Reduced Glyphosate Translocation Research Articles

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.

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.

Evolution of Target-Site Resistance to Glyphosate in an Amaranthus palmeri Population from Argentina and Its Expression at Different Plant Growth Temperatures.

The mechanism and expression of resistance to glyphosate at different plant growing temperatures was investigated in an Amaranthus palmeri population (VM1) from a soybean field in Vicuña Mackenna, Cordoba, Argentina. Resistance was not due to reduced glyphosate translocation to the meristem or to EPSPS duplication, as reported for most US samples. In contrast, a proline 106 to serine target-site mutation acting additively with EPSPS over-expression (1.8-fold increase) was respectively a major and minor contributor to glyphosate resistance in VM1. Resistance indices based on LD50 values generated using progenies from a cross between 52 PS106 VM1 individuals were estimated at 7.1 for homozygous SS106 and 4.3 for heterozygous PS106 compared with homozygous wild PP106 plants grown at a medium temperature of 24 °C day/18 °C night. A larger proportion of wild and mutant progenies survived a single commonly employed glyphosate rate when maintained at 30 °C day/26 °C night compared with 20 °C day/16 night in a subsequent experiment. Interestingly, the P106S mutation was not identified in any of the 920 plants analysed from 115 US populations, thereby potentially reflecting the difference in A. palmeri control practices in Argentina and USA.

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.

Environmental Factors Moderate Glyphosate-induced Antagonism of POST Herbicides on the Rapid Response Biotype of Glyphosate-Resistant Giant Ragweed (Ambrosia trifida)

AbstractIn the rapid response (RR) biotype of glyphosate-resistant (GR) giant ragweed (Ambrosia trifidaL.), exposure to glyphosate elicits H2O2production in mature leaves, resulting in foliage loss and reduced glyphosate translocation. When glyphosate is applied with POST herbicides intended to improve control ofA. trifida, the RR to glyphosate has the propensity to antagonize these herbicide combinations. This research documents how transient changes in air temperature, soil moisture, and light intensity during a 6-d period surrounding herbicide application regulate induction of the RR and the effect on POST herbicide interactions with glyphosate. Air temperature had the greatest influence on H2O2accumulation in leaf disks following treatment with glyphosate, as plants at 30 C produced more than twice the amount of H2O2at 2.5 h after treatment compared with 10 C. Plants under field capacity conditions accumulated nearly 50% more H2O2than those at one-third field capacity, while those under no shade had only 18% more H2O2compared with those in a shaded environment. Despite these initial results, dry weight reduction at 21 d after treatment never differed by more than 8% between levels of environmental factors, thus indicating a negligible influence on glyphosate efficacy. The magnitude of glyphosate-induced antagonism was generally greater at 30 C (12% to 21% less than expected control) versus 10 C (11% to 16%) on atrazine, cloransulam, dicamba, and topramezone and was greater at field capacity (20% to 24%) versus one-third field capacity (11% to 15%) on cloransulam and topramezone. These results indicate air temperatures and soil moisture levels conducive to optimal plant growth accelerate the RR to glyphosate, thereby increasing the likelihood of glyphosate-induced antagonism on several translocated herbicides.

Glyphosate resistance in Ambrosia trifida: Part 2. Rapid response physiology and non-target-site resistance.

The glyphosate-resistant rapid response (GR RR) resistance mechanism in Ambrosia trifida is not due to target-site resistance (TSR) mechanisms. This study explores the physiology of the rapid response and the possibility of reduced translocation and vacuolar sequestration as non-target-site resistance (NTSR) mechanisms. GR RR leaf discs accumulated hydrogen peroxide within minutes of glyphosate exposure, but only in mature leaf tissue. The rapid response required energy either as light or exogenous sucrose. The combination of phenylalanine and tyrosine inhibited the rapid response in a dose-dependent manner. Reduced glyphosate translocation was observed in GR RR, but only when associated with tissue death caused by the rapid response. Nuclear magnetic resonance studies indicated that glyphosate enters the cytoplasm and reaches chloroplasts, and it is not moved into the vacuole of GR RR, GR non-rapid response or glyphosate-susceptible A. trifida. The GR RR mechanism of resistance is not associated with vacuole sequestration of glyphosate, and the observed reduced translocation is likely a consequence of rapid tissue death. Rapid cell death was inhibited by exogenous application of aromatic amino acids phenylalanine and tyrosine. The mechanism by which these amino acids inhibit rapid cell death in the GR RR phenotype remains unknown, and it could involve glyphosate phytotoxicity or other agents generating reactive oxygen species. Implications of these findings are discussed. The GR RR mechanism is distinct from the currently described glyphosate TSR or NTSR mechanisms in other species. © 2017 Society of Chemical Industry.

Glyphosate Resistance in Giant Ragweed (<i>Ambrosia trifida</i> L.) from Mississippi Is Partly Due to Reduced Translocation

A giant ragweed population from a glyphosate-resistant (GR) soybean field in Mississippi, USA was suspected to be resistant to glyphosate. Greenhouse and laboratory studies were conducted to confirm and quantify the magnitude of glyphosate resistance in a resistant biotype selected from this population and to elucidate possible physiological and molecular mechanisms of glyphosate resistance. Glyphosate dose response studies indicated that ED50 (effective dose required to reduce plant growth by 50%) values for glyphosate-resistant (GR-MS) and glyphosate-susceptible (GS-MS) biotypes, based on percent injury, were 0.52 and 0.34 kg ae/ha glyphosate, respectively, indicating a 1.5-fold level of resistance in GR-MS. The absorption pattern of 14C-glyphosate in the two giant ragweed biotypes was similar throughout the measured time course of 168 h after treatment (HAT). The amount of 14C-glyphosate that translocated out of treated leaves of the GR-MS and GS-MS plants was similar up to 24 HAT. However, the GS-MS biotype translocated more (71% and 76% of absorbed at 48 and 96 HAT, respectively) 14C-glyphosate than the GR-MS biotype (44% and 66% of absorbed at 48 and 96 HAT, respectively) out of the treated leaf. No target site mutation was identified at the Pro106 location of the EPSPS gene of the GR-MS biotype. The mechanism of resistance to glyphosate in giant ragweed from Mississippi, at least, is due to reduced glyphosate translocation.

Reduced Glyphosate Translocation in Two Glyphosate-Resistant Populations of Rigid Ryegrass (Lolium rigidum) from Fence Lines in South Australia

Populations of rigid ryegrass with resistance to glyphosate have started to become a problem on fence lines of cropping fields of southern Australian farms. Seed of rigid ryegrass plants that survived glyphosate application were collected from two fence line locations in Clare, South Australia. Dose–response experiments confirmed resistance of these fence line populations to glyphosate. Both populations required 9- to 15-fold higher glyphosate dose to achieve 50% mortality in comparison to a standard susceptible population. The mechanism of resistance in these populations was investigated. Sequencing a conserved region of the gene encoding 5-enolpyruvyl-shikimate-3-phosphate synthase identified no differences between the resistant and susceptible populations. Absorption of glyphosate into leaves of the resistant populations was not different from the susceptible population. However, the resistant plants retained significantly more herbicide in the treated leaf blades than did the susceptible plants. Conversely, susceptible plants translocated significantly more herbicide to the leaf sheaths and untreated leaves than the resistant plants. The differences in translocation pattern for glyphosate between the resistant and susceptible populations of rigid ryegrass suggest resistance is associated with altered translocation of glyphosate in the fence line populations.

Rigid Ryegrass (Lolium rigidum) Populations Containing a Target Site Mutation in EPSPS and Reduced Glyphosate Translocation Are More Resistant to Glyphosate

Glyphosate is widely used for weed control in the grape growing industry in southern Australia. The intensive use of glyphosate in this industry has resulted in the evolution of glyphosate resistance in rigid ryegrass. Two populations of rigid ryegrass from vineyards, SLR80 and SLR88, had 6- to 11-fold resistance to glyphosate in dose-response studies. These resistance levels were higher than two previously well-characterized glyphosate-resistant populations of rigid ryegrass (SLR77 and NLR70), containing a modified target site or reduced translocation, respectively. Populations SLR80 and SLR88 accumulated less glyphosate, 12 and 17% of absorbed glyphosate, in the shoot in the resistant populations compared with 26% in the susceptible population. In addition, a mutation within the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) where Pro106had been substituted by either serine or threonine was identified. These two populations are more highly resistant to glyphosate as a consequence of expressing two different resistance mechanisms concurrently.

Glyphosate resistance in perennial Sorghum halepense (Johnsongrass), endowed by reduced glyphosate translocation and leaf uptake

In a large cropping area of northern Argentina, Sorghum halepense (Johnsongrass) has evolved towards glyphosate resistance. This study aimed to determine the molecular and biochemical basis conferring glyphosate resistance in this species. Experiments were conducted to assess target EPSPS gene sequences and (14)C-glyphosate leaf absorption and translocation to meristematic tissues. Individuals of all resistant (R) accessions exhibited significantly less glyphosate translocation to root (11% versus 29%) and stem (9% versus 26%) meristems when compared with susceptible (S) plants. A notably higher proportion of the applied glyphosate remained in the treated leaves of R plants (63%) than in the treated leaves of S plants (27%). In addition, individuals of S. halepense accession R(2) consistently showed lower glyphosate absorption rates in both adaxial (10-20%) and abaxial (20-25%) leaf surfaces compared with S plants. No glyphosate resistance endowing mutations in the EPSPS gene at Pro-101-106 residues were found in any of the evaluated R accessions. The results of the present investigation indicate that reduced glyphosate translocation to meristems is the primary mechanism endowing glyphosate resistance in S. halepense from cropping fields in Argentina. To a lesser extent, reduced glyphosate leaf uptake has also been shown to be involved in glyphosate-resistant S. halepense.

Glyphosate, paraquat and ACCase multiple herbicide resistance evolved in a Lolium rigidum biotype

Glyphosate is the world's most widely used herbicide. A potential substitute for glyphosate in some use patterns is the herbicide paraquat. Following many years of successful use, neither glyphosate nor paraquat could control a biotype of the widespread annual ryegrass (Lolium rigidum), and here the world's first case of multiple resistance to glyphosate and paraquat is confirmed. Dose-response experiments established that the glyphosate rate causing 50% mortality (LD(50)) for the resistant (R) biotype is 14 times greater than for the susceptible (S) biotype. Similarly, the paraquat LD(50 )for the R biotype is 32 times greater than for the S biotype. Thus, based on the LD(50 )R/S ratio, this R biotype of L. rigidum is 14-fold resistant to glyphosate and 32-fold resistant to paraquat. This R biotype also has evolved resistance to the acetyl-coenzyme A carboxylase (ACCase) inhibiting herbicides. The mechanism of paraquat resistance in this biotype was determined as restricted paraquat translocation. Resistance to ACCase-inhibiting herbicides was determined as due to an insensitive ACCase. Two mechanisms endowing glyphosate resistance were established: firstly, a point mutation in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene, resulting in an amino acid substitution of proline to alanine at position 106; secondly, reduced glyphosate translocation was found in this R biotype, indicating a co-occurrence of two distinct glyphosate resistance mechanisms within the R population. In total, this R biotype displays at least four co-existing resistance mechanisms, endowing multiple resistance to glyphosate, paraquat and ACCase herbicides. This alarming case in the history of herbicide resistance evolution represents a serious challenge for the sustainable use of the precious agrochemical resources such as glyphosate and paraquat.