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1.
An insect chitin synthetase (CS) is readily assayed using the microsomal fraction (～0.5 mg protein) from an homogenate of Tribolium castaneum larvae. This enzyme preparation is incubated at 22°C with uridine 5′-diphospho-N-acetyl[3H]glucosamine in 355 μl of 25 mM Tris-HCl buffer containing 10 mM MgCl2, 17 mM N-acetylglucosamine, and 1 mM dithiothreitol. Other divalent cations and amino sugars are less effective activators or are inhibitory. T. castaneum CS is strongly inhibited by polyoxin D and uridine 5′-diphosphate. These activation and inhibition properties of Tribolium castaneum gut CS are similar to those of fungal CS. The polymerization product formed by the Tribolium enzyme is stable in alkali but hydrolyzed by chitinase. Enzymes of Tribolium confusum, Tribolium brevicornis, Tenebrio molitor, and Galleria mellonella are also active under the same conditions. These enzymes are from the gut and probably from the peritrophic membrane. Integumental CS activity is not detected under the indicated assay conditions.  相似文献   

2.
Chitin polymerization is catalyzed by cell-free enzyme complexes from the integument of Trichoplusia ni larvae and wing tissue of developing Hyalophora cecropia pupae obtained on extraction of homogenates for 16 hr at 5°C in 25 mM Tris-HCl buffer, pH 7.2, containing 10 mM MgCl2, 1 mM dithiothreitol, 10 mg/ml bovine serum albumin, and 4 mg/ml digitonin. In contrast, integumental preparations from Boarmia selenaria, Earias insulana, Heliothis virescens, Oncopeltus fasciatus, Spodoptera exigua, and Tribolium castaneum exhibit little or no chitin synthetase (CS) activity. H. cecropia CS requires magnesium ions but not N-acetyl-d-glucosamine for normal activity and is almost insensitive to nikkomycin and polyoxin D. CS activity is not detected in diapausing H. cecropia pupae but synthesis of this enzyme is induced or its activity is stimulated by the molting hormone, ecdysterone, indicating possible hormonal control. T. ni CS requires magnesium ions and N-acetyl-d-glucosamine for optimal activity and is sensitive to inhibition by uridine di- and triphosphates, polyoxins B and D, and particularly nikkomycin. T. ni integumental CS activity decreases in starved larvae or those about to pupate. Both T. ni and H. cecropia CS enzymes as prepared and assayed are sensitive to captan but not to the potent insecticides diflubenzuron and BAY SIR 8514, two effective benzoylphenyl urea in vivo chitin synthesis inhibitors.  相似文献   

3.
A chitin synthetase (CS) preparation was obtained from mycelium of the plant pathogen Sclerotium rolfsii and partially characterised. The CS activity was stimulated by N-acetyl-D-glucosamine and magnesium cations with an optimum temperature of 30°C and an optimum pH of 6.4. The enzyme required proteolytic activation and so apparently existed in a zymogenic form. The fungal CS was strongly inhibited by the nucleoside-peptide antibiotics polyoxin-D and nikkomycin, and weakly inhibited by the terpenoyl benzimidazole, 1-geranyl-2-methylbenzimidazole. It was insensitive to captan and to insecticidal benzoylphenyl ureas. Disrupted chitin formation caused by polyoxin-D showed as swollen hyphae containing numerous depressions. The antibiotic also reduced formation of sclerotia and the few that did form were large, irregular in shape and light coloured.  相似文献   

4.
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   

5.
The effect of sublethal concentrations, 0.00141% (LC20), 0.00251% (LC30), and 0.0036% (LC40) of a dispersible formulation of an acylurea insect growth regulator, flufenoxuron (Cascade) on certain biochemical parameters in the larvae of Tribolium castaneum was investigated. When neonates were fed on diet treated with sublethal concentrations for 24 h, it was observed that at all concentrations tested, there was a significant reduction in chitin content on the 15th day of development. Total soluble protein content at LC20 and LC30 decreased with increasing age of the larvae. At LC20 and LC40 concentrations there was a progressive increase in the protein: chitin ratio as a function of increase in age of the larvae. SDS-PAGE analysis of the larval tissue extracts indicated gross quantitative changes in some of the protein bands (MW 50-97 kDa). Western blot analysis revealed significant increase in the level of HSP70 in the extracts of larvae fed on LC30 treated diet, on the 7th and 10th day of development in the decreasing order. Conversely, a significant decrease in the hyper-phosphorylated form of p34cdc2 kinase due to flufenoxuron treatment indicating modulation of cell cycle regulation was observed. Thus, sublethal concentrations of flufenoxuron alter expression of developmentally regulated proteins, HSP70 and p34cdc2 and chitin formation in a stage-specific manner thereby resulting developmental abnormalities in T. castaneum.  相似文献   

6.
The chitin synthesis inhibitors diflubenzuron and teflubenzuron have recently become ineffective for the control of Spodoptera exigua in floricultural crops. An extended laboratory test with second-instar larvae of S. exigua on Vicia faba plants was carried out to determine the influence of synergists on the biological activity of three benzoylphenyl ureas (BPUs). The co-application of piperonyl butoxide, an oxidase inhibitor, did not increase the activity of diflubenzuron, teflubenzuron or hexaflumuron. The best results were obtained with diethyl maleate, for suppressing glutathione S-transferase activity, and with dimethoate, as a hydrolase inhibitor. A joint application of diflubenzuron (at a concentration which resulted in 43% survival) with diethyl maleate or dimethoate gave only 6.2 and 8.9% surviving larvae, respectively. In addition, development to fourth-instar larvae was inhibited. The more stable teflubenzuron was synergized by both compounds to a much lesser extent than diflubenzuron. None of the synergists had a significant effect on the activity of hexaflumuron, which was the most potent insecticide of the three BPUs tested against S. exigua.  相似文献   

7.
Tetraphenylstibonium chloride (Ph4SbCl) at a dietary level of 500–2000 μmol/kg inhibits larval growth, gut invertase activity, pupation, and emergence of Tribolium confusum and Tribolium castaneum. Tetraphenylphosphonium chloride (Ph4PCl) in this concentration range inhibits growth and gut invertase activity without affecting pupation and emergence whereas tetraphenylarsonium chloride is essentially inactive in all respects. Triphenylstibine (Ph3Sb) but not the other triphenyl derivatives (amine, phosphine, and arsine) inhibits both larval growth and gut digestive enzyme activities (invertase and protease) in vivo. Protease but not invertase is inhibited in vitro by Ph4PCl and Ph4SbCl at 2 × 10?3M. Ph4PCl appears to act as an antifeeding agent and Ph4SbCl and Ph3Sb as more general toxicants.  相似文献   

8.
Two fungal metabolites, aspyrone (3-(1,2-epoxypropyl)-5-hydroxy-6-methyl-5,6-dihydropyran-2-one) and asperlactone (3-(1,2-epoxypropyl)-5-(1-hydroxyethyl)-5-furan-2-one) were isolated from an Aspergillus ochraceus Wilhelm strain showing IGR activity against Tribolium castaneum (Herbst). Synthetic derivatives of aspyrone were produced using published methods. These derivatives together with aspyrone and asperlactone were tested for insect growth-regulating activity against T. castaneum, and for ovicidal activity against Nezara viridula L. Of the compounds tested asperlactone appeared to be the most active.  相似文献   

9.
The effect of methyl bromide in admixture with 20%, 30% and 40% carbon monoxide (CO) onTribolium castaneum adults was tested. The combination of methyl bromide with 20% CO increased the toxicity of CH3Br 1.4-fold, with 30% CO-1.5-fold, and with 40% CO-1.7-fold. It is supposed that the inhibitory effect of CO on the insect’s microsomal mixed-function oxidase and on other enzymatic systems causes this effect.  相似文献   

10.
In a topical bioassay with pupae and adults of the mealwormTenebrio molitor (Coleoptera: Tenebrionidae), an important pest in stored products worldwide, three insect growth inhibitors (IGRs) were tested: the two chitin synthesis inhibitors diflubenzuron and flucycloxuron, and the ecdysone agonist halofenozide (RH-0345). To address differences in toxicity, we determined with the use of a14C-labeled isotope, the pattern of absorption through the cuticle and then the distribution in the different body tissues of gut, hemolymph, integument and reproductive organs (ovaries, testis). The rate of absorption through the cuticle was highest for flucycloxuron, and this concurred with its high toxicity. In general, the patterns of absorption followed a similar trend in young and old pupae and in the adult stage. Accumulation in the reproductive system of females and males was relatively high, which may explain the strong reproductive effects of the IGRs tested. In addition, we recorded large differences in the clearance of insecticide from the insect bodyvia the excrements, with the chitin synthesis inhibitors exhibiting higher rates than halofenozide.  相似文献   

11.
A study was conducted to determine the extent of resistance to malathion in field populations of insects collected from nine granaries located in different regions of Israel. The results showed that the maximum resistance factor calcuated from the LCso s of the different insect species tested was:Tribolium castaneum (Herbst), x 538.0;Oryzaephilus surinamensis (L.), x 8.0;Sitophilus oryzae (L.), x 1.2; andRhyzopertha dominica (F.), x 9.0. There were significant differences between the resistance level among strains collected from different locations in Israel. By using triphenyl phosphate (TPP), an inhibitor of carboxyesterase, it was shown that, in the case ofT. castaneum andR. dominica, the resistance is a malathion-specific type and that in the case ofO. surinamensis it is partially non-specific to malathion. The significance of these findings in selecting new insecticides to replace malathion as a grain protectant was considered.  相似文献   

12.
In this review an effort was made to summarize the up to date information on the knowledge on the action mechanism of diflubenzuron (DFB), a prototype chemical for the benzoylurea type insecticides, with respect to its molecular mechanism to inhibit insect chitin synthesis. The key problem in pinpointing the action site of this insecticide has been the lack of in vitro demonstration of its action to inhibit insect chitin synthesis under cell free conditions. This problem was solved when an approach using a intracellular vesicle preparation from the cuticle of newly molted Periplaneta americana was developed. Using this approach it has become possible to identify that DFB indeed inhibits the process of incorporation of N-acetylglucosamine into insect chitin. Recently there has been a breakthrough in this field, when a sulfonylurea receptor (SUR) was identified in Drosophila melanogaster. This information was instrumental in establishing that insect SUR in the above intracellular vesicular preparation from P. americana as well as Blattella germanica is likely the actual target site of DFB to cause inhibition of chitin synthesis. The role of SUR in this case has been determined, by using glibenclamide, a typical SUR specific inhibitor as an aid, to be helping the exocytotic movement of these vesicles as is the case of other members of the group of ABC-transporters to which insect SUR belongs. In this case both DFB and glibenclamide have been shown to cause the depolarization of the vesicle membrane through inhibition of the K+ channel, which leads to their inhibition of chitin synthesis.  相似文献   

13.
The effect of IKI-7899 and diflubenzuron on larval development and emergence ofTribolium castaneum and ofSpodoptera littoralis was determined. The LC50 of IKI-7899 at adult emergence ofT. castaneum larvae fed the compound in their diet and the ED50 of S.littoralis larvae treated topically were approximately one-sixth those of diflubenzuron.  相似文献   

14.
This study aimed to characterize nanoparticles loaded with clove (Syzygium aromaticum) essential oil-based polyethylene glycol (PEG) and to know their insecticidal activity against red flour beetle (Tribolium castaneum). The nanoparticles have irregular shapes in good dispersion. The nanoformulation could not enhance clove oil contact toxicity to T. castaneum, but could protect the oil from degradation and evaporation while simultaneously allowing sustained release, as indicated by the continued high toxicity for 16 weeks of storage.  相似文献   

15.
The antifeedant effect of several new withanolides on larvae ofSpodoptera littoralis, Epilachna varivestis andTribolium castaneum was investigated. 2,3-Dihydrowithanolide E (II) was an antifeedant forS. littoralis andE. varivestis; nicalbin A (XIV), and a mixture of withanicandrin (IX) and daturalactone A (X), were active against E. varivestis andT. castaneum. Some activity was shown by 6β,14α,17β,20αF-tetrahydroxy-1-oxowitha-2,4,24-trienolide (V) againstS. littoralis; 4β,5β-epoxy-6α,14α,17β,20αF-tetrahydroxy-l-oxowitha-2,24-dienolide (VII) againstE. varivestis andT. castaneum; 5α,6α,-epoxy-14α,17β,20αF-trihydroxy-1-oxowitha-2,24-dienolide (VI) againstE. varivestis; and nicalbin B (XV) againstT. castaneum.  相似文献   

16.
The effects of the chitin synthesis inhibitor (CSI) novaluron on egg hatch and on larval development ofTribolium castaneum (Herbst) concentrations of 1.0, 0.3, 0.2 or 0.1 ppm were tested. The effect of novaluron at low concentrations depended strongly on the exposure period. At 0.3 ppm, egg hatch ofT. castaneum was totally inhibited after 28 days; at 0.2 ppm the effect was much less but inhibition increased progressively to 66% in the 35-day experiment; at 0.1 ppm novaluron was ineffective. The viability of the larvae that hatched from the laid eggs and developed on untreated flour was also dependent on concentration of novaluron and exposure time: exposure ofT. castaneum adults to novaluron-treated flour at 0.3 ppm for 8 days, or at 0.2 ppm for 36 days, caused 100% or 97.5% mortality, respectively. At both 0.3 and 0.2 ppm, larval deaths were mainly in the first instar. Exposure ofT. castaneum adults to treated flour may serve as a good model for evaluating the effect of CSIs on internal feeders, especiallySitophilus oryzae (L.). The present study contributes to our understanding of CSI transovarial activity against internal stored product coleopterans whose larval stage develops inside the grain without contact with the toxicants. http://www.phytoparasitica.org posting Dec. 11, 2007.  相似文献   

17.
The mode of action has been investigated of the 1-(2,6-dichlorobenzoyl)-3-phenyl ureas, a new type of insecticide. A microautoradiographical study was made of the incorporation of glucose, tyrosine, and proline in endocuticle of fifth-instar larvae of Pieris brassicae L., both normal and treated with 1-(2,6-dichlorobenzoyl)-3-(3,4-dichlorophenyl)-urea (DU 19111). The results of this work clearly indicate that DU 19111 blocks the synthesis of cuticular chitin. The nature of this inhibition was investigated by comparison of the rates of incorporation of [14C]glucose into the ultimate chitin precursor, uridine diphosphate N-acetyl glucosamine (UDPAG), in both normal and DU 19111-treated Pieris larvae. It was found that these levels did not differ significantly.The finding that only in DU 19111-treated Pieris larvae were substantial amounts of labeled N-acetyl glucosamine present 1 hr after the injection of [14C]glucose is considered as a clue to the mechanism by which this insecticide inhibits chitin synthesis. Apparently the coupling of UDPAG to the chitin synthetase still proceeds, but the function of connecting N-acetyl glucosamine moieties to the chitin chain is disrupted.Tentative results with a structural analog of DU 19111 suggest that this compound induces accumulation of UDPAG, but not of N-acetyl glucosamine. This would imply that in the latter case the polycondensing enzyme is completely blocked.  相似文献   

18.
The potency of diflubenzuron is much greater in inhibiting growth and development of 1st instar larvae ofTribolium castaneum than of 4th instar larvae, as expressed by death at the apolytic stage and retardation of larval development. A dose-dependent decrease in the activity of trehalase, invertase and amylase was obtainedin vivo with the increase in diflubenzuron concentration. At 5 ppm dietary concentration, a reduction of 37 and 27% in invertase and trehalase activity, respectively, was obtained in 4th instar larvae fed for 3 days on treated diet. The amylase activity was affected to a lesser extent. The observed disturbances of trehalase activity might hamper the supply of glucose needed for chitin build-up and those of invertase and amylase activity might affect feeding. Diflubenzuron does not inhibit these enzymesin vitro; hence, thein vivo effect seems to result from general disturbances in carbohydrate metabolism.  相似文献   

19.
Insect growth regulators (IGRs) have been proposed as agents for the control of insect pests. These compounds disrupt the normal development of insects by mimicking juvenile hormone and the molting hormone, 20-hydroxyecdsyone, or by interfering with chitin synthesis. The effectiveness and selectivity of IGRs provide new tools for integrated pest management. The simultaneous advances in the chemistry of IGRs and the ability to study insect tissues in culture, have led to research on the mode of action of IGRs in vitro. Plodia interpunctella and Spodoptera frugiperda have been used to examine the effects of IGRs on wing imaginal discs in organ culture, as well as in hormonally responsive cell lines established from wing imaginal discs of these species. Our research has focused on the action of ecdysteroid mimics, chitin synthesis inhibitors and juvenile hormone mimics. The effects of the IGRs on chitin synthesis, uptake of amino-sugars, and cellular proliferation were studied in tissue culture. The results demonstrate the effectiveness of using organ cultures and hormonally responsive cell lines for investigating IGRs at the cellular and tissue level. © 1998 Society of Chemical Industry  相似文献   

20.
BACKGROUND: Environmental impacts and resistance to insecticides pose serious challenges to stored‐product insect and other types of pest control. Insect‐resistant transgenic grain is a potential alternative to fumigants, but candidate control proteins are needed, especially for coleopterans. Therefore, we evaluated the efficacy of a coleopteran‐active toxin, Bacillus thuringiensis Cry3Aa, with or without protease inhibitors, in laboratory feeding assays against coleopteran storage pests. RESULTS: In a comparison of the toxicity of Cry3Aa protoxin towards three species of coleopteran storage pests, Tenebrio molitor L. was found to be most sensitive, Tribolium castaneum (Herbst.) was most refractory and Rhyzopertha dominica F. displayed an intermediate response. For R. dominica, Cry3Aa combined with 3500 mg potato carboxypeptidase inhibitor or 5000 mg aprotinin kg?1 diet resulted in both delayed development and increased mortality. Potato carboxypeptidase inhibitor and bovine aprotinin reduced the LC50 of Cry3Aa for R. dominica two‐ and threefold respectively. Cry3Aa treatment resulted in fewer progeny from R. dominica, and progeny was further reduced when the protoxin was combined with potato carboxypeptidase inhibitor. CONCLUSIONS: These data support the hypothesis that a combination of Cry3Aa protoxin and protease inhibitors, particularly a potato carboxypeptidase inhibitor, may have applications in control strategies for preventing damage to stored products and grains by coleopteran pests. Published 2011 by John Wiley & Sons, Ltd.  相似文献   

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