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.     

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.     

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.     

Tolerance and accumulation of shikimic acid in response to glyphosate applications in glyphosate-resistant and nonglyphosate-resistant cotton (Gossypium hirsutum L.).

Measurement of shikimic acid accumulation in response to glyphosate inhibition of 5-enolpyruvylshikimate-3-phosphate synthase is a rapid and accurate assay to quantify glyphosate-induced damage in sensitive plants. Two methods of assaying shikimic acid, a spectrophotometric and a high-performance liquid chromatography (HPLC) method, were compared for their accuracy of recovering known amounts of shikimic acid spiked into plant samples. The HPLC method recovered essentially 100% of shikimic acid as compared with only 73% using the spectrophotometric method. Relative sensitivity to glyphosate was measured in glyphosate-resistant (GR) and non-GR cotton leaves, fruiting branches, and squares (floral buds) by assaying shikimic acid. Accumulation of shikimic acid was not observed in any tissue, either GR or non-GR, at rates of 5 mM glyphosate or less applied to leaves. All tissues of non-GR plants accumulated shikimic acid in response to glyphosate treatment; however, only fruiting branches and squares of GR plants accumulated a slight amount of shikimic acid. In non-GR cotton, fruiting branches and squares accumulated 18 and 11 times, respectively, more shikimic acid per micromolar of translocated glyphosate than leaf tissue, suggesting increased sensitivity to glyphosate of reproductive tissue over vegetative tissue. GR cotton leaves treated with 80 mM of glyphosate accumulated 57 times less shikimic acid per micromolar of translocated glyphosate than non-GR cotton but only 12.4- and 4-fold less in fruiting branches and squares, respectively. The increased sensitivity of reproductive structures to glyphosate inhibition may be due to a higher demand for shikimate pathway products and may provide an explanation for reports of fruit abortion from glyphosate-treated GR cotton.     

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.     

AMINO ACID APPLICATION CAN BE AN ALTERNATIVE TO PREVENT GLYPHOSATE INJURY IN GLYPHOSATE-RESISTANT SOYBEANS

Glyphosate-resistant (GR) soybeans have continuously increased; however, this expansion significantly increased the use of glyphosate and therefore, in some cases, has resulted in injury symptoms observed in GR soybean, known as “yellow flashing”. Previous reports of interference of glyphosate with nutrient availability and utilization by GR soybean may be linked to this injury symptom. Also, because glyphosate interferes with amino acid synthesis, supplementation with exogenous amino acids may help GR soybean recover from adverse effects of glyphosate. Therefore, an experiment was designed to evaluate different amino acid concentrations. Near-isogenic and GR soybean varieties were grown in the greenhouse in two soils with and without glyphosate at different rates and amino acids were foliarly applied with and without glyphosate. In general, the photosynthetic variables, nutrient contents, and shoot and root dry biomass parameters were affected by glyphosate, however, use of amino acid formulations suppressed harmful effects of glyphosate on these parameters.     

Glyphosate effects on photosynthesis,nutrient accumulation,and nodulation in glyphosate‐resistant soybean

Previous greenhouse studies have demonstrated that photosynthesis in some cultivars of first‐ (GR1) and second‐generation (GR2) glyphosate‐resistant soybean was reduced by glyphosate. The reduction in photosynthesis that resulted from glyphosate might affect nutrient uptake and lead to lower plant biomass production and ultimately reduced grain yield. Therefore, a field study was conducted to determine if glyphosate‐induced damage to soybean (Glycine max L. Merr. cv. Asgrow AG3539) plants observed under controlled greenhouse conditions might occur in the field environment. The present study evaluated photosynthetic rate, nutrient accumulation, nodulation, and biomass production of GR2 soybean receiving different rates of glyphosate (0, 800, 1200, 2400 g a.e. ha^–1) applied at V2, V4, and V6 growth stages. In general, plant damage observed in the field study was similar to that in previous greenhouse studies. Increasing glyphosate rates and applications at later growth stages decreased nutrient accumulation, nodulation, leaf area, and shoot biomass production. Thus, to reduce potential undesirable effects of glyphosate on plant growth, application of the lowest glyphosate rate for weed‐control efficacy at early growth stages (V2 to V4) is suggested as an advantageous practice within current weed control in GR soybean for optimal crop productivity.     

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.     

Isoflavone,glyphosate, and aminomethylphosphonic acid levels in seeds of glyphosate-treated,glyphosate-resistant soybean

The estrogenic isoflavones of soybeans and their glycosides are products of the shikimate pathway, the target pathway of glyphosate. This study tested the hypothesis that nonphytotoxic levels of glyphosate and other herbicides known to affect phenolic compound biosynthesis might influence levels of these nutraceutical compounds in glyphosate-resistant soybeans. The effects of glyphosate and other herbicides were determined on estrogenic isoflavones and shikimate in glyphosate-resistant soybeans from identical experiments conducted on different cultivars in Mississippi and Missouri. Four commonly used herbicide treatments were compared to a hand-weeded control. The herbicide treatments were (1) glyphosate at 1260 g/ha at 3 weeks after planting (WAP), followed by glyphosate at 840 g/ha at 6 WAP; (2) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied preemergence (PRE), followed by glyphosate at 1260 g/ha at 6 WAP; (3) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied PRE, followed by glyphosate at 1260 g/ha at full bloom; and (4) sulfentrazone at 168 g/ha plus chlorimuron at 34 g/ha applied PRE, followed by acifluorfen at 280 g/ha plus bentazon at 560 g/ha plus clethodim at 140 g/ha at 6 WAP. Soybeans were harvested at maturity, and seeds were analyzed for daidzein, daidzin, genistein, genistin, glycitin, glycitein, shikimate, glyphosate, and the glyphosate degradation product, aminomethylphosphonic acid (AMPA). There were no remarkable effects of any treatment on the contents of any of the biosynthetic compounds in soybean seed from either test site, indicating that early and later season applications of glyphosate have no effects on phytoestrogen levels in glyphosate-resistant soybeans. Glyphosate and AMPA residues were higher in seeds from treatment 3 than from the other two treatments in which glyphosate was used earlier. Intermediate levels were found in treatments 1 and 2. Low levels of glyphosate and AMPA were found in treatment 4 and a hand-weeded control, apparently due to herbicide drift.     

NUTRIENT ACCUMULATION AND PHOTOSYNTHESIS IN GLYPHOSATE-RESISTANT SOYBEANS IS REDUCED UNDER GLYPHOSATE USE

Global production of glyphosate-resistant (GR) soybean [Glycine max (L.) Merr.] continues to increase annually; however, there are no particular specific fertilizer recommendations for the transgenic varieties used in this system largely because reports of glyphosate effects on mineral nutrition of GR soybeans are lacking. Several metabolites or degradation products of glyphosate have been identified or postulated to cause undesirable effects on GR soybeans. In this work we used increasing glyphosate rates in different application on cv. ‘BRS 242 GR’ in order to evaluate photosynthetic parameters, macro- and micronutrient uptake and accumulation and shoot and root dry biomass production. Increasing glyphosate rates revealed a significant decrease in photosynthesis, macro and micronutrients accumulation in leaf tissues and also decreases in nutrient uptake. The reduced biomass in GR soybeans represents additive effects from the decreased photosynthetic parameters as well as lower availability of nutrients in tissues of the glyphosate treated plants.     

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。     

Shikimate accumulates in both glyphosate-sensitive and glyphosate-resistant horseweed (Conyza canadensis L. Cronq.)

Horseweed (Conyza canadensis) is a cosmopolitan weed that commonly grows throughout North America. Horseweed that is not completely controlled by normal applications of glyphosate has been reported in western Tennessee. This research had three objectives: (1) to develop and validate an analytical procedure for the quantitative determination of shikimate, an important indicator of glyphosate activity in plants; (2) to confirm resistance to glyphosate in a horseweed population; and (3) to examine the accumulation of shikimate in both glyphosate-resistant and glyphosate-susceptible horseweed plants. The analytical procedure to determine shikimate used extraction with 1 M HCl for 24 h, followed by liquid chromatography using photodiode array detection, and shikimate recoveries were >or=82%. Glyphosate applications of both 0.84 kg ae/ha (the standard application rate) and 3.8 kg ae/ha to susceptible plants caused complete plant death. The same glyphosate applications to putative resistant populations caused less than 15% growth reduction as determined by visual evaluations, and fresh weights of these resistant plants 17 days after glyphosate treatment (DAT) were reduced an average of 45% in one population and were not affected in a different population. This direct comparison conclusively confirms that horseweed plants collected in western Tennessee in 2002 are resistant to 4 times the normal application dosage of glyphosate. The glyphosate-resistant horseweed biotypes still exhibited some herbicidal effects from the glyphosate, such as yellowing in the most actively growing, apical shoot meristems. The yellowing in the shoot apexes was transitory, and the plants recovered from this damage. Shikimate concentrations in all untreated horseweed plants were less than 100 microg/g, which was significantly less than that in all plants which had been treated with 0.84 kg ae/ha of glyphosate. Unexpectedly, shikimate accumulated (>1000 microg/g) in both resistant populations and the susceptible population. However, there were differences in shikimate accumulation patterns between resistant and susceptible horseweed biotypes. Shikimate concentrations in resistant populations declined about 40% from 2 to 4 DAT, while shikimate concentrations in the susceptible horseweed plants increased about 35% from 2 to 4 DAT. The confirmed resistance of a widespread weed implies that alternative control strategies for glyphosate-resistant horseweed will be needed in those no-tillage production systems where it commonly occurs.     

Nitrogen Fixation,Ureide, and Nitrate Accumulation Responses to Soybean Aphid Injury in Glycine max

There is little information available about soybean aphid (Aphis glycines Matsumura) effects on the physiology and mineral nutrition of soybean (Glycine max [L.] merr.). Controlled-environment studies were conducted to measure soybean aphid infestation effects on dry weight, nitrogen (N) fixation, ureide-N, and nitrate-N concentration and accumulation. Plants grown in perlite using –N nutrient solution culture were infested at the 3rd trifoliolate (V3) stage and measured for N fixation, nodule characteristics, and ureide-N concentration at the full pod (R4) stage. When compared to uninfested control plants, aphid infestation reduced total nodule volume per plant by 34%, nodule leghemoglobin per plant by 31%, plant N fixation rate by 80% and shoot ureide-N concentration by 20%. Soil-grown plants were infested at the first trifoliolate (V1) stage and shoots were measured for dry weight, nitrate-N, and ureide-N at the full bloom (R2) stage. Infestation reduced shoot dry weight by 63%, increased nitrate-N concentration by 75%, but did not significantly affect ureide-N concentration. Because nutrient concentration is a single-point measurement that results from the integration of two dynamic processes, nutrient accumulation and dry matter production, we conclude that aphid-induced reductions in N fixation, coupled with decreased dry weight accumulation, caused shoot ureide-N concentration to remain unchanged in aphid-injured plants when compared to uninfested plants. Because nitrate-N concentration was greater in aphid-damaged shoot tissue, we further conclude that nitrate-N accumulation was less sensitive to aphid injury than dry weight accumulation.     

Microbial activity,community structure and potassium dynamics in rhizosphere soil of soybean plants treated with glyphosate

With the advent of glyphosate [N-(phosphonomethyl)glycine] tolerant crops, soils have now been receiving repeated applications of the herbicide for over 10 years in the Midwestern USA. There is evidence that long-term use of glyphosate can cause micronutrient deficiency but little is known about plant potassium (K) uptake interactions with glyphosate. The repeated use of glyphosate may create a selection pressure in soil microbial communities that could affect soil K dynamics and ultimately K availability for crops. Therefore, the objectives of this study were to characterize the effect of foliar glyphosate applied to GR (glyphosate resistant) soybeans on: (1) rhizosphere microbial community profiles using ester linked fatty acid methyl ester (EL-FAME) biomarkers, (2) exchangeable, non-exchangeable, and microbial K in the rhizosphere soil, and (3) concentrations of soybean leaf K. A greenhouse study was conducted in a 2 × 2 × 3 factorial design with two soil treatments (with or without long-term field applications of glyphosate), two plant treatments (presence and absence of soybean plants), and three rates of glyphosate treatments (0×, 1× at 0.87, and 2× at 1.74 kg ae ha^?1, the recommended field rate). After each glyphosate application, rhizosphere soils were sampled and analyzed for microbial community structure using ester linked fatty acid methyl ester biomarkers (EL-FAME), and exchangeable, plant tissue and microbial biomass K. Glyphosate application caused a significant decrease in the total microbial biomass in soybean rhizosphere soil that had no previous exposure to glyphosate, at 7 days after glyphosate application. However, no significant changes were observed in the overall microbial community structure. In conclusion, the glyphosate application lowered the total microbial biomass in the GR soybean rhizosphere soil that had no previous exposure to glyphosate, at 7 days after glyphosate application; caused no changes in the microbial community structure; and did not reduce the plant available K (soil exchangeable or plant tissue K).     

Simultaneous quantification of glyphosate, glufosinate, and their major metabolites in rice and soybean sprouts by gas chromatography with pulsed flame photometric detector

Procedures were developed for the simultaneous determination of glyphosate [N-(phosphonomethyl)glycine] and glufosinate [dl-homoalanin-4-yl-(methyl)phosphinic acid] and their major metabolites, aminomethylphosphonic acid (AMPA) and 3-(methylphosphinico)propionic acid (3-MPPA), in rice and soybean sprouts by gas chromatography (GC) equipped with a pulsed flame photometric detector (PFPD). Herbicides and their major metabolites were previously derivatized with TMOA (trimethyl orthoacetate (TMOA) in the presence of acetic acid, and their GC responses versus heating temperature (70-90 degrees C) and heating time (30-120 min) were optimized. It was found that increases in heating temperature and heating time were unfavorable for the derivatization of glyphosate or glufosinate, whereas high temperature and extended reaction time remarkably facilitated that of AMPA and 3-MPPA except at 90 degrees C for an extended reaction time (120 min). Combination of AG1-X8 anion-exchange chromatography with a Florisil cartridge cleanup process was favorable for the GC-PFPD analysis. Four types of derivatives spiked in rice and soybean sprout matrices were eluted, reaching a baseline separation, in a sequence of 3-MPPA, AMPA, glyphosate, and glufosinate within 14 min using a DB-608 capillary column. Recoveries of glyphosate, AMPA, glufosinate, and 3-MPPA (0.5 ppm) spiked in both sample matrices were determined to be 72-81, 71-86, 101-119, and 83-90%, respectively, whereas the coefficient of variation was determined to be <10% in three repeated determinations. The instrumental limits of detection for glyphosate, AMPA, glufosinate, and 3-MPPA in sample matrices were 0.02, 0.03, 0.02, and 0.01 ppm, respectively. The limits of quantification for glyphosate, AMPA, glufosinate, and 3-MPPA in sample matrices were 0.06, 0.10, 0.06, and 0.04 ppm, respectively.     

Response of selected horseweed (Conyza canadensis (L.) Cronq.) populations to glyphosate

Horseweed (Conyza canadensis (L.) Cronq.) seed was collected in Illinois, Indiana, Kentucky, Mississippi, Missouri, and Ohio to determine susceptibility of different horseweed biotypes to glyphosate. Horseweed resistant to glyphosate was found in Mississippi, Ohio, and western Tennessee. In a separate experiment examining Tennessee biotypes, a dose response curve demonstrated that four times as much glyphosate was needed to achieve a 50% fresh weight reduction (GR(50)) in resistant biotypes when compared to a susceptible biotype. Resistant biotypes from Tennessee displayed a GR(50) of 1.6 kg/ha as compared to a GR(50) of 0.4 kg/ha in a susceptible horseweed population. Although growth was reduced, the resistant plants did not completely die and could potentially produce seed. Variation in glyphosate resistance was found among the populations tested.     

Quantitative expression analysis of GH3, a gene induced by plant growth regulator herbicides in soybean

Symptoms resembling off-target plant growth regulator (PGR) herbicide injury are frequently found in soybean fields, but the causal agent is often difficult to identify. The expression of GH3, an auxin-regulated soybean gene, was quantified from soybean leaves injured by PGR herbicides using real-time RT-PCR. Expression of GH3 was analyzed to ascertain its suitability for use in a diagnostic assay to determine whether PGR herbicides are the cause of injury. GH3 was highly induced by dicamba within 3 days after treatment (DAT) and remained high at 7 DAT, but induction was much lower at 17 DAT. GH3 was also highly induced at 7 DAT by dicamba + diflufenzopyr, and to a lesser extent by the other PGR herbicides clopyralid and 2,4-D. The non-PGR herbicides glyphosate, imazethapyr, and fomesafen did not significantly induce GH3 expression above a low constitutive level. These results indicate that a diagnostic assay for PGR herbicide injury based on overexpression of auxin-responsive genes is feasible, and that GH3 is a potential candidate from which a diagnostic assay could be developed. However, time course analysis of GH3 expression indicates the assay would be effective for a limited time after exposure to the herbicide.     

Interaction of Glyphosate with Zinc for the Yield,Photosynthesis, Soil Fertility and Nutritional Status of Soybean

Application of glyphosate herbicide in genetically modified (GM) soybean [Glycine max (L.) Merrill] in soils with low zinc (Zn) concentration may interfere in the uptake of this and other nutrients, with negative impact on productivity. Thus, an experiment was conducted in greenhouse conditions on Ustoxix Quatzipsamment soil to investigate the effects of the interaction of glyphosate with Zn for the yield, photosynthesis, soil fertility and nutritional status of soybean. The treatments consisted of two soybean varieties [BRS 133 (conventional—NGM) and its essentially derived transgenic line BRS 245RR (GM) with and without glyphosate application] and five Zn rates (0, 5, 10, 20 and 40 mg kg^?1, source zinc sulfate (ZnSO₄)), with four replicates. Except for the copper (Cu) and iron (Fe) concentrations, the introduction of the herbicide-resistant gene is the predominant factor reducing nutrient uptake, photosynthetic (A) rate, stomatal conductance (Gs), leaf chlorophyll and ureide concentrations. The administration of Zn rates lowered the leaf phosphorus (P) concentration, and there was significant increase in Zn concentration in the soil and in the plant. Except for the 20 mg kg^?1 of Zn rate, the use of the herbicide did not affect the shoot dry weight (SDW) and seed yield, and on average, the maximum seed yield was obtained with Zn concentrations of 26.4 and 18.7 mg kg^?1 extracted by Mehlich 1 and diethylenetriaminepentaacetic acid-triethanolamine (DTPA-TEA), respectively.     

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。     

Physiological changes in soybean treated with ozone and molybdenum

Abstract

An open‐top field chamber experiment was conducted to evaluate the impact of Molybdenum (Mo) addition to soil on the physiological changes in soybean (Glycine max L. Merrill) exposed to ozone (O₃). Plants grown with Mo (0, 1.0, or 2.0 mg kg"¹ soil dry weight) were exposed to O₃ (O, 0.06, or 0.12 μmol mol^‐1) in open‐top field chambers for 12 h d^‐1 for 21 d with a N‐free fertilizer, during the sensitive growth stage (R2). The rate of photosynthesis (PN), specific root nodule nitrogenase activity (SNA), leaf nitrogen (N), chlorophyll (chl‐a, chl‐b) and biomass of soybean were measured. The increase in O₃ levels significantly reduced PN, SNA, leaf‐N, chl‐a, chl‐b, and biomass. Addition of Mo increased leaf‐N, shoot, root, and nodule dry weights but did not change PN, SNA, or chlorophyll. The addition of Mo (2 mg kg ^‐1) helped in significantly increasing PN and chlorophyll in the presence of 0.06 umol mol^‐1 O₃ but no change was observed in the presence of 0.12 μmol mol^‐1 O₃.