Alterations in glutathione pool and some related enzymes in leaves and roots of pea plants treated with the herbicide glyphosate

Our previous studies have demonstrated that application of glyphosate caused oxidative events in young pea and wheat plants. In this work, the changes in the endogenous level of glutathione (total and oxidized) and the activities of glutathione reductase (GR) and glutathione S-transferase (GST) after treatment with glyphosate were studied in pea plants (Pisum sativum L., cv. Skinado). Glyphosate was applied in two ways: (1) by leaf spraying with 10 mM solution; and (2) in nutrient medium as 0.01 mM solution. Measurements were made in both leaves and roots. Root and leaf treatments provoked the increase in both total and oxidized glutathione contents. Both types of herbicide application caused activation of GR in treated organs. Slight increase was detected also in untreated roots. It was found that glyphosate application to leaves provoked strong enhancement in the GST activity in leaves, while its root application stimulated the enzyme activity in the roots. We observed the higher GST activity in the organ directly treated with herbicide. Furthermore, we suggested that the activated isoforms of GST(s) participated in detoxification of hydrogen peroxide and lipid peroxides.     

Morphological and Physiological Responses of Cotton (Gossypium hirsutum L.) Plants to Salinity

Salinization usually plays a primary role in soil degradation, which consequently reduces agricultural productivity. In this study, the effects of salinity on growth parameters, ion, chlorophyll, and proline content, photosynthesis, antioxidant enzyme activities, and lipid peroxidation of two cotton cultivars, [CCRI-79 (salt tolerant) and Simian 3 (salt sensitive)], were evaluated. Salinity was investigated at 0 mM, 80 mM, 160 mM, and 240 mM NaCl for 7 days. Salinity induced morphological and physiological changes, including a reduction in the dry weight of leaves and roots, root length, root volume, average root diameter, chlorophyll and proline contents, net photosynthesis and stomatal conductance. In addition, salinity caused ion imbalance in plants as shown by higher Na⁺ and Cl⁻ contents and lower K⁺, Ca²⁺, and Mg²⁺ concentrations. Ion imbalance was more pronounced in CCRI-79 than in Simian3. In the leaves and roots of the salt-tolerant cultivar CCRI-79, increasing levels of salinity increased the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR), but reduced catalase (CAT) activity. The activities of SOD, CAT, APX, and GR in the leaves and roots of CCRI-79 were higher than those in Simian 3. CAT and APX showed the greatest H₂O₂ scavenging activity in both leaves and roots. Moreover, CAT and APX activities in conjunction with SOD seem to play an essential protective role in the scavenging process. These results indicate that CCRI-79 has a more effective protection mechanism and mitigated oxidative stress and lipid peroxidation by maintaining higher antioxidant activities than those in Simian 3. Overall, the chlorophyll a, chlorophyll b, and Chl (a+b) contents, net photosynthetic rate and stomatal conductance, SOD, CAT, APX, and GR activities showed the most significant variation between the two cotton cultivars.     

Glyphosate affects micro-organisms in rhizospheres of glyphosate-resistant soybeans

Aims: Glyphosate‐resistant (GR) soybean production increases each year because of the efficacy of glyphosate for weed management. A new or ‘second’ generation of GR soybean (GR2) is now commercially available for farmers that is being promoted as higher yielding relative to the previous, ‘first generation’ (GR1) cultivars. Recent reports show that glyphosate affects the biology and ecology of rhizosphere micro‐organisms in GR soybean that affect yield. The objective of this research was to evaluate the microbiological interactions in the rhizospheres of GR2 and GR1 soybean and the performance of the cultivars with different rates of glyphosate applied at different growth stages. Methods and Results: A greenhouse study was conducted using GR1 and GR2 soybean cultivars grown in a silt loam soil. Glyphosate was applied at V2, V4 and V6 growth stages at three rates. Plants harvested at R1 growth stage had high root colonization by Fusarium spp.; reduced rhizosphere fluorescent pseudomonads, Mn‐reducing bacteria, and indoleacetic acid–producing rhizobacteria; and reduced shoot and root biomass. Conclusions: Glyphosate applied to GR soybean, regardless of cultivar, negatively impacts the complex interactions of microbial groups, biochemical activity and root growth that can have subsequent detrimental effects on plant growth and productivity. Significance and Impact of the Study: The information presented here will be crucial in developing strategies to overcome the potential detrimental effects of glyphosate in GR cropping systems.     

Photosynthesis,antioxidant status and gas-exchange are altered by glyphosate application in peanut leaves

Glyphosate herbicide caused oxidative stress and exhibited negative effects on photosynthesis and gas exchange of peanut [Arachis hypogaea L. cv. Giza (G) 5 and 6] leaves. We demonstrated that glyphosate caused various morphological symptoms, such as chlorosis, yellowing, and appearance of curly edges in leaves treated with high doses of herbicide in both cultivars; however, the G5 cultivar was more sensitive and showed severer symptoms. Glyphosate lowered photosynthesis and reduced contents of pigments and proteins as well as free amino acids in both cultivars. The gas-exchange parameters, such as photosynthetic (P_N) and transpiration rate (E), were highly altered by the glyphosate application. For example, P_N and E were reduced by 65 and 61%, respectively, in G5 treated with high dose of glyphosate compared with control. Antioxidant enzymes, such as peroxidase, catalase, ascorbate peroxidase, and superoxide dismutase were induced by both low and high concentrations in the glyphosate-treated leaves. Moreover, the level of lipid peroxidation, indicated by a malondialdehyde content, as well as the hydrogen peroxide content increased in the glyphosate-treated leaves. However, an increase in total antioxidant activity was detected in leaves and this reflected changes in the antioxidant status and accumulation of antioxidants as a defense mechanism against glyphosate toxicity in peanut.     

渗透胁迫对黑麦幼苗活性氧和抗氧化酶活性的影响

用20%聚乙二醇(PEG 6000)研究了渗透胁迫对黑麦(Secale cereale L.)幼苗活性氧(reactive oxygen species, ROS)和主要抗氧化酶—— 超氧化物歧化酶(superoxide dismutase, SOD)、过氧化氢酶(catalase, CAT)、抗坏血酸过氧化物酶(ascorbate peroxidase, APX)和谷胱甘肽还原酶(glutathione reductase, GR)活性的影响。结果表明, 与对照相比, PEG处理明显提高了叶子和根中丙二醛(malondialdehyde, MDA)的含量、ROS的水平和以上4种抗氧化酶的活性。渗透胁迫下,叶子和根中MDA和ROS水平变化的规律基本相似, 但抗氧化酶活性在2种器官中表现不完全相同, 叶子中CAT的活性在对照和处理中无显著差异, 但在根中差异明显, 表明叶子中SOD、APX和GR在植物应答渗透胁迫中起重要作用, 而根中这4种抗氧化酶都参与植物对胁迫的反应。GR活性随PEG处理变化幅度显著高于其它抗氧化酶, 表明GR在黑麦应答渗透胁迫中所起作用可能强于其它抗氧化酶。     

Allele exchange at the EPSPS locus confers glyphosate tolerance in cassava

Effective weed control can protect yields of cassava (Manihot esculenta) storage roots. Farmers could benefit from using herbicide with a tolerant cultivar. We applied traditional transgenesis and gene editing to generate robust glyphosate tolerance in cassava. By comparing promoters regulating expression of transformed 5‐enolpyruvylshikimate‐3‐phosphate synthase (EPSPS) genes with various paired amino acid substitutions, we found that strong constitutive expression is required to achieve glyphosate tolerance during in vitro selection and in whole cassava plants. Using strategies that exploit homologous recombination (HR) and nonhomologous end‐joining (NHEJ) DNA repair pathways, we precisely introduced the best‐performing allele into the cassava genome, simultaneously creating a promoter swap and dual amino acid substitutions at the endogenous EPSPS locus. Primary EPSPS‐edited plants were phenotypically normal, tolerant to high doses of glyphosate, with some free of detectable T‐DNA integrations. Our methods demonstrate an editing strategy for creating glyphosate tolerance in crop plants and demonstrate the potential of gene editing for further improvement of cassava.     

Nickel elicits a fast antioxidant response in Coffea arabica cells.

The antioxidant responses of coffee (Coffea arabica L.) cell suspension cultures to nickel (Ni) were investigated. Ni was very rapidly accumulated in the cells and the accumulation could be directly correlated with the increase of NiCl(2) concentration in the medium. At 0.05 mM NiCl(2) growth was stimulated, but at 0.5 mM NiCl(2), the growth rate was reduced. An indication of alterations in the presence of reactive oxygen species was detected by an increase in lipid peroxidation at 0.5 mM NiCl(2). Catalase (CAT; EC 1.11.1.6), glutathione reductase (GR; EC 1.6.4.2), ascorbate peroxidase (APX; EC 1.11.1.11), guaiacol peroxidase (GOPX; EC 1.11.1.7) and superoxide dismutase (SOD; EC 1.15.1.1) activities were increased, particularly at earlier NiCl(2) exposure times and the activities were higher at 0.5 mM NiCl(2) for most of exposure times tested. Non-denaturing PAGE revealed one CAT isoenzyme, nine SOD isoenzymes and four GR isoenzymes. The SOD isoenzymes were differentially affected by NiCl(2) treatment and one GR isoenzyme was increased by NiCl(2). NiCl(2) at 0.05 mM did not induce lipid peroxidation and the main response appeared to be via the induction of SOD, CAT, GOPX and APX activities for the removal of the reactive oxygen species and through the induction of GR to ensure the availability of reduced glutathione.     

The effect of 2,4-dichlorophenol and pentachlorophenol on antioxidant system in the leaves of Phalaris arudinacea

The purpose of this work was to evaluate the effect of 2,4-dichlorophenol (2,4-DCP) and pentachlorophenol (PCP) on the activity of antioxidative system and lipid peroxidation in the leaves of reed canary grass (Phalaris arudinacea). The activity of catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), glutathione reductase (GR) and glutathione S-transferase (GST) as well as the content of glutathione, ascorbate and phenolic compounds were determined. An induced-increase in the APX, CAT, GPX and GR activities was stronger for PCP, while a significant increase in the GST activity was noted only for 2,4-DCP. Both compounds increased the content of phenolic compounds, oxidized and reduced glutathione as well as the content of ascorbic acid. PCP induced stronger increase in lipid peroxidation than 2,4-DCP. The observed changes revealed that chlorophenols induce oxidative stress and oxidative damage in the leaves of reed canary grass.     

Glyphosate reduces shoot concentrations of mineral nutrients in glyphosate-resistant soybeans

Although glyphosate-resistant (GR) technology is used in most countries producing soybeans (Glycine max L.), there are no particular fertilize recommendations for use of this technology, and not much has been reported on the influence of glyphosate on GR soybean nutrient status. An evaluation of different cultivar maturity groups on different soil types, revealed a significant decrease in macro and micronutrients in leaf tissues, and in photosynthetic parameters (chlorophyll, photosynthetic rate, transpiration and stomatal conductance) with glyphosate use (single or sequential application). Irrespective of glyphosate applications, concentrations of shoot macro- and micronutrients were found lower in the near-isogenic GR-cultivars compared to their respective non-GR parental lines Shoot and root dry biomass were reduced by glyphosate with all GR cultivars evaluated in both soils. The lower biomass in GR soybeans compared to their isogenic normal lines probably represents additive effects from the decreased photosynthetic parameters as well as lower availability of nutrients in tissues of the glyphosate treated plants.     

Cadmium-induced changes in antioxidant enzyme activities in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L. cv. Dongjin)

We studied how the relationship between cadmium (Cd) toxicity and oxidative stress influenced the growth, photosynthetic efficiency, lipid peroxidation, and activity of ntioxidative enzymes in the roots and leaves of rice(Oryza sativa L Dongjin). Plants were exposed to Cd for 21 d. Both seedling growth and photosynthetic efficiency decreased gradually with increasing cadmium concentrations. Lipid peroxidation increased slowly in both roots and leaves, causing oxidative stress. However, each tissue type responded differently to Cd concentrations with regard to the induction/ inhibition of antioxidative enzymes. The activity of Superoxide dismutase (SOD) increased in both roots and leaves. Ascorbate peroxidase (APX) activity increased in leaves treated with up to 0.25 μM Cd, then decreased gradually at higher concentrations. In contrast, APX activity in roots increased and remained constant between 0.25 and 25 μM Cd. Enhanced peroxidase (POD) activity was recorded for treatments with up to 25/M Cd, gradually decreasing at higher concentrations in the leaves but remaining unchanged in the roots. Catalase (CAT) activity increased in the roots, but decreased in the leaves, whereas the activity of glutathione reductase (GR) was enhanced in both roots and leaves, where it remained elevated at higher Cd concentrations. These results suggest that rice seedlings tend to cope with free radicals generated by Cd through coordinated, enhanced activities of the antioxidative enzymes involved in detoxification.     

Changes in the isozyme composition of antioxidant enzymes in response to aminotriazole in leaves ofArabidopsis thaliana

The changes in isozyme profiles of catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR) during severe deactivation of total CAT activity by aminotriazole (AT) treatment were investigated in the leaves ofArabidopsis thaliana (Columbia ecotype) in relation to H₂O₂-mediated oxidative stress. In spite of striking deactivation of total CAT activity by 0.1 mM AT, there were no significant differences in H₂O₂ levels or total leaf soluble protein contents including a Rubisco in both the control and AT-treated leaves. On the other hand, one specific protein band (molecular mass, 66 kD) was observed on the SDS-gel from leaf soluble proteins whose staining intensity was strikingly enhanced by AT treatment for 6 h. However, this band disappeared at 12 h. In the native-gel assays of CAT, POD, APX and GR isozymes, AT remarkably inhibited the expression of the CAT1 isozyme with no effects on CAT2 and CAT3, and generally had no effect on POD isozyme profiles. However, AT stimulated the intensity of activities of pre-existing APX1 and GR1 isozymes. In particular, it induced a new synthesis of one GR isozyme. Therefore, these results collectively suggest that a striking deactivation of total CAT activity by AT inA. thaliana leaves largely results from the suppression of CAT1 isozyme, and that APX1, GR1, and a newly synthesized GR isozyme could complement the role of CAT1 to metabolize H₂O₂ into non-toxic water.     

The effects of boron toxicity on root antioxidant systems of two chickpea (Cicer arietinum L.) cultivars

Significant differences in the antioxidant systems of the roots of two chickpea (Cicer arietinum L.) cultivars differing in tolerance to drought were observed in under toxic boron (B) conditions. Three-week-old chickpea seedlings were subjected to 0.05 mM (control), 1.6 mM or 6.4 mM B in the form of boric acid (H₃BO₃) for 7 days. At the end of the treatment period, root length, dry weight, boron concentration, malondialdehyde (MDA) content, and the activities of antioxidant enzymes—superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), ascorbate peroxidase (APOX) and glutathione reductase (GR)—were measured. Root length of the drought-tolerant Gökce cultivar did not change under 1.6 mM B but increased under 6.4 mM B. On the contrary, root length decreased in the drought-sensitive Küsmen cultivar under both B concentrations. While root dry weight was unaffected in Gökce, it decreased in Küsmen under both B concentrations. Boron concentration was significantly higher in Küsmen than in Gökce at both B levels. Significant increases in SOD and POX activities were observed in roots of both cultivars under 1.6 and 6.4 mM B. Root extracts exhibited three SOD and three POX activity bands in both cultivars under B stress when compared to control groups. Although CAT activity in Gökce was increased, it decreased in Küsmen at the highest B concentration as compared to control groups. Roots of both cultivars showed no significant change in APOX activity under B toxicity (except in 1.6 mM B treated roots of Küsmen) when compared to control groups. GR activity in the roots of Küsmen decreased significantly with increasing B concentration. However, a significant increase in GR activity was found in Gökce under 1.6 mM B stress. In addition, lipid peroxidation levels of drought-sensitive Küsmen increased, indicating more damage to membrane lipids due to B toxicity. Lipid peroxidation did not change in the drought-tolerant Gökce cultivar at either B concentration. These results suggest that roots of Gökce are better protected from B-stress-induced oxidative stress due to enhanced SOD, CAT and POX activities under high B levels.     

Response of antioxidant enzymes in rice (<Emphasis Type="Italic">Oryza sauva</Emphasis> L. cv. Dongjin) under mercury stress

We studied the effects of different concentrations of mercury (0.0 to 100 μM) on growth and photosynthetic efficiency in rice plants treated for 21 d. In addition, we investigated how this metal affected the malondialdehyde (MDA) content as well as the activity of five antioxidant enzymes — superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), guaiacol peroxidase (POD), and catalase (CAT). Photosynthetic efficiency (Fμ/F_m) and seedling growth decreased as the concentration of Hg was increased in the growth media. Plants also responded to Hg-induced oxidative stress by changing the levels of their antioxidative enzymes. Enhanced lipid peroxidation was observed in both leaves and roots that had been exposed to oxidative stress, with leaves showing higher enzymatic activity. Both SOD and APX activities increased in treatments with up to 50 μM Hg, then decreased at higher concentrations. In the leaves, both CAT and POD activities increased gradually, with CAT levels decreasing at higher concentrations. In the roots, however, CAT activity remained unchanged while that of POD increased a bit more than did the control for concentrations of up to 10 μM Hg. At higher Hg levels, both CAT and POD activities decreased. GR activity increased in leaves exposed to no more than 0.25 μM Hg, then decreased gradually. In contrast, its activity was greatly inhibited in the roots. Based on these results, we suggest that when rice plants are exposed to different concentrations of mercury, their antioxidative enzymes become involved in defense mechanisms against the free radicals that are induced by this stress.     

Postharvest chilling induces oxidative stress response in the dwarf tomato cultivar Micro-Tom

Oxidative stress is involved in the response of Lycopersicon esculentum fruits (cultivar Micro-Tom) to chilling. Changes in activated oxygen scavenging enzymes, superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), and glutathione reductase (GR, EC 1.6.4.2) were examined during ripening after postharvest chilling. Also, lipid peroxidation, respiration, and pigment contents were determined. These parameters were affected by chilling, especially the lycopene content and the respiration rate that showed a high value when the fruits were transferred to higher temperatures. CAT activity increased the day after the fruits were re-warmed, while the activity of GR was higher in the chilled than in the non-chilled green fruits. Lipid peroxidation was more evident at the 'pre-chilled' yellow and red fruits. APX and SOD were not affected by previous chilling in ripening fruits. These results indicate that oxidative stress is generated by conservation at 4°C. The antioxidant response of tomato fruit could be mediated by CAT and GR but not by SOD or APX. Moreover, CAT seemed to respond to the increase in the respiration rate.     

Differences in the Induction of Defence Responses in Resistant and Susceptible Muskmelon Plants Infected with Colletotrichum lagenarium

Defence reactions occurring in resistant (cv. Gankezaomi) and susceptible (cv. Ganmibao) muskmelon leaves were investigated after inoculating with Colletotrichum lagenarium. Lesion restriction in resistant cultivars was associated with the accumulation of hydrogen peroxide (H₂O₂). The activity of antioxidants catalase (CAT) and peroxidase (POD) significantly increased in both cultivars after inoculation, while levels of both CAT and POD activity were significantly higher in the resistant cultivar. Ascorbate peroxidase (APX) increased in both cultivars after inoculation, and level of APX activity was significantly higher in the resistant cultivar. Glutathione reductase (GR) activity significantly increased in both cultivars following inoculation, but was higher in the resistant cultivar, resulting in higher levels of ascorbic acid (AsA) and reduced glutathione (GSH). Phenylalanine ammonia lyase (PAL) significantly increased in inoculated leaves of both cultivars, resulting in higher levels of total phenolic compounds and flavonoids. The pathogenesis‐related proteins chitinase (CHT) and β‐1, 3‐glucanase (GLU) significantly increased following inoculation with higher activity in the resistant cultivar. These findings show that resistance of muskmelon plants against C. lagenarium is associated with the rapid accumulation of H₂O₂, resulting in altered cellular redox status, accumulation of pathogenesis‐related proteins, activation of phenylpropanoid pathway to accumulation of phenolic compounds and flavonoids.     

NaCl胁迫对菜用大豆种子膨大过程中抗氧化系统及渗透调节物质的影响

采用蛭石栽培试验,在100mmol.L-1 NaCl胁迫下,对耐盐性不同的两个菜用大豆品种种子膨大、抗氧化系统及渗透调节物质进行了研究。结果显示:(1)耐盐品种‘绿领特早’种子干质量及大小的增加在NaCl胁迫15d时未受显著影响,而盐敏感品种‘理想高产95-1’种子的膨大受到了显著抑制。(2)NaCl胁迫导致菜用大豆种子超氧阴离子(O2.-)、过氧化氢(H2O2)及丙二醛(MDA)的过量积累,但‘绿领特早’种子的相对积累量均低于‘理想高产95-1’。(3)与‘理想高产95-1’相比,‘绿领特早’在NaCl胁迫期间维持了相对较高的超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)及谷胱甘肽还原酶(GR)活性,以及较高的抗坏血酸(AsA)、可溶性蛋白和可溶性糖含量;胁迫中期维持了相对较高的过氧化物酶(POD)活性,胁迫后期维持了相对较高的过氧化氢酶(CAT)活性;胁迫前中期维持了相对较高的谷胱甘肽(GSH)含量。研究发现,‘绿领特早’种子在NaCl胁迫期间能够保持较高且协调平衡的保护酶SOD、POD、CAT活性和较高的AsA-GSH循环效率,同时维持了较强的渗透调节能力,有效抑制了O2.-和H2O2的过量积累,进而有效降低了细胞膜质过氧化程度,这可能是其耐盐性较强的重要原因之一。     

Oxidative stress is not related to the mode of action of herbicides that inhibit acetolactate synthase

Imazethapyr (IM) is an imidazolinone herbicide which inhibits the biosynthesis of branched chain amino acids, by blocking acetolactate synthase (ALS; EC 4.1.3.18), the first common enzyme of the pathway. To study new aspects of the mode of action of ALS-inhibiting herbicides, pea plants grown in hydroponic cultures were supplied with IM and were analysed with reference to the antioxidant system and oxidative markers. A slight lipid peroxidation was detected in leaves after IM treatment, but no changes were noted in electrolyte leakage or carbonyl content. The ascorbate pool of leaves was oxidized under IM treatment. The analysis of the antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), catalase (CAT) and guaiacol peroxidase (GPX), showed that IM treatment only caused an enhancement of GPX activity in leaves. In roots, the herbicide caused a decrease in lipid peroxidation. The enhancement of the reduced glutathione content detected in IM-treated roots can be related to the detected increase of GR activity. The lack of more noticeable effects on antioxidant enzymatic activities could be explained by the inability of IM-treated plants to respond to oxidative stress with modifications in their protein synthesis. Our results suggest that oxidative stress is not related to the mode of action of ALS-inhibitors. The slight changes detected in the antioxidative status of treated plants are too secondary in time and intensity to be related to the lethality caused by ALS-inhibitors     

Glyphosate affects lignin content and amino acid production in glyphosate-resistant soybean

Farmers report that some glyphosate-resistant soybean varieties are visually injured by glyphosate. Glyphosate is the main herbicide that directly affects the synthesis of secondary compounds. In this work, we evaluated the effect of increasing rates of glyphosate on lignin and amino acid content, photosynthetic parameters and dry biomass in the early maturity group cultivar BRS 242 GR soybean. Plants were grown in half-strength complete nutrient solution and subjected to various rates of glyphosate either as a single or in sequential applications. All parameters evaluated were affected by increasing glyphosate rates. The effects were more pronounced as glyphosate rates increased, and were more intense with a single total application than sequential applications at lower rates.     

A new type of class I bacterial 5-enopyruvylshikimate-3-phosphate synthase mutants with enhanced tolerance to glyphosate

Glyphosate or Roundup^® is the most extensively used herbicide for broad-spectrum control of weeds. Glyphosate inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), a key enzyme in the aromatic amino acid biosynthetic pathway in microorganisms and plants. Applying the staggered extension process, we randomly mutated and recombined the aroA genes of Salmonella typhimurium and Escherichia coli to obtain four variants that exhibit significantly enhanced tolerance to glyphosate. All four mutants are chimeras of the two parental genes and, in addition, three of them carry one or more de novo point mutations. None of the amino acid substitutions in the mutants was in a position previously known to be important for catalysis or substrate binding. Kinetic analysis of EPSPS activity from these mutants indicated that the tolerance was attributed to a 2–10-fold increased specific activity, 0.4–8-fold reduced affinity to glyphosate, and 2.5–19-fold decreased K_m for phosphoenolpyruvate. Such mutants will be instrumental for the structural and function study of the enzyme and for the generation of transgenic crops resistant to the herbicide.     

铝胁迫对海莲幼苗保护酶系统及脯氨酸含量的影响

为探讨Al~(3+)胁迫对海莲的影响,研究了10～50 mmol/L Al~(3+)处理下海莲幼苗叶片和根系的过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)的活性以及可溶性蛋白质、丙二醛(MDA)和游离脯氨酸(Pro)含量的变化。结果表明,海莲幼苗能耐受50 mmol/L的Al~(3+)胁迫处理,具有较高的耐铝性。但在50 mmol/L Al~(3+)处理时,海莲幼苗叶片和根系的质膜系统膜脂过氧化加重,MDA含量增加;细胞活性氧代谢失衡。在保护酶系统中,Al~(3+)处理促进了叶片中APX和POD活性的提高,降低了CAT的活性,SOD的活性呈下降趋势;海莲根部POD和SOD活性均显著提高,而CAT活性下降。25～50 mmol/LAl~(3+)处理下,海莲叶片和根部可溶性蛋白质含量均显著下降;Pro的含量在叶片和根均有显著增加。