Molecular Basis of Resistance to ALS-Inhibitor Herbicides in Greater Beggarticks

Weed Science ◽  
2009 ◽  
Vol 57 (5) ◽  
pp. 474-481 ◽  
Author(s):  
Fabiane P. Lamego ◽  
Dirk Charlson ◽  
Carla A. Delatorre ◽  
Nilda R. Burgos ◽  
Ribas A. Vidal

Soybean is a major crop cultivated in Brazil, and acetolactate synthase (ALS)-inhibiting herbicides are widely used to control weeds in this crop. The continuous use of these ALS-inhibiting herbicides has led to the evolution of herbicide-resistant weeds worldwide. Greater beggarticks is a polyploid species and one of the most troublesome weeds in soybean production since the discovery of ALS-resistant biotypes in 1996. To confirm and characterize the resistance of greater beggarticks to ALS inhibitors, whole-plant bioassays and enzyme experiments were conducted. To investigate the molecular basis of resistance in greater beggarticks theALSgene was sequenced and compared between susceptible and resistant biotypes. Our results confirmed that greater beggarticks is resistant to ALS inhibitors and also indicated it possesses at least three isoforms of theALSgene. Analysis of the nucleotide and deduced amino acid sequences among the isoforms and between the biotypes indicated that a single point mutation, G–T, in oneALSisoform from the resistant biotype resulted in an amino acid substitution, Trp574Leu. Two additional substitutions were observed, Phe116Leu and Phe149Ser, in a second isoform of the resistant biotype, which were not yet reported in any other herbicide-resistantALSgene; thus, their role in conferring herbicide resistance is not yet ascertained. This is the first report ofALSmutations in an important, herbicide-resistant weed species from Brazil.

Weed Science ◽  
2010 ◽  
Vol 58 (3) ◽  
pp. 244-251 ◽  
Author(s):  
Suzanne I. Warwick ◽  
Connie A. Sauder ◽  
Hugh J. Beckie

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.


2012 ◽  
Vol 92 (2) ◽  
pp. 303-309 ◽  
Author(s):  
Kee Woong Park ◽  
Judith M. Kolkman ◽  
Carol A. Mallory-Smith

Park, K. W., Kolkman, J. M. and Mallory-Smith, C. A. 2012. Point mutation in acetolactate synthase confers sulfonylurea and imidazolinone herbicide resistance in spiny annual sow-thistle [Sonchus asper (L.) Hill]. Can. J. Plant Sci. 92: 303–309. Suspected thifensulfuron resistant spiny annual sow-thistle was identified near Colfax, Washington, in two fields with a winter wheat and lentil rotation. Therefore, studies were conducted to examine resistance of spiny annual sow-thistle to thifensulfuron and cross-resistance to other acetolactate synthase inhibitors and to determine the physiological and molecular basis for herbicide resistance. Whole-plant bioassay confirmed that the biotype was highly resistant to the sulfonylurea (SU) herbicides, thifensulfuron, metsulfuron, and prosulfuron. The resistant (R) biotype was also highly resistant to the imidazolinone (IMI) herbicides, imazamox and imazethapyr. An in vivo acetolactate synthase (ALS) assay indicated that the concentrations of SU and IMI herbicides required for 50% inhibition (I50) were more than 10 times greater for R biotype compared with susceptible (S) biotype. Analysis of the nucleotide and predicted amino acid sequences for ALS genes demonstrated a single-point mutation from C to T at the als1 gene, conferring the substitution of the amino acid leucine for proline in the R biotype at position197. The results of this research indicate that the resistance of spiny annual sow-thistle to SU and IMI herbicides is due to on altered target site and caused by a point mutation in the als1 gene.


Weed Science ◽  
2005 ◽  
Vol 53 (5) ◽  
pp. 631-639 ◽  
Author(s):  
Suzanne I. Warwick ◽  
Connie Sauder ◽  
Hugh J. Beckie

Multiple cases of ALS inhibitor-resistant weed biotypes are reported for many species, including wild mustard. The physiological extent and molecular basis of resistance to ALS inhibitors was compared in four biotypes of wild mustard from western Canada: a sulfonylurea (SU)-resistant (R) biotype from Manitoba detected in 1992; an SU (ethametsulfuron)-R biotype from Alberta detected in 1993 (metabolism-based resistance); an SU-R biotype from Manitoba detected in 2002; and a SU- and imidazolinone (IMI)-R biotype from Saskatchewan detected in 2002. Herbicide dose-response experiments confirmed that the two Manitoba biotypes were resistant to the SU herbicides ethametsulfuron and tribenuron : thifensulfuron mixture, whereas the Saskatchewan biotype was resistant to both SU herbicides and to imazethapyr, an IMI herbicide. Sequence analysis of theALSgene detected target site mutations in three of the four R biotypes, with amino acid substitutions Pro197(CCT) to Ser (TCT) [Domain A of the gene] in the two SU-R Manitoba biotypes and Trp574(TGG) to Leu (TTG) [Domain B] in the Saskatchewan biotype. The Alberta SU-R biotype had the sameALSnucleotide and amino acid sequence as the susceptible population at these two positions. Two heterozygous individuals [Trp574(Tt/gG)] were detected in the Saskatchewan biotype, and genetic segregation for nucleotide bases and resistance phenotype was consistent with single gene control. Nucleotide variation in neutral regions of theALSgene varied with biotype, with no variation in the two Manitoba biotypes, two variants in the Saskatchewan biotype, and 16 neutral nucleotide polymorphisms (0.9%) in the Alberta biotype. The occurrence of at least three different ALS inhibitor-R biotypes in this important weed species is likely to impact negatively on the use of ALS inhibitors, such as the IMIs, and serves as a warning for strict implementation of herbicide rotations to prevent or delay the evolution and spread of such populations.


PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0258685
Author(s):  
Christopher E. Buddenhagen ◽  
Trevor K. James ◽  
Zachary Ngow ◽  
Deborah L. Hackell ◽  
M. Phil Rolston ◽  
...  

To estimate the prevalence of herbicide-resistant weeds, 87 wheat and barley farms were randomly surveyed in the Canterbury region of New Zealand. Over 600 weed seed samples from up to 10 mother plants per taxon depending on abundance, were collected immediately prior to harvest (two fields per farm). Some samples provided by agronomists were tested on an ad-hoc basis. Over 40,000 seedlings were grown to the 2–4 leaf stage in glasshouse conditions and sprayed with high priority herbicides for grasses from the three modes-of-action acetyl-CoA carboxylase (ACCase)-inhibitors haloxyfop, fenoxaprop, clodinafop, pinoxaden, clethodim, acetolactate synthase (ALS)-inhibitors iodosulfuron, pyroxsulam, nicosulfuron, and the 5-enolpyruvyl shikimate 3-phosphate synthase (EPSPS)-inhibitor glyphosate. The highest manufacturer recommended label rates were applied for the products registered for use in New Zealand, often higher than the discriminatory rates used in studies elsewhere. Published studies of resistance were rare in New Zealand but we found weeds survived herbicide applications on 42 of the 87 (48%) randomly surveyed farms, while susceptible reference populations died. Resistance was found for ALS-inhibitors on 35 farms (40%) and to ACCase-inhibitors on 20 (23%) farms. The number of farms with resistant weeds (denominator is 87 farms) are reported for ACCase-inhibitors, ALS-inhibitors, and glyphosate respectively as: Avena fatua (9%, 1%, 0% of farms), Bromus catharticus (0%, 2%, 0%), Lolium spp. (17%, 28%, 0%), Phalaris minor (1%, 6%, 0%), and Vulpia bromoides (0%, not tested, 0%). Not all farms had the weeds present, five had no obvious weeds prior to harvest. This survey revealed New Zealand’s first documented cases of resistance in P. minor (fenoxaprop, clodinafop, iodosulfuron) and B. catharticus (pyroxsulam). Twelve of the 87 randomly sampled farms (14%) had ALS-inhibitor chlorsulfuron-resistant sow thistles, mostly Sonchus asper but also S. oleraceus. Resistance was confirmed in industry-supplied samples of the grasses Digitaria sanguinalis (nicosulfuron, two maize farms), P. minor (iodosulfuron, one farm), and Lolium spp. (cases included glyphosate, haloxyfop, pinoxaden, iodosulfuron, and pyroxsulam, 9 farms). Industry also supplied Stellaria media samples that were resistant to chlorsulfuron and flumetsulam (ALS-inhibitors) sourced from clover and ryegrass fields from the North and South Island.


Weed Science ◽  
1992 ◽  
Vol 40 (4) ◽  
pp. 670-677 ◽  
Author(s):  
Mary J. Guttieri ◽  
Charlotte V. Eberlein ◽  
Carol A. Mallory-Smith ◽  
Donald C. Thill ◽  
David L. Hoffman

The DNA sequence of a 196 base pair (bp) region of the acetolactate synthase (ALS) genes of three weed species, kochia, prickly lettuce, and Russian thistle, was determined. This region encompasses the coding sequence for Domain A, a region of the amino acid sequence previously demonstrated to play a pivotal role in conferring resistance to herbicides that inhibit ALS. The Domain A DNA sequence from a chlorsulfuron-resistant (R) prickly lettuce biotype from Idaho differed from that of a chlorsulfuron-susceptible (S) biotype by a single point mutation, which substituted a histidine for a proline. The Domain A DNA sequence from an R kochia biotype from Kansas also differed from that of an S biotype by a single point mutation in the same proline codon. This point mutation, however, conferred substitution of threonine for proline. Two different ALS-homologous sequences were isolated from an R biotype of Russian thistle. Neither sequence encoded amino acid substitutions in Domain A that differed from the consensus S sequence. The DNA sequence variation among the R and S kochia biotypes was used to characterize six Ada County, Idaho, kochia collections for correlation between phenotypic chlorsulfuron susceptibility and restriction digest patterns (RFLPs) of polymerase chain reaction amplification products. Most collections showed excellent correspondence between the RFLP patterns and the phenotypic response to chlorsulfuron application. However, one entirely R collection had the RFLP pattern of the S biotype, suggesting that resistance was not due to mutation in the proline codon.


2020 ◽  
pp. 4-9
Author(s):  
V. Schwartau ◽  
L. Mykhalska

Goal. Investigate the possibility to use 1,8-naphthalic anhydride metabolism inductor to control acetolactate synthase (ALS) inhibitor-resistant biotype of common graminicides of aryloxyphenoxypropionic acid class in rice. Methodology. The interaction of 1,8-naphthalic anhydride and fenoxaprop-p-ethyl on variety Vikont rice plants was studied under laboratory aseptic conditions. The data were statistically processed. Results. In Ukraine we have identified the biotype of resistant to herbicide ALS inhibitors Echinochloa crus-galli, which is cross-resistant to widely used herbicides — ALS inhibitors of the following chemical classes: imidazolinones (imazamox, imazapyr), sulfonylurea (nicosulfuron), triazolopyrimidines (penoxsulam). The possibilities of chemical control of weeds in rice, corn, sunflower, etc. crops are significantly limited. Multi-resistance of this weed biotype to herbicides — inhibitors of photosynthesis, mitotic cycle, 5-enolpyruvylshikimate-3-phosphate synthase, acetyl-CoA-carboxylase, protein synthesis — has not been detected. Therefore, the use of graminicides of aryloxyphenoxypropionate class is promising for the control of this ALS-resistant biotype of Echinochloa crus-galli. To increase the selectivity of fenoxaprop-P-ethyl application to rice plants, we propose to treat the seeds of the crop with the inductor of xenobiotics metabolism in plants — 1.8-naphthalic anhydride before sowing. When using 1.8-naphthalic anhydride in concentrations of 10-5 M, phytotoxicity of fenoxaprop-P-ethyl in concentrations of 10-6 and 10-5 M to rice plants is effectively reduced. Conclusions. The use of 1.8-naphthalic anhydride is promising for increasing the selectivity of fenoxaprop-P-ethyl for rice plants and allows the development of technologies using graminicides of aryloxyphenoxypropionate class to control ALS-resistant biotype of Echinochloa crus-galli in crops. Also, it is necessary to pay attention to the problem of ALS-resistant weed biotype proliferation control in agrophytocenoses in regions of Ukraine.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Maria J. García ◽  
Candelario Palma-Bautista ◽  
José G. Vazquez-Garcia ◽  
Antonia M. Rojano-Delgado ◽  
María D. Osuna ◽  
...  

Abstract Amaranthus hybridus is one of the main weed species in Córdoba, Argentina. Until recently, this weed was effectively controlled with recurrent use of glyphosate. However, a population exhibiting multiple resistance (MR2) to glyphosate and imazamox appeared in a glyphosate resistant (GR) soybean field, with levels of resistance up to 93 and 38-fold higher to glyphosate and imazamox, respectively compared to the susceptible (S) population. In addition to imidazolinones, MR2 plants showed high resistance levels to sulfonylamino-carbonyl (thio) benzoates and moderate resistance to sulfonylureas and triazolopyrimidines. Multiple amino acid substitutions were found in both target genes, acetolactate synthase (ALS) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), responsible for conferring high herbicides resistance levels in this A. hybridus population. In the case of EPSPS, the triple amino acid substitution TAP-IVS was found. In addition, MR2 plants also showed increased EPSPS gene expression compared to susceptible plants. A Ser653Asn substitution was found in the ALS sequence of MR2, explaining the pattern of cross-resistance to the ALS-inhibitor herbicide families found at the ALS enzyme activity level. No other mutations were found in other conserved domains of the ALS gene. This is the first report worldwide of the target site resistance mechanisms to glyphosate and ALS inhibitors in multiple herbicide resistance Amaranthus hybridus.


2021 ◽  
pp. 1-7
Author(s):  
Jatinder S. Aulakh ◽  
Parminder S. Chahal ◽  
Vipan Kumar ◽  
Andrew J. Price ◽  
Karl Guillard

Abstract Palmer amaranth is the latest pigweed species documented in Connecticut; it was identified there in 2019. In a single-dose experiment, the Connecticut Palmer amaranth biotype survived the field-use rates of glyphosate (840 g ae ha−1) and imazaquin (137 g ai ha−1) herbicides applied separately. Additional experiments were conducted to (1) determine the level of resistance to glyphosate and acetolactate synthase (ALS) inhibitors in the Connecticut-resistant (CT-Res) biotype using whole-plant dose-response bioassays, and (2) evaluate the response of the CT-Res biotype to POST herbicides commonly used in Connecticut cropping systems. Based on the effective dose required for 90% control (ED90), the CT-Res biotype was 10-fold resistant to glyphosate when compared with the Kansas-susceptible (KS-Sus) biotype. Furthermore, the CT-Res biotype was highly resistant to ALS-inhibitor herbicides; only 18% control was achieved with 2,196 g ai ha−1 imazaquin. The CT-Res biotype was also cross-resistant to other ALS-inhibitor herbicides, including chlorimuron-ethyl (13.1 g ai ha−1), halosulfuron-methyl (70 g ai ha−1), and sulfometuron-methyl (392 g ai ha−1). The CT-Res Palmer amaranth was controlled 75% to 100% at 21 d after treatment (DAT) with POST applications of 2,4-D (386 g ae ha−1), carfentrazone-ethyl (34 g ai ha−1), clopyralid (280 g ae ha−1), dicamba (280 g ae ha−1), glufosinate (595 g ai ha−1), lactofen (220 g ai ha−1), oxyfluorfen (1,121g ai ha−1), and mesotrione (105 g ai ha−1) herbicides. Atrazine (2,240 g ai ha−1) controlled the CT-Res biotype only 52%, suggesting the biotype is resistant to this herbicide as well. Here we report the first case of Palmer amaranth from Connecticut with multiple resistance to glyphosate and ALS inhibitors. Growers should proactively use all available weed control tactics, including the use of effective PRE and alternative POST herbicides (tested in this study), for effective control of the CT-Res biotype.


Weed Science ◽  
2016 ◽  
Vol 64 (2) ◽  
pp. 240-247 ◽  
Author(s):  
William T. Molin ◽  
Vijay K. Nandula ◽  
Alice A. Wright ◽  
Jason A. Bond

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.


Agronomy ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1057 ◽  
Author(s):  
Rafael R. Mendes ◽  
Hudson K. Takano ◽  
Rubem S. Oliveira ◽  
Fernando S. Adegas ◽  
Todd A. Gaines ◽  
...  

Wild poinsettia (Euphorbia heterophylla L.) is an important weed species in southern Brazil, especially due to the evolution of multiple herbicide resistance (e.g., acetolactate synthase (ALS)- inhibitors, protoporphyrinogen oxidase inhibitors, and glyphosate). The mechanism of resistance to imazamox was investigated in two wild poinsettia populations (R1 and R2) from southern Brazil and compared to a known susceptible (S) population. Imazamox dose-response experiments revealed high levels of resistance: 45-fold and 224.5-fold based on dry biomass reduction, for R1 and R2, respectively. Extremely high concentrations of imazamox (20,000 µM) were not sufficient to provide 50% inhibition of ALS enzyme activity (I50) for R1 or R2. Hence, resistance levels were estimated to be greater than 123-fold for both populations based on in vitro ALS assays. The ALS gene from all R1 and R2 plants had a Trp574Leu mutation. A genotyping assay was developed to discriminate resistant and susceptible alleles based on the Trp574Leu mutation.


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