Target-site Mutation and Fitness Cost of Acetolactate Synthase Inhibitor-resistant Annual Bluegrass

Annual bluegrass (Poa annua L.) is an annual weed that is particularly troublesome in managed turfgrass. It has been controlled conventionally with herbicides, including acetolactate synthase (ALS) inhibitors. However, resistance to ALS inhibitors has been documented throughout the southeastern United States since 2012. A rate–response trial was conducted to confirm and determine the resistance level of suspected resistant P. annua biotypes from Mississippi (Reunion), followed by DNA sequencing to determine whether the mechanism of resistance is a target-site mutatio n. In addition, a fitness assay was conducted together with a susceptible biotype to determine whether resistance to ALS inhibitors is associated with decreased fitness. Reunion was at least 45 times more resistant to foramsulfuron than the standard susceptible biotype based on I50 estimates [I50 is the rate of herbicide giving a 50% response (50% visual necrosis)], requiring a predicted 331 g a.i./ha foramsulfuron for 50% control. DNA sequencing results identified a Trp574-to-Leu mutation in the ALS gene of the Reunion biotype, which has been shown by other studies to confer resistance to ALS inhibitors. Measurement of fitness parameters among the Reunion and susceptible biotypes demonstrated reduced seed yield, tillering, and flowering time in the resistant Reunion biotype, suggesting that ALS inhibitor resistance is possibly correlated to decreased fitness in P. annua. Alternative methods to control P. annua need to be considered as a result of the evolution of herbicide-resistant biotypes. An integrated management strategy to control P. annua weeds will help prevent further evolution of resistance. Because this study evaluated only the target-site mechanism of resistance, it is also necessary to determine whether the resistant biotype has reduced uptake, translocation, or enhanced metabolism as additional mechanisms of resistance. Consequently, a fitness study encompassing a more comprehensive list of plant parameters will provide conclusions of the fitness costs associated with ALS inhibitor resistance in P. annua. Chemical names: Foramsulfuron {1-(4,6-dimethoxypyrimidin-2-yl)-3-[2-(dimethylcarbamoyl)-5-formamidophenylsulfonyl] urea}.

Download Full-text

Multiple herbicide resistance in California Italian ryegrass (Lolium perenne ssp. multiflorum): characterization of ALS-inhibiting herbicide resistance

Weed Science ◽

10.1017/wsc.2019.1 ◽

2019 ◽

Vol 67 (3) ◽

pp. 273-280 ◽

Cited By ~ 4

Author(s):

Parsa Tehranchian ◽

Vijay K. Nandula ◽

Maor Matzrafi ◽

Marie Jasieniuk

Keyword(s):

Lolium Perenne ◽

Herbicide Resistance ◽

Resistance Mechanism ◽

Acetolactate Synthase ◽

Target Site ◽

Italian Ryegrass ◽

Resistance Level ◽

Als Inhibitors ◽

Inhibitor Resistance ◽

Als Inhibitor

AbstractMultiple resistance to glyphosate, sethoxydim, and paraquat was previously confirmed in two Italian ryegrass [Lolium perenne L. ssp. multiflorum (Lam.) Husnot] populations, MR1 and MR2, in northern California. Preliminary greenhouse studies revealed that both populations were also resistant to imazamox and mesosulfuron, both of which are acetolactate synthase (ALS)-inhibiting herbicides. In this study, three subpopulations, MR1-A (from seed of MR1 plants that survived a 16X rate of sethoxydim), MR1-P (from seed of MR1 plants that survived a 2X rate of paraquat), and MR2 (from seed of MR2 plants that survived a 16X rate of sethoxydim), were investigated to determine the resistance level to imazamox and mesosulfuron, evaluate other herbicide options for the control of these multiple resistant L. perenne ssp. multiflorum, and characterize the underlying ALS-inhibitor resistance mechanism(s). Based on LD50 values, the MR1-A, MR1-P, and MR2 subpopulations were 38-, 29-, 8-fold and 36-, 64-, and 3-fold less sensitive to imazamox and mesosulfuron, respectively, relative to the susceptible (Sus) population. Only MR1-P and MR2 plants were cross-resistant to rimsulfuron, whereas both MR1 subpopulations were cross-resistant to imazethapyr. Pinoxaden (ACCase inhibitor [phenylpyrazoline 'DEN']) only controlled MR2 and Sus plants at the labeled field rate. However, all plants were effectively controlled (>99%) with the labeled field rate of glufosinate. Based on I50 values, MR1-A, MR-P, and MR2 plants were 712-, 1,104-, and 3-fold and 10-, 18-, and 5-fold less responsive to mesosulfuron and imazamox, respectively, than the Sus plants. Sequence alignment of the ALS gene of resistant plants revealed a missense single-nucleotide polymorphism resulting in a Trp-574-Leu substitution in MR1-A and MR1-P plants, heterozygous in both, but not in the MR2 plants. An additional homozygous substitution, Asp-376-Glu, was identified in the MR1-A plants. Addition of malathion or piperonyl butoxide did not alter the efficacy of mesosulfuron on MR2 plants. In addition, the presence of 2,4-D had no effect on the response of mesosulfuron on the MR2 and Sus. These results suggest an altered target site is the mechanism of resistance to ALS inhibitors in MR1-A and MR1-P plants, whereas a non–target site based resistance apparatus is present in the MR2 plants.

Download Full-text

ALS–Resistant Annual Sedge (Cyperus compressus) Confirmed in Turfgrass

Weed Science ◽

10.1614/ws-d-15-00094.1 ◽

2016 ◽

Vol 64 (1) ◽

pp. 33-41 ◽

Cited By ~ 9

Author(s):

Patrick E. McCullough ◽

Jialin Yu ◽

J. Scott McElroy ◽

S. Chen ◽

H. Zhang ◽

...

Keyword(s):

Laboratory Experiments ◽

Acetolactate Synthase ◽

Shoot Biomass ◽

Resistance Level ◽

Physiological Basis ◽

History Of ◽

Leaf Tissues ◽

Als Inhibitors ◽

Inhibitor Resistance ◽

Als Inhibitor

Acetolactate synthase (ALS) inhibitors are widely used for POST control of sedges in turfgrass. A suspected resistant (R) biotype of annual sedge was collected from a bermudagrass turf in Georgia with a history of exclusive use of halosulfuron. Research was conducted to evaluate the resistance level of this biotype to halosulfuron, efficacy of ALS-inhibiting herbicides and other mechanisms of action for control, and the molecular and physiological basis for resistance. In greenhouse experiments, the halosulfuron rate required to reduce shoot biomass 50% in comparison with the nontreated at 8 wk after treatment (WAT) were 8 and > 1,120 g ai ha−1for the S (susceptible) and R biotypes, respectively. Imazapic, sulfosulfuron, and trifloxysulfuron reduced biomass of the S biotype greater than 60% at 8 WAT, but biomass was reduced less than 20% for the R biotype. Glufosinate, glyphosate, MSMA, and sulfentrazone reduced shoot biomass of the R biotype by 93, 86, 97, and 45%, respectively. In laboratory experiments, the halosulfuron concentration required to inhibit ALS activity by 50% in excised leaf tissues was 5.8 and > 1,000 μM for the S and R biotypes, respectively. Gene sequencing of the R biotype revealed a Pro-197-Ser substitution that confers resistance to ALS inhibitors. This is the first report of ALS-inhibitor resistance in annual sedge and herbicide resistance in a sedge species from a turfgrass system.

Download Full-text

Controlling Herbicide-Resistant Annual Bluegrass (Poa annua) Phenotypes with Methiozolin

Weed Technology ◽

10.1017/wet.2017.13 ◽

2017 ◽

Vol 31 (3) ◽

pp. 470-476 ◽

Cited By ~ 3

Author(s):

James T. Brosnan ◽

Jose J. Vargas ◽

Gregory K. Breeden ◽

Sarah L. Boggess ◽

Margaret A. Staton ◽

...

Keyword(s):

Acetolactate Synthase ◽

Target Site ◽

Multiple Resistance ◽

Herbicide Resistant ◽

Annual Bluegrass ◽

Microtubule Inhibitors ◽

Effective Option ◽

Target Site Resistance ◽

Susceptible Control ◽

Als Inhibitors

Methiozolin is an isoxazoline herbicide being investigated for selective POST annual bluegrass control in managed turfgrass. Research was conducted to evaluate methiozolin efficacy for controlling two annual bluegrass phenotypes with target-site resistance to photosystem II (PSII) or enolpyruvylshikimate-3-phosphate synthase (EPSPS)-inhibiting herbicides (i.e., glyphosate), as well as phenotypes with multiple resistance to microtubule and EPSPS or PSII and acetolactate synthase (ALS)-inhibiting herbicides. All resistant phenotypes were established in glasshouse culture along with a known herbicide-susceptible control and treated with methiozolin at 0, 125, 250, 500, 1000, 2000, 4000, or 8000 g ai ha−1. Methiozolin effectively controlled annual bluegrass with target-site resistance to inhibitors of EPSPS, PSII, as well as multiple resistance to EPSPS and microtubule inhibitors. Methiozolin rates required to reduce aboveground biomass of these resistant phenotypes 50% (GR50 values) were not significantly different from the susceptible control, ranging from 159 to 421 g ha−1. A phenotype with target-site resistance to PSII and ALS inhibitors was less sensitive to methiozolin (GR50=862 g ha−1) than a susceptible phenotype (GR50=423 g ha−1). Our findings indicate that methiozolin is an effective option for controlling select annual bluegrass phenotypes with target-site resistance to several herbicides.

Download Full-text

In vitro and whole-plant magnitude and cross-resistance characterization of two imidazolinone-resistant sugarbeet (Beta vulgaris) somatic cell selections

Weed Science ◽

10.1017/s0043174500090123 ◽

1998 ◽

Vol 46 (1) ◽

pp. 24-29 ◽

Cited By ~ 8

Author(s):

Terry R. Wright ◽

Donald Penner

Keyword(s):

Somatic Cell ◽

Shoot Culture ◽

Acetolactate Synthase ◽

Cross Resistance ◽

Shoot Cultures ◽

Als Inhibitors ◽

Whole Plants ◽

Inhibitor Resistance ◽

Als Inhibitor

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.

Download Full-text

Acetolactate Synthase Inhibitor–Resistant False Cleavers (Galium spurium) in Western Canada

Weed Technology ◽

10.1614/wt-d-11-00075.1 ◽

2012 ◽

Vol 26 (1) ◽

pp. 151-155 ◽

Cited By ~ 11

Author(s):

Hugh J. Beckie ◽

Suzanne I. Warwick ◽

Connie A. Sauder ◽

Gina M. Kelln ◽

Chris Lozinski

Keyword(s):

Point Mutations ◽

Low Frequency ◽

Acetolactate Synthase ◽

Western Canada ◽

Resistance Factors ◽

Als Inhibitors ◽

Synthase Inhibitor ◽

Inhibitor Resistance ◽

Als Inhibitor ◽

Galium Spurium

Cleavers species (false cleavers and catchweed bedstraw) are among the top 10 most abundant weeds across the prairie region of western Canada, and are increasing in relative abundance at the fastest rate since the 1970s. In 2008, two false cleavers populations from Tisdale and Choiceland, Saskatchewan, were suspected of acetolactate synthase (ALS) –inhibitor resistance. Dose-response experiments were conducted with the use of imazethapyr and florasulam, both ALS inhibitors, as well as fluroxypyr, a synthetic auxin. Additionally, a 1,954–base-pair region of theALSgene including sites known to conferALSresistance were sequenced. Both populations were highly resistant to imazethapyr (resistance factors greater than 100), one population (Tisdale) was highly resistant to florasulam (Choiceland population susceptible, although a second, larger screening of 200 individuals indicated low frequency [2%] florasulam resistance), and both populations were susceptible to fluroxypyr. All sequenced Tisdale individuals screened with imazethapyr posessed the Trp574Leu mutation. In contrast, three point mutations were found for Choiceland individuals sequenced: Ser653Asn, Trp574Leu, and Asp376Glu. TheseALStarget-site mutations have not been documented previously in this species.

Download Full-text

Nontarget-Site Resistance to ALS Inhibitors in Waterhemp (Amaranthus tuberculatus)

Weed Science ◽

10.1614/ws-d-14-00139.1 ◽

2015 ◽

Vol 63 (2) ◽

pp. 399-407 ◽

Cited By ~ 31

Author(s):

Jiaqi Guo ◽

Chance W. Riggins ◽

Nicholas E. Hausman ◽

Aaron G. Hager ◽

Dean E. Riechers ◽

...

Keyword(s):

Acetolactate Synthase ◽

Enzyme Assays ◽

Whole Plant ◽

Site Of Action ◽

Amaranthus Tuberculatus ◽

Resistance Trait ◽

Als Inhibitors ◽

Moderate Resistance ◽

Inhibitor Resistance ◽

Als Inhibitor

A waterhemp population (MCR) previously characterized as resistant to 4-hydroxyphenylpyruvate dioxygenase and photosystem II inhibitors demonstrated both moderate and high levels of resistance to acetolactate synthase (ALS) inhibitors. Plants from the MCR population exhibiting high resistance to ALS inhibitors contained the commonly found Trp574Leu ALS amino acid substitution, whereas plants with only moderate resistance did not have this substitution. A subpopulation (JG11) was derived from the MCR population in which the moderate-resistance trait was isolated from the Trp574Leu mutation. Results from DNA sequencing and ALS enzyme assays demonstrated that resistance to ALS inhibitors in the JG11 population was not due to an altered site of action. This nontarget-site ALS-inhibitor resistance was characterized with whole-plant dose–response experiments using herbicides from each of the five commercialized families of ALS-inhibiting herbicides. Resistance ratios ranging from 3 to 90 were obtained from the seven herbicides evaluated. Nontarget-site resistance to ALS has been rarely documented in eudicot weeds, and adds to the growing list of resistance traits evolved in waterhemp.

Download Full-text

Acetolactate Synthase (ALS) Target-Site Mutations in ALS Inhibitor-Resistant Russian Thistle (Salsola tragus)

Weed Science ◽

10.1614/ws-d-09-00083.1 ◽

2010 ◽

Vol 58 (3) ◽

pp. 244-251 ◽

Cited By ~ 26

Author(s):

Suzanne I. Warwick ◽

Connie A. Sauder ◽

Hugh J. Beckie

Keyword(s):

Molecular Basis ◽

Genome Structure ◽

Acetolactate Synthase ◽

Molecular Techniques ◽

Target Site ◽

Weed Species ◽

Single Nucleotide ◽

Herbicide Resistant ◽

Inhibitor Resistance ◽

Als Inhibitor

ALS inhibitor-resistant biotypes are the fastest growing class of herbicide-resistant (HR) weeds. A Canadian ALS inhibitor-resistant biotype of Russian thistle was first reported in 1989. The molecular basis for ALS-inhibitor resistance is unknown for Canadian populations of this polyploid weed species, and was determined in this study for one Alberta and two Saskatchewan HR Russian thistle populations. HR plants survived spray application of the ALS-inhibitor mixture thifensulfuron : tribenuron in the greenhouse. All three HR Russian thistle populations were heterogeneous and contained both HR and herbicide-susceptible (HS) individuals. The molecular basis for resistance was determined by sequencing theALSgene and/or conducting a TaqMan genotyping assay for single nucleotide polymorphism (SNP) for the Trp574Leu mutation. Two target-site mutations were observed: Trp574Leu in all three biotypes (554 individuals) and Pro197Gln in one biotype (one individual), suggesting multiple-founding events for Russian thistle HR populations in western Canada. Segregation patterns among F1 and F2 progeny arrays of HR lines sprayed under greenhouse conditions varied; some segregated (i.e., had HR and HS progeny), whereas other lines were exclusively HR. In contrast, no segregation of molecular types, i.e., Trp574, Trp/Leu574and Leu574, as would be expected with heterozygosity at a single locus Trp/Leu574, was observed. Such lack of segregation is consistent with the polyploid genome structure of Russian thistle and the presence of two copies of theALSgene. The presence of more than oneALSgene confounded the ability of the molecular techniques to accurately identify “true” heterozygotes in this study.

Download Full-text

Transfer and Expression of ALS Inhibitor Resistance from Palmer Amaranth (Amaranthus palmeri) to anA. spinosus×A. palmeriHybrid

Weed Science ◽

10.1614/ws-d-15-00172.1 ◽

2016 ◽

Vol 64 (2) ◽

pp. 240-247 ◽

Cited By ~ 17

Author(s):

William T. Molin ◽

Vijay K. Nandula ◽

Alice A. Wright ◽

Jason A. Bond

Keyword(s):

Field Experiments ◽

Acetolactate Synthase ◽

Palmer Amaranth ◽

Enzyme Assays ◽

Amaranthus Palmeri ◽

Weed Species ◽

Als Inhibitors ◽

Inhibitor Resistance ◽

Als Inhibitor ◽

Interspecific Transfer

Transfer of herbicide resistance among closely related weed species is a topic of growing concern. A spiny amaranth × Palmer amaranth hybrid was confirmed resistant to several acetolactate synthase (ALS) inhibitors including imazethapyr, nicosulfuron, pyrithiobac, and trifloxysulfuron. Enzyme assays indicated that the ALS enzyme was insensitive to pyrithiobac and sequencing revealed the presence of a known resistance conferring point mutation, Trp574Leu. Alignment of the ALS gene for Palmer amaranth, spiny amaranth, and putative hybrids revealed the presence of Palmer amaranth ALS sequence in the hybrids rather than spiny amaranth ALS sequences. In addition, sequence upstream of the ALS in the hybrids matched Palmer amaranth and not spiny amaranth. The potential for transfer of ALS inhibitor resistance by hybridization has been demonstrated in the greenhouse and in field experiments. This is the first report of gene transfer for ALS inhibitor resistance documented to occur in the field without artificial/human intervention. These results highlight the need to control related species in both field and surrounding noncrop areas to avoid interspecific transfer of resistance genes.

Download Full-text

A Statewide Survey of PPO-Inhibitor Resistance and the Prevalent Target-Site Mechanisms in Palmer amaranth (Amaranthus palmeri) Accessions from Arkansas

Weed Science ◽

10.1017/wsc.2017.68 ◽

2017 ◽

Vol 66 (2) ◽

pp. 149-158 ◽

Cited By ~ 24

Author(s):

Vijay K. Varanasi ◽

Chad Brabham ◽

Jason K. Norsworthy ◽

Haozhen Nie ◽

Bryan G. Young ◽

...

Keyword(s):

Resistance Mechanism ◽

Acetolactate Synthase ◽

Resistance Mechanisms ◽

Palmer Amaranth ◽

Target Site ◽

Evolution Of Resistance ◽

Target Site Resistance ◽

Novel Target ◽

Als Inhibitors ◽

Inhibitor Resistance

Palmer amaranth is one of the most problematic weeds in the midsouthern United States, and the evolution of resistance to protoporphyrinogen oxidase (PPO) inhibitors in biotypes already resistant to glyphosate and acetolactate synthase (ALS) inhibitors is a major cause of concern to soybean and cotton growers in these states. A late-season weed-escape survey was conducted in the major row crop–producing counties (29 counties) to determine the severity of PPO-inhibitor resistance in Arkansas. A total of 227 Palmer amaranth accessions were sprayed with fomesafen at 395 g ha−1to identify putative resistant plants. A TaqMan qPCR assay was used to confirm the presence of the ΔG210 codon deletion or the R128G/M (homologous to R98 mutation in common ragweed) target-site resistance mechanisms in thePPX2gene. Out of the 227 accessions screened, 44 were completely controlled with fomesafen, and 16 had only one or two severely injured plants (≥98% mortality) when compared with the 1986 susceptible check (100% mortality). The remaining 167 accessions were genotypically screened, and 82 (49%) accessions were found to harbor the ΔG210 deletion in thePPX2gene. The R128G was observed in 47 (28%) out of the 167 accessions screened. The mutation R128M, on the other hand was rare, found in only three accessions. About 13% of the accessions were segregating for both the ΔG210 and R128G mutations. Sixteen percent of the tested accessions had mortality ratings <90% and did not test positive for the ΔG210 or the R128G/M resistance mechanisms, indicating that a novel target or non–target site resistance mechanism is likely. Overall, PPO inhibitor–resistant Palmer amaranth is widespread in Arkansas, and the ΔG210 resistance mechanism is especially dominant in the northeast corridor, while the R128G mutation is more prevalent in counties near Memphis, TN.

Download Full-text

Characterization of Acetolactate Synthase (ALS)-Inhibitor Resistance in Pennsylvania smartweed (Persicaria pensylvanica)

Weed Science ◽

10.1017/wsc.2018.44 ◽

2018 ◽

Vol 66 (6) ◽

pp. 710-714

Author(s):

Vijay K. Varanasi ◽

Jason K. Norsworthy ◽

Chad Brabham ◽

Robert C. Scott

Keyword(s):

United States ◽

Resistance Mechanism ◽

Resistance Index ◽

Acetolactate Synthase ◽

Reduced Tillage ◽

Tillage Systems ◽

Als Inhibitors ◽

Inhibitor Resistance ◽

Als Inhibitor

AbstractPennsylvania smartweed [Persicaria pensylvanica(L.) M. Gómez] is a common weed of rice (Oryza sativaL.) in the midsouthern United States and has recently become a concern for farmers because of reduced tillage systems. Acetolactate synthase (ALS) inhibitors have been extensively used for controlling smartweeds in imidazolinone-resistant and conventional rice. In the present study, we confirmed resistance to commonly used ALS inhibitors in rice and characterized the underlying resistance mechanism in aP. pensylvanicabiotype from southeast Missouri. A dose–response experiment was conducted in the greenhouse using bensulfuron-methyl, imazethapyr, and bispyribac-sodium to determine the resistance index (resistance/susceptibility [R/S]) based on GR50estimates. The target-siteALSgene was amplified from R and S plants, and sequences were analyzed for mutations known to confer ALS-inhibitor resistance. TheP. pensylvanicabiotype in question was found to be resistant to bensulfuron-methyl (R/S=2,330), imazethapyr (R/S=12), and bispyribac-sodium (R/S=6). Sequencing of theALSgene from R plants revealed two previously known mutations (Pro-197-Ser, Ala-122-Ser) conferring resistance to sulfonylureas and imidazolinones. This is the first report of ALS-inhibitor resistance inP. pensylvanica.

Download Full-text