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1.
A total of 185 hexanic, dichloromethanic, ethanolic and hydroethanolic extracts from 24 species of Cerrado plants, were tested against Zabrotes subfasciatus, Acanthoscelides obtectus, and human saliva α-amylases. Twelve crude extracts presented inhibition rates greater than 80% against digestive α-amylases of the insect pest Z. subfasciatus, at a concentration of 1 mg mL−1. These extracts were also tested against A. obtectus and human saliva α-amylases to verify their affinity and specificity of action. The hydroethanolic Kielmeyera coriacea stem bark extract presented a strong inhibitory potential, with IC50 values of 110 μg mL−1 for Z. subfasciatus and 272.12 μg mL−1 for A. obtectus, in addition to a 97.09% reduction in enzyme activity of human saliva α-amylases at 125 μg mL−1. The hexanic Aspidosperma macrocarpon root wood extract totally inhibited the activity of Z. subfasciatus α-amylases, reduced the enzyme activity of A. obtectus by 14.69% at 1 mg mL−1, but did not alter the activity of human saliva α-amylases, thus characterizing greater inhibition affinity and specificity. The results suggest that the application of plant extracts against insect α-amylases represent a promising biotechnological tool for development of new insect pest control strategies, with noticeable affinity and specificity of action against different target enzymes.  相似文献   

2.
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.  相似文献   

3.
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.  相似文献   

4.
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.  相似文献   

5.
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.  相似文献   

6.
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.  相似文献   

7.
The inhibitory effect of three proteinaceous inhibitors isolated from little and finger millet was examined on gut α-amylases for four stored grain and four phytophagous insect-pests. Additionally, using native PAGE, several α-amylases isozymes were observed in all insect-pests studied. Furthermore, thermostabilities and the pH optimum for insect-pests α-amylases, which varied from acidic to alkaline, were also determined. On the other hand, proteinaceous inhibitors from little millet seeds Panicum sumatrense (LMCO3) and from finger millet (FMCO11 and FMCO13) inhibited insect-pests α-amylases with different proportions. The highest inhibition percent was recorded for LMCO3 and FMCO13 against Callosobruchus chinensis α-amylase, where the inhibition percent was approximately 70 and 50%, respectively. Furthermore, millet α-amylase inhibitors also reduced significantly digestive α-amylolytic activities of Acaea janata, C. cephalonica, Sitophilus oryzae, and Tribolium castaneum, indicating that these α-amylase inhibitors could be used toward crop insect-pests.  相似文献   

8.
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.  相似文献   

9.
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.  相似文献   

10.
The ability to control soil-borne pathogens in agriculture is highly conditioned by the restricted use of synthetic pesticides. Allelopathy, the antimicrobial activity of plant extracts, is a promising option against crop pathogens. Extracts from Lycium spp. such as L. barbarum, L. chinense and L. intricatum possess biological and therapeutic properties. Individual methanolic extracts from leaves and stems of the Mediterranean medicinal species L. europaeum collected in two locations of Tunisia were each evaluated in vitro against Verticillium dahliae (Vd), Sclerotinia sclerotiorum (Ss) and Harpophora maydis (Hm). The mycelial growth of the three fungi was significantly reduced by all the extracts at doses of 10 and 30 μl mL?1 (equivalent to 1 and 3 mg plant tissue mL?1). The sporulation of Hm was almost completely inhibited in all the amendments, but that of Vd was stimulated by one of the leaf extracts when 1 and 3 mg dried plant tissue mL?1 were used. Sclerotia of Ss were formed in a smaller number, their total weight increasing at extract doses equivalent to 1 mg plant tissue mL?1 and higher. In greenhouse, the pathogenicity of Hm was confirmed as early as 6 weeks after inoculation, since it caused significant decreases of weights in both roots and aboveground parts of maize. The detrimental effect of Hm on maize root weight in greenhouse was significantly counteracted by one of the leaf extracts added by watering. In total, 11 phenolic compounds were separated in the four extracts. The hydroxycinnamic acid family, including chlorogenic acid as a major compound, represented more than 50% of the total content in all the samples. Rutin was the most abundant flavonoid. The results of this work show the detrimental effect of L. europaeum extracts against the soil-borne pathogens Hm, Ss and Vd, and highlight their potential in crop protection if adequately developed into final products and used in combination with other tools.  相似文献   

11.
The carmine spider mite is the most serious crop mite pests in China. Abamectin has been used to control insects and mites worldwide but carmine spider mites, Tetranychus cinnabarinus, had developed resistance to it. Genetic research on insecticide resistance has been fundamental for understanding the resistance development, studying resistance mechanisms, and designing appropriate resistance management strategies to control insect pests. A resistant colony of T. cinnabarinus, RRG42, was established to examine the inheritance of abamectin resistance in T. cinnabarinus. The females of T. cinnabarinus were selected for bioassay using a slide dip method. After 42 generations of selection, the RRG42 strain was 8.7-fold resistant to abamectin compared with the susceptible strain (SS). The logarithm (log) concentration–probit response curve for F1s from reciprocal crosses, of F1RS and F1SR, were inclined to that for SS and the degree of dominance (D) values for F1s were −0.81 and −0.17. There was a significant difference in values of LC50 and slope of log concentration–probit lines between F1RS and F1SR. The observed mortalities of BC1 (F1RS♀ × RRG42♂) and BC1′ (F1SR♀ × SS♂) were significantly different from the expected mortalities based on a monogenic resistance in the chi-square tests. The inheritance of abamectin resistance in T. cinnabarinus is incompletely recessive and may be controlled by more than one gene. The maternal or cytoplasmic effect may exist in the inheritance of resistance to abamectin in T. cinnabarinus.  相似文献   

12.
The generation and accumulation of reactive oxygen species (ROS), superoxide anion (O2) and hydrogen peroxide (H2O2), were studied in the interaction between wheat cv. ‘Suwon 11’ and two races of Puccinia striiformis f. sp. tritici (avirulent and virulent). Generation of O2 and H2O2 was analyzed histochemically using nitroblue tetrazolium (NBT) and 3,3-diamino-benzidine (DAB), respectively. At the pre-penetration stage during appressorium formation both stripe rust races induced H2O2 accumulation in guard cells. In the incompatible interaction, a rapid increase of O2 and H2O2 generation at infection sites was detected. The percentage of infection sites showing NBT and DAB staining was 36.1% and 40.0%, respectively, 12 h after inoculation (hai). At extended incubation time until 24 hai, percentage of infection sites showing H2O2 accumulation further increased, whereas those exhibiting O2 accumulation declined. The early infection stage from 12 to 24 hai coincided with primary haustoria formation in mesophyll cells. In contrast, in the compatible interaction, O2 and H2O2 generation could not be detected in most of the infection sites. In the incompatible interaction, intensive DAB staining was also determined in mesophyll cells, especially in cell walls, surrounding the infected cells 16–24 hai; thereafter, these cells contained fluorescing compounds and underwent hypersensitive response (HR). The number of necrotic host cells surrounding the infection sites increased continuously from 20 to 96 hai. It might be concluded that H2O2 accumulation during the early infection stage is associated with the occurrence of hypersensitive cell death and that resistance response is leading to arrest the avirulent race of the obligate stripe rust pathogen. In the compatible interaction at 96 hai, H2O2 accumulation was observed in mesophyll cells surrounding the rust lesion.  相似文献   

13.
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.  相似文献   

14.
Seven isolates of the pathogen Colletotrichum musae (Berk & Curt.) v. arx. were isolated from banana fruit. These isolates produced ethylene to varying degrees in methionine-amended Czapek Dox liquid medium as both shake and static cultures. Rates of ethylene production by C. musae were positively associated with the concentration of methionine in the growth medium. C. musae did not produce ethylene on basal medium containing l-glutamate, α -ketoglutarate or l-cysteine. Isolate CM 100 produced the highest cumulative amount of ethylene (2·27 μm g−1 dry wt) over 12 days on 35 mm methionine-amended shake cultures of basal medium. In the presence of methionine, ethylene biosynthesis by C. musae occurred via 2-keto-4-methylthiobutyric acid (KMBA). The capacity of C. musae to produce ethylene may have a role in its pathogenicity on climacteric banana fruit.  相似文献   

15.
Cowpea (Vigna unguiculata) is a subsistence crop for small and poor farmers from Latin America and Africa. This culture is commonly damaged by cowpea weevil (Callosobruchus maculatus), which burrow into stored seeds to fed on. Due to impact of larval predation, several plant defense studies have been developed, indicating that α-amylase inhibitors are able to impede and/or reduce bruchids digestive process. In this report, a novel α-amylase inhibitor from papaya seeds (Carica papaya) with activity against cowpea weevil enzymes was purified and biochemical characterized. Peeled seeds were macerated and extracted with a 0.6 M NaCl and 0.1% HCl solution. Crude extract was precipitated with ammonium sulphate (100%). After dialyses, this rich fraction was applied onto a CM-Cellulose column and retained peak was submitted to an analytic reversed-phase column HPLC (Vydac C-18TP) yielding several peaks. Only one fraction, with molecular mass of 4562 Da, showed significant inhibitory activity against C. maculatus α-amylases. Otherwise, no inhibitory activities against mammalian α-amylases were observed. Bioassays using artificial seeds containing C. papaya α-amylase inhibitor rich fraction (0.5% and 1.0%) were also conduced showing that α-amylase inhibitors were able to increase larval mortality (50%) and also decrease insect fecundity and adult longevity. These results showed the presence of an α-amylase inhibitor from C. papaya seeds with high specificity to insect enzymes, indicating that this inhibitor probably could be used, through genetic engineering, in the construction of transgenic plants with enhanced resistance toward cowpea weevil.  相似文献   

16.
The Mexican bean weevil Zabrotes subfasciatus (Coleoptera: Chrysomelidae: Bruchinae) is among the key pests of common bean Phaseolus vulgaris L. around the world. Identifying resistance sources and the resistance types involved is important for development of pest-resistant bean varieties. This study assessed the adult attractiveness and oviposition preference of Z. subfasciatus toward eight genotypes of common bean, including seven pre-selected resistant genotypes and one standard for susceptibility. Attraction to the genotypes was assessed at one and four days after infestation. Oviposition preference was tested under free- and no-choice (confinement) conditions. We observed that IAC 853 was the least oviposited genotype in the free-choice test. However, this finding was not confirmed by the no-choice test. Therefore, IAC 853 could not be characterized as resistant to Z. subfasciatus. Presence of the toxic protein arcelin does not appear to influence host selection by Z. subfasciatus.  相似文献   

17.
Cecropin B (CB) is a very efficient antimicrobial agent against Xylella fastidiosa. CB activity decreased after incubation with xylem fluid from Vitis vinifera cv. Chardonnay. SDS-PAGE demonstrated the existence of protein(s) in xylem fluid of V. vinifera cv. Chardonnay that interact with CB and inhibit CB activity. The antimicrobial activity of CB was dependent on incubation time and colony size of X. fastidiosa. X. fastidiosa colonies in periwinkle wilt medium plus (PW+), and chemically defined media (3G10-R and CHARD2) showed different patterns in colony size and colony number in these media. After 1 h of incubation in 10 μM CB, only large-sized colonies were observed in the three media. These results suggested that most of the bacterial cells or small aggregates in suspension were killed by CB. Aggregation of X. fastidiosa cells may serve as a mechanism of protection against CB.  相似文献   

18.
Three 2,4‐diaminopyrimidines were tested against several insect species. They were active against lepidopteran pests with LC50 values <3 mg liter−1 for most species tested. They were also active against two‐spotted spider mite, Tetranychus urticae, (LC50 10–40 mg liter−1). Folinate, but not hypoxanthine or thymidine was found to be an effective rescue agent, requiring a concentration of 100 mg liter−1 diet to rescue half of the intoxicated larvae. The results confirm dihydrofolate reductase to be the site of action for these insecticides and are consistent with the mode of action of folinate rescue in mammals. © 2000 Society of Chemical Industry  相似文献   

19.
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.  相似文献   

20.
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.  相似文献   

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