Evaluation of molluscicidal activities of Arisaema tubers extracts on the snail Oncomelania hupensis

Four extracts of Arisaema erubescens tubers by acetic acetal (AAE), benzinum (BZE), n-butanol (NBE) and chloroform (CFE) were obtained to evaluate their molluscicidal activities against the snail Oncomlania hupensis. The responses of choline esterase (ChE), alkaline phosphatase (ALP), esterase (EST), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) to the extracts (NBE) were also investigated. In the four extracts (AAE, BZE, NBE and CFE), NBE showed the highest toxicity on the snails after 48 h exposure. NBE also showed the time- and concentration-dependent effect, for example, the LC₉₀ values of the NBE were decreased from 365.5 mg/L (24 h) to 36.4 mg/L (96 h). At the end of exposure to NBE (LC₅₀ concentration), the activities of ChE and ALP in snail tissues (cephalopodium and liver) decreased significantly. Isozyme electrophoresis profiles indicated that responses of isozymes (EST, SOD and GSH-Px) to NBE were more intense in liver than in cephalopodium. After 72 h exposure to NBE, the EST activity in snail liver decreased and some enzyme bands (EST1 and EST4) disappeared. But the activities of SOD 1 and GSH 2 in liver increased after 48 h exposure. The results indicated that NBE was the highest toxic component in the four extracts. The decline of the detoxification ability and the oxidative damage in snail tissues might be the main reason for the molluscicidal activities.     

Comparative molluscicidal action of extract of Ginko biloba sarcotesta,arecoline and niclosamide on snail hosts of Schistosoma japonicum

In search for new local plant molluscicides for the control of the vectors of schistosomiasis, we compared the molluscicidal action of the extract of Ginkgo biloba sarcotesta by benzinum (EGSB) to that of arecoline (ARE) and niclosamide (NIC) against Oncomelania hupensis snails. NIC showed the highest toxicity on snails with 24 h LC₅₀ vales of 0.12 mg/L and LC₉₀ of 0.98 mg/L, while the LC₅₀ and LC₉₀ of EGSB were much lower than that of ARE. Sublethal in vivo 24 h exposure to 40% and 80% LC₅₀ of NIC, EGSB and ARE altered the activities of different enzymes in different body tissues of snails. EGSB could significantly inhibit Choline esterase (ChE), Alanine aminotransferase (ALT), Alkaline phosphatase (ALP) and Malic dehydrogenase (MDH) activities both in the cephalopodium and liver. ARE could significantly cause a reduction in ChE, ALP activities in the cephalopodium and ChE, ALT, ALP, Succinodehydrogenase (SDH), MDH activities in the liver. NIC significantly altered activities of ChE, ALT, ALP, SDH, and MDH in the cephalopodium and ChE, ALT, ALP, SDH activities in the liver. All molluscicides could not affect Lactate dehydrogenase (LDH) activity in the cephalopodium and the liver. Maximum inhibition of ALT and MDH activities was found in the cephalopodium and liver of snails treated with 80% of 24 h LC₅₀ of EGSB. However, NIC and ARE caused maximum reduction in ALP and SDH activities, respectively. The results indicated that molluscicidal action of EGSB was different to that of ARE and NIC in some extent.     

Comparative molluscicidal action of extract of Ginko biloba sarcotesta, arecoline and niclosamide on snail hosts of Schistosoma japonicum

In search for new local plant molluscicides for the control of the vectors of schistosomiasis, we compared the molluscicidal action of the extract of Ginkgo biloba sarcotesta by benzinum (EGSB) to that of arecoline (ARE) and niclosamide (NIC) against Oncomelania hupensis snails. NIC showed the highest toxicity on snails with 24 h LC₅₀ vales of 0.12 mg/L and LC₉₀ of 0.98 mg/L, while the LC₅₀ and LC₉₀ of EGSB were much lower than that of ARE. Sublethal in vivo 24 h exposure to 40% and 80% LC₅₀ of NIC, EGSB and ARE altered the activities of different enzymes in different body tissues of snails. EGSB could significantly inhibit Choline esterase (ChE), Alanine aminotransferase (ALT), Alkaline phosphatase (ALP) and Malic dehydrogenase (MDH) activities both in the cephalopodium and liver. ARE could significantly cause a reduction in ChE, ALP activities in the cephalopodium and ChE, ALT, ALP, Succinodehydrogenase (SDH), MDH activities in the liver. NIC significantly altered activities of ChE, ALT, ALP, SDH, and MDH in the cephalopodium and ChE, ALT, ALP, SDH activities in the liver. All molluscicides could not affect Lactate dehydrogenase (LDH) activity in the cephalopodium and the liver. Maximum inhibition of ALT and MDH activities was found in the cephalopodium and liver of snails treated with 80% of 24 h LC₅₀ of EGSB. However, NIC and ARE caused maximum reduction in ALP and SDH activities, respectively. The results indicated that molluscicidal action of EGSB was different to that of ARE and NIC in some extent.     

Effects of trichlorfon and sodium dodecyl sulphate on antioxidant defense system and acetylcholinesterase of Tilapia nilotica in vitro

The effects of organophosphorus insecticide trichlorfon, surfactant sodium dodecyl sulphate (SDS), and the mixture of trichlorfon and SDS on the antioxidant defense system and acetylcholinesterase (AChE) in Tilapia nilotica were assessed in vitro. Various concentrations of trichlorfon (0, 0.0001, 0.001, 0.01, 0.1 and 1 g/L) and SDS (0, 0.0625, 0.125, 0.25, 0.5, 1 g/L) were incubated with homogenate of liver and muscle, respectively, at 25 °C for 0, 30, 60 and 90 min. Two concentrations of mixture of trichlorfon and SDS (0.0001 g/L trichlorfon + 0.5 g/L SDS, 0.1 g/L trichlorfon + 0.5 g/L SDS) and 0.0001 g/L trichlorfon, 0.1 g/L trichlorfon, 0.5 g/L SDS and control, were incubated simultaneously with homogenate of liver and muscle, respectively, at 25 °C for 60 min. After incubation, the content of reduced-glutathione (GSH) and the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) in homogenate of liver were determined, and the activities of AChE in homogenate of muscle were also measured.Treatment with trichlorfon caused a significant concentration-dependent and time-related inhibition of AChE activity at all treatment concentrations and times since trichlorfon is a cholinesterase inhibitor. For the same trichlorfon treatment, an apparent decrease in GSH content was found in concentration of 0.01, 0.1, 1 g/L, whereas no significant alteration in antioxidant enzyme activity were found at all experiment concentrations and times, which might indicate that antioxidant enzymes have not involved in the metabolism of trichlorfon. The depletion of GSH might indicate that ROS could be involved in the toxic effects of trichlorfon. Exposure of SDS can inhibit activities of AChE, GST and CAT at concentrations of 0.5 and/or 1 g/L, which could be due to the denaturing process of SDS to the enzymes. For the mixture exposure of trichlorfon and SDS, the effect of the mixture of 0.0001 g/L trichlorfon and 0.5 g/L SDS on inhibition of AChE shows synergistic other than simple additive of trichlorfon and SDS. The combined effects of chemicals and detergents deserve to be particularly noted. It should be noted that the toxicity experiments were made in tissue homogenates instead of whole organisms. The responses against the toxic compounds will not be the same in both systems.     

Toxic effects of acephate on Paramecium caudatum with special emphasis on morphology,behaviour, and generation time

The continuous increase in the number of new chemicals as well as the discharges of solid and liquid wastes triggered the need for simple and inexpensive bioassays for routine testing. In recent years, there has been increasing development of methods (particularly rapid tests) for testing environmental samples. This paper describes the quick toxic evaluation of an organophosphorus insecticide, acephate (O,S-dimethyl acetylphosphoramidothioate) on Paramecium caudatum for acute and sub-acute toxicity studies with reference to morphology, behaviour, and its generation time. The lethal concentrations for 10 min and 2 h were determined by probit method, as 500 mg L⁻¹ and 300 mg L⁻¹, respectively. Higher concentrations of 10 min exposure caused cell lysis with disintegration of cell membrane and precipitation of protoplasm. Combination of conventional light microscopy and computerized video tracking systems were used to study the locomotor behaviour of paramecia. The test organism was under stress and exhibited an initial increase and subsequent decrease in the swimming speed when exposed to 1/4, 1/2, 3/4, and LC₅₀ concentrations for 10 min (125, 250, 375, and 500 mg L⁻¹, respectively). Similar changes were also noticed when paramecia were exposed to LC₅₀ for 2 h. In a separate set of experiments, the number of generations and generation time in 24 h was evaluated with respect to the different sub-lethal concentrations (30, 60, 120, and 240 mg L⁻¹). The number of generations decreased and generation time extended significantly in a concentration dependent manner. The results indicate that the Paramecium toxicity assay could be used as a complimentary system to rapidly elucidate the cytotoxic potential of insecticides. The major advantages associated with these tests are: they are inexpensive, simple, user-friendly, space saving, and seem to be attractive alternatives to conventional bioassays.     

Biochemical,physiological and morfological responses in common carp (Cyprinus carpio L.) after long-term exposure to terbutryn in real environmental concentration

The effects of terbutryn at concentrations of 0.02 (reported concentration in Czech rivers), 0.2, and 2.0 μg l^?1 were assessed in one-year-old common carp (Cyprinus carpio L.) exposed for 90 days. Influence on biometric parameters, hematology, biochemistry, histology, and oxidative stress was investigated. Exposure to 0.02, 0.2 and 2.0 μg l^?1 showed significant differences oxidative stress biomarkers compared to controls but exposure to 0.2 and 2.0 μg l^?1 significantly affected biochemical and hematological profiles. Long-term exposure of terbutryn in carp resulted in slight alterations in internal organs and increased reactive oxygen species formation, resulting in oxidative damage to lipids and proteins and inhibition of antioxidant capacities.     

Oxygen consumption and ammonia excretion of the freshwater crab Trichodactylus borellianus exposed to chlorpyrifos and endosulfan insecticides

The effects of lethal and sublethal concentrations of chlorpyrifos and endosulfan on oxygen consumption and ammonia excretion rate of the crab Trichodactylus borellianus were evaluated. Oxygen consumption and energy expenditure had significant effect in relation to exposure times. Regarding endosulfan, a significant difference in consumption among times of exposure was registered in 625 μg L⁻¹. Moreover, at the highest concentration, energy expenditure rate was observed stabilized during 1–3 h. A significant increase in ammonia excretion was evidenced in 150 and 300 μg L⁻¹ of chlorpyrifos. The O:N ratio showed a decrease in chlorpyrifos and in 2500 μg L⁻¹ of endosulfan. This indicated a shift towards protein primary metabolism. An increment in the O:N ratio was observed in the lower endosulfan solutions. The relation oxygen:nitrogen showed a shift towards lipid and carbohydrate primary metabolism. This work indicated the complexity of the metabolism in the freshwater crab affected by xenobiotic elements.     

Induction of disease resistance against Botrytis cinerea by heat shock treatment in melon (Cucumis melo L.)

The present study investigated resistance against Botrytis cinerea after heat shock treatment in melon plants. Heat shock at 50 °C for 20 s 0–24 h before inoculation resulted in maximal B. cinerea symptom reduction and peroxidase gene expression, which peaked 12 and 72 h post-treatment and decreased 24–48 h post-treatment, suggesting pathogenesis-related protein expression priming. Hot water dipping did not directly inhibit mycelia growth. Plants treated with 2-benzisothiazol-3(2H)-one 1,1-dioxide, which induces systemic acquired resistance, demonstrated higher peroxidase gene expression but no B. cinerea resistance, indicating possible involvement of additional novel mechanisms in heat shock-activated resistance of melon against B. cinerea.     

Agrobacterium induces plant cell death in wheat (Triticum aestivum L.)

The symptoms of gall or hairy root do not occur in the interactions between wheat (Triticum aestivum L.) and other monocotyledonous plants, with Agrobacterium tumefaciens or Agrobacterium rhizogenes. However, both bacteria colonized wheat root surfaces at similar levels (2.0 × 10⁷ colony forming U g⁻¹ root) and grew without inhibition in suspension with intact or wounded wheat embryos or root segments present. Suspension-cultured wheat embryo cells grown in 7.4 m M O₂ displayed 23% cell death after 1 h exposure to Agrobacterium cells, while the extent of cell death with 2.1 m M O₂ averaged 8%. Cultured wheat embryo and root cells rapidly produced hydrogen peroxide (H₂O₂) when contacted with A. tumefaciens or A. rhizogenes. Production of H₂O₂ was lower at 2.1 m M O₂ than 7.4 mM O₂. Browning and autofluorescence of epidermal cells of callus derived from wheat embryos and wheat roots was observed after inoculation with Agrobacterium. An increase in ferulic acid was detected in the walls of roots exposed to Agrobacterium. However, neither lignin nor callose was detected by diagnostic staining methods. These findings suggest that Agrobacterium induced a resistance-like response in wheat that may reduce the efficacy of transformation and limit the normal symptom formation.     

A soluble carbohydrate elicitor from Blumeria graminis f. sp. tritici is recognized by a broad range of cereals

Wheat plants rapidly recognize pathogenic and non-pathogenic conidia of the powdery mildew fungusBlumeria (syn. Erysiphe)graminis on their leaf surfaces. This suggests that a chemical signal emanates from conidia at the pre-penetration stage of infection. Conidia of B. graminis f. sp. tritici were found to contain an elicitor that was easily washed off their surface. The elicitor activity is heat stable and could not be removed by phenol extraction. By contrast, elicitor activity is sensitive to periodate oxidation and partial acid hydrolysis suggesting that the elicitor activity resides in a carbohydrate moiety. Analysis of carbohydrates revealed mostly glucose, with smaller amounts of xylose and mannose. The glucosyl residues of the B. graminis elicitor were found to be linked (1 → 2)-, (1 → 4), and (1 → 6)-, with (1 → 4, 1 → 6)- branch point residues, and no 3-linked glucose residues were detected. As treatment with β -mannanase significantly reduced elicitor activity, mixed-linkage (1 → 4), (1 → 6)-mannosyl residues appeared to be important for elicitor activity. The B. graminis elicitor induced the expression of all defence-related genes tested in wheat and also induced resistance to subsequent attack by B. graminis f. sp. tritici. In contrast, a hypersensitive response was not induced by the elicitor in the absence or the presence of a challenging inoculum of B. graminis f. sp. tritici. The elicitor also induced the accumulation of thaumatin-like proteins in barley, oat, rye, rice and maize, but did not induce necrosis in any of these species. This suggests that the B. graminis elicitor represents a host non-specific determinant of non-self recognition in cereals activating general defence responses other than the hypersensitive reaction.     

Nile tilapia (Oreochromis niloticus), liver morphology,CYP1A activity and thyroid hormones after Endosulfan dietary exposure

Endosulfan is a worldwide used insecticide suspected to be highly toxic to aquatic organisms, including fish. Most of the available studies have focused in water exposures, although this pollutant can be transferred through food chain. Therefore, in the present study, the effects of Endosulfan on tilapia (Oreochromis niloticus), when administered through the diet. Fish were fed 21 days with diets containing 1 and 0.5 μg g⁻¹ of Endosulfan, after which qualitative histological liver analysis showed that Endosulfan induced hepatocyte destruction, vessel endothelium rupture and increased melanomacrophages aggregates. To test lower environmentally relevant doses of Endosulfan could induce hepatic damage, as well as other negative effects, such as altered phase I metabolism and plasma thyroid hormone levels. Hence, tilapia were orally exposed to 0.1 and 0.001 μg g⁻¹ for 35 days. Low environmentally realistic doses of Endosulfan were still able to induce liver histopathological damage such as increased hepatocyte vacuolization and increased eosinophil granular cell aggregates. Liver cytochrome P450 1A activity, evaluated through ethoxyresorufin-o-deethylase (EROD), was enhanced in tilapia exposed to 0.001 μg g⁻¹, whereas the highest dose had no measurable effects in this enzyme activity. Fish exposed to 0.1 μg g⁻¹ of Endosulfan had depressed T4 plasma levels. Overall, the results of the present study further demonstrate the toxic effects of Endosulfan in tilapia when administered in the diet at environmentally relevant concentrations, which indicates that in the field food chain transfer may also be an importance source of this pollutant.     

Neurophysiological alterations in Caenorhabditis elegans exposed to dichlorvos,an organophosphorus insecticide

AChE inhibition is widely regarded as a good biomarker of exposure to organophosphorus insecticides (OPIs), suggesting increase in the cholinergic transmission and consequent accumulation of acetylcholine in the organism. This might lead to behavioral changes and create widespread disturbances in the normal physiology of an organism. In the present study we have employed the model organism Caenorhabditis elegans to evaluate the biochemical and behavioral alterations induced by dichlorvos, a well known OPI. Exposure of worms to dichlorvos (at sublethal concentrations: 5, 50, 100, 150 and 200 μM) induced a concentration and time dependent AChE inhibition, and accumulation of acetylcholine. Further, we also observed cessation in feeding (by 72%), shutting of pharyngeal pumping, inhibition of egg laying (34–55%), contraction of nose (45%) and significant paralysis (50%) after 4 h of exposure. Significant correlation was observed between biochemical effects and behavioral parameters clearly suggesting the implications of sublethal concentrations of dichlorvos on non-target invertebrate organism such as C. elegans. These data further suggest that assessment of subtle neurophysiological parameters may serve as useful indicators of OPI exposure.     

In vivo growth and pathotoxin production by the maize pathogen Cochliobolus carbonum

Conidia of Cochliobolus carbonum secrete a toxin (HC-toxin) during appressorium formation on maize leaves. Plasma desorption mass spectrometry revealed that approximately 70 ng of toxin per 10⁶ conidia were secreted during the first 16 h of morphogenesis. Growth of the fungus was monitored microscopically. Extensive fungal growth occurred in the susceptible interaction by 24 h. In spite of a substantial amount of HC-toxin, the fungus failed to become established in the resistant host even after 36 h. Results suggest that the resistance conditioned by Hm1, which encodes a toxin reductase, causes inactivation of the toxin early in the interaction.     

Effect of miconazole,an antifungal agent,on allene oxide synthase: Inhibition,kinetics, and binding

We investigated the inhibition of allene oxide synthase (AOS), a key enzyme in jasmonic acid biosynthesis, by miconazole. Kinetic analysis indicated that miconazole was a mixed-type inhibitor of AOS with a K_i value of approximately 8.4 ± 0.2 μM. Analysis of the interactions between miconazole and AOS by optical difference spectroscopy revealed that miconazole binding induces type II binding spectra with a K_d value of approximately 6.0 ± 0.2 μM.     

Correlation between sensitivity of barley to Pyrenophora teres toxins and susceptibility to the fungus

Detached leaves of 25 barleys, ranging from highly susceptible to highly resistant to Pyrenophora teres f. teres and Pyrenophora teres f. maculata, were tested for their reaction to three phytotoxins isolated from cultures of the fungus: toxin A [ L, L - N -(2-amino-2-carboxyethyl)aspartic acid], toxin C (aspergillomarasmine A) and toxin B (anhydroaspergillomarasmine A). 0.75 m M toxin A caused mainly dark yellow chlorotic symptoms but little necrosis, whereas leaves treated with 0.25 m M toxin C developed distinct necrotic symptoms and zones of light yellow chlorosis. Toxin B is only weakly toxic, and toxin B and control solutions containing aspartic acid in the concentration of 0.75 m M did not cause any symptoms. The best differentiation between the barleys was obtained by scoring chlorosis after 120 h, and the optimal toxin concentrations for this differentiation were 0.75 m M toxin A and 0.25 m M toxin C, respectively. Results with different toxin concentrations inducing distinct variation in symptom expression indicate that the two toxins have different potencies as phytotoxins. The reaction of the barleys to toxins A and C correlated well with their reaction to infection by P. teres f. teres and P. teres f. maculata, suggesting that toxins A and C may be used to select resistant barley lines in the early stages of a breeding programme.     

Chlorpyrifos induced alterations in the levels of hydrogen peroxide,nitrate and nitrite in rat brain and liver

Organophosphate pesticides are among the most widely used synthetic chemicals for controlling a wide variety of pests. Chlorpyrifos (CPF) is among the leading organophosphate (OP) pesticides used extensively throughout the world including India. Besides being potent anticholinesterase compounds, these OP pesticides are known to generate oxidative stress. Present study was carried out to see the level of reactive oxygen species (ROS), hydrogen peroxide (H₂O₂), nitrate (NO₃⁻) and nitrite (NO₂⁻) in different parts of rat brain and liver after giving different doses of CPF intramuscularly for 3 days or for 1 month. Results of the present study clearly revealed that levels of H₂O₂, NO₃⁻ and NO₂⁻ were increased significantly in all the three parts of rat brain i.e., fore-, mid- and hind-brain as well as in liver of rats given for 3 days or 1 month due to exposure of different doses of CPF. The study clearly demonstrated that CPF generated oxidative stress in all the three brain regions as well as liver of rats and the ROS were accumulated. Accumulation of ROS in all the regions of brain and other tissues may disturb the normal physiological functions aggravating the toxicity symptoms of CPF.     

Physiological and molecular mechanisms of glyphosate tolerance in an in vitro selected cotton mutant

We have selected an upland cotton (Gossypium hirsutum L.) cell line (R1098) that is highly tolerant to glyphosate. This cell line was developed by in vitro selection with gradually increasing glyphosate concentrations, and its mechanisms conferring glyphosate tolerance were studied. Based on a whole-plant dose–response bioassay, R1098 plants were tolerant to glyphosate at a concentration of 1500 g ae ha⁻¹ glyphosate (1.5× the recommended field rate) whereas the control plants (Coker 312) were unable to survive at 150 g ae ha⁻¹ glyphosate. Coker 312 accumulated 13.1 times more shikimate in leaves at 5 days after glyphosate treatment (1500 g ae ha⁻¹) than that of R1098. Two distinct cDNAs for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), EPSPS-1 and EPSPS-2, were isolated from R1098. Both cDNAs were 97.7% identical within the common protein-coding region and the predicted sequences of the mature proteins were greater than 83% identical with EPSPS proteins from other known higher plants. In comparison to the glyphosate-susceptible cotton Coker 312, sequence analysis of the EPSPS-1 gene indicated that R1098 has an alanine insertion at nucleotide position 1216 resulting in frameshift. It leads to two copy functional EPSPS genes in R1098. There was no difference between R1098 and Coker 312 in EPSPS mRNA levels before glyphosate treatment. However, its treatment caused a 2–4 times increase in the basal EPSPS mRNA level in R1098.     

Subacute and delayed toxicity of iminoctadine liquid formulation,which contains iminoctadine triacetate as an antifungal component on a nontarget domesticated insect,the silkworm,Bombyx mori L. (Lepidoptera: Bombycidae)

We investigated insecticidal effects of iminoctadine liquid formulation, a fungicide containing iminoctadine triacetate, on the silkworm Bombyx mori that is an important domesticated insect. Iminoctadine liquid formulation was incorporated in an artificial diet and fed to silkworms for 24 h on the first day of either the third or fourth instar, thereafter reared without the fungicide. Third instar larvae that ingested the diet containing 1% (wt/wt) iminoctadine liquid formulation died within 24 h (subacute toxicity), and those that ingested either 0.1% or 0.3% iminoctadine liquid formulation developed to the pharate fourth stage and then became deficient in molting (delayed toxicity). In fourth instar larvae, iminoctadine liquid formulation exhibited subacute toxicity and delayed toxicity at 0.3% and >1%, respectively. Furthermore, we fed the larvae a diet with iminoctadine liquid formulation for 24 h at different intervals in the inter-molting period of the third and fourth instar. Iminoctadine liquid formulation administered at an earlier stage in the inter-molting period exerted a more severe effect than a later administration. Iminoctadine triacetate, an anti-fungal ingredient in the iminoctadine liquid formulation, exhibited toxicity similar to that of the liquid formulation.     

Role of sucrose in the development of Fusarium wilt in lupine embryo axes

The response of embryo axes of Lupinus luteus L. cv. Polo to invasion by Fusarium oxysporum Schlecht f.sp. lupini was studied 84, 96 and 108 h after inoculation and under various trophic conditions. Four variants were compared: these included inoculated embryo axes cultured with or without 60 mM sucrose (+Si and −Si) and non-inoculated embryo axes cultured with or without 60 mM sucrose (+Sn and −Sn). The exogenous sucrose caused an accumulation of sucrose, glucose and fructose in +Sn embryo axes. Pathogen invasion generally resulted in a sharp decline in soluble carbohydrate content of +Si embryo axes. Symptoms of Fusarium wilt included growth inhibition of the embryo axes and a decrease in their fresh weight, which was particularly strong in −Si axes. While the addition of exogenous sucrose to the medium 24 h prior to inoculation significantly alleviated the disease, its application 48 h after inoculation did not restrict disease development. In +Si embryo axes, the activity of peroxidases (EC 1.11.1.7), covalently and ionically bound to the cell walls, increased considerably, mainly up to 96 h after inoculation. This increase was significantly higher than that seen in +Sn embryo axes. These results are in contrast to those of an earlier study [Morkunas I, Marczak Ł, Stachowiak J, Stobiecki M. Sucrose-stimulated accumulation of isoflavonoids as a defense response of lupine to Fusarium oxysporum. Plant Physiol Biochem 2005;43:363–73], which showed a pronounced decrease in the flavonoid glycoside content of +Si tissues. Furthermore, starting 96 h after inoculation, a decrease in phenylalanine ammonialyase activity (PAL, EC 4.3.1.5) was observed in inoculated tissues, in comparison to non-inoculated tissues. The decrease was especially pronounced in +Si tissues. However, a significant increase in the free isoflavone aglycones content of +Si embryo exes, i.e. genistein, 2′-hydroxygenistein, wighteone and luteone. Simultaneously, a strong increase in the activity of β-glucosidase (EC 3.2.1.21), hydrolyzing isoflavone glycosides, was observed in inoculated tissues. The activity of these enzymes was much higher in −Si embryo axes than in +Si embryo axes. The results suggest that soluble carbohydrates may play a role in the restriction of Fusarium wilt development, as they may be related both to intensified lignification and to the antimicrobial role of free aglycones, which may be products of the hydrolysis of isoflavone glycosides generated prior to the invasion stage.     

Metabolic effects in rapeseed (Brassica napus L.) seedlings after root exposure to glyphosate

Metabolic effects in rapeseed (Brassica napus L.) seedlings after root exposure to sublethal concentrations of glyphosate were examined in order to evaluate the possibilities of using a response pattern in plants as a measure of exposure to glyphosate through the growth media, more sensitive than the well-known biomarker shikimate. Rapeseed seedlings were grown in hydroponic nutrient solutions containing varying sublethal concentrations of glyphosate (1–50 μM). After 9 days of glyphosate exposure, the shoots of the seedlings were analysed with respect to the effects on selected metabolites downstream from the primary affected metabolite shikimate, which accumulated linearly in response to glyphosate exposure (from 0 to ∼126 μmol/g DW). The selected metabolites analysed, comprising the free amino acids, and the glucosinolates derived therefrom, showed complex patterns in response to glyphosate exposure. Most noteworthy was though that they responded at the lowest concentrations of exposure to glyphosate (1 μM), where no visual effects, decrease in shoot DW or shikimate could be detected, indicating that a biomarker response more sensitive than that of shikimate can be established for plants exposed to glyphosate.