Level Of Glyphosate Resistance Research Articles

Glyphosate hormesis effects on the vegetative and reproductive development of glyphosate-susceptible and -resistant Conyza sumatrensis biotypes

Low glyphosate doses that produce hormesis may alter the susceptibility to herbicides of weeds or enhance their propagation and dispersal. The objective of this work was to evaluate the hormetic effects of glyphosate on the vegetative, phenological and reproductive development in resistant (R) and susceptible (S) Conyza sumatrensis biotypes. The glyphosate resistance level of biotype R was 11.2-fold compared to the S biotype. Glyphosate doses <11.25 g ae ha−1 induced temporary and permanent hormetic effects for the number of leaves, plant height and dry mass accumulation up to 28 d after application in both R and S biotypes. The S biotype required 15–19% fewer thermal units at 1.4 and 2.8 g ae ha−1 glyphosate than untreated plants to reach the bolting stage. Also, this biotype had less thermal units associated with the appearance (1225 vs 1408 units) and opening (1520 vs 1765 units) of the first capitulum than the R biotype. In addition, glyphosate affected reproductive traits of both biotypes compared to their controls, increasing the number of capitulum's and seeds per plant up to 37 and 41% (at 2.8 and 0.7 g ae h−1, respectively) in the S biotype, and by 48 and 114% (both at 5.6 g ae ha−1) in the R biotype. Depending on environmental parameters, glyphosate may or may not cause hormetic effects on the vegetative and phenological development of C. sumatrenis biotypes; however, this herbicide increases the speed and fecundity of reproduction, regardless of the glyphosate susceptibility level, which can alter the population dynamics and glyphosate susceptibility of future generations.

EPSPS gene amplification confers glyphosate resistance in Palmer amaranth in Connecticut

Abstract A Palmer amaranth biotype (CT-Res) with resistance to glyphosate was recently confirmed in a pumpkin field in Connecticut. However, the underlying mechanisms conferring glyphosate resistance in this biotype is not known. The main objectives of this research were 1) to determine the effect of plant height (10, 20, and 30 cm) on glyphosate resistance levels in CT-Res Palmer amaranth biotype, and 2) to investigate whether the target site–based mechanisms confer glyphosate resistance. To achieve these objectives, progeny seeds of the CT-Res biotype after two generations of recurrent selection with glyphosate (6,720 g ae ha−1) were used. Similarly, known glyphosate-susceptible Palmer amaranth biotypes from Kansas (KS-Sus) and Alabama (AL-Sus) were included. Results from greenhouse dose-response studies revealed that CT-Res Palmer amaranth biotype had 69-, 64-, and 54-fold resistance to glyphosate as compared with the KS-Sus biotype when treated at heights of 10, 20, and 30 cm, respectively. Sequence analysis of the EPSPS gene revealed no point mutations at the Pro106 and Thr102 residues in the CT-Res Palmer amaranth biotype. Quantitative polymerase chain reaction analysis revealed that the CT-Res biotype had 33 to 111 relative copies of the EPSPS gene compared with the AL-Sus biotype. All these results suggest that the EPSPS gene amplification endows a high level of glyphosate resistance in the GR Palmer amaranth biotype from Connecticut. Because of the lack of control with glyphosate, growers should adopt the use of effective alternative preemergence and postemergence herbicides in conjunction with other cultural and mechanical tactics to mitigate the further spread of GR Palmer amaranth in Connecticut.

Aldo-keto reductase may contribute to glyphosate resistance in Lolium rigidum.

We have previously demonstrated that an aldo-keto reductase (AKR) from Echinochloa colona (EcAKR4-1) can metabolize glyphosate and confers glyphosate resistance. This study aims to investigate if the EcAKR4-1 orthologs from Lolium rigidum also play a role in glyphosate resistance in non-target-site based, glyphosate-resistant (R) L. rigidum populations from Western Australia. The full-length L. rigidum AKR gene (LrAKR4C10) orthologous to EcAKR4-1, together with a distinct LrAKR1, were cloned from plants of a glyphosate-susceptible (S) (VLR1) and three glyphosate R L. rigidum populations (WALR50, WALR60 and WALR70). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) results showed that basal expression levels of the two LrAKR genes did not differ between the R and S populations, but their expression was significantly induced by glyphosate (up to 4.3-fold) or 2,4-D treatment (up to 3.4-fold) in R populations. Escherichia coli cells transformed respectively with LrAKR4C10 and LrAKR1 were more tolerant to glyphosate. Rice (Oryza sativa) seedlings overexpressing each of the two LrAKR gene survived glyphosate rates that were lethal to the green fluorescence protein (GFP) control plants. Structural modeling predicts a similar way of glyphosate binding and detoxification by LrAKR4C10 and EcAKR4-1, but an alternative way of glyphosate binding by LrAKR1. Relatively lower capacity of the two LrAKRs in conferring glyphosate resistance than the known EcAKR4-1 was discussed in relation to structural interaction. Glyphosate-induced higher expression of the two LrAKR genes in L. rigidum populations contributes to a moderate level of glyphosate resistance likely through enhanced glyphosate metabolism. The herbicide 2,4-D can also induce the LrAKR expression, indicating the potential antagonistic effect of 2,4-D to glyphosate. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Australia’s first case of glyphosate-resistant tall fleabane (Conyza sumatrensis) and alternative herbicide options for its control

AbstractTall fleabane is emerging as a problematic weed species in the eastern cropping region of Australia. Recently, growers indicated poor control of tall fleabane to the field rate of glyphosate in fallow fields. Pot studies were conducted in an open field at the Gatton farm of the University of Queensland, Queensland, Australia, to confirm the level of glyphosate resistance in a putative glyphosate-resistant (GR) tall fleabane population and to evaluate the performance of alternative postemergence herbicides to control GR tall fleabane. Compared with a glyphosate-susceptible (GS) population, the level of resistance in the GR population was 4-fold and 3.5-fold greater based on plant survival and biomass, respectively. The target-site resistance mechanism was not present because both the GR and GS populations had the same gene sequence. There were several effective alternative herbicides for the control of small (4-leaf stage) plants of tall fleabane, but to control large (12- to 14-leaf stage) plants, the sole application of saflufenacil + trifludimoxazin or its mixtures with glyphosate, glufosinate, or paraquat were the best herbicide treatments. This is the first published report on the occurrence of GR tall fleabane in Australia. Growers need to use integrated management strategies to mitigate the further spread of GR tall fleabane in fallow fields as well as glyphosate-resistant crops.

Glyphosate Efficacy in Chloris virgata Sw. in Response to Temperature and Tank Mixing

Glyphosate alone or a tank mixture of glyphosate and 2,4-D is commonly used for broad-spectrum weed control under fallow conditions in Australia. Air temperature or mixing glyphosate with 2,4-D, may influence the efficacy of glyphosate on feather fingergrass (Chloris virgata Sw.), a problematic summer-season weed of Australia. Dose-response studies were conducted with four populations of feather fingergrass under temperature-controlled glasshouse conditions (35/25 °C and 25/15 °C at 12 h/12 h) to assess the level of glyphosate resistance in relation to temperature regimes. Four parameter log-logistic models were used to develop dose-response curves. Based on plant mortality percentage, LD50 (lethal dose for 50% mortality) values of glyphosate at 25/15 °C for populations Ch, SGM2, SGW2, and CP2 were 137, 60, 650, and 1067 g ae ha-1, respectively. However, at 35/25 °C, the corresponding LD50 values were 209, 557, 2108, and 2554 g ae ha-1, respectively. A similar response was observed for the parameter GR50 (dose for 50% growth reduction) values of glyphosate. These results indicate that populations SGW2 and CP2 are highly glyphosate-resistant and in the summer season, it may be very difficult to control these populations due to poor glyphosate efficacy. These results further suggest that the efficacy of glyphosate for feather fingergrass control can be improved if applied during cooler temperatures (25/15 °C) or the spring season compared with warmer temperatures (35/25 °C) or the summer season. In another study, 2,4-D antagonized glyphosate remarkably in the CP2 (glyphosate-resistant) population but only marginally in the Ch (glyphosate-susceptible) population. Thus, it is not advisable to mix 2,4-D with glyphosate for the control of glyphosate-resistant feather fingergrass populations. The results further suggest that the use of this mixture is useful if the feather fingergrass is not glyphosate-resistant; however, the use of the mixture is to be avoided if the population is glyphosate-resistant in order to not exacerbate the potential resistance problem.

Glyphosate resistance in junglerice (Echinochloa colona) and alternative herbicide options for its effective control

AbstractControl of glyphosate-resistant (GR) junglerice is a challenging task in eastern Australia. There is limited information on the efficacy and reliability of alternate herbicides for GR populations of junglerice, especially when targeting large plants and when temperatures are high. A series of experiments were conducted to confirm the level of glyphosate resistance in three populations of junglerice and to evaluate the efficacy of alternate herbicides for the control of GR junglerice populations. The LD50 of glyphosate of B17/7, B17/34, and B17/35 populations was found to be 298, 2,260, and 1,715 g ae ha–1, respectively, suggesting that populations B17/34 and B17/35 were highly resistant to glyphosate. Glyphosate efficacy was reduced at high-temperature (35 C day/25 C night) compared with low-temperature conditions (25 C day/15 C night), suggesting that control of susceptible populations may also be reduced if glyphosate is sprayed under hot conditions. Preemergence herbicides dimethenamid-P (1,000 g ai ha–1) and pendimethalin (1,500 g ai ha–1) provided 100% control of GR populations (B17/34 and 17/35). Postemergence herbicides, such as clethodim (60 or 90 g ai ha–1), glufosinate (750 g ai ha–1), haloxyfop (52 or 78 g ai ha–1), and paraquat (400 or 600 g ai ha–1), applied at the four-leaf stage provided 100% control of GR populations. For larger junglerice plants (eight-leaf stage), postemergence applications of paraquat (400 or 600 g ai ha–1) provided greater weed control than clethodim, glufosinate, and haloxyfop. A mixture of either glufosinate or haloxyfop with glyphosate provided poor control of GR junglerice populations compared with application of glufosinate or haloxyfop applied alone. Efficacy of glufosinate and haloxyfop for the control of GR populations decreased when applied in the sequential spray after glyphosate application. This study identified alternative herbicide options for GR junglerice populations that can be used in herbicide rotation programs for sustainable weed management.

Fitness Cost Associated With Enhanced EPSPS Gene Copy Number and Glyphosate Resistance in an Amaranthus tuberculatus Population.

The evolution of resistance to pesticides in agricultural systems provides an opportunity to study the fitness costs and benefits of novel adaptive traits. Here, we studied a population of Amaranthus tuberculatus (common waterhemp), which has evolved resistance to glyphosate. The growth and fitness of seed families with contrasting levels of glyphosate resistance was assessed in the absence of glyphosate to determine their ability to compete for resources under intra- and interspecific competition. We identified a positive correlation between the level of glyphosate resistance and gene copy number for the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) glyphosate target, thus identifying gene amplification as the mechanism of resistance within the population. Resistant A. tuberculatus plants were found to have a lower competitive response when compared to the susceptible phenotypes with 2.76 glyphosate resistant plants being required to have an equal competitive effect as a single susceptible plant. A growth trade-off was associated with the gene amplification mechanism under intra-phenotypic competition where 20 extra gene copies were associated with a 26.5 % reduction in dry biomass. Interestingly, this growth trade-off was mitigated when assessed under interspecific competition from maize.

Distribution of Glyphosate-Resistance in Echinochloa crus-galli Across Agriculture Areas in the Iberian Peninsula

The levels of resistance to glyphosate of 13 barnyard grass (Echinochloa crus-galli) populations harvested across different agriculture areas in the Southern Iberian Peninsula were determined in greenhouse and laboratory experiments. Shikimate accumulation fast screening separated the populations regarding resistance to glyphosate: susceptible (S) E2, E3, E4, and E6 and resistant (R) E1, E5, E7, E8, E9, E10, E11, E12, and E13. However, resistance factor (GR50 E1–E13/GR50 E6) values separated these populations into three groups: (S) E2, E3, E4, and E6, (R) E1, E5, E7, E8, and E9, and very resistant (VR) E10, E11, E12, and E13. 14C-glyphosate assays performed on two S populations (E2 and E6) showed greater absorption and translocation than those found for R (E7 and E9) and VR (E10 and E12) populations. No previous population metabolized glyphosate to amino methyl phosphonic acid (AMPA) and glyoxylate, except for the E10 population that metabolized 51% to non-toxic products. The VR populations showed two times more 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity without herbicide than the rest, while the inhibition of the EPSPS activity by 50% (I50) required much higher glyphosate in R and VR populations than in S populations. These results indicated that different target-site and non-target-site resistance mechanisms were implicated in the resistance to glyphosate in E. crus-galli. Our results conclude that resistance is independent of climate, type of crop, and geographic region and that the level of glyphosate resistance was mainly due to the selection pressure made by the herbicide on the different populations of E. crus-galli studied.

Glyphosate Resistance in Amaranthus viridis in Brazilian Citrus Orchards

Glyphosate is the main tool for weed management in Brazilian citrus orchards, where weeds, such as Conyza bonariensis and Digitaria insularis, have been found with resistance to this herbicide. Field prospections have allowed the identification of a possible new case of glyphosate resistance. In this work, the susceptibility levels to glyphosate on three Amaranthus viridis L. populations, with suspected resistance (R1, R2, and R-IAC), collected in citrus orchards from the São Paulo State, Brazil, as well as their accumulation rates of shikimic acid, were determined. The fresh weight of the susceptible population (S) was reduced by 50% (GR50) with ~30 g ea ha−1 glyphosate, while the GR50 values of the R populations were between 5.4 and 11.3 times higher than that for S population. The LD50 (herbicide dose to kill 50% of individuals of a weed population) values of the S population were ≤150 g ea ha−1 glyphosate, while the LD50 of the R populations ranged from 600 to 920 g ea ha−1. Based on the reduction of fresh weight and the survival rate, the R1 population showed the highest level of glyphosate resistance, which had GR50 and LD50 values of 248 and 918 g ea ha−1 glyphosate, respectively. The S population accumulated 240 µg shikimic acid at 1000 µM glyphosate, while the R1, R2, and R-IAC populations accumulated only 16, 43, and 33 µg shikimic acid, respectively (between 5.6 to 15 times less than the S population). Enzyme activity assays suggested that at least one target site-type mechanism was involved in resistance. This result revealed the first report of glyphosate resistance in A. viridis reported in the world.

Growth behavior and glyphosate resistance level in 10 populations of Echinochloa colona in Australia.

Recently, poor control of Echinochloa colona with glyphosate has been reported in no-till agriculture systems of the northern grain region (NGR) of Australia. Two experiments were conducted using 10 populations of E. colona selected from the NGR of Australia to understand differences in their growth behavior and resistance pattern. Growth studies revealed that these populations differed in plant height (53-70 cm plant-1), tiller production (30-52 tillers plant-1), leaf production (124-186 leaves plant-1) and seed head production (37-65 seed heads plant-1). Days taken to seed heads and shoot biomass in these populations ranged between 40-48 d and 21-27 g plant-1, respectively. Seed production in these populations ranged between 5380 and 10244 seeds plant-1; lowest for population B17/25 and highest for population B17/13. Correlation studies revealed that seed number plant-1 had a positive correlation with tiller number plant-1 (r = 0.73) and negative relation with days taken to seed head initiation (r = - 0.65). The glyphosate dose-response study showed a wide range of responses in these populations and the glyphosate dose required to kill 50% plants (LD50 values) was estimated between 161 to 2339 g a.e. glyphosate ha-1. LD50 values of populations B17/16, B 17/34 and B17/35 were 1086, 2339 and 1153 g ha-1, respectively, making them 6.7, 15.1 and 7.2-fold resistant to glyphosate compared with the susceptible population B17/37. Growth behavior and seed production potential in these populations had no correlation with the resistance index. These results suggest that some populations of E. colona are highly problematic; for example, population B17/34 was not only highly glyphosate-resistant, but also produced a high seed number (9300 seeds plant-1). This study demonstrated that there is a possibility of great risk with the increased use of glyphosate for managing E. colona in the NGR of Australia. The results warrant integrated weed management strategies and improved stewardship guidelines are required for managing glyphosate-resistant populations of E. colona and to restrict further movement of resistant populations to other regions of Australia.

Distribution Differences in the EPSPS Gene in Chromosomes Between Glyphosate-Resistant and -Susceptible Goosegrass (Eleusine indica)

Abstract Glyphosate is a popular herbicide used to control goosegrass [Eleusine indica (L.) Gaertn.], one of the most troublesome weeds in cotton fields. However, high selection pressure has led to some populations being difficult to control in cotton fields in China. In this study, levels of glyphosate resistance were quantified and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) amplification was confirmed. In addition, distribution of the EPSPS gene among the chromosomes was also investigated using fluorescence in situ hybridization (FISH) methodology. One population (AHCZ) was confirmed to be glyphosate resistant with a resistance index of 3.4 and significantly reduced shikimate accumulation compared to susceptible population. All examined individuals exhibited no mutations in the EPSPS gene in AHCZ. Expression and copy numbers of EPSPS in the AHCZ population were 5.7 and 15.4 times higher, respectively, than in the susceptible population. A positive correlation was identified between signal intensities of primary anti-EPSPS antibody and copy numbers of the EPSPS protein, as indicated by immunoblot analysis. FISH results revealed that, in mitotic metaphase chromosomes, signals were observed dispersed across two sets of homologous chromosomes in a resistant individual (copy number = 31), while in susceptible individuals, signals were only partially detected in one set of homologous chromosomes. In interphase nuclei, EPSPS signals were brightest in resistant individual compared to those that were susceptible. In conclusion, one E. indica population from a cotton field in the Anhui Province has evolved resistance to glyphosate, and EPSPS gene amplification was confirmed as the resistant mechanism.

A novel TIPT double mutation in EPSPS conferring glyphosate resistance in tetraploid Bidens subalternans.

Bidens subalternans (greater beggarticks) is a tetraploid and troublesome weed infesting annual crops in most tropical regions of the world. A glyphosate-resistant (GR) B. subalternans biotype was detected in a soybean field from Paraguay. A series of physiological and molecular analyses were conducted to elucidate its resistance mechanisms. The GR biotype had a high level of resistance (> 15-fold LD50 ), relative to a glyphosate-susceptible (GS) biotype. Shikimate accumulation was up to ten-fold greater for GS compared with GR. We found no differences in sensitivity when plants were treated and kept under lower (10/4 °C) or higher temperatures (25/20 °C). GS and GR had the same relative EPSPS gene copy number, and similar glyphosate absorption and translocation rates. Neither biotype metabolized glyphosate. A double amino acid substitution (TIPT - Thr102Ile and Pro106Thr) was found in only one EPSPS allele from one of the two EPSPS homoeologs present in tetraploid GR B. subalternans. This is the first report of a TIPT double mutation conferring high levels of glyphosate resistance in a weed species. The presence of both wild-type and TIPT mutant EPSPS on the polyploid genome of GR B. subalternans may offset a potential fitness cost, requiring additional research to confirm the absence of deleterious effects. © 2019 Society of Chemical Industry.

Selection for glyphosate resistance in Conyza spp. occurring in the railway network of southern Spain

Conyza spp. are broadleaf weeds that occur in many crops but are also common in non-crop systems such as roadsides and railways. Conyza have selected for glyphosate resistance along railway tracks in southern Spain due to the misuse of this herbicide and the high seed dispersal rate of these species. Twenty-three samples of the genus Conyza (11 Conyza canadensis and 12 Conyza bonariensis) were collected from the margins of railways in different routes of the Andalusia railway network running adjacent to nearby crop fields. The glyphosate resistance level of Conyza populations was evaluated through GR50 (herbicide rate causing 50% growth reduction) and resistance factor (RF) values in every population collected. The highest GR50 were 1851.2 g a.e. ha−1 (RF = 52.53) in C. canadensis (Malaga–Cordoba route) and 1972.4 g a.e. ha−1 (RF = 35.20) in C. bonariensis (Seville–Cordoba route), and the lowest were 46.9 g a.e. ha−1 (RF = 1.33) in C. canadensis (Seville–Cordoba route) and 23.2 g a.e. ha−1 (RF = 0.41) in C. bonariensis (Seville–Cordoba route). The results showed that, among all the C. canadensis populations collected, 18.2% were glyphosate-resistant (RF &gt; 10), 45.5% showed a tendency to develop resistance (RF = 2.5–5), and 36.4% were susceptible (RF &lt; 2.5). Of the 25% of C. bonariensis populations that had resistance to glyphosate, 16.7% had moderate resistance (RF = 5–10) and 58.3% were susceptible. This study found that there are already glyphosate-resistant Conyza spp. along the railway network in southern Spain. This could lead to possible seed exchange between the railway and adjacent places. Therefore, it is vital to consider the railway network when planning control measures against resistance.

The Triple Amino Acid Substitution TAP-IVS in the EPSPS Gene Confers High Glyphosate Resistance to the Superweed Amaranthus hybridus.

The introduction of glyphosate-resistant (GR) crops revolutionized weed management; however, the improper use of this technology has selected for a wide range of weeds resistant to glyphosate, referred to as superweeds. We characterized the high glyphosate resistance level of an Amaranthus hybridus population (GRH)—a superweed collected in a GR-soybean field from Cordoba, Argentina—as well as the resistance mechanisms that govern it in comparison to a susceptible population (GSH). The GRH population was 100.6 times more resistant than the GSH population. Reduced absorption and metabolism of glyphosate, as well as gene duplication of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) or its overexpression did not contribute to this resistance. However, GSH plants translocated at least 10% more 14C-glyphosate to the rest of the plant and roots than GRH plants at 9 h after treatment. In addition, a novel triple amino acid substitution from TAP (wild type, GSH) to IVS (triple mutant, GRH) was identified in the EPSPS gene of the GRH. The nucleotide substitutions consisted of ATA102, GTC103 and TCA106 instead of ACA102, GCG103, and CCA106, respectively. The hydrogen bond distances between Gly-101 and Arg-105 positions increased from 2.89 Å (wild type) to 2.93 Å (triple-mutant) according to the EPSPS structural modeling. These results support that the high level of glyphosate resistance of the GRH A. hybridus population was mainly governed by the triple mutation TAP-IVS found of the EPSPS target site, but the impaired translocation of herbicide also contributed in this resistance.

Sequencing the Plastid Genome of Giant Ragweed (Ambrosia trifida, Asteraceae) From a Herbarium Specimen.

We report the first plastome sequence of giant ragweed (Ambrosia trifida); with this new genome information, we assessed the phylogeny of Asteraceae and the transcriptional profiling against glyphosate resistance in giant ragweed. Assembly and genic features show a normal angiosperm quadripartite plastome structure with no signatures of deviation in gene directionality. Comparative analysis revealed large inversions across the plastome of giant ragweed and the previously sequenced members of the plant family. Asteraceae plastid genomes contain two inversions of 22.8 and 3.3 kb; the former is located between trnS-GCU and trnG-UCC genes, and the latter between trnE-UUC and trnT-GGU genes. The plastid genome sequences of A. trifida and the related species, Ambrosia artemisiifolia, are identical in gene content and arrangement, but they differ in length. The phylogeny is well-resolved and congruent with previous hypotheses about the phylogenetic relationship of Asteraceae. Transcriptomic analysis revealed divergence in the relative expressions at the exonic and intronic levels, providing hints toward the ecological adaptation of the genus. Giant ragweed shows various levels of glyphosate resistance, with introns displaying higher expression patterns at resistant time points after the assumed herbicide treatment.

EPSPS duplication and mutation involved in glyphosate resistance in the allotetraploid weed species Poa annua L.

Poa annua is a widespread winter annual weed species in California. Recently, poor control of this species with glyphosate was reported by growers in an almond orchard in California with a history of repetitive glyphosate use. The objectives of this research were to evaluate the level of glyphosate resistance in a developed S4 P. annua line (R) and identify the mechanisms of resistance involved. Whole-plant dose-response experiments confirmed glyphosate resistance in R, which required 18-fold more glyphosate to achieve a 50% growth reduction compared with a susceptible line (S), results that were supported by the lower shikimate accumulation observed in R compared with S. No differences in glyphosate absorption, translocation, or metabolism were observed, suggesting that non-target-site mechanisms of resistance are not involved in the resistance phenotype. A missense single nucleotide polymorphism was observed in EPSPS coding position 106 in R, resulting in a leucine to proline substitution. This polymorphism was observed exclusively in P. supina EPSPS homeologs. A seven-fold increase in the number of copies of EPSPS alleles was observed in R compared with S. We report the first case of glyphosate resistance associated with both EPSPS duplication and target-site mutation at position 106, leading to high levels of glyphosate resistance in the allotetraploid weed species Poa annua L. Data obtained in this research will be useful for the development of diagnostic tools for rapid glyphosate resistance identification, monitoring and containment. © 2018 Society of Chemical Industry.

No Impact of Increased EPSPS Gene Copy Number on Growth and Fecundity of Glyphosate-Resistant Kochia (Bassia scoparia)

AbstractThe level of glyphosate resistance in kochia [Bassia scoparia(L.) A. J. Scott] was reported to be due to an increase in 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene copy number. A field study was conducted near Manhattan, KS, in 2014 and 2015 to evaluate the relationship between EPSPS gene copy number and growth and fecundity variables ofB. scopariaindividuals within suspected glyphosate-resistant (GR) populations from western Kansas. Initial assays of EPSPS gene copy and in vivo shikimate accumulation showed thatB. scopariapopulations from Finney (FN-R), Scott (SC-R), and Thomas (TH-R) counties were segregating for glyphosate resistance, with some individuals still being glyphosate susceptible (GS). A target-neighborhood competition approach was used to evaluate the competitive response of individual target plants with relatively low (classified as GS) and high (classified as GR) EPSPS gene copy number within the populations. There was no relationship observed between EPSPS gene copy number and vegetative or fecundity variables. There was no differential competitive response of target plant biomass to increasing neighbor density between individuals with low and high EPSPS gene copy number within each population. Lack of associated vegetative growth and fecundity cost to the increased EPSPS gene copy in the GRB. scopariaplants suggests that the plants are likely to persist in field populations, except when effective weed management strategies are adopted that would prevent their growth and seed production.

Alternative Herbicides for Control of Glyphosate-Resistant Giant Ragweed in Nebraska

Giant ragweed is an early emerging and one of the most competitive summer annual species found in many fields throughout North America. Extensive use of glyphosate in glyphosate-tolerant (GT) crops has evolved giant ragweed populations with glyphosate resistance. Field dose-response studies were conducted to determine the influence of growth stage on the level of glyphosate resistance in a suspected giant ragweed population. In addition, efficacy of alternative pre-plant, pre-emergence (PRE) and post-emergence (POST) herbicides were evaluated in corn and soybeans for glyphosate-resistant (GR) giant ragweed control. The field glyphosate dose-response studies confirmed that the suspected giant ragweed population were resistant ranging from 14- to 32-fold resistance depending on the growth stage of glyphosate application. The 10, 20 and 30 cm tall giant ragweed had 14, 17 and 32X resistance level, respectively. The dose-response studies indicated that the 10, 20, and 30 cm tall GR giant ragweed was controlled 90% with 214, 402 and 482 g ae ha-1 of dicamba, respectively, when tank-mixed with glyphosate (1060 g ae ha-1) 21 days after treatment (DAT). All evaluated pre-plant herbicides for corn provided &amp;ge; 90% control of the GR giant ragweed at 30 DAT; among which the best control (100%) was achieved with pre-plant application of atrazine (2240 g ai ha-1), isoxaflutole (90 g ai ha-1), and premix of flumioxazin/pyroxasulfone (315 g ai ha-1). Herbicide combinations of different site of action provided greater than 90% control of the GR giant ragweed population in a PRE followed by POST herbicide program in corn and soybean, suggesting that alternative herbicide for giant ragweed control are available.

High Levels of Glyphosate Resistance in Conyza canadensis from Agricultural and Non-Agricultural Sites in Ohio and Iowa

Glyphosate is an important herbicide worldwide, but its efficacy has been compromised where weed species have evolved glyphosate resistance. To better understand evolutionary outcomes of continued and strong selection from glyphosate exposure, we characterized variation in resistance in self-pollinating Conyza canadensis (horseweed) in Ohio and Iowa, where glyphosate resistance was first reported in 2002 and 2011, respectively. In 2015, we collected seeds from a total of 74 maternal plants (biotypes) from no-till soybean fields vs. non-agricultural sites in each state, using one representative plant per site. Young plants from each biotype were sprayed with glyphosate rates of 0x, 1x (840 g ae ha−1), 8x, 20x, or 40x. Resistant biotypes with at least 80% survival at each dosage were designated as R1 (1x), R2 (8x), R3 (20x), or R4 (40x). Nearly all Ohio agricultural biotypes were R4, as were 62% of biotypes from the non-agricultural sites. In Iowa, R4 biotypes were clustered in the southeastern soybean fields, where no-till agriculture is more common, and 45% of non-agricultural biotypes were R1–R4. Our results show that resistance levels to glyphosate can be very high (at least 40x) in both states, and that non-agricultural sites likely serve as a refuge for glyphosate-resistant biotypes.

The role of plant size in the selection of glyphosate resistance in Sorghum halepense.

The effect of plant size (seedlings versus young plants versus adult plants) on the phenotypic level of glyphosate resistance and selection intensity (SI) in Sorghum halepense with and without a reduced glyphosate translocation resistance mechanism was evaluated. Resistance parameters [the 50% lethal dose (LD50 ) and the dose required to cause a 50% reduction in plant growth (GR50 )] in adult plants were notably higher than in seedlings regardless of the resistance status. However, under similar plant size increases, populations comprised of glyphosate-resistant (R) individuals showed higher survival and growth when glyphosate treated compared with glyphosate-susceptible (S) plants. An increase in SI was always evident with increasing glyphosate doses. However, the rate of increase in SI was higher under glyphosate selection of young R and S plants, followed by seedlings and adult R and S plants. However, in conditions of R seedlings coexisting with adult S plants under glyphosate treatment (1000-4000 g ha-1 ), selection against glyphosate resistance was observed. Any increase in size from the seedling stage of R plants translates into an amplification of resistance. Depending on the particular size combinations of spatially coexisting R and S plants, selection for glyphosate resistance may be faster, slower or even not evident. © 2018 Society of Chemical Industry.