Glyphosate effects on phenolic metabolism of nodulated soybean (Glycine max L. merr.).

Glyphosate is a herbicide that blocks the shikimic acid pathway. Three Bradyrhizobium japonicum strains with different sensitivities to glyphosate were used to test the effect of this herbicide on the phenolic metabolism of nodulated soybeans and on the bacteroid nitrogenase activity. Glyphosate caused an inhibition in the bacteroid nitrogenase activity that was related with the sensitivity of the nodule-forming strains. Both leaves and nodules accumulated huge amounts of shikimate and phenolic acids (mainly protocatechuic acid), indicating that the herbicide was translocated to the nodule and disturbed phenolic metabolism. However, this accumulation was not clearly related to the sensitivity of the different strains. Bacteroids from control plants were incubated with the same concentration of shikimate, and phenolic acid accumulated in glyphosate-treated plants. Despite the high levels found in nodules, they were not responsible for the decrease of the nitrogenase activity. Glyphosate by itself caused a small inhibition of the bacteroid nitrogenase activity.     

Glyphosate-resistant and -susceptible soybean (Glycine max) and canola (Brassica napus) dose response and metabolism relationships with glyphosate

Experiments were conducted to determine (1) dose response of glyphosate-resistant (GR) and -susceptible (non-GR) soybean [Glycine max (L.) Merr.] and canola (Brassica napus L.) to glyphosate, (2) if differential metabolism of glyphosate to aminomethyl phosphonic acid (AMPA) is the underlying mechanism for differential resistance to glyphosate among GR soybean varieties, and (3) the extent of metabolism of glyphosate to AMPA in GR canola and to correlate metabolism to injury from AMPA. GR50 (glyphosate dose required to cause a 50% reduction in plant dry weight) values for GR (Asgrow 4603RR) and non-GR (HBKC 5025) soybean were 22.8 kg ae ha-1 and 0.47 kg ha-1, respectively, with GR soybean exhibiting a 49-fold level of resistance to glyphosate as compared to non-GR soybean. Differential reduction in chlorophyll by glyphosate was observed between GR soybean varieties, but there were no differences in shoot fresh weight reduction. No significant differences were found between GR varieties in metabolism of glyphosate to AMPA, and in shikimate levels. These results indicate that GR soybean varieties were able to outgrow the initial injury from glyphosate, which was previously caused at least in part by AMPA. GR50 values for GR (Hyola 514RR) and non-GR (Hyola 440) canola were 14.1 and 0.30 kg ha-1, respectively, with GR canola exhibiting a 47-fold level of resistance to glyphosate when compared to non-GR canola. Glyphosate did not cause reduction in chlorophyll content and shoot fresh weight in GR canola, unlike GR soybean. Less glyphosate (per unit leaf weight) was recovered in glyphosate-treated GR canola as compared to glyphosate-treated GR soybean. External application of AMPA caused similar injury in both GR and non-GR canola. The presence of a bacterial glyphosate oxidoreductase gene in GR canola contributes to breakdown of glyphosate to AMPA. However, the AMPA from glyphosate breakdown could have been metabolized to nonphytotoxic metabolites before causing injury to GR canola. Injury in GR and non-GR canola from exogenous application of AMPA was similar.     

Evaluation of spectrophotometric and HPLC methods for shikimic acid determination in plants: models in glyphosate-resistant and -susceptible crops

Endogenous shikimic acid determinations are routinely used to assess the efficacy of glyphosate in plants. Numerous analytical methods exist in the public domain for the detection of shikimic acid, yet the most commonly cited comprise spectrophotometric and high-pressure liquid chromatography (HPLC) methods. This paper compares an HPLC and two spectrophotometric methods (Spec 1 and Spec 2) and assesses the effectiveness in the detection of shikimic acid in the tissues of glyphosate-treated plants. Furthermore, the study evaluates the versatility of two acid-based shikimic acid extraction methods and assesses the longevity of plant extract samples under different storage conditions. Finally, Spec 1 and Spec 2 are further characterized with respect to (1) the capacity to discern between shikimic acid and chemically related alicyclic hydroxy acids, (2) the stability of the chromophore (t1/2), (3) the detection limits, and (4) the cost and simplicity of undertaking the analytical procedure. Overall, spectrophotometric methods were more cost-effective and simpler to execute yet provided a narrower detection limit compared to HPLC. All three methods were specific to shikimic acid and detected the compound in the tissues of glyphosate-susceptible crops, increasing exponentially in concentration within 24 h of glyphosate application and plateauing at approximately 72 h. Spec 1 estimated more shikimic acid in identical plant extract samples compared to Spec 2 and, likewise, HPLC detection was more effective than spectrophotometric determinations. Given the unprecedented global adoption of glyphosate-resistant crops and concomitant use of glyphosate, an effective and accurate assessment of glyphosate efficacy is important. Endogenous shikimic acid determinations are instrumental in corroborating the efficacy of glyphosate and therefore have numerous applications in herbicide research and related areas of science as well as resolving many commercial issues as a consequence of glyphosate utilization.     

Aminomethylphosphonic acid, a metabolite of glyphosate, causes injury in glyphosate-treated, glyphosate-resistant soybean

Glyphosate-resistant (GR) soybean [Glycine max (L.) Merr.] was developed by stable integration of a foreign gene that codes insensitive enzyme 5-enolpyruvylshikimate-3-phosphate synthase, an enzyme in the shikimate pathway, the target pathway of glyphosate. Application of glyphosate to GR soybean results in injury under certain conditions. It was hypothesized that if GR soybean is completely resistant to the glyphosate, injury could be caused by a metabolite of glyphosate, aminomethylphosphonic acid (AMPA), a known phytotoxin. Glyphosate and AMPA effects on one- to two-trifoliolate leaf stage (16-18-days old) GR and non-GR soybean were examined in the greenhouse. In GR soybean, a single application of glyphosate-isopropylammonium (1.12-13.44 kg/ha) with 0.5% Tween 20 did not significantly reduce the chlorophyll content of the second trifoliolate leaf at 7 days after treatment (DAT) or the shoot dry weight at 14 DAT compared with Tween 20 alone. A single application of AMPA (0.12-8.0 kg/ha) with 0.5% Tween 20 reduced the chlorophyll content of the second trifoliolate leaf by 0-52% at 4 DAT and reduced shoot fresh weight by 0-42% at 14 DAT in both GR and non-GR soybeans compared with Tween 20 alone. AMPA at 0.12 and 0.50 kg/ha produced injury in GR and non-GR soybean, respectively, similar to that caused by glyphosate-isopropylammonium at 13.44 kg/ha in GR soybean. AMPA levels found in AMPA-treated soybean of both types and in glyphosate-treated GR soybean correlated similarly with phytotoxicity. These results suggest that soybean injury to GR soybean from glyphosate is due to AMPA formed from glyphosate degradation.     

Aminomethylphosphonic acid accumulation in plant species treated with glyphosate

Aminomethylphosphonic acid (AMPA) is the most frequently detected metabolite of glyphosate in plants. The objective of this study was to determine if there is any correlation of metabolism of glyphosate to AMPA in different plant species and their natural level of resistance to glyphosate. Greenhouse studies were conducted to determine the glyphosate I 50 values (rate required to cause a 50% reduction in plant growth) and to quantify AMPA and shikimate concentrations in selected leguminous and nonleguminous species treated with glyphosate at respective I 50 rates. Coffee senna [ Cassia occidentalis (L.) Link] was the most sensitive ( I 50 = 75 g/ha) and hemp sesbania [ Sesbania herbacea (P.Mill.) McVaugh] was the most resistant ( I 50 = 456 g/ha) to glyphosate. Hemp sesbania was 6-fold and Illinois bundleflower [ Desmanthus illinoensis (Michx.) MacM. ex B.L.Robins. & Fern.] was 4-fold more resistant to glyphosate than coffee senna. Glyphosate was present in all plant species, and its concentration ranged from 0.308 to 38.7 microg/g of tissue. AMPA was present in all leguminous species studied except hemp sesbania. AMPA concentration ranged from 0.119 to 4.77 microg/g of tissue. Shikimate was present in all plant species treated with glyphosate, and levels ranged from 0.053 to 16.5 mg/g of tissue. Non-glyphosate-resistant (non-GR) soybean accumulated much higher shikimate than glyphosate-resistant (GR) soybean. Although some leguminous species were found to be more resistant to glyphosate than others, and there was considerable variation between species in the glyphosate to AMPA levels found, metabolism of glyphosate to AMPA did not appear to be a common factor in explaining natural resistance levels.     

Manganese in RR Cotton as Affected by Glyphosate

There is evidence that glyphosate application in soybean tolerant to herbicides could interfere in the manganese (Mn) nutrition of the crop, but there is no information on this effect in cotton plants. This study aimed at assessing manganese accumulation and distribution in cotton as affected by glyphosate application. The experiment was conducted in nutrient solution with four Mn concentrations and two cotton cultivars: conventional NuOpal and NuOpal tolerant to glyphosate (RR). Glyphosate was applied or not to the tolerant cultivar. The inclusion of the glyphosate resistance gene in cotton and herbicide, application increased shikimic acid (ShA) concentration in the plants. Glyphosate application decreased cotton leaf area and the dry matter production of the plant structures. The adverse effects of glyphosate were not overcome with higher Mn rates in the solution.     

Simulated glyphosate drift influences nitrate assimilation and nitrogen fixation in non-glyphosate-resistant soybean

Nontarget injury from glyphosate drift is a concern among growers using non-glyphosate-resistant (non-GR) cultivars. The effects of glyphosate drift on nitrate assimilation and nitrogen fixation potential, nodule mass, and yield of non-GR soybean were assessed in a field trial at Stoneville, MS. A non-GR soybean cultivar 'Delta Pine 4748S' was treated with glyphosate at 12.5% of use rate of 0.84 kg of active ingredient/ha at 3 (V2), 6 (V7), and 8 (R2, full bloom) weeks after planting (WAP) soybean to simulate glyphosate drift. Untreated soybean was used as a control. Soybeans were sampled weekly for 2 weeks after each glyphosate treatment to assess nitrate assimilation and N2 fixation potential. Nitrate assimilation was assessed using in vivo nitrate reductase assay in leaves, stems, roots, and nodules. Nitrogen fixation potential was assessed by measuring nitrogenase activity using the acetylene reduction assay (ARA). Nitrogen content of leaves, shoots, and seed and soybean yield were also determined. In the first sampling date (4 WAP), glyphosate drift caused a significant decrease in NRA in leaves (60%), stems (77%), and nodules (50%), with no decrease in roots. At later growth stages, NRA in leaves was more sensitive to glyphosate drift than stems and roots. Nitrogenase activity was reduced 36-58% by glyphosate treatment at 3 or 6 WAP. However, glyphosate treatment at 8 WAP had no effect on nitrogenase activity. Nitrogen content was affected by glyphosate application only in shoots after the first application. No yield, seed nitrogen, protein, or oil concentration differences were detected. These results suggest that nitrate assimilation and nitrogen fixation potential were significantly reduced by glyphosate drift, with the greatest sensitivity early in vegetative growth. Soybean has the ability to recover from the physiological stress caused by glyphosate drift.     

Glyphosate applied preharvest induces shikimic acid accumulation in hard red spring wheat (Triticum aestivum)

Glyphosate is a nonselective herbicide used as a harvest aid in a variety of crops. Glyphosate is absorbed into the foliage and translocated to metabolically active regions in the plant where it interferes with the shikimic acid pathway. Experiments were conducted to determine the accumulation and distribution of shikimic acid in wheat treated with glyphosate at soft and hard dough stages of kernel development and to determine the fate of shikimic acid during milling and bread making. Elevated levels of shikimic acid were detected throughout the wheat plant. Shikimic acid concentrations peaked 3-7 days after treatment and then declined until harvest. Shikimic acid content was 3-fold greater in flour and 2-fold greater in the bread derived from treated wheat than nontreated wheat. Similarly, elevated levels of shikimic acid were found in the crumbs and crust of bread made with flour from glyphosate treated wheat. Glyphosate applied preharvest resulted in shikimic acid accumulation in hard red spring wheat and subsequent end-use products.     

Management practices to control premature senescence in Bt cotton

Commercial cultivation of Bt cotton produced higher boll load which led to stiff inter-original competition for photosynthates, resulting in early cessation of growth (premature senescence) due to more availability of sink and less sources. To overcome this problem, field experiment was conducted during 2011 and 2012 using five treatments of plant growth manipulation viz. no fruiting branch removal (F₁), removal of first fruiting branch (F₂), removal of first and second fruiting branch (F₃), removal of all squares from first fruiting branch (F₄), removal of all squares from first and second fruiting branches (F₅), and three potassium (K) application rates viz. 50 kg ha^?1 (K₁), 100 kg ha^?1 (K₂), and 150 kg ha^?1 (K₃). More nodes above white flower were recorded in F₅, followed by F₃, while minimum were recorded in F₁. Among potassium levels, maximum nodes above white flower were recorded in K₃ followed by K₂ and K₁ during both years of study. Plant height recorded at physiological cutout stage or at maturity stage showed that plants gained more height with removal of all squares from first tosecond fruiting branches with higher potassium dose. Leaf K increased with increasing applied potassium and also with square/branch removal. So early removal of squares/fruiting branches along with higher potassium dose helped in delaying canopy senescence in Bt cotton.     

Effect of Fruiting Branch/Square Removal on Growth and Quality of Bt Cotton under Different Potassium Rates

Removal of early fruiting branches with greater potassium doses caused more source and no sink at early stages of growth, leading to improved yield, yield components, and fiber quality traits in Bt cotton. The study used manual alteration of plant architecture (F₁, no branch removal; F₂, removal of first fruiting branch; F₃, removal of first and second fruiting branches; F₄, removal of all squares from first fruiting branch; F₅, removal of all squares from first and second fruiting branches) and potassium rates (50, 100, and 150 kg ha^?1) in a randomized complete block design and was repeated for 2 years (2011 and 2012). Increasing potassium application increased total bolls per plant and cotton yield to the greatest levels in F₃ and F₅, against lowest level in the control. Ginning out turn, fiber length, seed oil, and seed protein content were influenced by fruiting branch or square removal but the difference was less. Increasing potassium improved seed and fiber quality.     

Influence of glyphosate on amino acid composition of Egyptian broomrape

The parasitic plant broomrape is entirely dependent on its host for reduced carbon and nitrogen and is also susceptible to inhibition by glyphosate that is translocated to the parasite through a host. Studies were conducted to examine the effect of broomrape parasitism on amino acid concentrations of two hosts: common vetch that is tolerant of low levels of glyphosate and oilseed rape that has been genetically engineered for glyphosate resistance. The influence of glyphosate on the amino acid content of broomrape and the two hosts was also examined. Amino acid concentrations in leaves and roots of parasitized common vetch plants were generally similar to those of the corresponding tissues of nonparasitized plants. Amino acid concentrations in broomrape were lower than those of the parasitized common vetch root. For common vetch, glyphosate applied at rates that selectively inhibited broomrape growth did not alter individual amino acid concentrations in the leaves, but generally increased amino acid levels at 0.18 kg ha-1. Glyphosate application also increased the amino acid concentrations, with the exception of arginine, of broomrape growing on common vetch and did not generally influence concentrations in leaves or roots of common vetch. In oilseed rape, parasitization by broomrape generally led to higher amino acid concentrations in leaves but lower concentrations in roots of parasitized plants. Broomrape had higher amino acid concentrations than roots of the parasitized oilseed rape. Glyphosate applied at 0.25 and 0.5 kg ha-1 generally increased the amino acid concentrations in oilseed rape leaves, but the 0.75 kg ha-1 application caused the amino acid concentrations to decrease compared to those of untreated plants. In oilseed rape root the general trend was an increase in the concentration of amino acids at the two highest rates of glyphosate. Individual amino acid concentrations in broomrape attachments growing on oilseed rape were generally increased following glyphosate application of 0.25 kg ha-1. These results indicate that low rates of glyphosate alter amino acid profiles in both host and broomrape and raise questions about the regulation of amino acid metabolism in the parasite.     

Glyphosate degradation in glyphosate-resistant and -susceptible crops and weeds

High levels of aminomethylphosphonic acid (AMPA), the main glyphosate metabolite, have been found in glyphosate-treated, glyphosate-resistant (GR) soybean, apparently due to plant glyphosate oxidoreductase (GOX)-like activity. AMPA is mildly phytotoxic, and under some conditions the AMPA accumulating in GR soybean correlates with glyphosate-caused phytotoxicity. A bacterial GOX is used in GR canola, and an altered bacterial glyphosate N-acetyltransferase is planned for a new generation of GR crops. In some weed species, glyphosate degradation could contribute to natural resistance. Neither an isolated plant GOX enzyme nor a gene for it has yet been reported in plants. Gene mutation or amplification of plant genes for GOX-like enzyme activity or horizontal transfer of microbial genes from glyphosate-degrading enzymes could produce GR weeds. Yet, there is no evidence that metabolic degradation plays a significant role in evolved resistance to glyphosate. This is unexpected, considering the extreme selection pressure for evolution of glyphosate resistance in weeds and the difficulty in plants of evolving glyphosate resistance via other mechanisms.     

New insights on glyphosate mode of action in nodular metabolism: Role of shikimate accumulation

The short-term effects of the herbicide glyphosate (1.25-10 mM) on the growth, nitrogen fixation, carbohydrate metabolism, and shikimate pathway were investigated in leaves and nodules of nodulated lupine plants. All glyphosate treatments decreased nitrogenase activity rapidly (24 h) after application, even at the lowest and sublethal dose used (1.25 mM). This early effect on nitrogenase could not be related to either damage to nitrogenase components (I and II) or limitation of carbohydrates supplied by the host plant. In fact, further exposure to increasing glyphosate concentrations (5 mM) and greater time after exposure (5 days) decreased nodule starch content and sucrose synthase (SS; EC 2.4.1.13) activity but increased sucrose content within the nodule. These effects were accompanied by a great inhibition of the activity of phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31). There were remarkable and rapid effects on the increase of shikimic and protocatechuic (PCA) acids in nodules and leaves after herbicide application. On the basis of the role of shikimic acid and PCA in the regulation of PEPC, as potent competitive inhibitors, this additional effect provoked by glyphosate on 5-enolpyruvylshikimic-3-phosphate synthase enzyme (EPSPS; EC 2.5.1.19) inhibition would divert most PEP into the shikimate pathway, depriving energy substrates to bacteroids to maintain nitrogen fixation. These findings provide a new explanation for the effectiveness of glyphosate as a herbicide in other plant tissues, for the observed differences in tolerance among species or cultivars, and for the transitory effects on glyphosate-resistant transgenic crops under several environmental conditions.     

冀东野生大豆对草甘膦的耐性鉴定及耐性机制初步研究

为筛选耐草甘膦野生大豆种质并了解其耐性机制,本试验对采集于冀东地区的862份野生大豆进行了草甘膦的耐性鉴定。在草甘膦处理后,测定了高耐和敏感材料的莽草酸、丙二醛和叶绿素含量,过氧化物酶(POD)、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活性,以及草甘膦相关基因EPSPS表达量。结果显示,喷施草甘膦后,862份野生大豆材料中,药害等级在4级以上的材料占82.84%,3级占9.51%,2级占4.87%,1级占2.78%。筛选到高耐草甘膦的野生大豆材料Yong-33,其在1.125 kg a.i·hm^-2 草甘膦处理后植株存活率达到96.67%。经草甘膦处理后,与对照相比,高耐材料的叶绿素、丙二醛和莽草酸含量在检测的各时间点均无显著差异,敏感材料叶绿素含量显著降低,丙二醛和莽草酸含量显著升高;高耐材料POD、CAT和SOD活性以及EPSPS基因表达量均显著升高,而敏感材料酶活性及EPSPS基因表达量无显著差异。以上结果表明,野生大豆中存在高耐草甘膦的种质资源,在草甘膦处理后其植株内活性氧清除酶系活性升高,EPSPE基因上调表达,推测这是野生大豆对草甘膦耐性较好的原因。本研究筛选到的耐草甘膦野生大豆材料可为培育耐草甘膦栽培大豆新品种提供种质资源。     

Dynamics of mineral nutrient element concentrations in developing cotton leaves,bracts, and floral buds in relation to position in the canopy

Plant mineral nutrient element status is an important factor influencing cotton (Gossypium hirsutum L.) growth, development, metabolism, and yield. A field study was conducted to determine changes in mineral nutrient element concentrations in leaves, bracts, and floral buds of field‐grown cotton plants during development of squares (floral bud with three bracts) as affected by fruiting position within the plant canopy. During square ontogeny, the nitrogen (N), phosphorous (P) and potassium (K) concentrations of sympodial leaves and floral bracts decreased, whereas the calcium (Ca) and magnesium (Mg) increased, and sulfur (S) concentration exhibited little change (leaves) or increased (bracts) with increasing square age. The N, P, Ca, and S concentrations in floral buds declined synchronously; K and Mg concentrations showed an increasing trend within the first 20 days, peaked at about 25‐day square age, and then sharply decreased three to five days before flowering. During square development, the effect of main‐stem node (MSN) and sympodial branch fruiting position in the plant canopy on mineral nutrient element concentrations of bracts was greater than on those of floral buds. Differences in the mineral nutrient element concentrations existed among the sympodial leaves, bracts, and floral buds. This study provides the patterns of mineral nutrient element concentrations in these plant tissues during the square development phase in relation to MSNs and branch fruiting positions in the cotton plant canopy.     

Partitioning of biomass in water‐ and nitrogen‐stressed cotton during pre‐bloom stage

The partitioning of biomass between aboveground parts and roots, and between vegetative and reproductive plant parts plays a major role in determining the ability of cotton (Gossypium hirsutum L.) to produce a crop in a given environment. We evaluated the single and combined effects of water and N supply on the partitioning of biomass in cotton plants exposed to two N supply levels, 0 and 12 mM of N, and two water regimes, well irrigated and water‐stressed at an early reproductive stage. The N treatments began when the third true leaf was visible, while water deficit treatments were imposed over the N treatments when the plants were transferred into controlled‐environment chambers at a leaf area near 0.05 m². Both water deficits and N deficits inhibited total biomass accumulation and its partitioning in cotton. Water deficit alone and N deficit alone inhibited the growth of leaves, petioles, and branches, but did not inhibit growth of the stem and enhanced the accumulation of biomass in squares. When water deficit was superimposed on N deficit, leaf growth was inhibited, although to a lesser extent than when it was the sole stress factor, and the accumulation of biomass in squares was also inhibited. Yet, the dry weight of squares in plants exposed to water and N deficits was greater than that of non‐stressed plants. Water and N deficits, either alone or in combination, did not inhibit the growth of the tap root. Growth of lateral roots was not inhibited either by water deficit alone or in combination with N deficit, but was enhanced when plants were exposed to N deficit alone. Exposure to water deficit alone or in combination with N deficit decreased the shoot:root ratio through the inhibition of shoot growth. Exposure to N deficit alone decreased the shoot:root ratio through the combination of shoot growth inhibition and root growth enhancement.     

干旱及复水条件下草甘膦对抗草甘膦大豆幼苗渗透调节物质和莽草酸含量的影响

为探明干旱胁迫及复水条件下不同剂量草甘膦对抗草甘膦大豆(RR1)幼苗叶片渗透调节物质、莽草酸(shikimic acid, SA)含量及根系活力的影响,采用盆栽试验,在大豆的第3复叶期进行水分胁迫5d和除草剂草甘膦处理,研究RR1幼苗叶片可溶性蛋白(soluble protein, SP)、可溶性糖(soluble sugar, SS)、游离脯氨酸(free praline, FP)、莽草酸(shikimic acid, SA)含量和根系活力(RA)的变化。结果表明,干旱胁迫前期RR1叶片的SP含量随草甘膦剂量的增加呈先升高后降低趋势,0.46kg/hm2叶片SP的含量最高,胁迫后期SP含量随草甘膦剂量的增加而降低;SS、FP和SA含量随草甘膦剂量的增加和胁迫时间的延长而增加,RA随草甘膦剂量的增加和胁迫时间的延长而降低。复水12d后,不同剂量草甘膦处理的各指标均有所恢复。干旱条件下,经草甘膦处理的RR1叶片的SP含量和RA低于草甘膦在正常水分条件下的处理,而SS、FP和SA含量相反。相关性分析表明,FP和SA含量与草甘膦剂量的相关关系最明显;而SS和SA含量与干旱胁迫时间的相关关系最明显。说明正常水分条件下,草甘膦对RR1幼苗造成的伤害经过一段时间后有所缓解;干旱胁迫加剧了草甘膦对RR1幼苗叶片渗透调节物质、莽草酸含量和根系活力的影响。抗草甘膦大豆主要通过积累FP、SS和SA对草甘膦和干旱胁迫做出响应。     

Nitrogen metabolism and seed composition as influenced by glyphosate application in glyphosate-resistant soybean

Previous research has demonstrated that glyphosate can affect nitrogen fixation or nitrogen assimilation in soybean. This 2-year field study investigated the effects of glyphosate application of 1.12 and 3.36 kg of ae ha(-1) on nitrogen metabolism and seed composition in glyphosate-resistant (GR) soybean. There was no effect of glyphosate application on nitrogen fixation as measured by acetylene reduction assay, soybean yield, or seed nitrogen content. However, there were significant effects of glyphosate application on nitrogen assimilation, as measured by in vivo nitrate reductase activity (NRA) in leaves, roots, and nodules, especially at high rate. Transiently lower leaf nitrogen or (15)N natural abundance in high glyphosate application soybean supports the inhibition of NRA. With the higher glyphosate application level protein was significantly higher (10.3%) in treated soybean compared to untreated soybean. Inversely, total oil and linolenic acid were lowest at the high glyphosate application rate, but oleic acid was greatest (22%) in treated soybean. These results suggest that nitrate assimilation in GR soybean was more affected than nitrogen fixation by glyphosate application and that glyphosate application may alter nitrogen and carbon metabolism.     

Shikimic acid accumulation in field-grown corn (Zea mays) following simulated glyphosate drift

Field studies were conducted in 2001 through 2003 to determine if shikimic acid accumulation could be used to accurately predict yield reductions in field corn exposed to sublethal rates of glyphosate. Glyphosate (0-0.32 kg ae/ha) was applied to corn at the V6 to V8 growth stage. Corn whorls were randomly collected up to 14 days after application (DAA), and shikimic acid accumulation in the whorls was determined using HPLC-UV. Maximum shikimic acid accumulation occurred 3-7 DAA in corn receiving 0.16 and 0.32 kg/ha. Shikimic acid accumulation 3, 5, and 7 DAA did correlate (r = 0.80-0.86) to yield losses from a sublethal application of glyphosate. Shikimic acid accumulation 3, 5, and 7 DAA was better correlated to visual injury at 14 DAA than to yield reductions. Visual injury ratings 14 DAA were a slightly better indicator of potential yield losses (r = 0.93) than shikimic acid accumulation in field-grown corn whorls (r = 0.8-0.86).     

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 annuus L.) 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.