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1.
An attempt was made to survey systems which carry out reduction of four pesticidal chemicals, heptachlor, parathion, 4,4-dichlorobenzophenone, and carbophenothion sulfoxide, in rat liver microsomes and in the supernatant in vitro. On the basis of the stimulatory effects of nicotinamide cofactors (NADPH and NADH), a flavin cofactor (FAD), localization of the systems, and their sensitivities to heat-inactivation treatment, four different reductive systems were recognized. They are (a) the mixed-function oxidase-coupled reductive systems, (b) an independent NADH-stimulated (instead of NADPH as above) microsomal system for parathion reduction, (c) FAD-stimulated system for reduction of carbophenothion sulfoxide and parathion, present specifically in the supernatant, and (d) a heat stable, nonenzymatic reduction system in the microsomes operated by flavoproteins and a flavin cofactor.  相似文献   

2.
This study was conducted to determine if metabolites of mexacarbate contribute to its residual toxicity to spruce budworm (Choristoneura fumiferana) larvae. Potted white spruce (Picea glauca) trees were treated with ‘Zectran’ UCZF # 19 at 100 g a.i. ha?1 and kept in a glasshouse. Mexacarbate residues declined by 98–99% within three days and reached non-detectable levels 10 days after treatment. Mortality of larvae fed on buds from these trees declined more gradually and was still 19–27% when exposed 10 days after treatment. The very low levels of mexacarbate (<0.07 μg g?1) found after three days did not produce such mortality. Gas chromatographic analysis of metabolites in needles revealed that after three days, 4-methylformamido-3,5-xylyl N-methylcarbamate was present at levels 20–30 times higher than the parent compound. This metabolite was about 50 times less toxic than mexacarbate to larvae when applied topically but was only 7 times less toxic when ingested. Two other methylcarbamate metabolites, the amino, and methylamino analogues were detectable for one day following treatment but not at later time points. They were as toxic as mexacarbate both topically and orally. Based on these findings, the methylformamido analogue could contribute to the residual toxicity of mexacarbate treatments of spruce.  相似文献   

3.
Homogenates of larvae, pupae, and adults of house flies (Musca domestica L.), flesh flies (Sarcophaga bullata Parker), and blow flies (Phormia regina (Meigen)) have been examined for enzymes which convert α- and β-ecdysone to apolar products. Most of the activity was found in the soluble fraction from house flies and flesh flies but none of the blow fly fractions was active. Two enzymes seem to be involved in the ecdysone metabolism, one requiring NADPH and the other functioning without this cofactor. The product of the latter enzyme is thought to be the 3-dehydro-ecdysone. This product is further converted to the 3α-hydroxy isomer of ecdysone by the NADPH-requiring enzyme. On feeding the insect growth regulator TH-6040 (1-(4-chlorophenyl)-3-(2,6-difluorobenzoyl)-urea) to larvae at dietary levels ranging from 0.3 to 10 ppm, the activity of the enzyme producing the 3-dehydro product is reduced by 20 to 82%. It is suggested that the growth regulator exerts its effect on pupal-adult ecdysis through its inhibition of ecdysone metabolism.  相似文献   

4.
p,p′-DDT was converted to DDD, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane, by unheated 12,000g supernate supplemented with NADPH, made from liver of rat, mouse, hamster, quail, chicken and pigeon. The additional presence of exogenous riboflavin did not augment reduction. If, however, riboflavin was added to unheated microsomes supplemented with NADPH or NADH the rate of reduction was more than doubled.Heated 12,000g supernate, unsupplemented or containing only exogenous riboflavin, did not reduce DDT. When NAD(P)H was present, big species differences in activity occurred, the pigeon and cockerel scarcely reducing any DDT in 2 hr, the Wistar rat supernate being most active. Addition to heated 12,000g supernate of 30 μg riboflavin as well as NAD(P)H resulted in increased reductive activity for all six species. The species difference in effect of exogenous riboflavin between the pigeon and rat was also observed using four DDT analogs as substrates. A sex-related difference in activity of preheated, supplemented 12,000g supernate from livers of the chicken and Norwegian hooded rat was not found when unheated preparations were used. In contrast to the activity possessed by preheated 12,000g supernate of rat liver supplemented with NAD(P)H, similarly supplemented preheated microsomes did not reduce DDT. On adding riboflavin as well as NAD(P)H, however, preheated hepatic microsomes of both pigeon and rat produced DDD, and this activity was further increased by addition of unheated microsomal (105,000g) supernate. The biocatalytic system functioning in preheated preparations appears to need at least one component of heated microsomes, NAD(P)H and one or more water-soluble components.  相似文献   

5.
覆膜条件下冬小麦根系生理特性及其空间分布变化   总被引:1,自引:0,他引:1  
覆膜条件下测定了冬小麦的根系活力与吸收面积,研究其空间分布情况。结果表明,在水平方向上,TTC还原量和TTC还原强度均随着离主茎距离的增加而减少;在垂直方向上,TTC还原量随着土层深度的加深而递减,TTC还原强度,返青期由上而下递减,开花期则上下土层无差异。根系总吸收面积和活跃吸收面积,在水平方向和垂直方向的变化与TTC还原量相同。覆膜处理并没有改变以上各指标的空间分布状态。但返青期水平方向的比吸收面积和比活跃吸收面积的变化趋势随覆盖地膜与否而不同,不覆膜处理由近到远递减,覆膜处理则无差异。返青期比吸收面积和比活跃吸收面积的垂直方向变化以及开花期二者的水平和垂直方向变化均无明显差异。根系活力与根系总吸收面积、活跃吸收面积呈极显著正相关。  相似文献   

6.
The temporal and spatial expression of three promoters was investigated in transgenic rice plants using promoter-β-glucuronidase (gusA) reporter gene fusions. The promoters studied were ubiquitin-1 (UBI-1) of Zea mays, Cauliflower Mosaic Virus 35S gene (CaMV35S) and a tubulin gene (TUB-1) of Arabidopsis thaliana. The TUB-1 promoter provided 7.32-fold more GUS activity in roots relative to tillers. This was significantly different from the corresponding value of 2.82-fold for CaMV35S but not from that of 4.55-fold for UBI-1, activity of both promoters was higher in the root tips and zone of elongation than mature roots. This younger root tissue represented a declining proportion of the expanding root system with time. Older tissue expressing GUS under control of the TUB-1 promoter showed a steeper decline in activity with time than occurred with the UBI-1 promoter. Nematode infection did not alter the overall pattern of expression from the two promoters, except that the giant cells induced by Meloidogyne incognita retained TUB-1 promoter activity as roots matured. Pratylenchus zeae invaded older root regions than M. incognita and no changes in promoter activity were detected where it fed. The results suggest the TUB-1 promoter has characteristics that favour its use for delivering anti-feedants, such as cysteine proteinase inhibitors, to M. incognita.  相似文献   

7.
The metabolism of etrimfos, O,O-dimethyl-O-(6-ethoxy-2-ethyl-4-pyrimidinyl) phosphorothioate was studied in vitro in a diazinon-resistant (Rutgers) and a susceptible (CSMA) strain of house flies. Practically no metabolism of etrimfos occurred without the addition of cofactors. However, the addition of the cofactor, reduced glutathione, resulted in a substantial amount of metabolism in both strains, the metabolism being higher in the resistant strain. The major route of metabolism was via the glutathione transferase system and the predominant metabolite was desmethyl etrimfos. Although the oxygen analog could not be isolated, microsomal oxidation of etrimfos resulted in the inhibition of acetylcholinesterase, suggesting the formation of the oxygen analog. Bovine serum albumin also degraded etrimfos yielding desmethyl etrimfos and 6-ethoxy-2-ethyl-4-hydroxypyrimidine.  相似文献   

8.
Microsomal sulfoxidation in fall armyworm (Spodoptera frugiperda) larvae was examined using phorate as substrate. The system required NADPH and was inhibited by CO and by piperonyl butoxide. Sulfoxidase activity was found in the alimentary canal, fat body, and Malpighian tubules, with the midgut being the most active. Microsomal substrates such as aldrin, heptachlor, biphenyl, and methyl parathion significantly inhibited the enzyme activity whereas p-nitroanisole and p-choloro-N-methylaniline had no effect. Enzyme activity increased during larval development, reaching a maximum shortly before pupation. Allelochemicals (monoterpenes, indoles, and flavones), drugs (phenobarbital and 3-methylcholanthrene), and host plants (corn cotton, parsnip, and parsley) significantly increased the enzyme activity. Increased phorate sulfoxidation through enzyme induction was found to decrease oral toxicity of phorate to the larvae. Analyses of internal insecticide revealed that, at various intervals, induced larvae retained less phorate and its oxidative metabolites than the controls. It was concluded that induction of phorate sulfoxidase activity results in an overall increase in the rate of phorate detoxication in this insect.  相似文献   

9.
Glutathione (GSH) content and GSH S-transferase activity are consistently increased in corn roots on 24-hr exposure of corn seedlings to part per million levels of N,N-diallyl-2,2-dichloroacetamide (R-25788) and related antidotes for thiocarbamate herbicide injury in susceptible corn varieties. This combined enhancement of enzyme activity and cofactor level leads to rapid detoxification of thiocarbamate sulfoxides, which are proposed to be the active herbicidal compounds formed on metabolic sulfoxidation. S-(N,N-Dipropylcarbamyl)-GSH is formed by this enzyme-catalyzed detoxification of EPTC sulfoxide. This hypothesis on antidote mode of action is supported by studies on 32 dichloroacetamides and related compounds and on the concentration- and time-dependent relationships of R-25788 action. The liver GSH content is normal in mice injected with high doses of R-25788, but the content is reduced when EPTC or EPTC sulfoxide is administered. EPTC sulfoxide also carbamoylates the thiol group of coenzyme A in neutral aqueous medium.  相似文献   

10.
11.
A procedure was developed to obtain non-embryogenic callus and regenerated lines from root segments of Zea mays grown in aseptic conditions. The activity of glutathione-S-transferases (GSTs), for non-embryogenic callus, was determined toward 1-chloro-2,4-dinitrobenzene (CDNB) and it was compared with that obtained for corn seedlings grown without hormones. For the callus masses, increases of specific activity toward CDNB and the kinetic parameter Vmax were observed with respect to corn seedlings. The procedure permitted the regenerating of tissues from callus explants, therefore the GST(CDNB) activity and the effect of the safener benoxacor on its expression were investigated for the regenerated tissues grown in agarized substrate and in liquid medium. These explants showed a constitutive GST(CDNB) activity higher than corn seedlings and this activity was increased, for both tissues, in response to the presence of the safener benoxacor in the growing medium. The GST activity for the above tissues was also assayed toward benoxacor and terbuthylazine, metolachlor and fluorodifen herbicides. Measurable GST activity was found toward some of the above chemicals and it was found to be significantly enhanced in response to benoxacor treatment.  相似文献   

12.
This study presents evidence for the dehydrogenation of lindane by a hepatic microsomal mixed-function oxidase system. Preliminary investigation established that the incubation of lindane with rat liver homogenates produces a chlorinated, nonpolar compound identified as hexachlorocyclohexene. Differential centrifugation resulted in the sedimentation of most of the dehydrogenase activity in the microsomal fraction. Optimum in vitro assay conditions were established and it was found that the dehydrogenase system required molecular oxygen and reduced pyridine nucleotide coenzyme for maximum activity. Inhibition by SKF 525-A and CO suggested that the enzyme was cytochrome P-450 dependent. Lack of inhibition by cyanide indicated that the cytochrome b5 desaturase system was probably not involved. Pretreatment of rats with DDT, which stimulates lindane metabolism, also induced significantly higher dehydrogenase activity. Both the in vivo and in vitro metabolism of hexachlorocyclohexene produced previously identified lindane metabolites. The existence of a cytochrome P-450 dependent mixed-function oxidase which catalyzes the dehydrogenation of lindane has not previously been reported and may be of importance in the metabolism of other xenobiotics.  相似文献   

13.
The amounts of p,p′-DDT and of five other trichloroethane derivatives decreased upon incubation under anaerobic conditions with 12,000g × 30 min pigeon liver supernatant fraction. The addition of an exogenous NADPH-generating system sometimes, but not invariably, increased the rate of metabolism. Only one hexane-soluble metabolite was detected in the postincubation reaction medium of each of the six trichloroethane derivatives. After isolation by tlc the six metabolites were shown by mass spectrometry to have molecular weights 34 units less than their parent compounds. Comparison of the isotope patterns in the spectra of each substrate and its metabolite reveals that in each case the metabolism involves the loss of a chlorine atom. From these data it is concluded that several substituted trichloroethanes undergo reductive dechlorination when they are incubated with liver preparations in an atomosphere of nitrogen. Two dichloroethane derivatives, tested in a similar manner, were unchanged and were recovered quantitatively. Mass spectrometric and chromatographic data of reactants and products are recorded.  相似文献   

14.
Studies were conducted to assess the contribution of the hepatic microsomal mixed function oxidase system to a 7.2-fold difference in susceptibility to the lethal effects of endrin between endrin-resistant and -susceptible pine voles, Microtus pinetorum. Evaluations of microsomal enzyme systems were conducted for basal and endrin-treated pine voles of both strains. The microsomal activity of ICR white mice was investigated to provide a species comparison. Maximal microsomal mixed function oxidase activities were determined in in vitro incubations for the model substrates ethylmorphine, aniline, and benzo(a)pyrene. Vmax values were estimated for the rate of disappearance of benzo(a)pyrene in in vitro incubations. No significant strain differences in basal microsomal enzyme activity were found for the model substrates investigated, although activity was invariably higher in the resistant strain. The concentration of cytochrome P-450 was significantly higher in the resistant vole though actually less than 20% different. The occurrence of significant strain differences in the levels of microsomal enzyme activity induced by endrin were rare. Significant endrin treatment effects on the levels of microsomal enzyme activity for the pine vole were observed but the degree and direction of change depended on the substrate used. A marked species difference in microsomal mixed function oxidase activity was noted between pine voles and white mice. This was particularly evident for endrin-treated animals. The microsomal activity of endrin-treated white mice was greatly induced relative to basal levels. The degree of induction depended on the substrate used. The small strain differences in microsomal enzyme activity for the systems investigated were judged to be insufficient to explain the strain difference in susceptibility to endrin.  相似文献   

15.
The activity of key biochemical defense mechanisms in seedlings of winter oilseed rape cultivars, differing in tolerance to infection with Leptosphaeria maculans, was investigated. The studied winter oilseed rape cultivars differed in their biochemical defense potential. In the resistant cultivar, a significant increase in the activity of chitinase and β-1,3-glucanase was observed that can be regarded as a symptom of a systemic defense reaction. In addition, the resistant cultivar had a more efficient antioxidant system, that is, higher activity of specific SOD isoforms and higher level of low-molecular antioxidants. Only the resistant cultivar had three Fe-SOD sub-isoforms. A correlation between the content of cell wall-bound phenolics and hydrogen peroxide in the resistant cultivar may indicate that an important method of H2O2 neutralization was its participation in the process of phenolics incorporation into the cell wall. Elevated content of cell wall-bound phenolics significantly enhances the cell wall integrity, which then becomes a more effective firewall against L. maculans. Moreover, high utilization of soluble sugars during synthesis of the phenolic compounds in the pathogenesis was demonstrated in the resistant cultivar. We conclude that the key factors contributing to a greater tolerance to L. maculans were an effective antioxidant system and higher activity of PR proteins, i.e., chitinase and β-1,3-glucanase.  相似文献   

16.
Antifungal activity of oligochitosan against nine phytopathogens was investigated in vitro. Oligochitosan was more effective than chitosan in inhibiting mycelial growth of Phytophthora capsici and its inhibition on different stages in life cycle of P. capsici was observed. Rupture of released zoospores induced by oligochitosan was reduced by addition of 100 mM glucose. The effects of oligochitosan on mycelial growth and zoospore release, but not zoospore rupture, were reduced largely when pH value was above 7. The ultrastructural study showed that oligochitosan caused distortion and disruption of most vacuoles, thickening of plasmalemma, and appearance of unique tubular materials. Plasmalemmasomes in hyphal tip cells were not found in the presence of oligochitosan. These results suggest polycationic nature of oligochitosan contributes only partly to its antifungal activity and multiple modes of action of oligochitosan exist including the disruption of endomembrane system.  相似文献   

17.
Experiments on the effect of various compounds with plant growth regulating activity led to the hypothesis that conditions inhibitory to indoleacetic acid (IAA) action or leading to a decrease in the IAA level in cucumber seedlings would be unfavourable for the development of cucumber scab, caused byCladosporium cucumerinum. Susceptibility decreased as the result of treatment with growth retardants, which cause an increase in IAA-oxidase activity in the plants, whereas application of indoleacetic acid or compounds expected to decrease the rate of oxidative breakdown of IAA in the seedlings resulted in an increase of susceptibility. Growing the plants under various periods of illumination also influenced both the susceptibility of the plants and the IAA-oxidase activity in the hypocotyl tissue.To obtain further information about the relation between the increase in resistance and the increase in the rate of IAA oxidation, the effect of one compound, viz. L-threo-β-phenylserine (abbr. phenylserine) was studied in more detail. The rate of oxidative breakdown of IAA was increased in extracts of hypocotyls and cotyledons of plants treated with phenylserine as compared with extracts of tissue of untreated plants. Extracts of phenylserine-treated plants were found to contain a higher cofactor and/or a lower inhibitor content than those of control plants. The substances involved were not identified, but this result may indicate a shift in the concentrations of various phenols in the tissue.The question must remain open whether a shift in the concentration of phenols some-how effected by phenylserine treatment determines both degree of susceptibility and rate of IAA oxidation independently, or whether the rate of IAA breakdown and the resulting change in IAA content in the plants is directly related to the degree of susceptibility.  相似文献   

18.
Resistance to organophosphorus (OP) insecticides in Lucilia cuprina arises from two mutations in carboxylesterase E3 that enable it to hydrolyse the phosphate ester of various organophosphates, plus the carboxlyester in the leaving group in the case of malathion. These mutations are not found naturally in the orthologous EST23 enzyme in Drosophila melanogaster. We have introduced the two mutations (G137D and W251L) into cloned genes encoding E3 and EST23 from susceptible L. cuprina and D. melanogaster and expressed them in vitro with the baculovirus system. The ability of the resultant enzymes to hydrolyse the phosphate ester of diethyl and dimethyl organophosphates was studied by a novel fluorometric assay, which also provided a sensitive titration technique for the molar amount of esterase regardless of its ability to hydrolyse the fluorogenic substrate used. Malathion carboxylesterase activity was also measured. The G137D mutation markedly enhanced (>30-fold) hydrolysis of both classes of phosphate ester by E3 but only had a similar effect on the hydrolysis of dimethyl organophosphate in EST23. Introduction of the W251L mutation into either gene enhanced dimethyl (23-30-fold) more than diethyl (6-10-fold) organophosphate hydrolysis and slightly improved (2-4-fold) malathion carboxylesterase activity, but only at high substrate concentration.  相似文献   

19.
The effects of p,p′-DDT and four of its analogs on electrical activity in the central nervous system of the cockroach, Periplaneta americana (L.), were investigated. Cockroaches were injected intraabdominally with an organochlorine compound at LD50 96-hr doses (except for p,p′-DDE). Extracellular recordings were made from the central nervous system at 1 hr, 24 hr, or 3 weeks postinjection. p,p′-DDT, methoxychlor, and p,p′-DDD induced behavioral changes (tremors, jitters, hyperexcitability) and repetitive firing in the central nervous system prior to 1 hr postinjection. By 24 hr postinjection, most behavioral signs of poisoning had disappeared, though repetitive firing could still be readily elicited in the central nervous system. Cockroaches injected with o,p′-DDT, however, usually required about 48 hr before overt signs of poisoning became apparent. Cockroaches treated with p,p′-DDT or o,p′-DDT behaved normally at 3 weeks postinjection but still displayed a significant occurrence of repetitive firing in the central nervous system. A mechanism is proposed to explain how a cockroach might recover behaviorally from a sublethal dose of an organochlorine compound but still display repetitive firing in its central nervous system. A direct “cause and effect” relationship between repetitive firing in the central nervous system and mortality (and external signs of poisoning) is therefore questioned.  相似文献   

20.
The GST cDNA was successfully cloned from an Oryza sativa cDNA library by PCR using oligonucleotide primers based on the OsGSTU5 (GenBank Accession No. AF309377) sequence. The cDNA was composed of a 687-bp open reading frame encoding for 228 amino acids. The deduced amino acid sequence of this gene shared over 60% sequence identity with the sequences of the tau class ZmGSTU6 and ZmGSTU19. This gene was expressed in Escherichia coli with the pET vector system and the gene product was purified to homogeneity by GSH-Sepharose affinity column chromatography. The expressed OsGSTU5 formed a homo-dimer composed of 25 kDa subunit and its pI value was approximately 7.5. The OsGSTU5 displays high activities towards 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy)propane. The activity of the OsGSTU5 was significantly inhibited by hematin and ethacrynic acid. The OsGSTU5 shows the highest activity towards chloro-s-triazine and acetanilide herbicides.  相似文献   

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