Foliar-applied glyphosate substantially reduced uptake and transport of iron and manganese in sunflower (helianthus annuus L.) plants

Evidence clearly shows that cationic micronutrients in spray solutions reduce the herbicidal effectiveness of glyphosate for weed control due to the formation of metal-glyphosate complexes. The formation of these glyphosate-metal complexes in plant tissue may also impair micronutrient nutrition of nontarget plants when exposed to glyphosate drift or glyphosate residues in soil. In the present study, the effects of simulated glyphosate drift on plant growth and uptake, translocation, and accumulation (tissue concentration) of iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) were investigated in sunflower ( Helianthus annuusL.) plants grown in nutrient solution under controlled environmental conditions. Glyphosate was sprayed on plant shoots at different rates between 1.25 and 6.0% of the recommended dosage (i.e., 0.39 and 1.89 mM glyphosate isopropylamine salt). Glyphosate applications significantly decreased root and shoot dry matter production and chlorophyll concentrations of young leaves and shoot tips. The basal parts of the youngest leaves and shoot tips were severely chlorotic. These effects became apparent within 48 h after the glyphosate spray. Glyphosate also caused substantial decreases in leaf concentration of Fe and Mn while the concentration of Zn and Cu was less affected. In short-term uptake experiments with radiolabeled Fe (59Fe), Mn (54Mn), and Zn (65Zn), root uptake of 59Fe and 54Mn was significantly reduced in 12 and 24 h after application of 6% of the recommended dosage of glyphosate, respectively. Glyphosate resulted in almost complete inhibition of root-to-shoot translocation of 59Fe within 12 h and 54Mn within 24 h after application. These results suggest that glyphosate residues or drift may result in severe impairments in Fe and Mn nutrition of nontarget plants, possibly due to the formation of poorly soluble glyphosate-metal complexes in plant tissues and/or rhizosphere interactions

Glyphosate-induced impairment of plant growth and micronutrient status in glyphosate-resistant soybean (Glycine max L.)

This investigation demonstrated potential detrimental side effects of glyphosate on plant growth and micronutrient (Mn, Zn) status of a glyphosate-resistant (GR) soybean variety (Glycine max cv. Valiosa), which were found to be highly dependent on the selected growth conditions. In hydroponic experiments with sufficient Mn supply [0.5 μM], the GR cv. Valiosa produced similar plant biomass, root length and number of lateral roots in the control treatment without glyphosate as compared to its non-GR parental line cv. Conquista. However, this was associated with 50% lower Mn shoot concentrations in cv. Conquista, suggesting a higher Mn demand of the transgenic cv. Valiosa under the selected growth conditions. Glyphosate application significantly inhibited root biomass production, root elongation, and lateral root formation of the GR line, associated with a 50% reduction of Mn shoot concentrations. Interestingly, no comparable effects were detectable at low Mn supply [0.1 μM]. This may indicate Mn-dependent differences in the intracellular transformation of glyphosate to the toxic metabolite aminomethylphosphonic acid (AMPA) in the two isolines. In soil culture experiments conducted on a calcareous loess sub-soil of a Luvisol (pH 7.6) and a highly weathered Arenosol (pH 4.5), shoot biomass production and Zn leaf concentrations of the GR-variety were affected by glyphosate applications on the Arenosol but not on the calcareous Loess sub-soil. Analysis of micronutrient levels in high and low molecular weight (LMW) fractions (80% ethanol extracts) of young leaves revealed no indications for internal immobilization of micronutrients (Mn, Zn, Fe) by excessive complexation with glyphosate in the LMW phase

Glyphosate resistance in annual ryegrass (Lolium rigidum Gaud.) with multiple resistance mechanisms.

Glyphosate (N-(phosphonomethyl)glycine) is a post-emergent, systemic and non-selective herbicide for the control of annual and perennial weeds. This herbicide has very low toxicity to the mammals. The target enzyme for glyphosate in plants is 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Glyphosate inhibits the biosynthesis of the aromatic amino acids phenylalanine, tyrosine and tryptophan in the plant. The first case of glyphosate resistance was reported in Lolium rigidum in Australia after 15 years of persistence use of this herbicide and the number weeds reported resistant to glyphosate has increased around the world. So far, two mechanisms known to be involved in resistance to glyphosate are target-site mutation and reduced herbicide translocation. Recently, two populations of L. rigidum from Australia have been discovered with very high levels of resistance to glyphosate. This project aims to determine the levels of glyphosate resistance in these populations, investigate glyphosate resistance mechanisms in the populations and finally assess the mode of inheritance of resistance. In this project, four resistant (NLR70, SLR77, SLR80 and SLR88) and one susceptible (VLR1) L. rigidum populations were evaluated for their response to glyphosate. From the dose response experiments, the susceptible population of VLR1 was completely controlled with the recommended rate of glyphosate (450 g a.e ha⁻ ¹). In contrast, the resistant populations were not fully controlled by this herbicide rate. There was considerable variation between the populations in their resistance to glyphosate. In comparison to the susceptible population VLR1, SLR77 was 2.2 to 3.5 fold resistant to glyphosate, NLR70 was 3.7 to 8.4 fold resistant to glyphosate, SLR88 was 5.6 to 11.4 fold resistant to glyphosate and SLR80 was 8.2 to 76.7 fold resistant to glyphosate. The mechanism of glyphosate resistance in the populations was investigated. ¹⁴ C-glyphosate was used to determine the absorption and translocation of glyphosate among the populations. There was no significant difference on the absorption of ¹⁴ C-glyphosate 48 hours after treatment in the population. However, the accumulation of ¹⁴ C-glyphosate in the stem region was higher in the susceptible VLR1 population (25.9%) and in resistant SLR77 (25%) than the other three populations. The resistant populations NLR70, SLR88 and SLR80 had about half the amount of glyphosate accumulating in the stem region. These three resistant populations appear to be resistant to glyphosate as a result of reduced translocation of glyphosate to the shoot meristem. Part of the EPSP synthase gene of the susceptible and four resistant populations was amplified and sequenced to identify any changes in the nucleotide sequence. The predicted amino acid sequence from the susceptible population VLR1 was the same as the consensus sequence from other plant species in the conserved region sequenced. However, the resistant populations of NLR70, SLR77, SLR80 and SLR88 showed polymorphisms within the nucleotide sequence in this region. Single nucleotide substitutions of A for C at codon 106 were observed in the resistant populations SLR77 and SLR80. This nucleotide change is predicted to substitute threonine for proline at position 106. In the resistant population SLR88, a nucleotide substitution of T for C was observed at the same codon. This nucleotide substitution is predicted to change the amino acid from proline 106 to serine. Therefore, these three populations appear to be resistant to glyphosate as a result of a target-site mutation. An inheritance study was conducted by cross pollinating the susceptible VLR1 and resistant SLR88 population. From the dose response, the parent susceptible was completely killed with the recommended rate of glyphosate and higher rates of glyphosate were required to control parental resistant and both F₁ progenies (maternal susceptible and resistant). Both F₁ progenies showed an intermediate response to glyphosate compared with the parental populations. This indicated that the resistance to glyphosate in population SLR88 is inherited by nuclear gene(s) through the transfer of pollen during the cross pollination. It is suggested that SLR88 and SLR80 population contain both glyphosate resistant mechanisms due to the cross pollination between individuals with different resistant mechanisms. Having two resistant mechanisms results in populations being highly resistant to glyphosate compared to those with one resistance mechanism. The higher level of glyphosate resistance in these multiple glyphosate resistance populations will likely make them harder to manage.Thesis (M.Ag.Sc.) -- University of Adelaide, School of Agriculture, Food and Wine, 201

Glyphosate inhibition of ferric reductase activity in iron deficient sunflower roots

Iron (Fe) deficiency is increasingly being observed in cropping systems with frequent glyphosate applications. A likely reason for this is that glyphosate interferes with root uptake of Fe by inhibiting ferric reductase in roots required for Fe acquisition by dicot and nongrass species. This study investigated the role of drift rates of glyphosate (0.32, 0.95 or 1.89 mM glyphosate corresponding to 1, 3 and 6% of the recommended herbicidal dose, respectively) on ferric reductase activity of sunflower (Helianthus annuus) roots grown under Fe deficiency conditions. Application of 1.89 mM glyphosate resulted in almost 50% inhibition of ferric reductase within 6 h and complete inhibition 24 h after the treatment. Even at lower rates of glyphosate (e.g. 0.32 mM and 0.95 mM), ferric reductase was inhibited. Soluble sugar concentration and the NAD(P)H oxidizing capacity of apical roots were not decreased by the glyphosate applications. To our knowledge, this is the first study reporting the effects of glyphosate on ferric reductase activity. The nature of the inhibitory effect of glyphosate on ferric reductase could not be identified. Impaired ferric reductase could be a major reason for the increasingly observed Fe deficiency in cropping systems associated with widespread glyphosate usage

Glyphosate reduced seed and leaf concentrations of calcium, manganese, magnesium, and iron in non-glyphosate resistant soybean

Greenhouse experiments were conducted to study the effects of glyphosate drift on plant growth and concentrations of mineral nutrients in leaves and seeds of non-glyphosate resistant soybean plants (Glycine max, L.). Glyphosate was sprayed on plant shoots at increasing rates between 0.06 and 1.2% of the recommended application rate forweed control. In an experiment with 3-week-old plants, increasing application of glyphosate on shoots significantly reduced chlorophyll concentration of the young leaves and shoots dry weight, particularly the young parts of plants. Concentration of shikimate due to increasing glyphosate rates was nearly 2-fold for older leaves and 16-fold for younger leaves compared to the control plants without glyphosate spray. Among the mineral nutrients analyzed, the leaf concentrations of potassium (K), phosphorus (P), copper (Cu) and zinc (Zn) were not affected, or even increased significantly in case of P and Cu in young leaves by glyphosate, while the concentrations of calcium (Ca), manganese (Mn) and magnesium (Mg) were reduced, particularly in young leaves. In the case of Fe, leaf concentrations showed a tendency to be reduced by glyphosate. In the second experiment harvested at the grain maturation, glyphosate application did not reduce the seed concentrations of nitrogen (N), K, P, Zn and Cu. Even, at the highest application rate of glyphosate, seed concentrations of N, K, Zn and Cuwere increased by glyphosate. By contrast, the seed concentrations of Ca, Mg, Fe and Mn were significantly reduced by glyphosate. These results suggested that glyphosatemay interfere with uptake and retranslocation of Ca, Mg, Fe and Mn, most probably by binding and thus immobilizing them. The decreases in seed concentration of Fe, Mn, Ca and Mg by glyphosate are very specific, and may affect seed quality

Turfgrass species response exposed to increasing rates of glyphosate application

To investigate the response of nine turfgrass species exposed to increasing rates of glyphosate application, the dry matter production, visual leaf injury symptoms (e.g., chlorosis and necrosis) and the concentrations of shikimate and mineral nutrients were determined in shoots. The rates of foliar glyphosate application were 0%, 5% (1.58 mM), and 20% (6.32mM) of the recommended application rate for weed control. In general, there was a negative and weak correlation between the intensity of visual injury and relative decreases in shoot dry matter production caused by glyphosate application. The decreases in shoot dry matter production and the severity of leaf damage pronounced by increasing glyphosate rate showed a substantial variation among the turfgrass species. Of the turfgrass species tested, Festuca arundinacea ‘Falcon’ and Buchloe dactyloides ‘Bowie’were selected as the most tolerant and sensitive species to applied sublethal rates of glyphosate as judged from visual injury ratings, respectively. At the highest glyphosate rate, shoot dryweightwas decreased by 4-fold in Bowie and only 1.6-fold in Falcon. When glyphosatewas not applied, shoot shikimate concentration of all species was very low and below 2.8mol g−1 FW (fresh weight). Glyphosate applications resulted in increases in shoot shikimate concentration with substantial variations among species. At 6.32mM glyphosate treatment, shikimate concentration ranged between 156.1mol g−1 (F. rubra, Ambrose) and 16.5mol g−1 FW (F. rubra, Cindy Lou). However, the highly sensitive and the tolerant genotypes were not different in shoot shikimate concentrations. Even, in the case of some genotypes, high glyphosate tolerance is accompanied by higher shoot concentrations of shikimate. Depending on the turfgrass species and mineral nutrients tested, increasing glyphosate application either did not affect or reduced mineral nutrient concentrations. In the case of decreases in shoot concentration of mineral nutrients, the decreases in Ca, Mg, Mn and Fe were most distinct. The results obtained indicate existence of a large genetic variation in tolerance to glyphosate toxicity among the turfgrass species. This differential variation in tolerance to glyphosate could not be explained by the changes in shoot concentrations of shikimate and mineral nutrients

Effect of a glyphosate-containing herbicide on Escherichia coli and Salmonella Ser. Typhimurium in an in vitro rumen simulation system

Glyphosate (N-(phosphonomethyl)glycine) is the most-used herbicide worldwide. Many studies in the past have shown that residues of the herbicide can be found in many cultivated plants, including those used as livestock feed. Sensitivity to glyphosate varies with bacteria, particularly those residing in the intestine, where microbiota is exposed to glyphosate residues. Therefore, less susceptible pathogenic isolates could have a distinct advantage compared to more sensitive commensal isolates, probably leading to dysbiosis. To determine whether the ruminal growth and survival of pathogenic Escherichia coli or Salmonella serovar Typhimurium are higher when glyphosate residues are present in the feed, an in vitro fermentation trial with a “Rumen Simulation System” (RUSITEC) and a glyphosate-containing commercial formulation was performed. Colony forming units of E. coli and Salmonella ser. Typhimurium decreased steadily in all fermenters, regardless of the herbicide application. Minimum inhibitory concentrations of the studied Salmonella and E. coli strains did not change, and antibiotic susceptibility varied only slightly but independent of the glyphosate application. Overall, application of the glyphosate-containing formulation in a worst-case concentration of 10 mg/L neither increased the abundance for the tested E. coli and Salmonella strain in the in vitro fermentation system, nor promoted resistance to glyphosate or antibiotics

Sorption and desorption of glyphosate in Mollisols and Ultisols soils of Argentina

In Argentina, glyphosate use has increased exponentially in the past years due to the widespread adoption of no-till management combined with genetically modified glyphosate-resistant crops. This massive use of glyphosate has created concern about its potential environmental impact. Sorption-desorption of glyphosate was studied in three Argentinean soils with contrasting characteristics. Glyphosate sorption isotherms were modeled using the Freundlich equation to estimate the sorption coefficient (Kf). Glyphosate sorption was high and the Kf varied from 115.6 to 1612 mg 1-1/nL1/n /Kg. Cerro Azul soil had the highest glyphosate sorption capacity due to a combination of factors such as higher clay content, CEC, total Fe, Al oxides and lower available phosphorous and pH. Desorption isotherms were also modeled using the Freundlich equation. In general, desorption was very low (<12%). The low values of hysteresis coefficient (H) confirm that glyphosate strongly sorbs to the soils and that it is almost an irreversible process. Anguil soil had a significant higher desorption coefficient (Kfd) than the other soils, associated with its lower clay content and higher pH and phosphorous. Glyphosate high sorption and low desorption to the studied soils may prevent groundwater contamination. However, it may also affect its bioavailability increasing its persistence and favoring its accumulation in environment. Results of this study contribute to the knowledge and characterization of glyphosate retention in different soils.Fil: Gómez Ortiz, Ana Maria. Universidad Nacional de Mar del Plata; ArgentinaFil: Okada, Elena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Bedmar, Francisco. Universidad Nacional de Mar del Plata; ArgentinaFil: Costa, Jose Luis. Instituto Nacional de Tecnología Agropecuaria; Argentin

Glyphosate in waters and soils from genetically modified canola cultivation in Parkes, NSW, Australia

Investigations were conducted of farmland from the Parkes region of New South Wales, Australia, cultivated with genetically modified canola, involving the determination of glyphosate (N-(phosphonomethyl)glycine) concentrations in water and soils, and its sorption. The soils are classified as loam under the USDA system (clay 13.8-15.8%, silt 39-43%, sand 41.2-47.2%). Firstly, a low-cost fluorometric method was developed for the analysis of glyphosate in waters and soils, calibrated against analytical standards and spectrophotometric and enzyme-linked immunosorbent assay (ELISA) methods. Soil and water samples were then collected using the NEPM sampling protocol into glass containers, chilled and analysed within two weeks. The samples were collected in multiple episodes, taking account of glyphosate and pesticide crop applications. The soil and water physical and chemical properties were characterised, and glyphosate levels were determined. Field concentrations of glyphosate ranged between 0.01 - 0.067 mg/L in water and 0.10 - 0.575 mg/kg in soil. The aqueous levels lie below Australian and international drinking water guidelines, but reach a Canadian freshwater guideline. Glyphosate levels varied with time of application and rainfall events. Glyphosate sorption isotherms were also constructed by batch tests on several soils, and were fitted with Freundlich and Langmuir isotherms. Desorption tests indicated 25% to 58% of soil glyphosate is extractable by 0.1M KH2PO4