Mechanism Of Glyphosate Resistance Research Articles

ESSENTIAL OIL IN THE MANAGEMENT OF THE SEED BANK OF INFESTANT PLANTS

The weed seed bank causes damage to agriculture, due to competition with cultivated plants and raising production costs, Cenchrus echinatus has a high seed multiplication and dissemination mechanism and Conyza bonariensis glyphosate resistance mechanism. The objective was to evaluate the potential of Mentha arvensis essential oil by means of fumigation in the weed seed bank. Soil samples (2 kg per treatment) were collected, homogenized and subjected to fumigation for 24 and 48 hours at concentrations of 2.5% and 5.0% of essential oil. The soil was solarized (control) and dried in the shade for three days as a witness. Then, 500 g of soil were distributed in four trays in a completely randomized design, where they received irrigation, identification and registration of emerging seedlings. Data were checked for normality and homogeneity of variance, transformed when necessary and submitted to analysis of variance at a 5% significance level. The averages were performed by Tukey and Dunnet tests at 5% probability. It was concluded that fumigation reduced grass and horseweed emergence regardless of dose and exposure time, with results similar to solarization.

Genomic adaptation of Burkholderia anthina to glyphosate uncovers a novel herbicide resistance mechanism.

Glyphosate (GS) specifically inhibits the 5-enolpyruvyl-shikimate-3-phosphate (EPSP) synthase that converts phosphoenolpyruvate (PEP) and shikimate-3-phosphate to EPSP in the shikimate pathway of bacteria and other organisms. The inhibition of the EPSP synthase depletes the cell of the EPSP-derived aromatic amino acids as well as of folate and quinones. A variety of mechanisms (e.g., EPSP synthase modification) has been described that confer GS resistance to bacteria. Here, we show that the Burkholderia anthina strain DSM 16086 quickly evolves GS resistance by the acquisition of mutations in the ppsR gene. ppsR codes for the pyruvate/ortho-Pi dikinase PpsR that physically interacts and regulates the activity of the PEP synthetase PpsA. The mutational inactivation of ppsR causes an increase in the cellular PEP concentration, thereby abolishing the inhibition of the EPSP synthase by GS that competes with PEP for binding to the enzyme. Since the overexpression of the Escherichia coli ppsA gene in Bacillus subtilis and E. coli did not increase GS resistance in these organisms, the mutational inactivation of the ppsR gene resulting in PpsA overactivity is a GS resistance mechanism that is probably unique to B. anthina.

Gene Amplification of EPSPS with a Mutation in Conserved Region: The Evolved Glyphosate Resistance Mechanism in Eleusine indica

Goosegrass (Eleusine indica (L.) Gaertn.) is one of the most vicious weeds worldwide, competing with crops and greatly reducing their yields. Glyphosate, a non-selective, broad spectrum, post-emergence herbicide has inevitably induced severe resistance in many weeds owing to its intensive use. Additionally, control strategies rely on the clarity of resistance mechanisms. In this study, we aimed to investigate the resistance levels and potential resistance mechanisms of two goosegrass populations collected from orchards. Results showed that the resistance indexes of LL and SS populations were 3.8 and 1.9, respectively. A single nucleotide change led to a Pro106Leu (P106L) mutation in the LL population and the SS population had a Pro106Ser (P106S) amino acid substitution. The EPSPS expression in both populations was 2.6 times that of the wild-type population. However, the relative copy number and EPSPS protein content in the LL population were higher than those of the SS population, as indicated by immunoblot analysis and enzyme-linked immunosorbent assays. Overall, we confirmed EPSPS amplification with a P106L mutation, resulting in overproduction of this mutated EPSPS protein, which conferred moderate glyphosate resistance. This study details a case of simultaneous evolution of mutation and amplification in EPSPS of glyphosate resistance weeds.

Investigations into differential glyphosate sensitivity between two horseweed (Conyza canadensis) growth types

AbstractHorseweed [Conyza canadensis (L.) Cronquist] grows in one of two distinct growth phenotypes, “rosette” and “upright” growth types, and they have recently been observed co-occurring in Michigan fields. Previous research found that upright plants from two glyphosate-resistant populations were 3- and 4-fold less sensitive to glyphosate than their rosette siblings. Further experiments were conducted to investigate whether differential glyphosate sensitivity of the growth types was due to glyphosate retention, absorption, or translocation. The total amount of glyphosate retained on the C. canadensis leaf surface was similar for both growth types; however, on a per-weight and per-area bases, the upright growth type retained 21% and 18% less glyphosate, respectively. Glyphosate absorption was up to 85% at 168 h after treatment (HAT), and was not different between the rosette and upright growth types or between the susceptible (S) and resistant (R) biotypes. Additionally, there was no difference in translocation between the two growth types within each biotype at any time point. Interestingly, at 168 HAT, [14C]glyphosate translocation was higher in the S rosette compared with the two growth types from the R biotype; however, the S upright type was similar to both R growth types. Thus, glyphosate resistance in the R biotype may be due to an alternative mechanism rather than impaired translocation, which has been cited as the primary mechanism of glyphosate resistance in C. canadensis. These results suggest that reduced glyphosate retention on a per-weight and per-area bases of the upright growth type may contribute to increased glyphosate tolerance due to a diluted concentration of glyphosate in the plant. However, another factor is likely related to the mechanism of resistance within the R biotype, which is contributing to a 3-fold difference in glyphosate sensitivity between the two growth types, such as alterations in EPSPS gene expression or changes in undescribed metabolism genes.

Antioxidant Enzyme and Cytochrome P450 Activities Are Involved in Horseweed (Conyza Sumatrensis) Resistance to Glyphosate

The intensive global use of glyphosate has led to the evolution of glyphosate resistant (GR) weed species, including the economically damaging horseweed (Conyza sumatrensis). We evaluated the glyphosate resistance mechanisms of C. sumatrensis. While 5-enolpyruvylshikimate-3-phosphate synthase activity was similar between the glyphosate resistant (GR) and nonresistant biotypes, plants from the GR population accumulated lower shikimate levels than susceptible ones, suggesting the absence of target-site resistance mechanisms. Decreases over time in glyphosate concentrations in GR leaves were not accompanied by increases in glyphosate concentrations in their stem and roots, indicating lower glyphosate distribution rates in GR plants. The early appearance of aminomethylphosphonic acid (the main glyphosate metabolite) in leaves, as well as its presence only in the stems and roots of GR plants, suggests faster glyphosate metabolism in GR plants than in susceptible ones. GR plants treated with glyphosate also showed greater antioxidant (ascorbate peroxidase [APX] and catalase [CAT]) and cytochrome P450-enzyme activities, indicating their great capacity to avoid glyphosate-induced oxidative stress. Three non-target mechanisms (reduced glyphosate translocation, increased metabolism, and increased antioxidant activity) therefore confer glyphosate resistance in C. sumatrensis plants. This is the first time that APX, CAT and P450-enzyme activities are related to GR in C. sumatrensis.

Constitutive overexpression of EPSPS by gene duplication is involved in glyphosate resistance in Salsola tragus.

Glyphosate-resistant Salsola tragus accessions have been identified in the USA and Argentina; however, the mechanisms of glyphosate resistance have not been elucidated. The goal of this study was to determine the mechanism/s of glyphosate resistance involved in two S.tragus populations (R1 and R2) from Argentina. Both glyphosate-resistant populations had a six-fold lower sensitivity to glyphosate than the S population (i.e. resistance index). No evidence of differential absorption, translocation or metabolism of glyphosate was found in the R1 and R2 populations compared to a susceptible population (S). No 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) mutations were detected, but S.tragus R1 and R2 plants had ≈14-fold higher EPSPS gene relative copy number compared to the S counterpart. In R1 and R2, EPSPS duplication entailed a greater constitutive EPSPS transcript abundance by approximately seven-fold and a basal EPSPS activity approximately three-fold higher than the S population. The current study reports EPSPS gene duplication for the first time as a mechanism of glyphosate resistance in S.tragus populations. The increase of glyphosate dose needed to kill R1 and R2 plants was linked to the EPSPS transcript abundance and level of EPSPS activity. This evidence supports the convergent evolution of the overexpression of the EPSPS gene in several Chenopodiaceae/Amaranthaceae species adapted to drought environments and the role of gene duplication as an adaptive advantage for plants to withstand stress. © 2022 Society of Chemical Industry.

In-field assessment of EPSPS amplification on fitness cost in mixed glyphosate-resistant and glyphosate-sensitive populations of Palmer amaranth (Amaranthus palmeri)

AbstractComparing fitness of herbicide-resistant and herbicide-susceptible weed biotypes is important for managing herbicide resistance. Previous research suggests there is little to no fitness penalty from amplification of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene (a mechanism of glyphosate resistance) in Palmer amaranth (Amaranthus palmeri S. Watson) in controlled studies in the greenhouse or growth chamber. A field study was conducted in North Carolina at three locations naturally infested with A. palmeri to determine vegetative, reproductive, and germination fitness of plants with and without EPSPS amplification grown season-long with cotton (Gossypium hirsutum L.). Seed number was not correlated with EPSPS copy number. However, when plants were binned into two groups, those having an EPSPS copy number ≥2 (relative to reference genes) and those having an EPSPS copy number <2, plant fresh weight and seed number were 1.4 and 1.6 times greater, respectively, for plants with fewer than 2 EPSPS copies. Amaranthus palmeri height and seed germination, and yield of cotton, did not differ when comparing the two binned groups. These data suggest that A. palmeri plants with EPSPS amplification are relatively less fit in the absence of glyphosate, but this reduced fitness does not translate into differences in interference with cotton.

Mechanisms of glyphosate resistance in common ragweed (Ambrosia artemisiifolia): patterns of absorption, translocation, and metabolism

AbstractGlyphosate’s efficacy is influenced by the amount absorbed and translocated throughout the plant to inhibit 5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS). Glyphosate resistance can be due to target-site (TS) or non–target site (NTS) resistance mechanisms. TS resistance includes an altered target site and gene overexpression, while NTS resistance includes reduced absorption, reduced translocation, enhanced metabolism, and exclusion/sequestration. The goal of this research was to elucidate the mechanism(s) of glyphosate resistance in common ragweed (Ambrosia artemisiifolia L.) from Ontario, Canada. The resistance factor for this glyphosate-resistant (GR) A. artemisiifolia biotype is 5.1. No amino acid substitutions were found at positions 102 or 106 of the EPSPS enzyme in this A. artemisiifolia biotype. Based on [14C]glyphosate studies, there was no difference in glyphosate absorption or translocation between glyphosate-susceptible (GS) and GR A. artemisiifolia biotypes. Radio-labeled glyphosate metabolites were similar for GS and GR A. artemisiifolia 96 h after application. Glyphosate resistance in this A. artemisiifolia biotype is not due to an altered target site due to amino acid substitutions at positions 102 and 106 in the EPSPS and is not due to the NTS mechanisms of reduced absorption, reduced translocation, or enhanced metabolism.

Inheritance and mechanism of glyphosate resistance in annual bluegrass (Poa annua L.).

In initial screening, glyphosate was ineffective in controlling five Poa annua populations. These populations were tested for resistance, and studies undertaken to determine resistance mechanisms and inheritance pattern. Dose-response studies conducted at 16/12°C and 27/20°C on the five putative resistant populations showed low-level resistance (1.4- to 2.5-fold) to glyphosate. Shikimic acid accumulation in response to glyphosate confirmed differences among the populations, with greater shikimic acid accumulation in the susceptible population. The EPSPS gene copy number was 0.5- to 5.2-fold greater in one resistant population (HT) than in the susceptible (S) population, but not in the others. EPSPS gene expression was five- to tenfold higher in HT compared with the susceptible population. Target site mutations, differences in glyphosate absorption or translocation or altered expression of aldo-keto reductase (AKR) were not identified in any of the resistant populations. Crosses were successful between one resistant population and the susceptible population (P262-16♂ ✕ S♀) and inheritance of glyphosate resistance appears to be controlled by a single, nuclear dominant gene in this population. Our study identified EPSPS gene amplification in a South Australian glyphosate-resistant P. annua population (HT). This mechanism of resistance was not identified in the other four glyphosate-resistant populations, and other common mechanisms were excluded. Although the resistance mechanism in some P. annua populations remains unknown, inheritance studies with one population suggest the involvement of a single dominant gene. © 2021 Society of Chemical Industry.

Coexistence of target-site and non-target-site mechanisms of glyphosate resistance in Amaranthus palmeri populations from Argentina

Amaranthus palmeri S. Waston is currently one of the most problematic weeds worldwide. Biotypes with resistance to herbicides such as glyphosate and ALS inhibitors are now present in almost all Argentinian cultivable areas. In this work, we studied glyphosate resistance in three different populations, some of them previously characterized as resistant to ALS inhibitors. Dose-response curves were conducted in order to assess the effect of glyphosate on the survival and dry biomass of the populations. Subsequently, the presence of target-site resistance (TSR) was studied. Results confirmed the glyphosate resistance in the three populations, showing different levels of resistance, being R2 and R3 significantly more resistant than r1 population. A high prevalence of the P106S substitution was detected in the three resistant populations, while none increase in the relative EPSPS copy number was noticed. Some surviving plants without any of the TSR mechanisms for glyphosate were detected in R3 population, suggesting the presence of non-target-site resistance (NTSR).

Non-Target Site Mechanisms Endow Resistance to Glyphosate in Saltmarsh Aster (Aster squamatus).

Of the six-glyphosate resistant weed species reported in Mexico, five were found in citrus groves. Here, the glyphosate susceptibility level and resistance mechanisms were evaluated in saltmarsh aster (Aster squamatus), a weed that also occurs in Mexican citrus groves. The R population accumulated 4.5-fold less shikimic acid than S population. S plants hardly survived at 125 g ae ha−1 while most of the R plants that were treated with 1000 g ae ha−1, which suffered a strong growth arrest, showed a vigorous regrowth from the third week after treatment. Further, 5-enolpyruvylshikimate-3-phosphate basal and enzymatic activities did not diverge between populations, suggesting the absence of target-site resistance mechanisms. At 96 h after treatment, R plants absorbed ~18% less glyphosate and maintained 63% of the 14C-glyphsoate absorbed in the treated leaf in comparison to S plants. R plants metabolized twice as much (72%) glyphosate to amino methyl phosphonic acid and glyoxylate as the S plants. Three non-target mechanisms, reduced absorption and translocation and increased metabolism, confer glyphosate resistance saltmarsh aster. This is the first case of glyphosate resistance recorded for A. squamatus in the world.

The First Report of Target-Site Resistance to Glyphosate in Sweet Summer Grass (Moorochloa eruciformis)

Sweet summer grass is a problematic weed in the central Queensland region of Australia. This study found glyphosate resistance in two biotypes (R1 and R2) of sweet summer grass. The level of resistance in these biotypes was greater than 8-fold. The glyphosate dose required to reduce dry matter by 50% (GR50) for the resistant populations varied from 1993 to 2100 g ha−1. A novel glyphosate resistance double point mutation in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene was identified for the first time in sweet summer grass. Multiple mutations, including multiple amino acid changes at the glyphosate target site, as well as mutations involving two nucleotide changes at a single amino acid codon, were observed. Both resistant biotypes exhibited a nucleotide change of CAA to ACA in codon 106, which predicts an amino acid change of proline to a threonine (Pro-106-Thr). In addition, the R1 biotype also possessed a mutation at codon 100, where a nucleotide substitution of T for G occurred (GCT to TCT), resulting in a substitution of serine for alanine (Ala-100-Ser). Understanding the molecular mechanism of glyphosate resistance will help to design effective management strategies to control invasive weeds.

Evolution of multiple target-site resistance mechanisms in individual plants of glyphosate-resistant Eleusine indica from China.

Glyphosate has been used for weed control in South China in various situations for four decades, and most Eleusine indica populations are suspected to have evolved resistance to glyphosate. This research investigated underling target-site glyphosate resistance mechanisms in six field-collected, putative glyphosate-resistant (R) E. indica populations. The six R E. indica populations were confirmed to be low (1.8 to 2.6-fold) to moderately (5.6- to 8.4-fold) resistant to glyphosate relative to the susceptible (S) population. Sixty-seven glyphosate-surviving plants from the six R populations were used to examine target-site resistance mechanisms. Target-site 5-enolpyruvylshikimate3-phosphate synthase (EPSPS) overexpression (OE) (plus further induction by glyphosate treatment) and gene copy number variation (CNV) occurred in 94% R plants, and among them, 16% had the P106A mutation and 49% had the heterozygous double TIPS (T102I + P106S) mutation (plus P381L). In addition, a low number of R plants (6%) only had the homologous TIPS (plus P381L) mutation. The (CT)6 insertion mutation in the EPSPS 5†-UTR always associates with EPSPS OE and CNV. Progeny plants possessing EPSPS OE/CNV (and P106A) displayed low level (up to 4.5-fold) glyphosate resistance. In contrast, plants homozygous for the TIPS mutation displayed higher (25-fold) resistance to glyphosate and followed by plants heterozygous for this mutation plus EPSPS OE/CNV (12-fold). Target-site glyphosate resistance in E. indica populations from South China is common with prevalence of EPSPS OE/induction/CNV conferring low level resistance. Individual plants acquiring both the TIPS mutation and EPSPS OE/CNV are favored due to evolutionary advantages. The role of (CT)6 insertion mutation in EPSPS CNV is worth further investigation. © 2021 Society of Chemical Industry.

Herbicide resistant weeds: the case of resistance to glyphosate

Glyphosate has become the most widely used herbicide worldwide since 1974 with a global use of 8.6 billion kg (glyphosate active ingredient) between 1974 and 2014. This study reports on glyphosate resistant (GR) weeds and their resistance mechanisms based on global scientifically reported cases. Forty-nine different weed species have evolved resistance to glyphosate in 29 countries with a total of 318 identified cases worldwide. Fifty percent of these resistance cases were found in glyphosate-resistant cropping systems. There were 255 identified cases (80.2%) of glyphosate resistance in the top five countries (in terms of number of cases and species), namely USA, Australia, Argentina, Brazil, and Canada. The five most popular weed species (in terms of number of cases) found to be resistant to glyphosate were Conyza canadensis, Amaranthus palmeri, Amaranthus tuberculatus, Lolium perenne ssp. Multiflorum,and Ambrosia artemisiifolia with 42, 42, 29, 26, and 21 reported cases, respectively. Out of 49 weed species, 19 GR weed species were found to not only be resistant to glyphosate but also to other herbicide sites of action (multiple herbicide resistance). Glyphosate resistance mechanisms in weeds include (1) target-site alterations: target-site mutation and target-site gene amplification; and (2) non-target-site mechanisms involving different modes of exclusion from the target site: reduced glyphosate uptake, reduced glyphosate translocation, and enhanced glyphosate metabolism. It is essential to have an integrated weed management program that includes not only smart herbicide mixtures and rotations, but also cultural, manual, mechanical, and crop-based weed management methods.

Molecular mechanisms underlying glyphosate resistance in bacteria.

Glyphosate is a nonselective herbicide that kills weeds and other plants competing with crops. Glyphosate specifically inhibits the 5-enolpyruvyl-shikimate-3-phosphate (EPSP) synthase, thereby depleting the cell of EPSP serving as a precursor for biosynthesis of aromatic amino acids. Glyphosate is considered to be toxicologically safe for animals and humans. Therefore, it became the most-important herbicide in agriculture. However, its intensive application in agriculture is a serious environmental issue because it may negatively affect the biodiversity. A few years after the discovery of the mode of action of glyphosate, it has been observed that bacteria evolve glyphosate resistance by acquiring mutations in the EPSP synthase gene, rendering the encoded enzyme less sensitive to the herbicide. The identification of glyphosate-resistant EPSP synthase variants paved the way for engineering crops tolerating increased amounts of the herbicide. This review intends to summarize the molecular mechanisms underlying glyphosate resistance in bacteria. Bacteria can evolve glyphosate resistance by (i) reducing glyphosate sensitivity or elevating production of the EPSP synthase, by (ii) degrading or (iii) detoxifying glyphosate and by (iv) decreasing the uptake or increasing the export of the herbicide. The variety of glyphosate resistance mechanisms illustrates the adaptability of bacteria to anthropogenic substances due to genomic alterations.

Mechanism(s) of Glyphosate Resistance in a Selected Plantago lanceolata (L.) R Biotype

In 2003, a glyphosate-resistant plantago (Plantago lanceolata L.) population located in the Robertson district of South Africa was subjected to different glyphosate dosages and the highest dosage (7200 g a.e. ha−1) gave no acceptable levels of control. Here we reconfirm resistance and investigate the mechanism of glyphosate resistance. Dose-response curves indicated that the glyphosate dosage rate causing 50% survival (LD50) for the resistant (R) biotype is 43 times greater than for the susceptible (S) biotype, i.e., 43-fold resistant to glyphosate. Investigation into the molecular mechanism of plantago showed shikimate accumulation of the R biotype was lower than that of the S biotype. The reported 31P and 13C nuclear magnetic resonance (NMR) spectra show rapid glyphosate translocation into the young untreated leaves of the S biotype. No glyphosate translocation was observed in the R biotype. A point mutation in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene, resulting in an amino acid substitution was also observed, indicating two unique glyphosate resistance mechanisms within the R biotype. The rapid evolution of glyphosate-resistant weeds threatens the usage of the world’s most important herbicide (glyphosate), which is essential in world food production and further limits grower options for weed control. New weed management strategies will be necessary to combat plantago R biotypes.

First Case of Glyphosate Resistance in Bromus catharticus Vahl.: Examination of Endowing Resistance Mechanisms.

Bromus catharticus Vahl. has been used as a valuable forage crop, but it has also been noted as a weed of winter crops and an invader in several countries. In Argentina, a putative glyphosate-resistant population of B. catharticus was identified as a consequence of the lack of effective control with glyphosate in the pre-sowing of wheat. Plant survival and shikimate accumulation analysis demonstrated a lower glyphosate-sensitivity of this population in comparison to a susceptible B. catharticus population. The resistant population was 4-fold more resistant to glyphosate than its susceptible counterpart. There was no evidence of target-site mechanisms of glyphosate resistance or an enhanced capacity to metabolize glyphosate in the resistant population. However, the resistant plants showed a lower foliar retention of glyphosate (138.34 μl solution g−1 dry weight vs. 390.79 μl solution g−1 dry weight), a reduced absorption of 14C-glyphosate (54.18 vs. 73.56%) and lower translocation of 14C-glyphosate from the labeled leaf (27.70 vs. 62.36%). As a result, susceptible plants accumulated a 4.1-fold higher concentration of 14C-glyphosate in the roots compared to resistant plants. The current work describes the first worldwide case of glyphosate resistance in B. catharticus. A reduced foliar retention of herbicide, a differential rate of glyphosate entry into leaves and an altered glyphosate translocation pattern would be the most likely mechanisms of glyphosate exclusion.

A PCR plus restriction enzyme-based technique for detecting target-enzyme mutations at position Pro-106 in glyphosate-resistant Lolium perenne.

The first step in managing herbicide-resistant weeds is to confirm their resistance status. It is, therefore, crucial to have a rapid, reliable and cost-effective technique to assess samples for herbicide resistance. We designed and evaluated three derived cleaved amplified polymorphic sequence (dCAPS) markers for detecting glyphosate resistance in Lolium perenne. conferred by non-synonymous mutations at codon-106 in the enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene. The dCAPS markers involve amplification of the target region, digestion of the amplified products with restriction enzymes and gel-based visualisation of the digested products. The results showed that all three dCAPS markers could successfully detect mutations at codon-106 in the target enzyme. The dCAPS markers can also inform us of the zygosity state of the resistance allele and was confirmed by sequencing the target region of the EPSPS gene. The markers described here are effective quick tests for the monitoring and evaluation of the target-enzyme mechanism of glyphosate resistance in Lolium perenne.

Evolution of Glyphosate-Resistant Weeds.

Widespread adoption of glyphosate-resistant crops and concomitant reliance on glyphosate for weed control set an unprecedented stage for the evolution of herbicide-resistant weeds. There are now 48 weed species that have evolved glyphosate resistance. Diverse glyphosate-resistance mechanisms have evolved, including single, double, and triple amino acid substitutions in the target-site gene, duplication of the gene encoding the target site, and others that are rare or nonexistent for evolved resistance to other herbicides. This review summarizes these resistance mechanisms, discusses what is known about their evolution, and concludes with some of the impacts glyphosate-resistant weeds have had on weed management.

Distribution and validation of genotypic and phenotypic glyphosate and PPO-inhibitor resistance in Palmer amaranth (Amaranthus palmeri) from southwestern Nebraska – ERRATUM

Failure to control Palmer amaranth with glyphosate and protoporphyrinogen IX oxidase (PPO)-inhibitor herbicides was reported across southwestern Nebraska in 2017. The objectives of this study were to 1) confirm and 2) validate glyphosate and PPO-inhibitor (fomesafen and lactofen) resistance in 51 Palmer amaranth accessions from southwestern Nebraska using genotypic and whole-plant phenotypic assay correlations and cluster analysis, and 3) determine which agronomic practices might be influencing glyphosate resistance in Palmer amaranth accessions in that location. Based on genotypic assay, 88% of 51 accessions contained at least one individual with amplification (>2 copies) of the 5-enolypyruvyl-shikimate-3-phosphate synthase (EPSPS) gene, which confers glyphosate resistance; and/or a mutation in the PPX2 gene, either ΔG210 or R128G, which endows PPO-inhibitor resistance in Palmer amaranth. Cluster analysis and high correlation (0.83) between genotypic and phenotypic assays demonstrated that EPSPS gene amplification is the main glyphosate resistance mechanism in Palmer amaranth accessions from southwestern Nebraska. In contrast, there was poor association between genotypic and phenotypic responses for PPO-inhibitor resistance, which was attributed to segregation for PPO-inhibitor resistance within these accessions and/or the methodology that was adopted herein. Genotypic assays can expedite the process of confirming known glyphosate and PPO-inhibitor resistance mechanisms in Palmer amaranth from southwestern Nebraska and other locations. Phenotypic assays are also a robust method for confirming glyphosate resistance but not necessarily PPO-inhibitor resistance in Palmer amaranth. Moreover, random forest analysis of glyphosate resistance in Palmer amaranth indicated that EPSPS gene amplification, county, and current and previous crops are the main factors influencing glyphosate resistance within that geographic area. Most glyphosate-susceptible Palmer amaranth accessions were found in a few counties in areas with high crop diversity. Results presented here confirm the spread of glyphosate resistance and PPO-inhibitor resistance in Palmer amaranth accessions from southwestern Nebraska and demonstrate that less diverse cropping systems are an important driver of herbicide resistance evolution in Palmer amaranth.Nomenclature: fomesafen; glyphosate; lactofen; Palmer amaranth, Amaranthus palmeri S. Watson