Topical Application Of Juvenile Hormone Research Articles

The BTB transcription factor, Abrupt, acts cooperatively with Chronologically inappropriate morphogenesis (Chinmo) to repress metamorphosis and promotes leg regeneration

Many insects undergo the process of metamorphosis when larval precursor cells begin to differentiate to create the adult body. The larval precursor cells retain stem cell-like properties and contribute to the regenerative ability of larval appendages. Here we demonstrate that two Broad-complex/Tramtrack/Bric-à-brac Zinc-finger (BTB) domain transcription factors, Chronologically inappropriate morphogenesis (Chinmo) and Abrupt (Ab), act cooperatively to repress metamorphosis in the flour beetle, Tribolium castaneum. Knockdown of chinmo led to precocious development of pupal legs and antennae. We show that although topical application of juvenile hormone (JH) prevents the decrease in chinmo expression in the final instar, chinmo and JH act in distinct pathways. Another gene encoding the BTB domain transcription factor, Ab, was also necessary for the suppression of broad (br) expression in T. castaneum in a chinmo RNAi background, and simultaneous knockdown of ab and chinmo led to the precocious onset of metamorphosis. Furthermore, knockdown of ab led to the loss of regenerative potential of larval legs independently of br. In contrast, chinmo knockdown larvae exhibited pupal leg regeneration when a larval leg was ablated. Taken together, our results show that both ab and chinmo are necessary for the maintenance of the larval tissue identity and, apart from its role in repressing br, ab acts as a crucial regulator of larval leg regeneration. Our findings indicate that BTB domain proteins interact in a complex manner to regulate larval and pupal tissue homeostasis.

Monoterpenoid signals and their transcriptional responses to feeding and juvenile hormone regulation in bark beetle Ips hauseri.

Hauser's engraver beetle, Ips hauseri, is a serious pest in spruce forest ecosystems in Central Asia. Its monoterpenoid signal production, transcriptome responses and potential regulatory mechanisms remain poorly understood. The quality and quantity of volatile metabolites in hindgut extracts of I. hauseri were found to differ between males and females and among three groups: beetles that were newly emerged, those with a topical application of juvenile hormone III (JHIII) and those that had been feeding for 24 h. Feeding males definitively dominated monoterpenoid signal production in I. hauseri, which uses (4S)-(-)-ipsenol and (S)-(-)-cis-verbenol to implement reproductive segregation from Ipstypographus and Ipsshangrila. Feeding stimulation induced higher expression of most genes related to the biosynthesis of (4S)-(-)-ipsenol than JHIII induction, and showed a male-specific mode in I. hauseri. JHIII stimulated males to produce large amounts of (-)-verbenone and also upregulated the expression of several CYP6 genes, to a greater extent in males than in females. The expression of genes involved in the metabolism of JHIII in females and males was also found to be upregulated. Our results indicate that a species-specific aggregation pheromone system for I. hauseri, consisting of (4S)-(-)-ipsenol and S-(-)-cis-verbenol, can be used to monitor population dynamics or mass trap killing. Our results also enable a better understanding of the bottom-up role of feeding behaviors in mediating population reproduction/aggregation and interspecific interactions.

Methyl farnesoate regulatory mechanisms underlying photoperiod‐dependent sex determination in the freshwater crustacean Daphnia magna

Freshwater zooplankton Daphnia magna has been widely used in ecotoxicology studies. During the last 20 years, it has been demonstrated that the topical application of juvenile hormone (JH) or JH analogs to mother daphnids induce male offspring production. Based on this finding, an in vivo screening validation method for chemicals with JH agonistic effect has developed. Although this screening system successfully identified a number of JH-like chemicals, molecular mechanisms underlying the male sex-determining process remain largely unknown. To address this issue, we established a reliable male- or female-producing system using Daphnia pulex WTN6 strain by changing the rearing photoperiod. Taking advantage of this rearing system, we successfully found several factors involving male sex determination such as ionotropic glutamate receptors, protein kinase C and pantothenate. Here, we used two D. magna strains that can also control the production of female or male offspring by photoperiod differences as model species for ecotoxicology studies. We demonstrated that either treatment of antagonist of ionotropic glutamate receptors or inhibitor of protein kinase C strongly suppressed male offspring production even under male-producing conditions. Moreover, we revealed that male sex-determining processes are likely diverged between D. magna and D. pulex based on the current experiment. This study provides a fine experimental method for in vivo screening not only JH agonists but also JH antagonists. Moreover, using daphnids with photoperiod-dependent sex determination manner will hugely contribute to understanding the mode-of-action of JH in daphnids.

Insulin signaling mediates previtellogenic development and enhances juvenile hormone-mediated vitellogenesis in a lepidopteran insect, Maruca vitrata

BackgroundInsulin/insulin-like growth peptide signaling (IIS) down-regulates hemolymph sugar level and facilitates larval growth in the soybean pod borer, Maruca vitrata. The objective of this study is to determine whether IIS of M. vitrata can mediate ovarian development of adult females.ResultsA pair of ovaries consists of 8 ovarioles, each of which is separated into distal germarium and proximal vitellarium in M. vitrata. In the germarium, oocyte development occurred with active mitotic activity which was visible by incorporating bromodeoxyribose uridine. Previtellogenic development and subsequent vitellogenesis began soon after adult emergence. They continued with increase of female age. Oocyte development was facilitated by up-regulation of vitellogenin (Vg) and Vg receptor (VgR) gene expression. Larval diets significantly influenced on ovarian development of M. vitrata because oocyte development varied with pupal size derived from larvae treated with different nutritional diets. Its ovarian development was dependent on endocrine signal(s) from the head because decapitation soon after adult emergence prevented oogenesis and subsequent vitellogenesis along with marked reduction of Vg and VgR expression. Topical application of juvenile hormone (JH) significantly recovered its ovarian development whereas farnesoic acid (a precursor of JH biosynthesis) or 20-hydroxyecdysone treatment did not. JH stimulated vitellogenesis and choriogenesis, but not previtellogenic development. In contrast, insulin injection to decapitated females stimulated oocyte differentiation and vitellogenesis along with increase of Vg and VgR expression. To further analyze the effect of insulin on ovarian development, expression of four IIS components (InR, FOXO, Akt, and TOR) genes was manipulated by RNA interference. Hemocoelic injection of gene-specific double stranded RNAs significantly reduced their target gene mRNA levels and interfered with ovarian development. An addition of insulin to JH treatment against decapitated females enhanced the gonadotropic effect of JH by stimulating oogenesis.ConclusionsIIS plays crucial role in mediating previtellogenic development of M. vitrata in response to nutrient signal. It also enhances the gonadotropic effect of JH II on vitellogenesis.

Molecular characterization and RNA interference analysis of vitellogenin receptor from Nilaparvata lugens (Stål)

Vitellogenin receptors (VgRs), members of the low-density lipoprotein receptor (LDLR) superfamily, are responsible for taking vitellogenin (Vg) into developing oocytes. Here the first full-length VgR cDNA from a hemipteran insect, the brown planthopper (Nilaparvata lugens), was cloned and sequenced. The complete mRNA sequence was 6174bp in length with an open reading frame (ORF) of 5796bp encoding 1931 amino acid residues. N. lugens VgR (NlVgR) contained two ligand-binding domains with five LDLR Class A cysteine-rich repeats in the first domain and eight in the second domain, which was similar to other insect VgRs. NlVgR was specifically expressed in the ovary, and the mRNA level started to increase after adult female emergence, with a peak on day 7 in the adult stage, and then declined. Western blot analysis of NlVgR protein revealed an ovary-specific expression pattern, which was consistent with NlVgR transcript detection. Injection with NlVgR double-stranded RNA (dsRNA) significantly disturbed NlVgR, which led to a decrease in NlVg protein content in the ovaries, an accumulation of NlVg protein in the hemolymph, the arrested development of ovaries, and the failure of insects to reproduce. Besides, NlVgR expression was significantly upregulated after the topical application of juvenile hormone (JH) III. These results suggest that VgR is critical for Vg uptaking of oocytes and it plays an important role in insect fecundity.

Analysis of Genes Expression of Spodoptera exigua Larvae upon AcMNPV Infection

BackgroundThe impact of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) infection on host gene expression in Spodoptera exigua 4th instar larvae was investigated through the use of 454 sequencing-based RNA-seq of cDNA libraries developed from insects challenged with active AcMNPV or heat-inactivated AcMNPV.Methodology/Principal FindingsBy comparing the two cDNA libraries, we show that 201 host genes are significantly up-regulated and 234 genes are significantly down-regulated by active AcMNPV infection. Down-regulated host genes included genes encoding antimicrobial peptides, namely three gloverin isoforms and an attacin, indicating that the viral infection actively repressed the expression of a portion of the host immune gene repertoire. Another interesting group of down-regulated host genes included genes encoding two juvenile hormone binding proteins and a hexamerin, all of which are involved in juvenile hormone regulation. The expression of these genes was enhanced by the topical application of Juvenile Hormone III (JHIII) in the insects challenged with heat-inactivated AcMNPV. However, infection with the active virus strongly suppresses the expression of these three genes, regardless of the absence or presence of JHIII.Conclusions/SignificanceUsing RNA-seq, we have identified groups of immune-regulated and juvenile hormone-regulated genes that are suppressed by infection with active AcMNPV. This information and further studies on the regulation of host gene expression by AcMNPV will provide the tools needed to enhance the utility of the virus as an effective protein expression system and as an insecticide.

Molecular cloning, transcriptional regulation, and differential expression profiling of vitellogenin in two wing‐morphs of the brown planthopper, Nilaparvata lugens Stål (Hemiptera: Delphacidae)

The brown planthopper, Nilaparvata lugens, is a serious pest of rice crops throughout Asia and exhibits wing dimorphism, with brachypterous adults having reduced wings and macropterous adults possessing fully developed wings. To understand the reproductive strategies in two wing-morphs of this insect, the transcript encoding the major yolk protein precursor, vitellogenin (Vg), was cloned. The complete mRNA transcript was 6314 bp, which encodes a protein of 2063 residues including an 18-residue putative signal peptide. Analysis of the mature protein revealed two vitellogenin-N (or lipoprotein amino-terminal) domains near the N-terminus and a von Willebrand factor type D domain near the C-terminus. In addition, a highly conserved motif GL/ICG, and a number of cysteine residues were identified near the C-terminus. Northern blot analysis identified a ∼6.8 kb Vg gene transcript that was expressed exclusively in the adult female fat body cells. The expression profile revealed that the Vg gene starts to be expressed earlier (on day 3) in brachypters as compared to macropters where the mRNA transcript was observed on day 4. However, in both morphs, the amount of Vg mRNA increased to reach high levels during vitellogenic periods [from day 4 (in brachypters) and day 5 (in macropters) and onwards]. Reflecting the RNA transcription pattern, the Vg signal was detected by immunoblotting on day 3 and day 4 in haemolymph of brachypterous and macropterous females, respectively, and that was increased every day and remained high during the vitellogenic periods. Furthermore, the topical application of juvenile hormone (JH) III had up-regulated the Vg gene expression suggesting that the Vg gene is regulated by JH in N. lugens. In addition, it was demonstrated by Southern blot analysis that there exists a single copy of the gene in the N. lugens genome. A delayed trend in expression (of both the transcript and the protein) demonstrated by macropterous females in the present studies supports the hypothesis of prereproductive long distance migration in this wing-dimorphic species.

The four hexamerin genes in the honey bee: structure, molecular evolution and function deduced from expression patterns in queens, workers and drones

BackgroundHexamerins are hemocyanin-derived proteins that have lost the ability to bind copper ions and transport oxygen; instead, they became storage proteins. The current study aimed to broaden our knowledge on the hexamerin genes found in the honey bee genome by exploring their structural characteristics, expression profiles, evolution, and functions in the life cycle of workers, drones and queens.ResultsThe hexamerin genes of the honey bee (hex 70a, hex 70b, hex 70c and hex 110) diverge considerably in structure, so that the overall amino acid identity shared among their deduced protein subunits varies from 30 to 42%. Bioinformatics search for motifs in the respective upstream control regions (UCRs) revealed six overrepresented motifs including a potential binding site for Ultraspiracle (Usp), a target of juvenile hormone (JH). The expression of these genes was induced by topical application of JH on worker larvae. The four genes are highly transcribed by the larval fat body, although with significant differences in transcript levels, but only hex 110 and hex 70a are re-induced in the adult fat body in a caste- and sex-specific fashion, workers showing the highest expression. Transcripts for hex 110, hex 70a and hex70b were detected in developing ovaries and testes, and hex 110 was highly transcribed in the ovaries of egg-laying queens. A phylogenetic analysis revealed that HEX 110 is located at the most basal position among the holometabola hexamerins, and like HEX 70a and HEX 70c, it shares potential orthology relationship with hexamerins from other hymenopteran species.ConclusionsStriking differences were found in the structure and developmental expression of the four hexamerin genes in the honey bee. The presence of a potential binding site for Usp in the respective 5' UCRs, and the results of experiments on JH level manipulation in vivo support the hypothesis of regulation by JH. Transcript levels and patterns in the fat body and gonads suggest that, in addition to their primary role in supplying amino acids for metamorphosis, hexamerins serve as storage proteins for gonad development, egg production, and to support foraging activity. A phylogenetic analysis including the four deduced hexamerins and related proteins revealed a complex pattern of evolution, with independent radiation in insect orders.

RNA interference directed against ribosomal protein S3a suggests a link between this gene and arrested ovarian development during adult diapause in Culex pipiens

Arrested ovarian development is a key characteristic of adult diapause in the mosquito Culex pipiens. In this study we propose that ribosomal protein S3a (rpS3a), a small ribosomal subunit, contributes to this shutdown. RpS3a is consistently expressed in females of C. pipiens that do not enter diapause, but in females programmed for diapause, expression of the rpS3a transcript is dramatically reduced for a brief period in early diapause (7-10 days after adult eclosion). RNA interference directed against rpS3a in nondiapausing females arrested follicle development, mimicking the diapause state. The effect of the dsRNA injection faded within 10 days, allowing the follicles to grow again, thus the suppression of rpS3a caused by RNAi did not permanently block ovarian development, implying that a brief suppression of rpS3a is not the only factor contributing to the diapause response. The arrest in development that we observed in dsRNA-injected females could be reversed with a topical application of juvenile hormone III, an endocrine trigger known to terminate diapause in this species. Though we speculate that many genes contribute to the diapause syndrome in C. pipiens, our results suggest that a shut down in the expression of rpS3a is one of the important components of this developmental response.

Morphometric changes on honeybee Apis mellifera L. workers fat body cells after juvenile hormone topic application at emergence

The effect of topical application of juvenile hormone (JH) over the lifetime of worker bees was evaluated in Apis mellifera, by measuring the area of the two cell types, trophocytes and oenocytes, found in the fat body. Topical application of 1 μl of a 1 μg/μl solution of JH in acetone to the abdomens of newly emerged workers produced an increase in cell size, in both types of cell of 5-day-old treated workers in relation to the untreated control. The treatment was more effective on the oenocytes, since there were significant differences compared to the averages of the treatments and the interaction of the treatments with the age of the workers. The developmental pattern seemed to differ from the treated group. However, subsequent effects were probably dependent on different, natural variations in hormonal levels.

Ultrastructural modifications in the venom glands of workers of Apis mellifera L. (Hymenoptera: Apidae) promoted by topical application of juvenile hormone

The present study analyzed, the influence of the treatment with juvenile hormone on the ultrastructure of Apis mellifera L. workers' venom glands. Newly emerged workers received topical application of 1 microl of juvenile hormone diluted in hexane, in the concentration of 2 microg/pl. Two controls were used; one control received no treatment (group C1) and other received topical application of 1 microl of hexane (group C2). The aspect of the glandular cells, in not treated newly emerged workers, showed that they are not yet secreting actively. Cellular modifications happened according to the worker age and to the glandular area considered. The most active phase of the gland happened from the emergence to the 14th day. At the 25th day the cells had already lost their secretory characteristic, being the distal area the first to suffer degeneration. The treatment with juvenile hormone and hexane altered the temporal sequence of the glandular cycle, forwarding the secretory cycle and degeneration of the venom gland.

Changes in ornithine decarboxylase activity in response to temperature stress and stimulation of juvenile hormone in Anastrepha fraterculus (Diptera, Tephritidae)

Ornithine decarboxylase (ODC) activity was analyzed in Anastrepha fraterculus (Diptera) females (4 days old) submitted to temperature stress (6 degrees C and 20/6 degrees C) and the topical application of juvenile hormone (JH). ODC activity and ejaculatory apodeme measurements (length and width) were made in males (15 days old) after 6 degrees C stress. JH dose of 500 ng and incubation of 3, 7, and 18 h increased ODC activity. Females reared at 6 degrees C and 20/6 degrees C had higher ODC activity than those reared at 25 degrees C. The treatment of 6 degrees C and JH incubation for 1 h increased ODC activity when compared to 6 degrees C treatments only. However, the treatment of 20/6 degrees C only after 3 or 18 h of JH incubation resulted in higher ODC activity than controls (20/6 degrees C) or 20/6 degrees C plus 1 h of JH incubation. Males did not undergo differences in ODC activity when reared at 6 degrees C or 25 degrees C but the ejaculatory apodeme measurements was higher in those reared at 25 degrees C than in those reared at 6 degrees C. The results can be considered an adaptive process to environmental changes.

Tissue localization and regulation by juvenile hormone of human allergen Bla g 4 from the German cockroach,Blattella germanica(L.)

The German cockroach, Blattella germanica (L.), produces several potent protein aeroallergens, including Bla g 4, a approximately 20 kDa lipocalin. RT-PCR, Northern analyses and in situ hybridization showed that Bla g 4 is expressed only in the adult male reproductive system. Western blotting and ELISA with rBla g 4 antiserum detected immunoreactivity in the utricles and the conglobate gland, but not in other tissues of the male reproductive system. The Bla g 4 protein content of males increased from adult emergence to day 14, but during copulation Bla g 4 was depleted in the male and transferred to the female within the spermatophore. Topical application of juvenile hormone III stimulated Bla g 4 production by both conglobate gland and utricles.

Breaking a paradigm: male-produced aggregation pheromone for the Colorado potato beetle.

A male-produced aggregation pheromone was identified for the Colorado potato beetle Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). While male beetles produced only minor amounts of the pheromone, its production could be enhanced by topical application of juvenile hormone III (JH III) (eightfold), by antennectomy (40-fold) or by the combined treatment of JH III and antennectomy (almost 200-fold); this enhancement enabled the identification of the compound as (S)-3,7-dimethyl-2-oxo-oct-6-ene-1,3-diol [(S)-CPB I], a unique structure for an insect pheromone. Antennal receptors of both sexes responded selectively to the (S)-enantiomer. Both male and female Colorado potato beetles were attracted to serial source loads of (S)-CPB I in laboratory bioassays; (R)-CPB I was inactive or inhibitory, as demonstrated by the inactivity of the racemate. This is the first identification of a pheromone for the Colorado potato beetle and differs from the paradigm of a female-produced pheromone for this insect. The attractant is also the first male-produced pheromone identified for the Chrysomelidae. The discovery that both JH III and antennectomy increase levels of the pheromone (S)-CPB I indicates the existence of a feedback system involving antennal input, and this system may be under hormonal control.

Mating experience and juvenile hormone enhance sexual signaling and mating in male Caribbean fruit flies

Young mated male Caribbean fruit flies [Anastrepha suspensa (Loew)] have greater sexual prowess than their virgin counterparts. After mating for the first time, 6- to 7-day-old males released twice as much sex pheromone and acquired another mate in less than half the time required by virgin males of the same age. Mass spectroscopic analysis of extracts of hemolymph from mated and virgin 7-day-old males resulted in identification of juvenile hormone III bisepoxide and juvenile hormone III in a ratio of 2.5:1. Extracts from mated males contained 3-fold more juvenile hormone than did extracts from virgins. Enhancement of sexual signaling, pheromone release, and mating was induced by topical application of juvenile hormone, methoprene, or fenoxycarb. Newly eclosed adult males treated with juvenoids engaged in sexual signaling, released pheromone, and mated at significantly earlier ages than control males. We conclude that juvenile hormone mediated a positive feedback system that imparted a competitive advantage, guaranteeing that males who mated at an early age would out-compete virgins of the same age for mating opportunities. Additionally, the results support the hypothesis that juvenile hormone is a pivotal hormone coordinating the development of sexual signaling and reproductive maturity in these flies.

Mating experience and juvenile hormone enhance sexual signaling and mating in male Caribbean fruit flies

Young mated male Caribbean fruit flies [ Anastrepha suspensa (Loew)] have greater sexual prowess than their virgin counterparts. After mating for the first time, 6- to 7-day-old males released twice as much sex pheromone and acquired another mate in less than half the time required by virgin males of the same age. Mass spectroscopic analysis of extracts of hemolymph from mated and virgin 7-day-old males resulted in identification of juvenile hormone III bisepoxide and juvenile hormone III in a ratio of 2.5:1. Extracts from mated males contained 3-fold more juvenile hormone than did extracts from virgins. Enhancement of sexual signaling, pheromone release, and mating was induced by topical application of juvenile hormone, methoprene, or fenoxycarb. Newly eclosed adult males treated with juvenoids engaged in sexual signaling, released pheromone, and mated at significantly earlier ages than control males. We conclude that juvenile hormone mediated a positive feedback system that imparted a competitive advantage, guaranteeing that males who mated at an early age would out-compete virgins of the same age for mating opportunities. Additionally, the results support the hypothesis that juvenile hormone is a pivotal hormone coordinating the development of sexual signaling and reproductive maturity in these flies.

Characterization of the esterase isozymes ofIps typographus (coleoptera, scolytidae)

Four esterase isozymes hydrolyzing α-naphthyl acetate (α-NA) were detected screening whole body homogenates of larvae and adults of Ips typographus by electrophoresis. Two of the four isozymes (isozymes 3 and 4) were not detected by α-NA staining in the pupal stage, but topical application of juvenile hormone III (JH III) on the pupa induced these isozymes. The JH esterase (JHE) activity on the gel was associated with the proteins of isozyme 2. The compounds OTFP, PTFP, and DFP inhibited this catalytic activity of isozyme 2 on the gel at low concentrations, whereas the proteins of isozyme 3 and 4 were affected only at higher concentrations. A quantitative developmental study was performed to characterize which of the esterases hydrolyzed JH III, using a putative surrogate substrate for JH (HEXTAT) and α-NA. The I50 of several esterase inhibitors and the JH metabolites were also defined. All findings supported the results that a protein associated with isozyme 2 is catabolizing JH and that isozymes 3 and 4 are the main contributors to the general esterase activity on α-NA. The JHE from Tenebrio molitor was purified by affinity chromatography. Although the recovery was low, an analytical isoelectric focusing gel showed that the JHE activity of the purified enzyme. T. molitor cochromatographed at the same pl as the JHE activity of I. typographus. Arch. Insect Biochem. Physiol. 34:203–221, 1997. © 1997 Wiley-Liss, Inc.

Effects of Juvenile Hormone and Head Ligations on the Succession of Cryptic Larval Color Patterns in Cerura vinula (Lepidoptera: Notodontidae)

The color patterns of the different larval instars of Cerura vinula Linnaeus 1758 are described and their protective function is discussed. The cuticular melanization is stepwise reduced, in different parts of the pattern at different molts. In the wandering stage, a temporary appearance of epidermal ommochrome results in completely red animals. It is investigated whether the succession of different larval colorations is controlled by the Corpora allata through different levels of juvenile hormone (JH). Head ligation at the end of a young larval instar, eliminating the Corpora allata, causes reddening of the whole epidermis without further development. Injection or topical application of JH before a larval molt causes uniform reduction of all melanin in the new cuticle. This effect does not mimic the stepwise reduction in different cuticular areas which occurs during the normal development. It is concluded that the ommochrome formation at the wandering stage depends on the absence of JH. Head ligation inhibits, in young larvae, the distribution of JH but, at the same time, interrupts that of prothoracicotropic hormone [PTTH], so that a small ecdysone peak results in the absence of JH, similar to the situation at the wandering stage, when it evokes ommochrome formation, but not a complete molt. Excessive JH causes an unspecific melanin inhibition, which cannot, by itself, explain the normal sequence of larval melanization patterns.

Involvement of juvenile hormone in the induction of antifreeze protein production by the fat body of larvae of the beetle Dendroides canadensis

AbstractLarvae of the Pyrochroid beetle Dendroides canadensis produce hemolymph antifreeze proteins (AFPs) as an adaptation to subzero overwintering temperatures. The AFPs are produced in early autumn in response to various environmental cues (short photoperiod, low temperature, short thermoperiod). The study presented here reinforces and extends the initial finding (Horwath and Duman, '83b) that juvenile hormone is involved in stimulating production of AFPs. Dose response curves showed seasonal variation in the sensitivity of the larvae to topical application of Juvenile Hormone (JH), with late summer larvae being more sensitive than early summer ones. The time response curve for induction of AFP activity in late summer larvae demonstrated that ∼14 days are required after JH treatment for the appearance of a significant increase in AFP activity. The timing of this response to JH induction is similar to that seen previously when larvae were acclimated to a short photoperiod.Studies with cultured fat bodies showed that the fat body is a probable site of production of the AFP, and further emphasized a role for JH in AFP induction. However, stimulation of AFP production by fat body only occurred when the fat bodies were removed from larvae that had been pretreated with JH. The presence of JH in the culture medium did not stimulate fat bodies taken from larvae that had not been pre‐treated. This suggests that an additional hormone(s), or other factor, may be required to induce fat body production of AFPs.

Juvenile Hormone and host‐plant colonization by the black bean aphid, Aphis fabae

Abstract Host‐plant colonization by winged, summer forms of Aphis fabae Scop, involves the resumption of embryogenesis and larviposition, leading eventually to flight muscle degeneration. Topical application of Juvenile Hormone I to young adults which (a) had access to the host plant, (b) were starved, (c) were starved and treated with precocene III, or (d) were decapitated shortly after the final ecdysis suggests that embryogenesis and flight muscle histolysis may be stimulated by an increase in Juvenile Hormone titres after settling. The duration of the pre‐reproductive period and the initial reproductive rate were not significantly affected, possibly because other neuroendocrine factors are involved in parturition.