Characterization and plant expression of a glyphosate-tolerant enolpyruvylshikimate phosphate synthase

BACKGROUND: Glyphosate tolerance is a dominant trait in modern biotech crops. RESULTS: A gene encoding a glyphosate-tolerant EPSP synthase (aroA(1398)) from bacterial strain ATX1398 was cloned and characterized. The protein is initiated at a GTG translational start codon to produce a protein that provides robust glyphosate resistance in Escherichia coli (Mig) Cast & Chalm. The aroA(1398) protein was expressed and purified from E. coli, and key kinetic values were determined (K(i) = 161 microM; K(m)(PEP) = 11.3 microM; k(cat) = 28.3 s(-1)). The full-length enzyme is 800-fold more resistant to glyphosate than the maize EPSP synthase while retaining high affinity for the substrate phosphoenol pyruvate. To evaluate further the potential of aroA(1398), transgenic maize events expressing the aroA(1398) protein were generated. T(0) plants were screened for tolerance to glyphosate sprays at 1.3x commercial spray rates, and T(1) plants were selected that completely resisted glyphosate sprays at 1x, 2x and 4x recommended spray rates in field trials. CONCLUSION: These data suggest that aroA(1398) is a suitable candidate for conferring glyphosate tolerance in transgenic crop plants.     

Isolation and biochemical diagnosis of cell lines of groundnut (Arachis hypogaea L) selected on glyphosate†

Three cell lines of groundnut (Arachis hypogaea L), an important oilseed legume, were selected on glyphosate using in-vitro culture techniques. The cell lines isolated through single as well as stepwise selection procedures showed c 20-fold increase in glyphosate tolerance as compared to the unselected control cell line. Studies on the biochemical mechanism of glyphosate tolerance in these cell lines showed a significant increase in the total extractable activity of the target enzyme, 5-enolpyruvyl shikimate-3-phosphate (EPSP) synthase (EC 2.5.1.19), which was further confirmed with immunological data. The over-expressed EPSP synthase activity was, however, subject to inhibition by glyphosate in vitro. Two other key regulated enzymes of the shikimic acid pathway, 3-deoxy-D -arabino heptulosonate 7-phosphate (DAHP) synthase (EC 4.1.2.15) and chorismate mutase (CM) (EC 5.4.99.5) did not show any change in specific activity in the selected cell lines. The enhanced activity of EPSP synthase in the tolerant cell lines was found to be stably inherited in the absence of selection pressure. © 1999 Society of Chemical Industry     

In vitro expression of variation of glyphosate tolerance in Sorghum halepense

The in vitro response of five different Sorghum halepense biotypes against the non-selective, broad-spectrum herbicide glyphosate was assessed. Seeds from donor plants (collected in various sites all over Greece) were aseptically germinated on a hormone-free liquid Murashige and Skoog (MS) medium and emerging plantlets were inoculated on a solid MS medium supplemented with 13.6 μM 2.4-dichlorophenoxyacetic acid and 4.6 μM kinetin for callus induction. Exponentially growing calli were initially subcultured twice on induction medium and then transferred to a selection medium containing 10⁻³ M or 10⁻⁴ M (a.i.) glyphosate. The fresh weight of the cultured calli and the callus viability (expressed as callus dehydrogenase activity) were reduced as glyphosate concentration increased. Significant differences were observed among different biotypes. Regenerated plantlets were submitted to a conventional evaluation for glyphosate tolerance. The observed in vitro response of S. halepense to glyphosate was directly related to the in vivo herbicide tolerance observed both on donor and on regenerant plants.     

Titanium dioxide photoinduced degradation of some pesticide/fungicide precursors

Five-membered nitrogen heterocycles (pyrrole, imidazole and 1,2,4-triazole) have been degraded using titanium dioxide and simulated solar radiation at pH = 8. The degradations followed a simple Langmuir-Hinshelwood mechanism. Accordingly, the adsorption equilibrium constants K of the heterocycles on the titanium dioxide surface and the rate constants k of degradation of the heterocycle-catalyst adduct have been obtained experimentally. While the K values decrease with heterocycle pKa, the k values increase with increasing pKa. Therefore, apparently, the rate constant depends on the availability of the electron pair on nitrogen, but at the same time the electron pair repulsion induced by the negatively charged titanium dioxide surface at pH = 8 causes a reverse effect in the adsorption equilibrium constant. Only in the case of imidazole, where the adsorption equilibrium constant is low enough (K = 0.013 M(-1)), can the rate constant be approximated to a pseudo-first-order rate expression: k(obs) = Kk. In all other cases, k(obs) = Kk/(1 + K(heterocycle)).     

耐草甘膦菜豆耐性酶学机理的研究

感性、耐性菜豆从萌芽到24d的EPSP合成酶比活性是不同的,但变化规律相似。出苗时比活性较高,10d时降至最低点,随后上升,16d时达到最高点,然后缓慢下降;两种菜豆在生长的同一天酶比活性差异小,最大倍数仅为1.1;用草甘膦处理两种三出复叶时的菜豆1d后酶比活性上升15%～25%。实验测定了经4步纯化,比活性达6219.4nmol· min-1· mg-1蛋白以上的两种菜豆EPSP合成酶的一些酶学性质。PEP是酶的最适底物,感性、耐性 Km(PEP)分别为3.5和7.1μmol· L-1,两者相差2倍,亲和力Ki(草甘膦)分别是6.2和 16.7μmol· L-1,两者相差2.7倍。两种菜豆对草甘膦耐性不同的酶学机理在于耐性菜豆的EPSP合成酶与草甘膦的亲和力小,与酶底物亲和力大;而感性菜豆EPSP酶则与草甘膦的亲和力大,与其底物亲和力小。     

Natural tolerance of Cuscuta campestris to herbicides inhibiting amino acid biosynthesis

The response of Cuscuta campestris Yuncker, a non‐specific above‐ground holoparasite, to amino acid biosynthesis inhibitor (AABI) herbicides, was compared with other resistant and sensitive plants in dose–response assays carried out in Petri dishes. Cuscuta campestris was found to be much more resistant to all AABI herbicides tested. The I₅₀ value of C. campestris growth inhibition by glyphosate was eightfold higher than that of transgenic, glyphosate‐resistant cotton (RR‐cotton). The I₅₀ value for C. campestris shoot growth inhibition by sulfometuron was above 500 μM, whereas that of sorghum roots was only 0.004 μM. Cuscuta campestris exposed to glyphosate gradually accumulated shikimate, confirming herbicide penetration into the parasite and interaction with an active form of the target enzyme of the herbicide, 5‐enolpyruvylshikimate‐3‐phosphate synthase. More than half of the C. campestris plants associated with transgenic, glyphosate‐resistant sugarbeet (RR‐sugarbeet) treated with glyphosate or with transgenic, sulfometuron‐resistant tomato (SuR‐tomato) treated with sulfometuron recovered and resumed regular growth 20–30 days after treatment. New healthy stems developed, followed by normal flowering and seed setting. The results of the current study demonstrate the unique capacity of C. campestris to tolerate high rates of AABI. The mechanism of this phenomenon is yet to be elucidated.     

EPSP 合成酶的特性及新抑制剂的研究进展

概述了EPSP 合成酶的生理作用。高等植物前体EPSP 合成酶进入叶绿体后, 经剪切而成成熟EPSP 合成酶并定位于叶绿体内质膜上; EPSP 合成酶的三维结构上存在三个活性区域, 改变活性区域的氨基酸残基可对草甘膦产生抗性, 同时表明了酶与草甘膦的结合位点; 不同生物的EPSP 合成酶有高的同源性。EPSP 合成酶催化底物反应机理为: PEP 首先与酶形成过渡态, 而后和S3P 形成缩酮四面体, 最后生成EPSP。草甘膦与EPSP 合成酶、EPSP形成三元复合物而阻断了EPSP 合成酶的催化作用; 以EPSP 和S3P ·glypho sate 为分子模型, 设计合成的化合物对EPSP 合成酶的活性有抑制作用。     

A mechanistic analysis of penetration of glyphosate salts across astomatous cuticular membranes

Penetration of glyphosate salts across isolated poplar (Populus canescens (Aiton) Sm) cuticular membranes (CM) was studied using Na+, K+, NH4+, trimethylsulfonium+ (TMS) and isopropylamine+ (IPA) as cations. After droplet drying, humidity over the salt residues on the outer surfaces of the CM was kept constant, and cuticular penetration was monitored by sampling the receiver solution facing the inner surfaces of the CM. Glyphosate salts disappeared exponentially with time from the surfaces of the CM. This first-order process could be quantitatively described using rate constants (k) or half-times (time for 50% penetration; t1/2). Humidity strongly affected the velocity of penetration, as k increased by factors of 5.3 (K-glyphosate), 6.9 (TMS-glyphosate), 7.1 (NH4-glyphosate), 8.5 (Na-glyphosate) and 10.5 (IPA-glyphosate) when humidity was increased from 70 to 100%. Depending on the type of cation and humidity, t1/2 varied between 4 and 70h, but the humidity effect was statistically significant only at 100% humidity, when half-times were highest with IPA-glyphosate and lowest with TMS-glyphosate. Glyphosate acid penetration was measured only at 90% humidity and found to be extremely slow (t1/2 = 866 h). Adding 0.2 g litre-1 of a wetter (alkylpolyglucoside) to the donor increased IPA-glyphosate rate constants by about four times, but increasing concentration produced no further increase in k. When donors contained 0.2 g litre-1 wetter, further additions of 4 g litre-1 Ethomeen T25 did not change rate constants measured with IPA-glyphosate at 90% humidity, while Genapol C-100 and diethyl suberate increased k by only 35%. Concentration of IPA-glyphosate (1, 2 and 4 g litre-1) did not influence k at 90% humidity, and pH of donor solutions (4.0, 7.7, 9.5) had no effect on k of K-glyphosate at 90% humidity. Temperature (10 to 25 degrees C) had only a small influence on velocity of penetration of IPA-glyphosate and K-glyphosate, as energies of activation amounted to only 4.26 and 2.92 kJ mole-1, respectively. These results are interpreted as evidence for penetration of glyphosate salts in aqueous pores.     

Expression of a wheat cytochrome P450 monooxygenase in yeast and its inhibition by glyphosate

Glyphosate is a non-selective herbicide which acts by inhibiting 5-enolpyruvylshikimate-3-phosphate synthase. Wheat cytochrome P450 monooxygenase specifically catalyzes the metabolism of some sulfonylurea herbicides such as chlorsulfuron and triasulfuron. Here we report that glyphosate is an inhibitor of a wheat cytochrome (CYP71C6v1), the cDNA of which was amplified by RT-PCR and heterologously expressed in yeast. The microsomal fractions derived from this strain had a Soret peak at 502 nm in the reduced carbon monoxide difference spectrum, which is a typical spectral characteristic. The addition of glyphosate to the microsomal protein resulted in a Type II spectrum indicative of binding via the nitrogen group to haem of cytochrome P450 as a sixth ligand. A spectral dissociation constant, K(s) of 70 micromol litre(-1) was observed and an IC50 of 11 micromol litre(-1) was found for glyphosate inhibition of CYP71C6v1 P450 activity.     

Effects of pesticides on 1,3-β-glucanase and urease activities in soil in the presence and absence of fertilisers,lime and organic materials

The effect of four herbicides (2, 4-D, di-allate, benzoylprop-ethyl and glyphosate) and one insecticide (malathion) on 1, 3-β-glucanase and urease activities in soil was investigated in the laboratory. Concentrations equivalent to five times the recommended field application rates of the pesticides, applied as their formulations, had no effect on the activity of either enzyme, under a variety of incubation conditions, which included various moisture regimes and soil treatments. The soil enzyme systems could only be disrupted by unrealistically high dosage rates of the active ingredients (0.1–1.0 g kg^?1 of soil), Thus 1, 3-β-glucanase activity was enhanced by 2, 4-D, inhibited by di-allate, benzoylprop-ethyl and malathion, but unaffected by glyphosate; whereas urease activity was inhibited by 2, 4-D, but unaffected by di-allate, benzoylprop-ethyl and glyphosate.     

Distribution of glyphosate and of its target enzyme inside wheat plants

Enolpyruvylshikimate phosphate (EPSP) synthase activity has been measured in different organs of 1–14-day-old wheat seedlings growing in culture chambers. Enzyme activity was first detected after two days, and reached its maximum value at day 4. It remained almost unchanged for 14 days thereafter. During germination, one of the most important sources of enzyme was the seed scutellum. Later, the developed leaves contained 43% of the whole stock of enzyme. Although the enzyme distribution was not identical under light or in the dark, there was no evident light induction of EPSP synthase. High enzyme activity was found in the coleoptile, continuing for the whole life-time of this organ, and which might be associated with its lignification. Five to 40% of the total enzyme was found in the roots, reaching a maximum at 14 days. At this stage, glyphosate applied to the first grown leaf was phloem-transported to the sink organs, with no clear relationship being shown between EPSP synthase activity and herbicide quantity in each organ. At day 4, glyphosate treatment, by soaking the seedlings in the herbicide solution, was followed by an active phloem transport from the scutellum to the growing parts. The average internal concentration of radiolabelled compounds (glyphosate plus its possible derivatives) which induced wheat seedling growth decrease, was shown to reach 50 to 100 μM under our conditions.     

Glyphosate: a once-in-a-century herbicide

Since its commercial introduction in 1974, glyphosate [N-(phosphonomethyl)glycine] has become the dominant herbicide worldwide. There are several reasons for its success. Glyphosate is a highly effective broad-spectrum herbicide, yet it is very toxicologically and environmentally safe. Glyphosate translocates well, and its action is slow enough to take advantage of this. Glyphosate is the only herbicide that targets 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPS), so there are no competing herbicide analogs or classes. Since glyphosate became a generic compound, its cost has dropped dramatically. Perhaps the most important aspect of the success of glyphosate has been the introduction of transgenic, glyphosate-resistant crops in 1996. Almost 90% of all transgenic crops grown worldwide are glyphosate resistant, and the adoption of these crops is increasing at a steady pace. Glyphosate/glyphosate-resistant crop weed management offers significant environmental and other benefits over the technologies that it replaces. The use of this virtually ideal herbicide is now being threatened by the evolution of glyphosate-resistant weeds. Adoption of resistance management practices will be required to maintain the benefits of glyphosate technologies for future generations.     

Effect of freezing and thawing on microbial activity and glyphosate degradation in two Norwegian soils

Little research has been done on pesticide dissipation in cold climates and there is a need to focus on the influence of climate on pesticide degradation in soil. Glyphosate, N-(phosphonomethyl)glycine, is a herbicide frequently used for controlling perennial weeds through application after harvest and was used as a model compound for this study. The effect of freeze-thaw activity on the availability of glyphosate in soil, and consequently its mineralization by soil microorganisms, was studied through laboratory incubations of repacked soil cores treated with 14C-labelled glyphosate and subjected to different freeze-thaw treatments. Winter simulation regimes applied were constant thaw (+5 degrees C), constant freezing (-5 degrees C), unstable conditions with short fluctuations (24 h of -5 degrees C followed by 24 h of +5 degrees C), and long duration fluctuations (3 weeks of -5 degrees C followed by 3 weeks of +5 degrees C). Distribution of 14C-glyphosate was followed during the incubation through measurements of the mineralized fraction (14CO2), soil water fraction, KOH extractable fraction, and non-extractable fraction. Microbial parameters used to characterize the soils were estimates of size of microbial biomass, overall microbial activity and microbial diversity. The constant freezing treatment exhibited the lowest amount of glyphosate mineralization. The constant thawed treatment and the treatments with fluctuating temperature exhibited significantly increased mineralization. These results were in accordance with the observed concentration of glyphosate in soil water; the higher the activity, the lower the concentration. The amount of glyphosate extractable with KOH and the resulting non-extractable fraction, however, were not significantly affected by soil type or temperature regime. The glyphosate mineralization pattern was comparable with the overall microbial activity in the soils. Observed different levels of diversity might explain some of the difference in total glyphosate mineralization between soils.     

Differential response of Amaranthus tuberculatus (Moq ex DC) JD Sauer to glyphosate

Midwest USA farmers have reported inconsistent control of Amaranthus tuberculatus (= rudis) (Moq ex DC) JD Sauer by glyphosate in glyphosate-resistant crops. The potential of selection for decreased A tuberculatus sensitivity to glyphosate was therefore investigated in a reportedly resistant Everly, IA population (P0-EV). Evaluation of six A tuberculatus populations from the Midwest USA estimated a seedling baseline sensitivity of 2.15 mM glyphosate. Based on these results, three generations of divergent recurrent selection were implemented on P0-EV to isolate resistant and susceptible populations. A seedling assay was developed to screen large amounts of seeds and thus expedite the selection process. Whole-plant and seedling rate responses of P0-EV and a known pristine A tuberculatus population from Paint Creek, OH (P0-WT) identified no significant difference in response to glyphosate; however, greater phenotypic variance was ostensibly evident in P0-EV. The first recurrent generation selected for resistance at 3.2 mM glyphosate (RS1-R) had a 5.9- and 1.7-fold resistance increase at the seedling and whole-plant levels, respectively, compared with the susceptible generation selected at 32 microM glyphosate. After three cycles of recurrent selection, 14.6-fold difference in resistance at the seedling level and 3.1-fold difference at the whole-plant level were observed when comparing the populations selected for resistance (RS3-R) and susceptibility (RS3-S). Overall, recurrent selection increased the frequency of resistant individuals and decreased the variability to glyphosate at the population level. Nevertheless, variability for glyphosate resistance was still evident in RS3-R. Results herein suggested that A tuberculatus is inherently variable to glyphosate and that selection decreased the sensitivity to glyphosate. We purport that evolved glyphosate resistance in A tuberculatus may require multiple cycles of selection under field conditions. Historic estimated use of glyphosate alludes to the evolution of tolerant weed populations.     

土壤高锰对空心莲子草生长与草甘膦耐性的影响

研究了空心莲子草Alternanthera philoxeroides在土壤中含高浓度锰(简称高锰)胁迫下的生长特性及对草甘膦的耐性。空心莲子草在锰添加量为8～128 mg/kg的土壤中培养20 d后,分枝数比对照(4.43个/株)增加了22.57％～48.31%, 主茎长度比对照(23.85 cm)增长了16.60％～19.87%,地下根茎数比对照(1.54个/株)增加29.87％～47.40%,全株干重比对照(0.23 g)增加22.45%～27.00%;植株叶面积在锰添加量为32～128 mg/kg范围内也显著高于对照(增加10.53%～11.10%)。用草甘膦(有效成分按草甘膦酸计) 68、102和136 g/hm2分别进行茎叶处理后21 d,锰添加量为128 mg/kg的处理,鲜重抑制率分别比不添加锰处理组下降27.39％、24.57％和30.06％。结果表明,土壤高锰条件下空心莲子草的生长和繁殖能力增强,这可能是土壤酸化背景下空心莲子草蔓延加剧的重要原因。土壤高锰条件下生长的空心莲子草对草甘膦的耐性显著增强,使得该杂草的防控形势更为严峻。     

The determination of glyphosate in soils with moderate to high clay content

An improved method for the quantitative analysis of the herbicide glyphosate [N-(phosphonomethyl)glycine] in soils containing moderate to high clay content is described. Critical evaluations of previously published methods have indicated that recoveries of glyphosate from soils with high clay content are often low. Where acceptable recovery estimates have been reported, these methods also report increased interferences and rarely include soils with clay content exceeding 30%. The proposed method was developed and characterized using six soils of different clay content (25–87% clay), with other physical and chemical properties as described. Recoveries of glyphosate from the soils were determined after duplicate extractions with 0.1 M potassium hydroxide. Clean-up of soil extract solutions was by cation-exchange column chromatography. Subsequent quantitation was by HPLC with post-column oxidation, followed by derivatization using OPA-MERC with fluorometric detection. No interferences were detected. Recovery estimates for each fortified sample were determined over a concentration range (0.56–11.25 mg glyphosate kg ?1 ) with all recoveries being greater than 80%. Detection limit for glyphosate in soil was 0.04 mg kg ?1, and instrument response was linear for solutions up to 50.0μg glyphosate ml ?1. Reproducibility relative confidence interval, for a single sample analysis, was determined as.     

有机硅喷雾助剂对草甘膦在空心莲子草上的沉积和生物活性的影响

研究了有机硅喷雾助剂(OSA)对草甘膦在空心莲子草Alternanthera philoxeroides上的沉 积与生物活性的影响。当采用较大喷雾雾滴,施药液量高于632.5 L/hm2时,添加OSA(0.35 g/L) 后,草甘膦药液在空心莲子草上的沉积量显著下降。最大稳定持留量(MRG)由未添加OSA的0.61~0.63 μ L/cm2下降到0.50~0.54 μ L/cm2。分别以33.7和67.4 μ g/株剂量的草甘膦点叶处理空心莲子草,发现添加OSA的处理对再生植株茎叶生长的抑制率分别比对照提高了8.89%和14.83%。草甘膦(有效成分199.3 g/hm2)施药后1 h进行人工模拟降雨处理,添加OSA后药剂对空心莲子草的生物活性比无OSA对照处理提高了20.5%。研究结果表明,添加有机硅喷雾助剂促进了草甘膦在空心莲子草中的向下传导性能,提高了草甘膦水剂在空心莲子草叶片的耐雨水冲刷性能,但会降低草甘膦药液在空心莲子草上的最大稳定持留量。     

Physiological effects of glyphosate over amino acid profile in conventional and transgenic soybean (Glycine max)

This paper compares the responses of conventional and transgenic soybean to glyphosate application in terms of the contents of 17 detectable soluble amino acids in leaves, analyzed by HPLC and fluorescence detection. Glutamate, histidine, asparagine, arginine + alanine, glycine + threonine and isoleucine increased in conventional soybean leaves when compared to transgenic soybean leaves, whereas for other amino acids, no significant differences were recorded. Univariate analysis allowed us to make an approximate differentiation between conventional and transgenic lines, observing the changes of some variables by glyphosate application. In addition, by means of the multivariate analysis, using principal components analysis (PCA), cluster analysis (CA) and linear discriminant analysis (LDA) it was possible to identify and discriminate different groups based on the soybean genetic origin.     

草甘膦铵盐对3种滩涂动物抗氧化酶、丙二醛及大弹涂鱼红细胞微核的影响

为评价草甘膦对河口及滩涂水域生物的影响,采用室内人工染毒法,研究了草甘膦铵盐对可口革囊星虫Phascolosoma esculenta、缢蛏Sinonovacula constricta Lamark和大弹涂鱼Boleophthalmus pectinirostris 3种滩涂动物超氧化物歧化酶 (SOD)、过氧化氢酶 (CAT)、丙二醛 (MDA) 及大弹涂鱼红细胞微核的影响。结果表明:经低浓度 (63.5 mg/L) 草甘膦短时间 (2 d) 染毒后,可口革囊星虫肠组织的SOD、CAT活性值及MDA含量分别为231.54、1.719 U/mg pro. 和0.60 nmol/mg pro.,3种指标均显著或极显著高于相应对照值 (204.88、1.521 U/mg pro. 和0.54 nmol/mg pro.),且随着药剂处理浓度增加,SOD、CAT活性及MDA含量均升高;但当染毒时间延长至8 d时,可口革囊星虫肠组织的SOD、CAT活性值及MDA含量均明显降低,分别为155.67、1.403 U/mg pro. 和0.53 nmol/mg pro.,显著低于对照,且随药剂处理浓度进一步增加,酶活性及MDA含量值降低更显著。草甘膦对缢蛏内脏团及大弹涂鱼肝脏SOD、CAT活性及MDA含量的影响表现出类似规律。此外,经低浓度 (53.5 mg/L) 草甘膦染毒8 d后,大弹涂鱼红细胞微核率及核异常率分别为17.00‰ 和21.33‰,均极显著高于相应对照值 (13.67‰ 和13.33‰),且随着草甘膦浓度增大,其红细胞微核率及核异常率均显著升高,表明大弹涂鱼对草甘膦有较高的敏感性。所得研究结果可为评价草甘膦对河口及滩涂水域生物的影响提供参考依据。     

高锰胁迫下草甘膦对空心莲子草体内莽草酸含量和抗氧化酶活性的影响

鉴于高锰胁迫下空心莲子草Alternanthera philoxeroides对草甘膦的耐药性增强,在水培条件下研究了不同浓度锰条件下草甘膦处理后该草体内莽草酸的积累和主要抗氧化酶系统的响应。次高浓度锰(0.31 mmol/L)条件下培养120 d后空心莲子草体内过氧化氢酶(CAT)活性显著高于常规浓度锰处理(0.009 1 mmol/L,对照);高浓度锰(2.45 mmol/L)条件下超氧化物歧化酶(SOD)活性升高,CAT活性下降。草甘膦(按草甘膦酸68 g/hm2)茎叶处理后6 d内,常规锰浓度培养的空心莲子草体内莽草酸含量比用草甘膦刚处理时增加了31.9％~226.0%,且显著高于同一时间次高锰和高锰的处理;不同锰浓度下培养的空心莲子草体内过氧化物酶(POD)和CAT、SOD活性均为先升高后再逐渐下降,但次高锰处理的该3种酶活性均高于对照,高锰处理的SOD和POD活性高于对照,而CAT活性与对照相当。上述结果表明,在较高锰浓度下空心莲子草能启动抗氧化酶系统而能有效地清除自由基;在草甘膦处理后初期,高锰条件下空心莲子草体内莽草酸途径受抑制程度较轻,抗氧化酶活性较高,这可能是空心莲子草耐高锰和高锰条件下该草耐草甘膦的部分机制。