The repetitive use of ALS inhibitors for smallflower umbrella sedge (Cyperus difformis L.) control has selected for herbicide-resistant (R) populations that threaten the sustainability of rice (Oryza sativa L.) production and demand alternative control measures be developed. A better understanding of seedling recruitment patterns at the field level is required to optimize the timing and efficacy of control measures. Therefore, a population-based threshold model was developed for optimizing germination prediction in multiple acetolactate synthase (ALS)-R and ALS-susceptible (ALS-S) C. difformis biotypes and applied to field-level emergence predictions. Estimated base temperatures (Tb) ranged from 16.5 to 17.6 C with no clear pattern between biotypes; such values are higher than Tb values of other important rice weeds, as well as for rice. Germination rates increased linearly from 16 to 33.7 C. ALS-R seeds germinate faster due to smaller median thermal times to germination (θT(50)) while also displaying lower germination synchronicity across water potentials. Interestingly, ALS-R biotypes were capable of germinating under lower moisture availability, as indicated by their lower (more negative) base water potential values (Ψb(50)) for seed germination; Ψb(50) values ranged from −0.24 to −1.13 MPa. In-field soil germination measurements found thermal times to emergence varied across three water regimes (daily water, flooded, or saturated). Seedling emergence under the daily water treatment was fastest; however, total seedling density was lower than for the other water regimes. In order to optimize springtime C. difformis seedling emergence, soil moisture should be kept around field capacity, as germination is hindered at lower moisture contents. By predicting when most of the seed population germinates, the thermal-time model can address issues regarding the optimal timing for herbicide applications, thereby allowing for improved C. difformis management in rice fields.

Eight triazolopyrimidine sulfonanilides were tested for metabolic stability in a number of crop and weed species. These data, along with in vitro determinations of activity (I50) against acetolactate synthase, successfully described the in vivo activity of these compounds in a two-parameter model. Whole plant activity increased with increasing compound stability and decreasing I50 (r2 =.78, N = 36). The difficulty in obtaining metabolic stability data during a structure optimization program prompted a study with substituent parameters in models of in vivo activity. Models describing whole plant activity in jimsonweed were developed using a series of 5-methyl triazolopyrimidine sulfonanilides that differed only in ortho and meta substituents on the aniline ring. The I50 term and clogP were most important to jimsonweed activity. Hence, in vitro activity (I50) may be a useful component of whole plant structure activity models to aid in identification of barriers to in vivo performance.

Acetohydroxyacid synthase was discovered as the site of action of imidazolinone and sulfonylurea herbicides over 6 yr ago. In recent years, advances have been made in the understanding of this enzyme as a herbicide target site. Derivatives of both imidazolinones and sulfonylureas have yielded new herbicide chemistry. AH of the herbicides display unusual “slow-binding” behavior with the enzyme, and this behavior may help explain efficacy of the herbicides. Resistance to these herbicides has been developed through a number of different procedures, and the mechanism of resistance is through changes in sensitivity of the enzyme to the herbicides. The changes are either selective to only one class of chemistry, or broad to a number of classes of chemistry. These data support the idea that binding sites for the herbicides on the enzyme are only partially overlapping. Progress in purification of AHAS from corn includes discovery of the existence of the enzyme in monomer and oligomer aggregation states. The interaction of the enzyme with the herbicides is affected by enzyme aggregation state.

Acetolactate synthase (ALS)-inhibiting herbicide carryover in soil can severely affect sugarbeets grown in the year(s) following application. Two newly developed imidazolinone-resistant (IMI-R) sugarbeet somatic cell selections (Sir-13 and 93R30B) were examined for magnitude of resistance and extent of cross-resistance to other classes of ALS inhibitors and compared to a previously developed sulfonylurea-resistant (SU-R) selection, Sur. In vitro shoot culture tests indicated Sir-13 resistance was specific to imidazolinone (IMI) herbicides at approximately a 100-fold resistance compared to the sensitive control sugarbeet. Sur was 10,000-fold resistant to the sulfonylurea (SU) herbicide, chlorsulfuron, and 40-fold resistant to the triazolopyrimidine sulfonanilide (TP) herbicide, flumetsulam, but not cross-resistant to the IMI herbicides. 93R30B was selected for IMI-R from a plant homozygous for the SU-R allele, Sur, and displayed similar in vitro SU-R and TP-R as Sur, but also displayed a very high resistance to various IMI herbicides (400- to 3,600-fold). Compared to the sensitive control, Sir-13 was 300- and > 250-fold more resistant to imazethapyr and imazamox residues in soil, respectively. Response by whole plants to postemergence herbicide applications was similar to that observed in shoot cultures. Sir-13 exhibited > 100-fold resistance to imazethapyr as well as imazamox, and 93R30B showed > 250-fold resistance to both herbicides. 93R30B showed great enough resistance to imazamox to merit consideration of imazamox for use as a herbicide in these sugarbeets. Sir-13 showed a two- to threefold higher level of resistance in the homozygous vs. heterozygous state, indicating that like most ALS-inhibitor resistance traits, it was semidominantly inherited.

‘Cargill 921′, ‘Great Lakes 422′, ‘Northrup King 9410′, ‘Pioneer 3901′, ‘Pioneer 3737′, and ‘Stauffer 5650’ corn hybrids were tested in the greenhouse for imazaquin tolerance. Imazaquin rates that reduced shoot growth 50%, when compared to the untreated hybrid check, ranged from 17 to 50 g/ha. When averaged together, the three most tolerant hybrids (Cargill 921, Pioneer 3901, and Great Lakes 422) were approximately one-half as sensitive to imazaquin as the three least tolerant hybrids (Northrup King 9410, Pioneer 3737, and Stauffer 5650). Studies were conducted to determine if the observed differential tolerance was caused by differences in acetolactate synthase (ALS, EC 4.1.3.18) levels and sensitivity of ALS to imazaquin. Differential imazaquin uptake, translocation, and/or metabolism were also studied as a basis for the tolerance range as was seed size and seedling growth. There were differences among hybrids in the physiological and growth parameters studied; however, these differences did not correlate with imazaquin tolerance. None of the factors studied could alone account for the differences in imazaquin tolerance.

Three sugarbeet selections differing in cross-resistance to three classes of acetolactate synthase (ALS)-inhibiting herbicides have been developed using somatic cell selection. Sugarbeet selections resistant to imidazolinone herbicides, Sir-13 and 93R30B, do not metabolize [14C]-imazethapyr any faster or differently than sensitive, wild-type sugarbeets or a sulfonylurea-resistant/imidazolinone-sensitive selection, Sur. ALS specific activity from the three herbicide-resistant selections ranged from 73 to 93% of the wild-type enzyme extracts in the absence of herbicide, indicating enzyme overexpression was not a factor in resistance. Acetolactate synthase from Sir-13 plants showed a 40-fold resistance to imazethapyr but no resistance to chlorsulfuron or flumetsulam. Polymerase chain reaction amplification and sequencing of two regions of the ALS gene spanning all known sites for ALS-based herbicide resistance in plants indicated a single nucleotide change in the Sir-13 gene (G337 to A337) resulting in a deduced substitution of threonine for alanine at position 113 in the sugarbeet amino acid sequence. Sur ALS was not significantly resistant to imazethapyr, but was 1,000- and 50-fold resistant to chlorsulfuron and flumetsulam, respectively. Sur gene sequencing indicated a single nucleotide change (C562 to T562) resulting in a serine for proline substitution at position 188 of the ALS primary structure. The 93R30B nucleotide sequence indicated two mutations resulting in two deduced amino acid substitutions: threonine for alanine at position 113 plus serine for proline at position 188. The 93R30B double mutant incorporated the changes observed in each of the single mutants above and correlated with higher resistance levels to imazethapyr (> 1,000-fold), chlorsulfuron (4,300-fold), and flumetsulam (200-fold) at the ALS level than observed in either of the single mutants. 93R30B represents the first double mutant derived by a two-step selection process that incorporates two class-specific ALS-inhibitor resistance mutations to form a single broad cross-resistance trait. The interaction of the two altered amino acids is synergistic with respect to enzyme resistance vs. the resistance afforded by each of the individual mutations.

Several weedy red rice populations have evolved resistance to imidazolinone herbicides worldwide. The understanding of the factors related to the herbicide resistance in weedy red rice is important to prevent its occurrence in new areas where imidazolinone-resistant rice cultivars are being used, and to manage the new rice cultivars resistant to herbicides with modes of action other than the acetolactate synthase (ALS)-inhibitors that are being developed. The objectives of this study were to analyze the relationship of weedy red rice populations from southern Brazil with rice cultivars and wild Oryza species and to evaluate the occurrence of introgression from rice cultivars and seed migration as the origin of resistance to imidazolinone herbicides in weedy rice. The study was based on 27 weedy red rice populations, seven rice cultivars, and four wild Oryza species that were genotyped with 24 simple sequence repeats and three ALS-specific single-nucleotide polymorphism markers. A large proportion of the genetic variation of the weedy red rice populations was found within (74%) rather than among populations (26%). The weedy red rice populations were more closely related to the newer rice cultivars that are imidazolinone-resistant than to the older cultivars. The South American native Oryza glumaepatula and the other wild Oryza species—Oryza rufipogon, Oryza longistaminata, and Oryza glaberrima—clustered separately from weedy red rice populations, indicating a low likelihood of introgression among weedy red rice and these wild species. Seed migration was an important factor in the genetic structure of the evaluated weedy red rice populations, although gene flow by pollen from resistant cultivars was the principal reason for the spread of herbicide resistance.

Over 90% of Canadian kochia populations are resistant to acetolactate synthase (ALS)– inhibiting herbicides. We questioned whether the target site–based resistance could affect plant growth and competitiveness. Homozygous F2 herbicide-resistant (HR) kochia plants with an amino acid substitution at Trp574 (sources: Alberta [AB], Saskatchewan [SK], and Manitoba [MB]), or Pro197 (MB, AB with two populations) were grown in replacement series with homozygous F2 herbicide-susceptible (HS) plants from the corresponding heterogeneous population (total: six populations). In pure stands, growth of HR plants from AB and SK was similar to that of HS plants, regardless of mutation; conversely, MB2-HR plants (Trp574Leu) developed more slowly and were taller than MB2-HS plants. Final dry weight of HR plants in pure stands was similar across all six populations, whereas that for HS plants in pure stands and HR–HS plants in mixed stands (50–50%) varied with population. Results for AB and SK populations suggest little impact of either ALS mutation on kochia growth, whereas those for MB lines would suggest an unidentified factor (or factors) affecting the HS, HR, or both biotypes. The variable response within and between lines, and across HS biotypes highlights the importance of including populations of various origins and multiple susceptible controls in HR biotype studies.

Inhibitors of acetolactate synthase (ALS) are important herbicides for control of wild mustard, a common weed of the north central United States and Canada. Wild mustard that survived treatments with the ALS inhibitors cloransulam, imazethapyr, and thifensulfuron was sampled from a North Dakota soybean field in 1999. The mechanism of resistance and response of this wild mustard biotype to ALS-inhibiting herbicides was investigated. In vitro enzyme-inhibition experiments confirmed a resistance mechanism associated with the ALS enzyme; imazethapyr or imazamox at 1 × 10−4 M caused only 10 to 11% and 12 to 16% reductions in ALS activity, respectively. ALS from a susceptible wild mustard biotype was inhibited 50% (I50) with imazethapyr at 8 × 10−7 M or imazamox at 1.1 × 10−6 M. Whole-plant greenhouse treatments confirmed cross-resistance across ALS-inhibitor classes. Treatment with twice-normal field rates of thifensulfuron, ethametsulfuron, triflusulfuron, imazamox, imazethapyr, flumetsulam, cloransulam, flucarbazone, and imazamethabenz reduced biomass of the susceptible biotype at least 96% 28 d after treatment. Biomass of the resistant biotype was reduced 49% by triflusulfuron and 35% by thifensulfuron, but was not reduced by other herbicides. DNA sequence analysis of ALS genes from resistant and susceptible biotypes revealed a point mutation inferring a Trp-to-Leu amino acid substitution in ALS of the resistant biotype. This mutation, corresponding to position 574 of the Arabidopsis ALS amino acid sequence, is known to confer cross-resistance to ALS-inhibiting herbicides and is the probable cause of resistance in the wild mustard biotype. Phylogenetic analysis of wild mustard and canola ALS sequences confirmed that the Trp574 mutation arose within wild mustard and was not derived via introgression from imidazolinone-resistant canola. The results of this research indicate a naturally occurring target-site point mutation responsible for conferring cross-resistance to ALS-inhibiting herbicides in this wild mustard biotype.

Common ragweed and common cocklebur plants were collected at two sites each in Illinois, Minnesota, and Ohio to analyze intraspecific variability of the gene encoding acetolactate synthase (ALS). A 385-nucleotide fragment within the coding sequence of ALS was compared among 24 plants of each of these two species from the six locations. Common ragweed ALS was highly variable, with polymorphisms observed at 48 (12.5%) of the 385 nucleotides among the 24 plants. Despite the numerous nucleotide polymorphisms, only two inferred amino acid polymorphisms were identified. No apparent population structure was suggested by the ALS sequence data, indicating widespread gene flow consistent with the wind-pollinated nature of common ragweed. In contrast to common ragweed, no ALS polymorphisms were identified among the common cocklebur plants used in this study. As a basis for comparing the extremes observed between common ragweed and common cocklebur, ALS intraspecific variability also was investigated in 10 plants each of tall waterhemp and smooth pigweed. Normalized to the number of plants analyzed, the number of nucleotide polymorphisms for both tall waterhemp and smooth pigweed was greater than that in common cocklebur but less than that observed in common ragweed. Information on variability of herbicide target-site genes may be useful in predicting the likelihood for herbicide-resistance development. However, all four of the species investigated in this study have evolved resistance to ALS-inhibiting herbicides, despite the different levels of ALS variability observed.

Herbicides that target the enzyme acetolactate synthase (ALS) are among the most widely used in the world. Unfortunately, these herbicides are also notorious for their ability to select resistant (R) weed populations. Now, there are more weed species that are resistant to ALS-inhibiting herbicides than to any other herbicide group. In most cases, resistance to ALS-inhibiting herbicides is caused by an altered ALS enzyme. The frequent occurrence of weed populations resistant to ALS inhibitors can be attributed to the widespread usage of these herbicides, how they have been used, the strong selection pressure they exert, and the resistance mechanism. In several cropping systems, ALS-inhibiting herbicides were used repeatedly as the primary mechanism of weed control. These herbicides exert strong selection pressure because of their high activity on sensitive biotypes at the rates used and because of their soil residual activity. Several point mutations within the gene encoding ALS can result in a herbicide-resistant ALS. From investigations of numerous R weed biotypes, five conserved amino acids have been identified in ALS that, on substitution, can confer resistance to ALS inhibitors. Substitutions of at least 12 additional ALS amino acids can also confer herbicide resistance in plants and other organisms but, to date, have not been found in R weed populations. Mutations in ALS conferring herbicide resistance are at least partially dominant, and because the gene is nuclear inherited, it is transmitted by both seed and pollen. Furthermore, in many cases there is apparently a negligible fitness cost of the resistance gene in the absence of herbicide selection. Although resistance to ALS-inhibiting herbicides has been a bane to weed management, it has spurred many advances within and beyond the weed science discipline. As examples, resistance to ALS-inhibiting herbicides has been exploited in the development of herbicide-resistant crops, studies of weed population dynamics, and in developing protocols for targeted gene modification. Resistance to ALS-inhibiting herbicides has greatly affected weed science by influencing how we view the sustainability of our weed management practices, what we consider when developing and marketing new herbicides, and how we train new weed scientists.

Weed biotypes putatively resistant to acetolactate synthase (ALS)–inhibiting herbicides were reported by Iowa farmers from 1997 to 2001. Greenhouse studies confirmed cross-resistance to triazolopyrimidine sulfonanilide and sulfonylurea (SU) herbicides in giant ragweed from Scott County, IA (Werner Farm), which corresponded to resistance to susceptibility (R:S) GR50 (50% growth reduction) ratios of 21 and 28 to cloransulam and primisulfuron + prosulfuron, respectively. At the enzyme level, this represented a 49- and 20-fold I 50 (50% enzyme inhibition) increase. Cross-resistance to imidazolinone (IMI) and SU herbicides was also observed in common sunflower from Cherokee, IA. Compared with a susceptible biotype, the resistant common sunflower biotype demonstrated GR50 R:S ratios of 36 and 43 to imazethapyr and chlorimuron, respectively. Shattercane from Malvern, IA, was susceptible to nicosulfuron but was resistant to imazethapyr (GR50 R:S ratio = 29). The woolly cupgrass biotypes from Union County, IA (Pettit Farm and Travis Farm), were reportedly resistant but were identified susceptible to both IMI and SU herbicides. Using an in vivo ALS assay, extractable endogenous 2,3-diketone concentrations ranged from 25 to 71 nmol g−1 fresh weight for all species. Compared with susceptible biotypes, 2,3-diketone levels accumulated to at least twofold higher levels in treated resistant plants 120 h after herbicide application. Field history data suggested that resistance evolved independently in three environments where ALS-inhibiting herbicides represented an important component of the selection pressure.

A number of redroot pigweed and Powell amaranth populations from various locations in Ontario, Canada, have distinct patterns of resistance to the acetolactate synthase–inhibiting herbicides imazethapyr and thifensulfuron. This suggested the presence of diverse ALS gene mutations among these populations. Seven polymerase chain reaction primer pairs were used to amplify the gene to obtain full sequence information and to determine the identity of resistance-conferring mutations. There was a high degree of similarity in the ALS gene of the two species with only five nucleotides and one amino acid differing. A total of four herbicide resistance-conferring mutations were identified in the two species. The Ala122Thr, Ala205Val, and Trp574Leu amino acid substitutions were found in redroot pigweed whereas Ala122Thr, Trp574Leu, and Ser653Thr were detected in Powell amaranth. The pattern of resistance known to be conferred by the mutations concurred with the resistance level observed at the whole plant level. Distinct mutations being found in geographically separated populations suggest that selection for resistance occurred simultaneously in different locations. It reinforces the fact that resistance to ALS inhibitors is easily selected and that growers need to take this into account when formulating weed management strategies.