a low-affinity binding site. For specific binding of i-dinoseb a binding constant K_b = 6.9 × 10^?8M has been determined. The number of binding sites corresponds to one molecule i-dinoseb per 830 molecules of chlorophyll, i.e., one molecule per two electron transport chains. i-Dinoseb can be displaced from the thylakoid membrane by DCMU-type inhibitors, and inhibitory uncouplers, but not by DBMIB-type inhibitors and uncouplers of oxidative phosphorylation. An extensive analysis of displacement by DCMU-type and phenolic herbicides indicates that DCMU-type herbicides interfere noncompetetively but phenolic herbicides interfere competetively with the specific binding of i-Dinoseb. It is concluded, therefore, that the binding sites of both types of herbicides are not identical although they are located on the same protein. The specific binding constant of i-dinoseb does not change in trypsin-treated chloroplasts whereas the number of binding sites is slightly reduced.     

psbA Mutation (valine219 to isoleucine) in Poa annua resistant to metribuzin and diuron

The herbicide‐binding region of the chloroplast psbA gene from a total of 20 biotypes of Poa annua L resistant and susceptible to metribuzin and diuron was selectively amplified using PCR. Sequence analysis of the fragment from six herbicide‐resistant biotypes of P annua exhibited a substitution from valine to isoleucine at position 219 of the D1 protein encoded by the psbA gene. This is the same mutation as reported for Chlamydomonas and Synechococcus through site‐directed mutagenesis and in cell cultures of Chenopodium rubrum L. To our knowledge this is the first report of a higher plant exhibiting resistance in the field to photosystem II inhibitors due to a psbA mutation other than at position 264. The existence of additional biotypes of P annua resistant to diuron or metribuzin but lacking mutation in the herbicide‐binding region indicates that resistance to these herbicides can also be attained by other mechanisms. © 2000 Society of Chemical Industry     

Increased herbicidal activity of triazine herbicides by piperonyl butoxide

Piperonyl butoxide (PB) is a known Synergist which enhances the activity of insecticides by inhibiting their biotransformation to less active products. We have evaluated the possible use of PB as a herbicide synergist using triazine herbicides in sensitive, tolerant, and resistant plants. The effects of PB, triazine herbicides, and their combinations were examined in whole plants as well as in chloroplasts isolated from triazine-sensitive (S) and -resistant (R) weed biotypes. PB itself, applied postemergence (0.1–0.5%, v/v), was slightly toxic to the plants tested. However, foliar application of PB combined with atrazine, terbutryn or prometryn to maize seedlings significantly increased the phytotoxicity of the herbicides. Low rates of atrazine, prometryn, and terbutryn in a tank-mixture with PB, effectively controlled Solatium nigrum L. and Abutilon theophrasli Medik. PB enhanced atrazine efficacy in both S and R biotypes of Lolium rigidum Gaud. The synergistic effect of PB was evident also in vitro when atrazine and methabenzthiazuron were used to inhibit photosystem II electron transport in chloroplasts isolated from resistant weeds. These data demonstrate the potential of PB as a herbicide synergist and its possible utilization as an aid for improving the activity of triazine herbicides in sensitive, tolerant and resistant plants.     

Differential atrazine interference with the Hill reaction of isolated chloroplasts from Chenopodium album L. biotypes

Atrazine [2-chloro-4-(ethylaminol-6-(isopropyl-amino)-s-triazine] resistant biotypes of lamb's quarters (Chenopodium athum L) were reported in the maize growing areas of Ontario, where the herbicide had been used fur a number of years. Field samples from four locations proved tolerant to higber than recommended rates of atrazine in controlled environment screening trials. A resistant biotype was not killed with up to 40 kg/ha atrazine. Diuron at 5 x10-5 M inhibited the Hill reaction with isolated chloroplasts of resisiant and susceptible biotypes of lamb's-quarters. However, with 10-4 M atrazine, the photochemical activity was inhibited in chloroplasts isolated from the susceptible biotype but not in chloroplasts from the resisiant biotype. With maize chloroplasts, inhibition with 10-4 M atrazine was the same as with the susceptible biotype of lamb's-quarters. These studies suggested that a new mechanism of intraspecific resistance in lamb's quarters to atrazine was involved, other than differences in uptake, translocation and metabolism reported with interspecific comparisons involving the s-triazines and other herbicides, It was also concluded that atrazine and diuron did not have precisely the same mechanism of action as photosynthetic inhibitors with lamb's-quarters, and that external and or internal structure or function of chloroplasts in relation to atrazine inhibition can vary significantly even in biotypes of the same species.     

Studies into the action of the diphenyl ether herbicides acifluorfen and oxyfluorfen. Part II: The interaction with photosynthetic electron transport reactions

The effects of acifluorfen and oxyfluorfen on photosynthetic electron transport reactions of pea chloroplasts were compared with those induced by paraquat and monuron. Monuron inhibited electron flow between photosystems I and II, and paraquat acted as an electron acceptor for photosystem I, promoting superoxide formation by illuminated chloroplasts. Neither acifluorfen nor oxyfluorfen at concentrations up to 50 μM affected non-cyclic electron flow or promoted superoxide formation. Both herbicides were shown to repress ferredoxin-dependent NADP⁺ reduction by illuminated chloroplasts. Further experiments showed that, in the presence of ferredoxin-NADP⁺ reductase and chloroplast membranes maintained in the dark, p-nitro diphenyl ether (DPE) herbicides promoted the rate of ferredoxin-dependent oxidation of NADPH, implying that these herbicides can accept electrons from reduced ferredoxin. The interaction between acifluorfen, ferredoxin and chloroplast membranes was examined further by following the effect of this herbicide on the peroxidation of illuminated thylakoids. Lipid peroxidation was promoted by acifluorfen, although this effect was abolished if thylakoids were washed prior to use. The effect of washing could be reversed by adding exogenous ferredoxin. These data demonstrate that interaction of DPE herbicides with photosynthetic electron transport in the vicinity of ferredoxin is necessary for light-dependent herbicide activation.     

Inhibition of acetolactate synthase in susceptible and resistant biotypes of Stellaria media

Acetolactate synthase (ALS) from one susceptible and two chlorsulfuronresistant biotypes of Stellaria media(L.) Vill. was assayed in the presence of eight known ALS inhibitors. As expected, ALS from the chlorsulfuronresistant biotypes (R1 and R2) showed reduced sensitivity to chlorsulfuron and other sulfonylurea herbicides. The patterns of cross-resistance varied, however, indicating that the alteration in ALS that confers chlorsulfuron resistance does not confer the same level of resistance to other sulfonylurea herbicides. The resistant biotypes were highly cross-resistant to sulfometuron-methyl and DPX-A7H81, but less cross-resistant to triasulfuron. Both R1 and R2 were highly cross-resistant to DTPS (N-[2,6-dichlorophenyl]-5,7-dimethyl-1,2,4-iriazolo[1,5a]pyrimidine-2-siilfoiiamide), but only slightly cross-resistant to imazamethahenz, an imidazolinone herbicide. The differences in the patterns of cross-resistance observed presumably reflect differences in the binding affinity of the herbicides for the altered ALS. The data presented suggest, but do not confirm, that R1 and R2 contain the same ALS mutation.     

Resistance of dicot weeds to acetolactate synthase (ALS)-inhibiting herbicides in Australia

A biotype of Sonchus oleraceus L. and two bio types of Sisymbrium orientate Torn., SSO 3 and NSO 1, are the first dicot weeds in Australia to develop resistance to ALS-inhibiting herbicides. The resistant biotypes had been exposed to va rying periods of selection with sulfonylurea her bicides. All three biotypes are resistant to a range of sulfonylurea and imidazolinone herbicides. The S. orientale biotypes are also resistant to the triazolopyrimidine herbicide, flumetsulam. LD₅₀ ratios of resistant Sonchus oleraceus for sulfony lurea and imidazolinone herbicides are greater than 64-fold and 4.5-fold, respectively. GR₅₀ ratios are greater than 9 for sulfonylureas and 7.4 for imazapyr. The LD₅₀ ratios for both S. orien tale biotypes for chlorsulfuron, sulfometuron methyl, metsulfuron-methyl, flumetsulam and imazethapyr are greater than 110-, 15-, 7-, 24- and 29-fold, respectively. All resistant biotypes are susceptible to MCPA, diuron and diflufenican, herbicides which do not inhibit ALS.     

Possible involvement of herbicide sequestration in the resistance to diclofop-methyl in Italian biotypes of Lolium spp.

The mechanism of resistance to diclofop-methyl in three Italian populations of Lolium spp. (two resistant and one susceptible) was investigated. The major proportion of R-1 (Tuscania 1997) and R-2 (Roma 1994) plants (approximately 80%) survived after herbicide treatment by emitting new tillers from the crown. Both resistant (R-1 and R-2) and susceptible (Vetralla 1994) Lolium spp. populations were target-site sensitive. No difference in diclofop-methyl absorption by shoots of resistant and susceptible biotypes was observed. At the dose corresponding to 1× the recommended field rate, a relatively higher metabolism was found in R-2 biotype. In contrast, at the doses 2× and 10× the field rate no difference in herbicide metabolism between susceptible and resistant biotypes was observed. At all the three herbicide doses (1×, 2×, and 10× the field rate) 48 h after the treatment (HAT), the total amount of metabolites produced by wheat was more than three times higher than that produced by resistant and susceptible ryegrass biotypes. At the doses 1× and 2× the field rate, the herbicide translocation was different in the susceptible biotypes compared to resistant biotypes. The total amount of the radiolabel found 48 HAT in culm and root was approximately twice in susceptible biotype than in resistant biotypes. Susceptible and resistant ryegrass biotypes differed in the capability of their roots to acidify the external medium. Susceptible biotype acidified the external solution at approximately 6 times the rates of the resistant biotypes. In the present study, the mechanism responsible for resistance in the investigated resistant biotypes was not univocally identified. Indirect evidence supports the possible involvement of herbicide sequestration or immobilization.     

Sulfonylurea herbicide resistance in Sonchus asper biotypes in Alberta, Canada

Summary Two Sonchus asper (spiny annual sow-thistle) biotypes, suspected of being resistant to the sulfonylurea herbicide metsulfuron-methyl, were collected in 1996 from two barley ( Hordeum vulgare ) fields in central Alberta, Canada. Both fields had received at least six applications of acetolactate synthase (ALS)-inhibiting herbicide(s). The responses of the two resistant (R) biotypes and two susceptible (S) biotypes to several sulfonylurea herbicides, and to herbicides and herbicide mixtures with other mechanisms of action, were compared. Both R biotypes were highly resistant to all sulfonylurea herbicides, but their control with other herbicides and mixtures was effective and comparable to that of the S biotypes. ALS extracted from an R biotype was about 440 times more resistant to metsulfuron-methyl than that of an S biotype, indicating that resistance was conferred by an ALS enzyme that was less sensitive to inhibition by the herbicide. Competitiveness and seed production of S. asper varied among biotypes, but the differences were probably the result of ecotype differences rather than resistance or susceptibility to sulfonylurea herbicides. This is the first reported occurrence of target site-based S. asper resistance to ALS-inhibiting herbicides.     

Inhibition of photosynthesis in intact plants of biotypes resistant or susceptible to atrazine and cross-resistance to other herbicides

A concentration of atrazine of 0?1 mm 1^?1 in the nutrient solution resulted in complete inhibition of photosynthesis in intact leaves of susceptible biotypes of Amaranthus retroflexus L., Polygonum lapathifolium L., Chenopodium album L., Solanum nigrum L., Poa annua L. and Stellaria media (L.) Vill. within a few hours of treatment, whereas the inhibition of the resistant biotypes by the same concentration varied from small to moderate. In contrast, diuron (10 or 20 μm 1^?1) produced only minor differences between resistant and susceptible biotypes. The influence of some other herbicides on photosynthesis of these resistant biotypes was also smaller than that on the susceptible biotypes. This cross-resistance was evaluated with the resistance factor for intact leaves of Brassica napus L., A. retroflexus, and S. nigrum. This factor is equal to the ratio of the herbicide concentration in leaves of the resistant biotype to that in leaves of the susceptible biotype with inhibition to half-maximum rate of photosynthesis. This concentration in the leaves was calculated from the concentration in the nutrient solution, and the total transpiration divided by leaf area from the beginning of the herbicide treatment until the moment of half-maximum of photosynthesis. The resistance factors for intact leaves of A. retroflexus, S. nigrum and B. napus were 26–30 for atraton, 3–7 for metamitron, 2–9 for bromacil, 3–5 for monolinuron, 1 for diuron and < 1 for bentazone. For isolated chloroplasts much higher values have been reported. The reason for this discrepancy is not clear. A somewhat higher resistance factor (around 50–60) was derived after infiltration of detached leaves of these species with atraton solutions.     

Molecular basis of resistance to acetolactate synthase-inhibiting herbicides in Sisymbrium orientale and Brassica tournefortii

Three Australian Sisymbrium orientale and one Brassica tournefortii biotypes are resistant to acetolactate synthase (ALS)-inhibiting herbicides due to their possession of an ALS enzyme with decreased sensitivity to these herbicides. Enzyme kinetic studies revealed no interbiotypic differences within species in K_m (pyruvate) (the substrate concentration at which the reaction rate is half maximal) but a greater V_max (the rate when the enzyme is fully saturated with substrate) for two of the resistant S orientale biotypes over susceptible levels. F₁ hybrids from reciprocal crosses between resistant and susceptible biotypes of S orientale showed an intermediate response to chlorsulfuron compared to the parental plants. ALS herbicide resistance in S orientale segregated in a 3:1 (resistant:susceptible) ratio in F₂ plants with a single rate of chlorsulfuron, indicating that resistance is inherited as a single, incompletely dominant nuclear gene. Two regions of the ALS structural gene known to vary in ALS-resistant biotypes were amplified and sequenced. Resistant S orientale biotypes NS01 and SS03 contained a single nucleotide substitution in Domain B, predicting a Trp (in susceptible) to Leu (in resistant) amino acid change. Two adjacent nucleotide substitutions (CC T to AT T) predicting a Pro (in susceptible) to Ile (in resistant) change in the primary amino acid sequence were identified in Domain A of resistant S orientale biotype SS01. Likewise, a single nucleotide substitution at the same site in the resistant B tournefortii biotype predicts a Pro (in susceptible) to Ala (in resistant) substitution. No other interbiotypic nucleotide differences predicted amino acid changes in the sequenced regions, suggesting that the amino acid substitutions reported above are responsible for resistance to ALS-inhibiting herbicides in the respective biotypes. © 1999 Society of Chemical Industry     

Resistance of barnyardgrass (Echinochloa crus-galli) to atrazine and quinclorac

Two populations of Echinochloa crus-galli (R and I) exhibited resistance to quinclorac. Another population (X) exhibited resistance to quinclorac and atrazine. The R and I populations were collected from monocultures of rice in southern Spain. The X population was collected from maize fields subjected to the application of atrazine over several years. The susceptible (S) population of the same genus was collected from locations which had never been treated with herbicides. The quinclorac ED₅₀ value (dose causing 50% reduction in shoot fresh weight) for the R and I biotypes were 26- and 6-fold greater than for the S biotype. The X biotype was 10 times more tolerant to quinclorac than the S biotype and also showed cross-resistance to atrazine, being 82-fold more resistant to atrazine than the R, I and S biotypes. Chlorophyll fluorescence and Hill reaction analysis supported the view that the mechanism of resistance to atrazine in the X biotype was modification of the target site, the DI protein. Quinclorac at 20 mg litre^-1 did not inhibit photosynthetic electron transport in any of the test biotypes. The quinclorac I₅₀ values (herbicide dose needed for 50% Hill reaction reduction) of the S population was over 50000-fold higher than the atrazine I₅₀ value for the same S population, indicating that quinclorac is not a PS II inhibiting herbicide. Propanil at doses greater than 0·5 kg ha^-1 controlled all the biotypes. © 1997 SCI     

Herbicide binding in various mutants of the photosystem II D1 protein of Chlamydomonas reinhardtii

Various mutants of the D1 protein of photosystem II were generated by transforming a Chlamydomonas reinhardtii strain specifically engineered to lack a 0.4 kb fragment in the psbA gene. This mutant is unable to grow photosynthetically. The following mutants were obtained: Asp₁₇₀Glu, Gly₁₇₈Ser, Leu₂₁₈Ser or Thr, Arg₂₃₈Val, Thr₂₄₅Ala or Ser and Ser₂₆₄Lys or Ile. In these mutants, the following herbicides were assayed for their inhibitory activity: triazines, triazinones, ureas, biscarbamates, and phenols. The results indicate that besides Leu₂₁₈ and Ser₂₆₄ also Gly₁₇₈ influences herbicide binding, probably via long-range effects. The herbicide resistances of Ser₂₆₄ mutants have been recognized since a long time. However, substitution of Ser₂₆₄ specifically by Lys exhibit extreme resistance to triazines and triazinones, but no or slight resistance to ureas.     

Resistance to glyphosate in Lolium rigidum

Annual ryegrass (Lolium rigidum) is a widespread and important weed of Australia and populations of this weed have developed resistance to most major herbicides, including glyphosate. The possible mechanisms of resistance have been examined in one glyphosate-resistant Lolium population. No major differences were observed between resistant and susceptible biotypes in respect of (i) the target enzyme (EPSP synthase), (ii) DAHP synthase, the first enzyme of the target (shikimate) pathway, (iii) absorption of glyphosate, or (iv) translocation. Following treatment with glyphosate, there was greater accumulation of shikimate (derived from shikimate-3-Pi) in susceptible than in resistant plants. In addition, the resistant population exhibited cross-resistance to 2-hydroxy-3-(1,2,4-triazol-1-yl)propyl phosphonate, a herbicide which, although structurally similar to glyphosate, acts at an unrelated target site. On the basis of these observations we speculate that movement of glyphosate to its site of action in the plastid is involved in the resistance mechanism. © 1999 Society of Chemical Industry     

杂草对AHAS抑制剂的抗药性分子机理研究进展

除草剂在田间的重复及不合理使用,导致了杂草抗药性的发生和发展。其中AHAS抑制剂由于靶标单一,抗性发展十分迅速。截至2009年,已有103种杂草对AHAS抑制剂产生了抗药性,占19类化学除草剂总抗药性杂草生物型的近1/3。从AHAS基因突变位点及种类与杂草抗药性水平的关系、AHAS基因突变与AHAS酶活性的关系、AHAS基因拷贝数与杂草抗药性的关系以及AHAS酶与除草剂结合前后的三维结构等方面,综述了杂草对AHAS抑制剂产生抗药性的机理,旨在为AHAS抑制剂抗性研究提供参考。并对自然种群目标基因的等位基因检测技术(ECOTILLING)和衍生型酶切扩增多态性序列(dCAPS)两种通过检测等位基因多态性的手段快速诊断抗药性杂草的新技术进行了介绍,讨论了延缓杂草抗药性发生和发展的策略。     

Mechanisms of herbicide sorption in microalgae and the influence of environmental factors

The sorption kinetics of photosystem II herbicides in the unicellular microalgae Ankistrodesmus braunii are independent of the herbicide concentration. While diuron-type herbicides attain an equilibrium state of sorption after at most 5 min, it takes up to 60 min for phenol-type herbicides. The kinetics of the inhibition of photosynthetic electron transport display a good correlation to the kinetics of sorption. For both types of herbicides, a phase of prevalent partitioning into the lipid phase of the cell membranes preceeds binding to thylakoid membranes. With phenol-type herbicides this phase is much more extended than with diuron-type compounds. As a consequence, there is no displacement of previously bound herbicides of this type from algal cells by addition of an excess of other photosystem II herbicides. The sorption of phenol-type herbicides to microalgae is under strong influence of environmental factors, such as light, temperature, pH, or oxygen concentration. Thus, besides the lipophilicity of these biocides, the response of the target cells to environmental factors is at least as important for herbicide sorption in the cells. From these results it is concluded that a prediction of the biocide accumulation in aquatic microorganisms can only be partly deduced from the properties of the pesticide molecule. The target cells and their complex responses to their environment strongly relativize correlations of chemical properties, e.g., lipophilicity and biological activity.     

Sulfonylurea resistance in Stellaria media [L.] Vill.

A sulfonylurea resistant biotype of common chickweed (Stellaria media L. Vill.) was found in a field treated with chlorsulfuron or metsulfuron for eight consecutive years. In pot experiments the biotype was resistant to postemergence treatments with the following acetolactate synthase (ALS) inhibitors: chlorsulfuron, metsulfuron, tribenuron, triasulfuron, rimsulfuron, sulfometuron, flumetsulam and imazapyr. The level of resistance to chlorsulfuron and sulfometuron was higher than to the other sulfonylurea herbicides. Whereas the level of cross resistance to the triazolopyrimidine herbicide, flumetsulam was comparable to that of metsulfuron, that of imazapyr was significantly lower. In contrast to imazapyr the biotype was not resistant to imazethapyr, an other imidazolinone herbicide. ALS in vitro assays revealed that resistance was due to an ALS enzyme that was less sensitive to ALS inhibiting herbicides. Herbicides with different modes of action were equally effective on the susceptible and resistant biotypes.     

Comparison of inhibitory activity of amides derivatives in triazine-resistant and -susceptible chloroplasts from Chenopodium album and Brassica campestris

Half-inhibitory concentrations of some substituted ureas and other amide derivatives were measured in chloroplasts extracted from triazine-resistant and -susceptible biotypes of Chenopodium album and Brassica campestris. It was shown that the binding contribution weakened in triazineresistant chloroplasts was not hydrophobic. The existence of two independent interactions could be deduced from their differential behavior toward triazine resistance. A nitrogen-dependent interaction, possibly involving an acidic proton, is preferentially affected by triazine resistance, whereas a carbonyl-dependent interaction maintains its contribution to binding. Analogies with triazine behavior could be shown, which are compatible with an identity of electronic binding components at the site of action of amide derivatives and triazines.     

Mechanisms of Resistance to Aryloxyphenoxypropionate Herbicides in Two Resistant Biotypes ofAlopecurus myosuroides(blackgrass): Herbicide Metabolism as a Cross-Resistance Mechanism

The mechanisms of AOPP herbicide resistance in twoAlopecurus myosuroidesbiotypes were investigated. Resistant biotype Peldon A1, which is highly resistant to the phenyl-urea chlorotoluron, is moderately resistant to the AOPP herbicides diclofop-methyl, fenoxaprop-ethyl, fluazifop-P-butyl, and the CHD tralkoxydim. Resistant biotype Lincs. E1, which is only moderately resistant to chlorotoluron, is highly resistant to the AOPP herbicide fenoxaprop-ethyl, and moderately resistant to diclofop-methyl, fluazifop-P-butyl, and the CHD tralkoxydim. There is no clear evidence of resistance to the CHD sethoxydim in either biotype. Both Peldon A1 and Lincs. E1 exhibited moderately enhanced metabolism of diclofop-methyl. The approximate half life of diclofop was 8 and 9 HAT, respectively, compared to 17 HAT for the susceptible Rothamsted biotype. Peldon A1 showed moderately enhanced metabolism of fenoxaprop-P-ethyl. However, in the highly resistant Lincs. E1, fenoxaprop-P-ethyl metabolism rates were intermediate between Peldon A1 and the susceptible biotype. Fenoxaprop-P-ethyl metabolism inA. myosuroideswas not significantly reduced by inhibitors of cytochrome P450: PBO, tetcyclasis, or ABT. While enhanced herbicide metabolism can account for the moderate AOPP/CHD resistance observed in Peldon A1in vivo, it cannot account in total for fenoxaprop-ethyl resistance in Lincs. E1. Lincs. E1 may possess one or more additional resistance mechanism.     

Herbicidal activity on acetolactate synthase‐resistant barnyardgrass (Echinochloa crus‐galli) in Arkansas,USA

The intensive use of the acetolactate synthase (ALS)‐inhibiting herbicides, imazethapyr, penoxsulam and bispyribac‐sodium, in imidazolinone‐resistant (Clearfield) rice increases the risk of the evolution of ALS‐resistant barnyardgrass. In 2009, imazethapyr failed to control barnyardgrass that was collected from a field in Arkansas, USA, following the failure of the herbicide in 2008. A greenhouse experiment was conducted to confirm and document the level of resistance of the biotype against three ALS‐inhibiting herbicides that currently are labeled in rice. The level of control of the resistant biotype at the labeled rate of bispyribac‐sodium of 35 g ai ha^?1 was 10%, penoxsulam at 22 g ai ha^?1 was 0% and imazethapyr at 70 g ai ha^?1 was 25%. The level of mortality of the susceptible biotype was 100% with all the herbicides at the labeled rate. The dose needed to kill 50% of the resistant plants was 49 g ha^?1 of bispyribac‐sodium, 254 g ha^?1 of penoxsulam and 170 g ha^?1 of imazethapyr. For the susceptible biotype, bispyribac‐sodium at 6 g ha^?1, penoxsulam at 10 g ha^?1 and imazethapyr at 12 g ha^?1 killed 50% of the treated plants. Based on these findings, it was confirmed that a barnyardgrass population has evolved cross‐resistance to three ALS‐inhibiting herbicides in rice culture in Arkansas. Furthermore, an experiment was conducted to determine if the ALS‐resistant biotype could be controlled using other mechanisms of action. The results indicated that propanil, a photosystem II inhibitor, and quinclorac, a synthetic auxin, failed to control the resistant biotype at the labeled rates, whereas all the other evaluated herbicides provided effective control of both biotypes.