Point Mutations as Main Resistance Mechanism Together With P450-Based Metabolism Confer Broad Resistance to Different ALS-Inhibiting Herbicides in Glebionis coronaria From Tunisia (original) (raw)
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Weed Research, 2012
Acetolactate synthase (ALS)-inhibiting herbicide resistance is common in Raphanus raphanistrum (wild radish) populations across the Western Australian (WA) grain belt. This study investigates the molecular and biochemical basis of ALS herbicide resistance in five R. raphanistrum populations. Five known ALS herbicide resistance-endowing mutations were identified, and their resistance spectrum to ALS-inhibiting herbicides was determined using purified populations individually homozygous for each mutation (except for Pro-197-Ala). Plants homozygous for ALS mutations at Pro-197 were found to be cross-resistant to ALS-inhibiting sulfonylurea (SU) and triazolopyrimidine (TP) herbicides, while plants homozygous for Trp-574-Leu were resistant to SU, TP and imidazolinone (IMI) ALS herbicide classes. The Asp-376-Glu mutation is reported here for the first time in R. raphanistrum populations and characterised at both the whole-plant and enzyme level. Plants homozygous for Asp-376-Glu were highly resistant to SU and TP herbicides, based on LD 50 R ⁄ S ratios (>130 and 128 respectively) and I 50 R ⁄ S ratios (170 and >110 respectively). In contrast, these plants were moderately resistant to the IMI imazamox (LD 50 R ⁄ S ratio of 8, I 50 R ⁄ S ratio of 3) and imazethapyr (I 50 R ⁄ S ratio of 8) and susceptible to imazapyr (I 50 R ⁄ S ratio of 0.76). A novel observation in this study is that resistance of homozygous Glu-376 plants is associated with a remarkable growth reduction in the presence of the ALS herbicides tested, making early resistance diagnosis and management difficult.
Journal of Agricultural Science and Technology A, 2015
The good understanding of the mechanisms of resistance to herbicides in weeds is a necessity to implement sustainable weed management strategies. Here, a study was conducted to characterize the molecular bases of resistance to acetyl coenzyme A carboxylase (ACCase) and acetolactate synthase (ALS) inhibiting herbicides in Lolium rigidum populations from Tunisia. Nine Lolium rigidum (ryegrass) populations collected in wheat fields from Northern Tunisia were investigated for their resistance to two ACCase-inhibiting herbicides and an ALS-inhibiting herbicide. All populations were tested in the greenhouse in pots using the commercial dose to determine resistance status. Survival plants were also tested for the presence of two ACCase (L1781 and N2041) and two ALS (P197 and W574) mutant resistant alleles using molecular markers. Resistance to ACCase-inhibiting herbicides was found in all tested populations. Comparison of the results from herbicide sensitivity bioassays with genotyping indicated that more than 80% of the plants resistant to ACC-inhibiting herbicides would be resistant via increased herbicide metabolism. However, ALS-inhibiting herbicides are still more or less controlling ACCase resistant populations, so indicating that the selection process of resistance is ongoing. Target-site resistance appears to be the major mechanism for these early cases of ALS inhibitor resistance. This study reported the first case of resistance to ALS-inhibiting herbicides in ryegrass in Tunisia, and investigated the molecular bases of this resistance. It establishes the clear importance of non target-site resistance to ACCase-and/or ALS-inhibiting herbicides.
Molecular Biology Reports, 2019
Acetolactate synthase (ALS)-inhibiting herbicides have been widely used for effective management and control of wild mustard (Sinapis arvensis) biotypes in Iran. The resistance of the ALS inhibitor to weeds is attributed to either target site alteration or enhanced herbicide degradation. Molecular and genetic characterization of the resistance mechanism is relevant to the evolution and management of herbicide resistance. The aims of this research were (a) to characterize the mechanism molecular suspected to Granstar (tribenuron methyl) and Atlantis (Mesosulfuron + Iodosulfuron) resistance in S. arvensis biotypes in the greenhouse and laboratory (b) to investigate the organization of the target-site loci in field selected S. arvensis populations and (c) instantly recognize the mutations that cause resistance to ALS inhibitors. Eighty resistant populations of S. arvensis were carefully collected from fields repeatedly treated with Granstar and Atlantis. The resistance level and pattern of the population were determined through a greenhouse dose-response experiment by applying the above-mentioned herbicides. Extraction of genomic DNA was carried out for PCR and ALS gene analysis. Our results showed that by greenhouse experiment across 80 biotypes suspected to resistance collected in the fields of whole Kermanshah Province, 30 biotypes (37.5%) conferred S. arvensis resistance species reported in the farm. Among 30 biotypes screened in a greenhouse experiment, six biotypes (20%), No. 9, 14, 17, 19, 23 and 28 revealed a mutation in the ALS gene that was detected by PCR-based method. Biotype No. 9 in the position 376 (Asp376-Gly, GAC to GGC), biotypes 14 and 19 in the position 197 (Pro197-Ala, CCT to GCT), biotypes 17, 23 and 28 in the position 574 (Trp574-Leu, TGG to TTG) and biotype No. 23 in the position 122 (Thr-122-Ala, ACA to GCA) showed herbicide resistance. The specific mutation in the position of 122 of the ALS gene in S. arvensis is the first report. Other biotypes showed resistance in the greenhouse but didn't indicate any mutation by PCR-based method. Most of the resistance to Granstar and Atlantis are genetic and are induced by mutations in the ALS gene. The resistance to herbicides may contain a non-mutagenic and non-genetic origin. The reason of herbicide resistance as non-target-site in some of the biotypes may relate to the activity of the herbicide-metabolizing enzyme(s) or transporter proteins that will naturally lead to an increase in herbicide degradation or compartmentation away from its active site.
Molecular Basis of Resistance to ALS-Inhibitor Herbicides in Greater Beggarticks
Weed Science, 2009
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 sin...
Weed Research, 2008
Acetolactate synthase (ALS) inhibitors are the most resistance-prone herbicide group. Rapid resistance diagnosis is thus of importance for their optimal use. We formulate rules to use the derived cleaved amplified polymorphic sequence method to develop molecular tools detecting a change at a given codon, the nature of which is unknown. We applied them to Alopecurus myosuroides (black grass) to develop assays targeting ALS codons A122, P197, A205, W574 and S653 that are crucial for herbicide sensitivity. These assays detected W574L or P197T, or both substitutions, in most plants analysed from a field where ALS inhibitors failed after 3 years of use. Similar assays can easily be set up for any species. Given the rapidity of selection for resistance to ALS inhibitors, these assays should be very useful in proactive herbicide resistance diagnosis.
Scientia Agricola, 2003
Bidens pilosa and Amaranthus quitensis are major weeds infesting soybean [Glycine max L (Merrill)] fields in Brazil and Argentina. The repetitive use of acetolactate synthase (ALS EC 4.1.3.18) inhibiting herbicides in São Gabriel do Oeste, MS, Brazil and in the provinces of Córdoba and Tucumã, Argentina, has selected for resistant (R) biotypes of these weeds. Research work was developed to study the management, growth, biochemistry, and genetics of these R weed biotypes. In a field experiment it was found that chlorimuron-ethyl and imazethapyr at recommended rates (both ALS inhibitor herbicides), did not control R B. pilosa, but the alternative lactofen, fomesafen and bentazon were effective, either sprayed alone or mixed with the ALS inhibitor herbicides. Greenhouse studies confirmed the cross-resistance of both R biotypes to the imidazolinone and sulfonylurea herbicides, and these alternative herbicides, when sprayed alone or mixed with the ALS inhibitor, efficiently controlled bo...
Pest Management Science
BACKGROUND: Herbicide resistant weeds are a serious problem worldwide. Recently, two populations of Amaranthus palmeri with suspected cross-resistance to ALS-inhibiting herbicides (R1 and R2) were found by farmers in two locations from Argentina (Vicuña Mackenna and Totoras, respectively). We conducted studies to confirm and elucidate the mechanism of resistance. RESULTS: We performed in vivo dose-response assays, and confirmed both populations had a strong resistance to chlorimuron-ethyl, diclosulam and imazethapyr when compared to a susceptible population (S). In vitro ALS activity inhibition tests only indicated a considerable resistance to imazethapyr and chlorimuron-ethyl, indicating that other non-target mechanisms could be involved in diclosulam resistance. Subsequently, molecular analysis of als nucleotide sequences revealed three single base-pair mutations conferring substitutions in amino acids previously associated with resistance to ALS inhibitors, A122, W574, and S653 CONCLUSION: This is the first report of als resistant alleles in Amaranthus palmeri from Argentina. The data support the involvement of a target-site mechanism of resistance to ALS inhibiting herbicides.
Plant Science, 2011
We investigated the diversity of mechanisms conferring resistance to herbicides inhibiting acetolactate synthase (ALS) in corn poppy (Papaver rhoeas L.) and the processes underlying the selection for resistance. Six mutant ALS alleles, Arg 197 , His 197 , Leu 197 , Ser 197 , Thr 197 and Leu 574 were identified in five Italian populations. Different alleles were found in a same population or a same plant. Comparison of individual plant phenotype (herbicide sensitivity) and genotype (amino-acid substitution(s) at codon 197) showed that all mutant ALS alleles conferred dominant resistance to the field rate of the sulfonylurea tribenuron and moderate or no resistance to the field rate of the triazolopyrimidine florasulam. Depending on the allele, dominant or partially dominant resistance to the field rate of the imidazolinone imazamox was observed. Putative non-target-site resistance mechanisms were also likely present in the populations investigated. The derived Cleaved Amplified Polymorphic Sequence assays targeting ALS codons crucial for herbicide sensitivity developed in this work will facilitate the detection of resistance due to mutant ALS alleles. Nucleotide variation around codon 197 indicated that mutant ALS alleles evolved by multiple, independent appearances. Resistance to ALS inhibitors in P. rhoeas clearly evolved by redundant evolution of a set of mutant ALS alleles and likely of non-target-site mechanisms.
Plant Science, 2010
The genetic control of non-target-site-based resistance (NTSR) to three herbicides inhibiting acetylcoenzyme A carboxylase (ACCase) and one inhibiting acetolactate-synthase (ALS) was investigated in Alopecurus myosuroides (black-grass). Hundred controlled pairings were used to determine the minimum number of loci involved in NTSR to each herbicide and their associated resistance pattern. Resistant parental plants survived herbicide doses ranging from 1 to 12 times the field rate. In a single plant, NTSR to a given herbicide involved at least one to three dominant loci or one recessive locus. Accumulation of up to at least three NTSR loci in a single plant could be necessary to confer resistance. Most NTSR loci endowing resistance to one herbicide did not confer detectable resistance to any other herbicide assayed. This is the first study demonstrating that NTSR is a quantitative trait. It also revealed three nested levels of complexity in NTSR: the loci, which can confer resistance at the whole plant level depending on the herbicide and on the dose used; the individual plants, which can accumulate various sets of NTSR loci via sexual reproduction; and the populations, which are made of various frequencies of genotypes each containing different numbers and combinations of NTSR loci.
Weed Research, 2009
Lolium species (ryegrasses) are genetically highly variable plants that are both forage crops and major weeds across the globe. As weeds, they rapidly evolve resistance under the selective pressure of acetolactatesynthase (ALS) inhibitors, the most resistance-prone herbicide group. Quick and accurate diagnosis is therefore of importance to prevent resistance spread in ryegrass. To develop proactive molecular tools for the detection of mutant, resistant ALS alleles, we assessed variation in the ryegrass ALS gene. Sequencing the full 1929-bp ALS coding sequence in 59 plants from six distant locations revealed a total of 208 polymorphic nucleotide positions (one every 9.3 nucleotides). The heterogeneous distribution of synonymous and non-synonymous substitutions along the ALS coding sequence suggested that nucleotide variation of ALS is shaped by purifying and background selection. Using regions of the ALS coding sequence with a low number of polymorphic nucleotide sites, five derived cleaved amplified polymorphic sequence (dCAPS) assays were developed targeting codons crucial for herbicide sensitivity. These enabled the first detection in ryegrass of a Pro-197-Thr substitution that confers herbicide resistance. Most assays could also be used to genotype Festuca and Vulpia plants. These dCAPS assays should prove powerful tools for both resistance diagnosis and population genetics studies.