EJE, vol. 92 (1995), issue 1

Contributions from Eleventh Ecdysone Workshop.

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Eur. J. Entomol. 92 (1): 1995

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On the use of ecdysteroid nomenclature

KARLSON P.

Eur. J. Entomol. 92 (1): 7-8, 1995

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Edcdysteroid biosynthesis and inactivation in relation to function

REES H.H.

Eur. J. Entomol. 92 (1): 9-39, 1995

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Analysis of ecdysteroid conjugates: Chromatographic characterization of sulfates, phosphates and glucosides

PIS J., GIRAULT J.P., GRAU V., LAFONT R.

Eur. J. Entomol. 92 (1): 41-50, 1995

Ecdysone monosulfates (at C-2, C-3 and C-22) and ecdysone 2,22-disulfate were synthesised using sulfur trioxide triethylamine complex as sulfation agent. Direct sulfation of ecdysone afforded a mixture of various sulfates. Regioselective sulfation of ecdysone was achieved by utilizing temporary protection of hydroxy groups. The chromatographic behaviour of synthetic sulfates was compared with that of various other polar conjugates of ecdysone and 20-hydroxyecdysone. General rules to characterise ecdysteroid conjugates are proposed.

Characterization of the glycosylated ecdysteroids in the hemolymph of baculovirus-infected gypsy moth larvae and cells in culture

KELLY T.J., PARK E.J., MASLER C.A., BURAND J.P.

Eur. J. Entomol. 92 (1): 51-61, 1995

Fourth-instar gypsy moth (Lymantria dispar; Lepidoptera: Lymantriidae) larvae, infected with the gypsy moth baculovirus (LdNPV), show an elevated and prolonged extension of the hemolymph ecdysteroid titer peak associated with molting. The ecdysteroid immunoreactivity associated with this peak elutes as two peaks following HPLC on a C18  reverse-phase column. Both peaks elute in a region more polar than 20-hydroxyecdysone, but less polar than the highly polar ecdysteroid immunoreactivity associated with the apolysis peak of control animals. Glycosylated ecdysteroid standards, produced by in vitro incubation of UDP-glucose with...

Metabolism of 3-dehydroecdysone in the crayfish Orconectes limosus (Crustacea: Decapoda)

BOCKING D., DAUPHIN-VILLEMANT C., LAFONT R.

Eur. J. Entomol. 92 (1): 63-74, 1995

3-Dehydroecdysone (3DE) is the major secretory product of crayfish moulting glands. The metabolic fate of 3DE in the crayfish Orconectes limosus was investigated after in vivo injection of [3H]3DE and after incubation of various tissues with [3H]3DE in vitro. Metabolites in whole body and tissue extracts were analysed by HPLC. In vivo [3H]3DE is efficiently reduced and hydroxylated to [3H]20-hydroxyecdysone (20E), which is one of the major metabolites observed in our study and also the major circulating ecdysteroid. 3 beta-reduction and 20-hydroxylation were the predominant enzymatic activities obtained...

Three new antisera with high sensitivity to ecdysone, 3-dehydroecdysone and other A-ring derivatives: Production and characterization

VON GLISCYNSKI U., DELBECQUE J.P., BOCKING D., SEDLMEIER D., DIRCKSEN H., LAFONT R.

Eur. J. Entomol. 92 (1): 75-79, 1995

Three new antisera have been produced in rabbits immunized with E-2/3-hemisuccinate conjugates. In a competitive enzyme immunoassay (EIA) with 20-ecdysone-peroxidase as a tracer, the antisera exhibit high specificity towards the side chain of the ecdysone (E) molecule. Modifications of the A-ring do result in no or small changes of sensitivity.

One antiserum (RUD-2) allows the direct quantification of total ecdysteroids in samples containing E and 3-dehydroecdysone in varying ratios, without further sophisticated sample treatment. At least one of the other antisera allows the direct quantification of mixtures of E and 25-deoxyecdysone (RUD-1)....

Ecdysteroid release and ecdysteroid titer during larval-adult development of the Mediterranean field cricket, Gryllus bimaculatus (Ensifera: Gryllidae)

GERSTENLAUER B., HOFFMANN K.H.

Eur. J. Entomol. 92 (1): 81-92, 1995

Female penultimate instar larvae of Gryllus bimaculatus show a maximum in haemolymph ecdysteroid titer as well as in hormone release in vitro by prothoracic glands on day 4, two days before final larval moult. Ovary and abdominal integument of the larvae contain considerable amounts of ecdysteroids, but no significant hormone release is detectable during a 16 h in vitro incubation of these tissues.

In last instar larvae, the prothoracic glands reach their maximal ecdysteroid release on day 5. This peak is followed by a maximal release of hormones by the ovary as well as by the abdominal integument on day 6. The highest titer of ecdysteroids...

The ovary is a source of circulating ecdysteroids in Blattella germanica (Dictyoptera: Blattellidae)

ROMANA I., PASCUAL N., BELLES X.

Eur. J. Entomol. 92 (1): 93-103, 1995

The developmental fluctuations in immunoreactive ecdysteroids in the haemolymph of adult female Blattella germanica (L.) are reported. There is a relatively low titre in freshly ecdysed females, an increase during vitellogenesis to a peak at choriogenesis, and a decrease after ovulation. These fluctuations parallel those of ecdysteroid content in the ovary. In addition, haemolymph ecdysteroid levels are reduced following ovariectomy, and single ovarioles incubated in vitro effectively release ecdysteroids into the medium. The data strongly suggest that the ovaries are a major source of circulating ecdysteroids in adult females of this species....

Autoradiographic and morphological investigations of the defensive ecdysteroid glands in adult Pycnogonum litorale (Arthropoda: Pantopoda)

TOMASCHKO K.H.

Eur. J. Entomol. 92 (1): 105-112, 1995

Pycnogonum litorale (Ström) is a primitive marine chelicerate. When mechanically irritated, it discharges a mixture of eight ecdysteroids (ES) that acts as a chemical defense against predatory decapod crustaceans. Injection of [3H]-20-hydroxyecdysone into the hemolymph or into the gut and subsequent autoradiography of frozen sections revealed that the majority of the ES in the pycnogonids is concentrated in epidermal glands. A novel autoradiographic technique for the localization of water soluble radioactive substances at uneven surfaces visualized the ES secretion through cuticular pores. Scanning electron microscopy shows that each...

Non-cerebral ecdysiotropic and gonadotropic activities from the mosquito Aedes aegypti (Diptera: Culicidae)

MASLER E.P., KELLY T.J.

Eur. J. Entomol. 92 (1): 113-122, 1995

The head is traditionally considered to be the site of production and release of the egg development neurosecretory hormone (EDNH), involved in ovarian maturation in dipterans. We find, however, that the thorax and abdomen of the mosquito Aedes aegypti each possess factors which resemble EDNH in both physiological and biochemical properties. Extracts of thoraces or abdomens each stimulated the dose-dependent appearance of ecdysteroid in incubations of A. aegypti ovaries in vitro. In addition, each extract is capable of stimulating ovarian maturation in vivo in decapitated Aedes atropalpus. The head contains higher levels of activity,...

Low molecular weight ecdysiotropins in proctodaea of fifth instars of the tobacco hornworm, Manduca sexta (Lepidoptera: Sphingidae), and hosts parasitized by the braconid wasp Cotesia congregata (Hymenoptera: Braconidae)

GELMAN D.B., BECKAGE N.E.

Eur. J. Entomol. 92 (1): 123-129, 1995

Using an in vitro prothoracic gland assay (PTG) to assess production of ecdysteroids, a low molecular weight ecdysiotropin(s) (500-1,500 daltons) was detected in proctodaea (hindguts) of unparasitized and parasitized Manduca sexta 5th instars. Dose-response curves for the three groups of insects tested were similar with maximum stimulation being observed in PTGs incubated with 0.00125-0.005 organ equivalent. However, when results were adjusted for differences in wet weight of the proctodaea, extracts from hosts which initially were parasitized in the first instar, possessed 2.6 times more ecdysiotropic activity than extracts from day-3 unparasitized...

Low molecular weight ecdysiotropins in the hemolymph of 5th instars of the European corn borer, Ostrinia nubilalis (Lepidoptera: Pyralidae), and the gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae)

GELMAN D.B., BELL R.A.

Eur. J. Entomol. 92 (1): 131-141, 1995

Fluctuations of low molecular weight (500-1,500 daltons) ecdysiotropin titers in the hemolymph of the European corn borer, Ostrinia nubilalis, and of the gypsy moth, Lymantria dispar, were measured. For hemolymph from nondiapause-bound European corn borer 5th instars, dose/response curves suggest that there exist at least two ecdysiotropins capable of stimulating prothoracic glands to produce ecdysteroid, one (ECTR I) with maximum activity at 0.5 µl hemolymph/25 µl drop of incubation medium and one (ECTR II) with maximum activity at 4 µl hemolymph/25 µl drop of incubation medium. ECTR I was detected on days 2,...

Pleiotropic effects of trypsin modulating oostatic factor (Neb-TMOF) of the fleshfly Neobellieria bullata (Diptera: Calliphoridae)

BYLEMANS D., HUA Y.J., CHIOU S.J., KOOLMAN J., BOROVSKY D., DE LOOF A.

Eur. J. Entomol. 92 (1): 143-149, 1995

Trypsin Modulating Oostatic Factor (TMOF) of the grey fleshfly Neobellieria bullata is a hexapeptide that has been isolated from vitellogenic ovaries. Its precursor is a 75 kDa protein that is abundantly present in oocytes but absent from hemolymph. TMOF inhibits trypsin biosynthesis in the gut, it lowers yolk polypeptide concentration in the hemolymph and strongly inhibits ecdysone biosynthesis by larval ring glands. Evidence is presented that a molecule with identical chromatographic behaviour and physiological effects is also present in the larvae of Neobellieria at the time when they stop feeding, Ecdysone inhibition does not suppress...

Drosophila \"elastin-type\" gene identified with antibody to Bombyx PTTH

LI W., FILIPPOV V.A., FILIPPOVA M.A., JINDRA M., SEHNAL F.

Eur. J. Entomol. 92 (1): 151-153, 1995

Screening of two Drosophila nervous system cDNA libraries with DNA probes derived from lepidopteran prothoracicotropin (PTTH) did not yield any hybridizing clone. Monoclonal antibody to Bombyx PITH recognized in the expression cDNA library several clones but their sequencing revealed relationship to elastins and collagens rather than to PTTH. We conclude that Drosophila PTTH cannot be identified with probes derived from lepidopteran PTTH.

The different effects of ecdysone and 20-hydroxyecdysone on the induction of larval ecdysis in the silkworm, Bombyx mori (Lepidoptera: Bombycidae)

TANAKA Y.

Eur. J. Entomol. 92 (1): 155-160, 1995

The silkworm, Bombyx mori was reared on artificial diets supplemented with 20-hydroxyecdysone (20E) during the 4th instar and on diets supplemented with various concentrations of either 20E or ecdysone (E) in the 5th (last) instar. The larvae transferred to diet containing E molted to the 6th instar earlier than the control larvae and their mortality was within norm. On the other hand, most of the larvae that were transferred to the diet supplemented with 300 ppm 20E continued to feed until they died without apolysis. The amount of 20E in their hemolymph was higher than in the larvae fed 200 ppm E. Larvae transferred to a diet containing both...

Colour pattern specific melanin synthesis is controlled by ecdysteroids via dopa decarboxylase in wings of Precis coenia (Lepidoptera: Nymphalidae)

KOCH P.B.

Eur. J. Entomol. 92 (1): 161-167, 1995

The synthesis of dopamine melanin in the wing scales of Precis coenia depends on pattern specific activity of dopa decarboxylase (DDC) at the end of pupal development when hemolymph ecdysteroids decrease. DDC activity in forewings increased in 6 days old pupae and is maximal on day 6 at midnight. DDC activity increased also in isolated wings incubated in Grace's medium on day 5 at midnight, following the same time profile as in vivo. The wings developed their typical colour pattern. However, at most time points DDC activity in vitro was lower than in vivo. Wings incubated in medium containing 10 µg/ml 20-hydroxyecdysone (20E) did not melanize....

Genetics of ecdysteroid-regulated central nervous system metamorphosis in Drosophila (Diptera: Drosophilidae)

RESTIFO L.L., ESTES P.S., DELLO RUSSO C.

Eur. J. Entomol. 92 (1): 169-187, 1995

We are interested in identifying members of the generic pathway through which 20-hydroxyecdysone (20HE) mediates reorganization of the central nervous system (CNS) during metamorphosis. Our entry point is the Drosophila Broad-Complex (BR-C), an early 20HE-inducible locus with three genetic subfunctions, each represented by a lethal complementation group. Our previous analysis of mutants demonstrated that all three BR-C subfunctions are necessary for CNS morphogenesis and one is essential for visual system organization. We believe the mutant phenotypes result from faulty expression of genes normally regulated by the BRC family of...

Ovary-specific interaction of ecdysone 22-phosphate with proteins in adult Drosophila melanogaster (Dipetra: Drosophilidae)

GRAU V., PIS J., LAFONT R.

Eur. J. Entomol. 92 (1): 189-196, 1995

Radiotracer experiments were performed to analyze the interaction of metabolites of ecdysone with proteins in adult Drosophila melanogaster females. [3H]Ecdysone was injected into the abdomen and after 4 h of incubation the flies were dissected. PBS-soluble material of the ovaries, haemolymph, gut and abdominal carcass were separated by gel-filtration. In all samples, two peaks containing radioactivity eluted with macromolecules (probably proteins) of an apparent molecular mass of more than 1,000 kDa (peak I) and about 500 kDa (peak II). In the extracts of the ovaries an additional peak in the molecular weight range of about 50 kDa...

Influence of ovary and ecdysteroids on pheromone biosynthesis in Drosophila melanogaster (Diptera: Drosophilidae)

WICKER C., JALLON J.M.

Eur. J. Entomol. 92 (1): 197-202, 1995

The influence of ovary and ecdysteroids on pheromone biosynthesis was investigated in D. melanogaster. Strains mutant for ovo produced enhanced amounts of cuticular hydrocarbons in both sexes. Female sex pheromone production was not affected by the mutation, except homozygous females for ovoDlrsl which exhibited a higher proportion of the pheromone, 7,11 heptacosadiene.

The amount of hydrocarbons in wild-type (CS) flies was stable at 21 and 25°C but dramatically increased at 29°C whereas the cuticular hydrocarbon pattern was unchanged. In contrast, ecdysone-less 1ls (ecd-1) flies...

Developmental variability of the antibacterial response in larvae and pupae of Calliphora vicina (Diptera: Calliphoridae) and Drosophila melanogaster (Diptera: Drosophilidae)

CHERNYSH S.I., SIMONENKO N.P., BRAUN A., MEISTER M.

Eur. J. Entomol. 92 (1): 203-209, 1995

The inducibility of antibacterial peptides in Calliphora is strongly reduced during pupal development. Similarly, a downregulation of the inducibility of a diptericin reporter transgene is observed in the pupae of transgenic Drosophila, indicating existence of a negative correlation between inducibility and ecdysteroid titre. In the present paper we show that the level of antibacterial activity induced by the injection of bacteria into the diapausing larvae of Calliphora is reduced if the insects simultaneously receive 20-hydroxyecdysone. The same effect is observed in ligated Calliphora larvae deprived of their brain-retrocerebral...

Cuticular protein genes in Tenebrio molitor (Coleoptera: Tenebrionidae)

RONDOT I., BOUHIN H., CHARLES J.P., MATHELIN J., DELACHAMBRE J.

Eur. J. Entomol. 92 (1): 211-214, 1995

We have previously isolated from the beetle Tenebrio molitor, cDNAs coding for two glycine-rich cuticular proteins named ACP-20, ACP-22 and ACP-17 and an alanin-rich cuticular protein named LPCP-22. The ACP-20, ACP-22, ACP-17 mRNAs are detected by Northern blot and in situ hybridization analysis only in epidermal regions secreting heavily sclerotized cuticle during the pharate adult stage. The LPCP-22 mRNA is detected in most epidermal regions during the secretion of larval and pupal cuticles. Then, its presence is restricted to the epidermal zones secreting intersegmental soft cuticle in the newly ecdysed pupa. This stage- and tissue-specific...

Stage specific ecdysteroid response of larval hemolymph protein genes in Galleria mellonella (Lepidoptera: Pyralidae)

WANG C., DUTTA-GUPTA A., KUMARAN A.K.

Eur. J. Entomol. 92 (1): 215-216, 1995

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Effect of 20-hydroxyecdysone on synthesis and uptake of arylphorin by the larval fat body of Calliphora vicina (Diptera: Calliphoridae)

BURMESTER T., MATZNER U., SCHELLER K.

Eur. J. Entomol. 92 (1): 217-227, 1995

During the final larval instar of the blowfly Calliphora vicina the storage protein arylphorin is stage-specifically produced and re-absorbed by the fat body cells. In this study the hormonal regulation of arylphorin biosynthesis and endocytosis has been investigated in vitro and in vivo. Application of 10 to 50 ng 20-hydroxyecdysone (20E) in vivo or of 10-7 to 10-6 M 20E in vitro specifically inhibits arylphorin translation without altering the arylphorin mRNA level. After termination of arylphorin synthesis in vivo, it can be artificially re-induced by neck-ligation, an effect which can be reversed by application of 50...

Correlations between ecdysteroid titre and silk gene expression in the last instar larvae of Galleria mellonella (Lepidoptera: Pyralidae)

YANG C., KRISHNAN M., ZUROVEC M., SEHNAL F., SCHELLER K.

Eur. J. Entomol. 92 (1): 229-234, 1995

Developmental profiles of total RNA and four silk protein mRNAs exhibit different correlations with ecdysteroid titre changes in course of the 180 hr-long last larval instar. Total RNA and heavy-chain fibroin mRNA increase gradually until 168 hr and 156 hr, respectively, dropping sharply afterwards. The content of sericin1 mRNA rises for 120 hr, remains high until 168 hr and then drops. The amounts of ligh-chain fibroin mRNA and sericin2 mRNA increase sharply at the start of the instar; the former rises until 168 hr, whereas the latter begins to decline already at 120 hr. The pupation-inducing ecdysteroid surge at 144-168 hr causes a drop in all examined...

Ecdysteroid-induced expression of the GmE75 and GHR3 orphan receptor genes in Galleria mellonella (Lepidoptera: Pyralidae) larvae and cultured silk glands

JINDRA M., SEHNAL F., RIDDIFORD L.M.

Eur. J. Entomol. 92 (1): 235-236, 1995

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Regulation of gene expression by 20-hydroxyecdysone in the fat body of Aedes aegypti (Diptera: Culicidae)

DEITSCH K.W., DITTMER N., KAPITSKAYA M.Z., CHEN J.S., CHO W.L., RAIKHEL A.S.

Eur. J. Entomol. 92 (1): 237-244, 1995

In response to a blood meal, the fat body of the female mosquito produces several yolk proteins (YP) which are accumulated by the developing oocytes. 20-hydroxyecdysone (20E) stimulates high levels of YP gene expression in fat bodies cultured in vitro, but initiation of this expression was eliminated with cycloheximide. In the 1.5 kb upstream region of the gene encoding vitellogenic carboxypeptidase, a mosquito YP, there are several putative regulatory sequences that resemble steroid response elements (SRE), but they do not match the ecdysteroid response element (EcRE) consensus. A cDNA encoding a mosquito ecdysteroid receptor (AaEcR) has recently...

Endogenous ecdysteroid receptor binds non-specific DNA as a heteromer, the formation of which is enhanced by ligand and DNA

BRADBROOK D.A., KOOLMAN J.

Eur. J. Entomol. 92 (1): 245-250, 1995

Chromatography of nuclear extracts from Calliphora vicina on heparin-agarose separates two fractions binding ponasterone A. One does not bind calf-thymus (ct) DNA, has a molecular mass of 100 kDa, and may represent an ecdysteroid receptor monomer. The other binds to ctDNA and is 180 kDa, indicating this to be a receptor complex of two or more heteromeric partners. The formation of this DNA-binding complex is promoted by the presence of high-affinity ligand and of DNA itself.

Ecdysteroid receptor in Chironomus thummi (Dipetra: Chironomidae)

DEAK P., LAUFER H.

Eur. J. Entomol. 92 (1): 251-257, 1995

An ecdysteroid receptor protein has been identified and characterized from whole body extracts of Chironomus thummi larvae. The kinetic and physical characteristics of this binding are consistent with the criteria established for steroid hormone receptors and are similar to those exhibited by the Drosophila ecdysteroid receptor. The binding data of cytosols obtained from different developmental stages (third and fourth stage larvae, pupae and adults) indicate only a single type of binding site which exists throughout the life cycle of the insect with an average Kd value of 5 × 10-9 mol/g of protein. Furthermore,...

Hormonal regulation of the Drosophila melanogaster ng-genes

D'AVINO P.P., CRISPI S., FURIA M.

Eur. J. Entomol. 92 (1): 259-261, 1995

Intermoult puffs constitute the first set of ecdysone-responsive puffs detected on salivary gland chromosomes of D. melanogaster third instar larave. In order to analyse the hormonal regulation of this puff set we have investigated the genetic regulatory hierarchy governing the expression of three genes mapping at the 3C polytene region, from which one of the most prominent intermoult puff originates. The results obtained reveal that the expression of the genes studied, named ng-1, ng-2 and ng-3, is coordinate and specifically restricted to the Puffing Stage 1 (PS 1). The repression of these genes appears to be related to...

Structure and expression of the br-c locus in Drosophila melanogaster (Diptera: Drosophilidae)

ZHIMULEV I.F., BELYAEVA E.S., MAZINA O.M., BALASOV M.L.

Eur. J. Entomol. 92 (1): 263-270, 1995

We review data on the structure and expression of the Broad Complex, a key gene in the response to ecdysterone (20E) in Drosophila cells.

The 2B3-7 X chromosome region, containing an early (20E) inducible puff, was saturated with mutations in the genes dor, hfw (swi) and BR-C that cause disturbances in the normal ecdysterone-inducible puffing sequence. By analysis of chromosome rearrangements localized on the genetic, cytological and physical maps, the BR-C locus has been mapped exactly within the 2B3-5 puff. Homozygous mutations for BR-C result in the multiple distortions of metamorphosis and...

A strategy for the identification of ecdysteroid receptor agonists and antagonists from plants

DINAN L.

Eur. J. Entomol. 92 (1): 271-283, 1995

A strategy is described for receptor-based phytochemical screening of plant extracts for ecdysteroid agonists and antagonists. Milligram amounts of seed are methanol extracted. Lipids and pigments are removed by hexane partioning. Agonist and antagonist activities are detected with a microplate-based specific bioassay using the Drosophila melanogaster ecdysteroid-responsive BII cell line. Extracts are also screened with ecdysteroid-specific RIAs to identify extracts containing phytoecdysteroids. Over 1,700 species of plant have been screened in this way so far. Extracts are being sought which (i) contain large amounts of phytoecdysteroid, (ii)...

Preparative RP-HPLC of ecdysteroids

GILDENGORN V.D., TYUKAVIN Y.A., VOLODIN V.V.

Eur. J. Entomol. 92 (1): 285, 1995

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Tissue and cell culture of phytoecdysone-producing plants as a perspective source of ecdysteroids

ANUFRIEVA A.N., KOVLER L.A., KOLEGOVA N.A., VOLODIN V.V.

Eur. J. Entomol. 92 (1): 286, 1995

N/A

Acetylated ecdysteroids from Ajuga reptans var. atropurpurea (Lamiales: Lamiaceae)

CALCAGNO M.P., CAMPS F., COLL J., MELE E.

Eur. J. Entomol. 92 (1): 287-293, 1995

The structural elucidation of acetylated ecdysteroids isolated from dried roots of Ajuga reptans var. atropurpurea, based on spectroscopic procedures, is reported. Among them 2-O-acetyl- and 3-O-acetyl-20-hydroxyecdysone (20E2Ac, 20E3Ac) and 3-O-acetyl-29-norcyasterone (29NCY3Ac) had previously been only partially described and 3-O-acetylcyasterone (CY3Ac) was heretofore unreported.

Distribution and levels of phytoecdysteroids within individual plants of species of the Chenopodiaceae

DINAN L.

Eur. J. Entomol. 92 (1): 295-300, 1995

Radioimmunoassay was used to quantify the levels of ecdysteroids in extracts of portions of single plants of several members of the Chenopodiaceae. Species were chosen because previous studies had shown that seeds of these species were ecdysteroid-containing and because they represent several tribes within the Chenopodiaceae. Total ecdysteroid levels varied considerably between species and between different parts of the same plant. Phytoecdysteroid-containing members of the genus Chenopodium appear to possess the same distribution as previously found for C. album, as does Spinacia oleracea. Rhagodia candolleana is characterized...

In vitro incorporation of radiolabelled cholesterol and mevalonic acid into ecdysteroid by hairy root cultures of a plant, Serratula tinctoria

DELBECQUE J.P., BEYDON P., CHAPUIS L., CORIO-COSTET M.F.

Eur. J. Entomol. 92 (1): 301-307, 1995

After the transformation of stems with Agrobacterium rhizogenes, hairy root cultures have been obtained from Serratula tinctoria (Asteraceae), a plant containing very high amounts of 20-hydroxyecdysone (20E) and numerous other phytoecdysteroids. These cultures were found to grow regularly in vitro, either in liquid or solid media, and to maintain a high ecdysteroid production (c.a. 0.1-0.2% dry weight). 20E was the predominant ecdysteroid, together with the corresponding 3-acetate (20E3Ac). Ecdysteroids were not secreted in the medium but remained in root tissues. Moreover, a concentration gradient was observed in transformed hairy roots,...

Comparison between the effects of 20-hydroxyecdysone and phytohormones on growth and development in plants

MACHACKOVA I., VAGNER M., SLAMA K.

Eur. J. Entomol. 92 (1): 309-316, 1995

Insect moulting hormone, 20-hydroxyecdysone or ecdysterone (E), was tested in a series of bioassays that are commonly used for evaluation of the activity of phytohormones. The assays included: auxin coleoptile bioassay in wheat, gibberellin bioassays in dwarf maize and dwarf rice; cytokinin bioassay on tobacco callus; brassinolide related ethylene formation assay in dwarf maize, alfalfa and Chenopodium, flowering assay on Chenopodium and special assay on somatic embryogenesis in cell cultures of alfalfa.

The 20-hydroxyecdysone has no biological activity in tests used for detecting phytohormone activity, excepting for slight gibberellin-like...

Hormonal status of RH-5849 and RH-5992 synthetic ecdysone agonists (ecdysoids) examined on several standard bioassays for ecdysteroids

SLAMA K.

Eur. J. Entomol. 92 (1): 317-323, 1995

Effects of the synthetic ecdysone agonists RH-5849 and RH-5992 have been compared with the action of 20-hydroxyecdysone in several standard bioassays that are currently used for determination of the moulting hormone activity. In the assay on ligated larvae of Galleria, these synthetic hormone mimics, ecdysoids according to the endocrinological terminology, stimulated the extra-larval moult cycles with all developmental consequences similar to the action of 20-hydroxyecdysone. The ED-50 dosages were 1.3 µg for 20-hydroxyecdysone, 1.75 µg for RH-5849 and 0.9 µg/specimen for RH-5992. In the assays on diapausing pupae of Manduca...

Age-specific effects of a non-steroidal ecdysteroid agonist, RH-5992, on the spruce budworm, Choristoneura fumiferana (Lepidoptera: Tortricidae)

PALLI S.R., PRIMAVERA M., TOMKINS W., LAMBERT D., RETNAKARAN A.

Eur. J. Entomol. 92 (1): 325-332, 1995

The non-steroidal ecdysone agonist, RH-5992, is effective in inducing an incomplete molt in the spruce budworm, Choristoneura fumiferana (Clemens), only when it is fed to the larvae prior to the appearance of the endogenous ecdysteroid peak. When this compound is administered after the ecdysteroid peak, the larvae molt normally into the next stage. However, because of its persistence, the agonist induces an incomplete molt in the subsequent larval stage. The effect does not appear to be due to ecdysone responsive tissues becoming refractory after the ecdysteroid peak. Choristoneura hormone receptor 3 (CHR3), a homologue of Manduca...

Biological activity and receptor-binding of ecdysteroids and the ecdysteroid agonists RH-5849 and RH-5992 in imaginal wing discs of Spodoptera exigua (Lepidoptera: Noctuidae)

SMAGGHE G., DEGHEELE D.

Eur. J. Entomol. 92 (1): 333-340, 1995

The effects of 20-hydroxyecdysone (20E), ponasterone A (PoA) and both dibenzoylhydrazine-based ecdysteroid agonists RH-5849 and RH-5992, were tested on in vitro-cultured imaginal wing discs of last-instar larvae of the beet armyworm, Spodoptera exigua (Hübner). In each case, the response was qualitatively similar to that induced by 20E, although the concentrations required to induce response varied widely. The EC50 of both ecdysteroids 20E and PoA to elicit wing disc evagination in 50% of the discs was 89.8 and 2.57 nM, respectively; for both nonsteroid agonists RH-5849 and RH-5992 this was 870 and 11.7 nM, respectively. Binding competition...

Receptor affinities and biological responses of nonsteroidal ecdysteroid agonists on the epithelial cell line from Chironomus tentans (Diptera: Chironomidae)

QUACK S., FRETZ A., SPINDLER-BARTH M., SPINDLER K.D.

Eur. J. Entomol. 92 (1): 341-347, 1995

The two nonsteroidal ecdysteroid agonists, RH 5849 and RH 5992 were tested in the epithelial cell line from Chironomus tentans. Both compounds induce all effects elicited by 20-OH-ecdysone in this cell line, namely differentiation, arrest of cell growth and regulation of chitin metabolism. Usually, the cell line forms multicellular vesicles consisting of a squamous monolayer. Treatment with ecdysteroids or RH compounds leads to the formation of a stratified columnar epithelium. This differentiation is accompanied by changes in protein pattern. Ecdysteroids and RH compounds also interfere with chitin metabolism. Chitin synthesis is inhibited,...

Stimulation of growth and development in Japanase quails after oral administration of ecdysteroid-containing diet

KOUDELA K., TENORA J., BAJER J., MATHOVA A., SLAMA K.

Eur. J. Entomol. 92 (1): 349-354, 1995

Freshly hatched Japanese quails of the strain Faraon were fed for 50 days by diets containing graded amounts of pulverized seeds of an Asiatic plant Leuzea carthamoides Iljin. The dry seeds of this plant contained 1.8%-2.1% of 20-hydroxyecdysone, together with some smaller amounts of other ecdysteroids. In additional experiments the quails were also offered the standard diet plus whole seeds of Leuzea ad libitum and the standard diet enriched with certain commercially available biostimulating preparations for birds, Biostrong and Ecovit.

The effects of the diets were evaluated first at the age of 37 days and then at the age of...

Insect hormones - ecdysteroids: their presence and actions in vertebrates

SLAMA K., LAFONT R.

Eur. J. Entomol. 92 (1): 355-377, 1995

N/A

Functional transfer of an elementary ecdysone gene regulatory system to mammalian cells: Transient transfections and stable cell lines

YANG G., HANNAN G.N., LOCKETT T.J., HILL R.J.

Eur. J. Entomol. 92 (1): 379-389, 1995

A 3.1 Kb fragment of a Drosophila melanogaster ecdysone receptor (EcR) cDNA (splice product, EcR B1) comprising the 2.6 Kb coding region with 218 base pairs of 5' and 258 base pairs of 3'-untranslated sequence, was cloned into the mammalian expression vectors pH beta APr-1 and pSG5 (which place EcR under the control of a human beta-actin and a SV40 early promoter, respectively). Chinese hamster ovary cells have been stably transfected with the beta-actin promoter construct. Antiserum, prepared against an EcR-fusion protein has been used to demonstrate the synthesis of an apparently complete ecdysone receptor in a stable cell line...

Effect of feeding on a diet containing varying amounts of Rhaponticum carthamoides hay meal on selected morphological parameters in rats

SELEPCOVA L., SOMMER A., VARGOVA M.

Eur. J. Entomol. 92 (1): 391-397, 1995

Effects of diet meal containing 10, 20, 30 and 50% of Rhaponticum carthamoides hay meal on the increase of body weight, growth and development of selected internal organs and behaviour of animals were examined. The 10% diet, which is preferable from both health and economical viewpoints, stimulated the growth of testes, while the body growth curve was similar to that of animals fed on a conventional diet. The diet with 20% Rhaponticum resulted in high variability of the weight of testes. Effects of tested diets on the body weight and food consumption were different in males and females.