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Ecdysone 20-hydroxylase activity has been detected in pupal wing discs of Pieris brassicae. This activity is due to an enzyme system located in microsomal fractions. Its apparent Km is 58 nM for ecdysone. The enzyme is inhibited by the reaction product 20-hydroxyecdysone with an apparent Ki of 2.6 μM. Its activity varied during pupal-adult development with a maximum on day 4, when ecdysone levels are the highest in the animal. Although low, the peak activity is sufficient to assure 25% of the conversion of endogenous ecdysone into 20-hydroxyecdysone in pupae. Ecdysone and 20-hydroxyecdysone levels were measured in hemolymph and whole animals; ecdysone appears to be mainly located in hemolymph, whereas 20-hydroxyecdysone seems to be equally distributed between hemolymph and tissues. All these findings are discussed in relation to the roles of ecdysone and 20-hydroxyecdysone during pupal-adult development.  相似文献   

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Impatiens glandulifera (Himalayan balsam) is an invasive riparian plant species that can outcompete native perennials. Population genetic data on dispersal may aid in the management of invasive species, so we have developed microsatellite markers for this significant invader using an intersimple sequence repeat (ISSR)‐based cloning method. Eight polymorphic markers displayed between two and five alleles, with overall levels of observed and expected heterozygosities ranging from 0.0500 to 0.7500 and from 0.1449 to 0.7692, respectively.  相似文献   

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Wild type (Oregon R) and non-pupariating as well as late-pupariating mutant larval tissues were cultured in vitro up to 5 weeks with and without 20-hydroxy ecdysone (1 μg/ml). The following responses were elicited by the hormone: in the case of wild type tissues detachment of the larval epidermis and muscles from the cuticle; puparial tanning and sclerotization of the larval cuticle; dissociation of the fat body into single cells; inhibition of the movement of the hind intestine. Most of these responses developed within 1 week of culturing. Of the 4 mutants tested, 3 behaved like the wild type. In cultures of ?(1)npr-1, however, puparial tanning, disc evagination, and inhibition of the movement of the hind intestine was abnormally weak and the dissociation of fat body was not observed at all. Detachment of the epidermis and muscles as well as formation of the pupal cuticle by disc tissue occurred normally. The results are discussed with respect to the ecdysteroid-induced metamorphosis of the tissues and the autonomy of mutant gene action.  相似文献   

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We conducted genomic characterization based on SNP and SilicoDArT markers on the invasive Himalayan balsam (Impatiens glandulifera) plants originating from native and non‐native regions of their distribution. When genetic relationships were explored by PCoA using SNP and SilicoDArT marker data, the first, second, and third principal coordinates explained altogether 37.4% and 31.0% of the variability, respectively. Samples from the UK, Canada, and Pakistan were grouped together, while Indian plants were clearly distinct based on SNP markers but relatively close to the UK–Canada–Pakistan group based on SilicoDArT markers. Constructed trees differentiated individuals into clusters resembling the PCoA patterns. The Bayesian BAPS analysis performed for the SNP data revealed that the individuals were distributed in seven clusters, representing samples from each of the four Finnish populations, India, Pakistan, and the combination of the UK and Canada. Similar clustering was visible in the UPGMA tree. The Indian cluster did not display any ancestral gene flow with the others, while the Pakistani cluster showed ancestral gene flow only with the combined UK and Canada cluster. Furthermore, the latter cluster displayed ancestral gene flow with the Finnish populations varying from 0% to 3.1%. The BAPS analyses conducted for the SilicoDArT data differ slightly: The individuals were distributed in nine clusters, and the Indian cluster exhibited ancestral gene flow with the mixed cluster including Canadian, Pakistani, and UK samples, and one Finnish sample. The AMOVA showed that 45% and 26% of variation was present among the I. glandulifera groups/populations and the rest within them based on SNP and SilicoDArT markers, respectively. The Bayesian BAPS analyses and the gene flow networks were the most informative tools for resolving relationships among native and introduced plants. It is notable that the small sample sizes for non‐Finnish plant materials may affect the accuracy of the gene flow and other estimates.  相似文献   

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  总被引:1,自引:0,他引:1  
Recombinant Escherichia coli JM101 was used for the in vivo biocatalytic synthesis of 3-tert-butyl- catechol. The bacterial strain synthesized the laboratory-evolved variant HbpA(T2) of 2-hydroxybiphenyl 3-monooxygenase (HbpA, EC 1.14.13.44) from Pseudomonas azelaica HBP1. The mutant enzyme HbpA(T2) is able to hydroxylate 2-tert-butylphenol to the corresponding catechol, a reaction that is not catalyzed by the wild-type enzyme. The biotransformation was performed in a 3-L bioreactor for 24 h. To mitigate the toxicity of the 2-tert-butylphenol starting material, we applied a limited substrate feed. Continuous in situ product removal with the hydrophobic resin Amberlite XAD-4 was used to separate the product from culture broth. In addition, binding to the resin stabilized the product, which was important because 3-tert-butylcatechol is very labile in aqueous solution. The productivity of the process was 63 mg L(-1) h(-1) so that after 24 h, 3.0 g of 3-tert-butylcatechol were isolated. Down-stream processing consisted of two steps. First, bound 2-tert-butylphenol and 3-tert-butylcatechol were eluted from Amberlite XAD-4 with methanol. Second, the two compounds were separated over neutral aluminum oxide, which selectively binds the produced catechol but not the phenol substrate. The final purity of 3-tert-butylcatechol was greater than 98%.  相似文献   

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Total ecdysteroid levels as well as concentrations of several individual ecdysteroids were determined for hemolymph and testes of fifth instars, pupae, and pharate adults of the European corn borer, Ostrinia nubilalis (Hubner). For total levels, the patterns of fluctuation in hemolymph and testes were similar, but the concentrations in testes were lower than those in hemolymph. In both hemolymph and testes there were two ecdysteroid peaks: the first just prior to the formation of the pharate pupa, the second just prior to the formation of the pharate adult. An examination of ecdysteroid profiles revealed some important differences. Ecdysone was either absent or present at extremely low levels in larval testes, whereas in hemolymph there was a premolt ecdysone peak. In pupal testes, ecdysone was present, but levels of 26-hydroxyecdysone were much lower than those in hemolymph. Thus, in regard to ecdysteroids, testes have the ability to control their own internal milieu.  相似文献   

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After exposure to thermal stress or a control temperature, the relative abundance of ecdysone (E) and 20-hydroxyecdysone (20E) was measured in a wild-type line of Drosophila virilis (101) that is stress responsive and in a mutant line (147) that is not stress responsive. In line 101, the 20E content was higher and E content lower in females than in males. The abundance of E and 20E in females of line 147 was significantly higher than that in females of line 101. Females of line 101 were found to respond to 60 min of heat stress (38°C) by an increase in the abundance of both E and 20E, while in males of this line the amount of 20E increased and that of E declined. A role of the ecdysteroids in the control of reproduction of D. virilis under stress is discussed.  相似文献   

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The insecticide cyromazine, classified as an insect growth regulator, inhibits the growth and development of Diptera. The precise mode of cyromazine action remains unknown. We investigated mortality and developmental time effects after exposing first instar Drosophila melanogaster larvae to cyromazine for 8-h intervals. Significant increases in mortality were only observed for cyromazine exposure within the first 16 h of the first instar stage, suggesting that cyromazine accumulates in the larvae over time and that there is a delay between cyromazine exposure and effect. We observed that exposure to cyromazine during the early first instar stage resulted in earlier eclosion of adults when compared to cyromazine exposure at later first instar stages. The presence of 20-hydroxyecdysone during cyromazine exposure significantly reduced the lethal effect of cyromazine. We raise the possibility that the mode of cyromazine action is related to the development hormone, 20-hydroxyecdysone.  相似文献   

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Ecdysteroid levels throughout ovarian development and in newly-laid eggs of S. gregaria have been determined. A simple method for the separation of free and conjugated ecdysteroids is described. Both free and polar conjugated ecdysteroids are present at the end of oögenesis and in newly-laid eggs, but the polar conjugated ecdysteroids always predominate; 95% of the total ecdysteroid in newly-laid eggs is in the conjugated form. Ecdysone, 2-deoxyecdysone and 20-hydroxyecdysone have been fully characterized from both the ‘free’ and ‘conjugated’ fractions. The presence of traces of 26-hydroxyecdysone in the ‘conjugate’ fraction was indicated by HPLC analyses. The levels of ecdysteroid released from the conjugates of newly-laid eggs were 35 μg/egg pod (44 μg/g wet weight) for ecdysone, 16 μg/egg pod (19.4 μg/g) for 2-deoxyecdysone and 5 μg/egg pod (6.1 μg/g) for 20-hydroxyecdysone. The level of free ecdysone found in newly-laid eggs was 2 μg/egg pod (2.6 μg/g).  相似文献   

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BACKGROUND AND AIMS: The summer annual Impatiens glandulifera can reach 3 m in height within deciduous woodland. The primary objective was to determine if NO(3)(-) accumulation, and hence its osmotic effect, is an important physiological mechanism allowing Impatiens to achieve substantial height under low irradiance. METHODS: Stem extension, concentrations of K(+) and NO(3)(-) in leaves and concentrations of K(+), NO(3)(-) and other inorganic anions, malate, sugars, total N and total osmoticum in stem were measured in I. glandulifera sampled at different irradiance levels in deciduous woodland and in a glasshouse. Also, the energetic costs, as absorbed photons, of generating osmolarity in stem cell vacuoles with KNO(3), K(2)malate or hexose sugar were determined. KEY RESULTS: Results were similar in the woodland and glasshouse. At 50-100 % relative irradiance (Ir; open ground PAR = 100 % Ir) and 2-10 % Ir, plant height increased from 7-14 cm to 130-154 cm in 64-67 d. Leaf and stem NO(3)(-) concentrations were negligible at 50-100 % Ir while K(+), malate(2-) and sugars, respectively, accounted for 33.2-50.1 %, 19.3-20.8 % and 2.0-2.6 % of total osmoticum in stems. At 2-10 % Ir, NO(3)(-) concentrations were four to eight times greater in stems than leaves. Here, NO(3)(-) constituted 26.7-34.3 % of the total osmotic concentration in the stem and NO(3)(-)-N constituted 69-81 % of total N in stem tissue. Also at 2-10 % Ir, K(+) comprised 44.9-45.9 % and malate plus sugars 2.2-3.1 % of total osmotic concentration. The energy cost of osmoticum as KNO(3) was calculated as less than half that of malate and less than one-seventh that for hexose. Further calculations suggest that use of KNO(3), K(2)malate or glucose as osmoticum at low irradiance would, respectively, cost approx. 7 %, 16 % and 50 % of the total construction cost of the stem. CONCLUSIONS: It is concluded that accumulation of NO(3)(-) in place of organic molecules in stems is an important mechanism allowing I. glandulifera to achieve substantial height at low irradiance.  相似文献   

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2‐Hydroxybiphenyl 3‐monooxygenase (HbpA; EC 1.14.13.44) from Pseudomonas azelaica HBP1 was produced in Escherichia coli both as native and SeMet‐labelled protein. The two enzymes were purified to homogeneity and crystallized by the hanging‐drop vapour‐diffusion method. The crystals belong to the monoclinic space group C2, with unit‐cell parameters a = 108.6, b = 196.8, c = 79.3 Å, β = 97.7° for the native protein and a = 108.3, b = 196.8, c = 79.0 Å, β = 97.8° for SeMet HbpA. Crystal‐packing considerations led to the assumption of two HbpA subunits per asymmetric unit, which corresponds to a VM value of 3.3 Å3 Da−1 and a solvent content of 62%. The crystals were radiation‐sensitive and only had a lifespan of about 120 s when exposed to synchrotron radiation on an undulator beamline. To obtain complete data sets, data were collected from 23 native and 26 derivative crystals. The high‐resolution limit was 2.0 Å for native and 2.25 Å for SeMet HbpA.  相似文献   

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应用无细胞放射实验技术,逐日测定了家蚕末龄幼虫发育期间前胸腺和脂肪体匀浆液内蜕皮素20单氧酶(E-20-M)的活性。结果发现前胸腺内E-20-M活性特点是:从蜕皮到第3天幼虫前胸腺缺乏或具较低水平E-20-M活性;第4天前胸腺E-20-M活性开始升高;到第5天前胸腺E-20-M活性达到高峰。然而,与前胸腺相比,脂肪体E-20-M活性要高的多。  相似文献   

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The roots of Chenopodium bonus-henricus and the seeds of Spinacia oleracea contain 20-hydroxyecdysone and polypodine B. The seeds of S. oleracea also contain a compound with properties similar to those of 24(28)-dehydromakisterone-A and may contain small amounts of ecdysone.  相似文献   

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Abstract Using cell free radioassay, activities of ecdysone 20-monooxygenase (E-20-M) were determined in homogenates of prothoracic glands (PGs) and fat bodies at 24 h intervals during last instar larval development of Bombyx mori. It was found that the profile of E-20-M activity in PG homogenates was characterized by a basal line from day 0 to day 3 which begins to rise on 4th day and reaches a peak on 5th day. Nevertheless, in comparison with PGs, E-20-M activity in fat body was much higher.  相似文献   

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Recently we have shown that BhSGAMP‐1 is a developmentally regulated reiterated gene that encodes an antimicrobial peptide (AMP) and is expressed exclusively in the salivary glands, at the end of the larval stage. We show, for the first time, that a gene for an AMP is directly activated by 20‐OH ecdysone. This control probably involves the participation of short‐lived repressor(s). We also found that the promoter of BhSGAMP‐1 is not equipped with elements that respond to infection, provoked by the injection of microorganisms, in the salivary glands or in the fat body. We produced polyclonal antibodies against the synthetic peptide and found that the BhSGAMP‐1 peptide is secreted in the saliva. The BhSGAMP‐1 gene was also activated during the third larval molt. These facts confirm our hypothesis that this preventive system of defense was selected to produce an environment free of harmful microorganisms in the insect's immediate vicinity, during molts. genesis 47:847–857, 2009. © 2009 Wiley‐Liss, Inc.  相似文献   

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