High Juvenile Hormone Research Articles

Investigating the role of aae-miR-34-5p in the regulation of juvenile hormone biosynthesis genes in the mosquito Aedes aegypti

Juvenile hormone (JH) controls the development and reproduction of insects. Therefore, a tight regulation of the expression of JH biosynthetic enzymes is critical. microRNAs (miRNAs) play significant roles in the post-transcriptional regulation of gene expression by interacting with complementary sequences in target genes. Previously, we reported that several miRNAs were differentially expressed during three developmental stages of Aedes aegypti mosquitoes with different JH levels (no JH, high JH, and low JH). One of these miRNAs was aae-miR-34-5p. In this study, we identified the presence of potential target sequences of aae-miR-34-5p in the transcripts of some genes encoding JH biosynthetic enzymes. We analysed the developmental expression patterns of aae-miR-34-5p and the predicted target genes involved in JH biogenesis. Increases in miRNA abundance were followed, with a delay, by decreases in transcript levels of target genes. Application of an inhibitor and a mimic of aae-miR-34-5p led respectively to increased and decreased levels of thiolase transcripts, which is one of the early genes of JH biosynthesis. Female adult mosquitoes injected with an aae-miR-34-5p inhibitor exhibited significantly increased transcript levels of three genes encoding JH biosynthetic enzymes, acetoacetyl-CoA thiolase (thiolase), farnesyl diphosphate phosphatase, and farnesal dehydrogenase. Overall, our results suggest a potential role of miRNAs in JH production by directly targeting genes involved in its biosynthesis.

Juvenile hormone antagonistic activity of secondary metabolites from Streptomyces lactacystinicus and their insecticidal activity against Plutella xylostella

• IMBL-1412 isolate showed high JHAN and insecticidal activity against P. xylostella . • IMBL-1823 isolate showed high JHAN and insecticidal activity against P. xylostella . • Both IMBL-1412 and IMBL-1823 were identified as Streptomyces lactacystinicus . • Ethyl acetate fraction of them showed high JHAN and insecticidal activities. Due to their specificity to target insects and low toxicity to non-target organisms, insect growth regulators (IGRs) have been promising alternatives to neurotoxic insecticides. Actinobacteria produce a wide range of secondary metabolites with insecticidal and insect growth regulatory activities. In this study, the culture media of 25 actinobacteria isolates showing high juvenile hormone antagonist (JHAN) activity were assessed for their insecticidal activity to identify novel IGR compounds toxic to Plutella xylostella . Among them, four isolates exhibited high insecticidal activity against 3rd instar larvae of P. xylostella . Two isolates of IMBL-1412 and IMBL-1823 showing relatively high insecticidal activities (greater than90% mortality) were identified as Streptomyces lactacystinicus based on colony color on various International Streptomyces Project (ISP) media and nucleotide sequences of the 16S rRNA gene. The ethyl acetate fractions of both isolates showed high JHAN and insecticidal activities against P. xylostella larvae at a concentration of 100 ppm when the culture media of these two isolates were extracted sequentially using hexane, ethyl acetate, and butyl alcohol. These results suggested that secondary metabolites of these actinobacterial isolates could be efficiently applied to develop novel IGR insecticides for the control of P. xylostella .

Juvenile Hormone III R Stereoisomer Is Specifically Synthesized by Honeybees (Apis mellifera ligustica) and Shows a Higher Biological Activity in Regulating Their Social Behavior.

The juvenile hormone (JH) plays a key role in the development of honeybee larvae and the alteration of adult behavior. Verification methods of types and stereoisomers of JHs in honeybees were established in this study. The regulatory modes of different stereoisomers of JH III on the social behaviors of honeybees were revealed by the disparity of interaction and RNA-seq. This result represented the first assessment of the effects of R-JH III and S-JH III in honeybee interactions; the former (367 times in total) was significantly higher than the latter (235 times in total); honeybees with high JH titers are always welcome in the colony because the effect of JH III on bees involves the sensing and signaling of hormones, and R-JH III is much more active than S-JH III in this regulation. Efficient R-JH III may be the insurance for bees to establish their social system advantages.

Humoral and Cellular Defense Mechanisms in Rebel Workers of Apis mellifera.

Simple SummaryImmune mechanisms in insects include cellular and humoral defenses. One of these cellular defense mechanisms is phagocytosis. The humoral defense components are produced in the fat body cells, the development of which depends on the juvenile hormone (JH) titers and vitellogenin (Vg) concentration. These mechanisms also regulate caste formation. In a colony, a queen, a worker, or a rebel can develop from eggs of the same genome. Rebels are reproductive workers that can lay eggs while maintaining tasks inside and outside the colony the same as sterile workers. The aim of our studies was to determine the phagocytic index, JH titer, Vg concentration, the number of oenocytes (fat body cells), and their size in the rebels and to compare them to those in normal workers. The rebels were characterized by high phagocytic indices, JH and Vg levels, and increased numbers and sizes of oenocytes in the fat body cells in comparison to the normal workers. These characteristics can be viewed as an evolutionary adaptation of these reproductive workers to life in a eusocial society.The physiological state of an insect depends on efficiently functioning immune mechanisms such as cellular and humoral defenses. However, compounds participating in these mechanisms also regulate reproductive caste formation and are responsible for reproductive division of labor as well as for labor division in sterile workers. Divergent reaction of the same genotype yielding reproductive queens and worker castes led to shaping of the physiological and behavioral plasticity of sterile or reproductive workers. Rebels that can lay eggs while maintaining tasks inside and outside the colony exhibit both queen and worker traits. So, we expected that the phagocytic index, JH3 titer, and Vg concentration would be higher in rebels than in normal workers and would increase with their age. We also assumed that the numbers of oenocytes and their sizes would be greater in rebels than in normal workers. The rebels and the normal workers were collected at the age of 1, 7, 14, and 21 days, respectively. Hemolymph and fat bodies were collected for biochemical and morphological analyses. The high levels of JH, Vg, and the phagocytic index, as well as increased numbers and sizes of oenocytes in the fat body cells demonstrate the physiological and phenotypic adaptation of rebels to the eusocial life of honeybees.

Juvenile hormone functions as a metabolic rate accelerator in bumble bees (Bombus terrestris)

Juvenile hormone (JH) is a modulator of many physiological transitions in insects, including molting, metamorphosis, diapause, and reproduction. These processes often include metabolic changes. Here we show that JH accelerates metabolic rate in bumble bees (Bombus terrestris). We reduced JH levels in worker bumble bees by removing their corpora allata (allatectomy) and elevated JH levels in queens through a topical application of JH-III. Natural and high JH levels increased the metabolic rate in both workers and queens and triggered an increased protein turnover rate. Following the treatments, JH also caused an increase in food consumption and a reduction in lipid levels and flight muscle mass of queens, and a reduction in lipids levels in workers. Furthermore, the topical application of a JH analog to queens prior to their diapause caused a decline in their survival of diapause. These findings support the hypothesis that JH acts as a metabolic rate accelerator, initiating a resource shift in bumble bees, and thereby reducing diapause survival in queens. Based on previous studies on JH we suggest that, additional to its behavioral or physiological effects, JH's function as an accelerator of metabolic processes is conserved across different life stages and insect species.

How Can Termites Achieve Their Unparalleled Postembryonic Developmental Plasticity? A Test for the Role of Intermolt-Specific High Juvenile Hormone Titers

Termites are “social cockroaches” and amongst the most phenotypically plastic insects. The different castes (i.e., two types of reproductives, workers, and soldiers) within termite societies are all encoded by a single genome and present the result of differential postembryonic development. Besides the default progressive development into winged sexuals of solitary hemimetabolous insects, termites have two postembryonic, non-terminal molts (stationary and regressive; i.e., molts associated, respectively, with no change or reduction of size/morphological differentiation) which allow them to retain workers, and two terminal developmental types to become soldiers and replacement reproductives. Despite this unique plasticity, especially the mechanisms underlying the non-terminal development are poorly understood. In 1982, Nijhout and Wheeler proposed a model how this diversity might have evolved. They proposed that varying juvenile hormone (JH) titers at the start, mid-phase, and end of each intermolt period account for the developmental diversity. We tested this rarely addressed model in the lower termiteCryptotermes secundususing phase-specific pharmacological manipulations of JH titers. Our results partially support the Nijhout and Wheeler model. These data are supplemented with gene expression studies of JH-related genes that characterize different postembryonic developmental trajectories. Our study provides new insights into the evolution of the unique postembryonic developmental plasticity of termites that constitutes the foundation of their social life.

Reproduction and signals regulating worker policing under identical hormonal control in social wasps

In social Hymenoptera, fertility and fertility signalling are often under identical hormonal control, and it has been suggested that such hormonal pleiotropies can help to maintain signal honesty. In the common wasp Vespula vulgaris, for example, fertile queens have much higher juvenile hormone (JH) titers than workers, and JH also controls the production of chemical fertility cues present on the females’ cuticle. To regulate reproductive division of labour, queens use these fertility cues in two distinct ways: as queen pheromones that directly suppress the workers’ reproduction as well as to mark queen eggs and enable the workers to recognize and police eggs laid by other workers. Here, we investigated the hormonal pleiotropy hypothesis by testing if experimental treatment with the JH analogue methoprene could enable the workers to lay eggs that evade policing. In support of this hypothesis, we find that methoprene-treated workers laid more eggs, and that the chemical profiles of their eggs were more queen-like, thereby causing fewer of their eggs to be policed compared to in the control. Overall, our results identify JH as a key regulator of both reproduction and the production of egg marking pheromones that mediate policing behaviour in eusocial wasps.

Juvenile hormone regulates brain-reproduction tradeoff in bumble bees but not in honey bees

Gonadotropic hormones coordinate processes in diverse tissues regulating animal reproductive physiology and behavior. Juvenile hormone (JH) is the ancient and most common gonadotropin in insects, but not in advanced eusocial honey bees and some ants. To start probing the evolutionary basis of this change, we combined endocrine manipulations, transcriptomics, and behavioral analyses to study JH regulated processes in a bumble bee showing a relatively simple level of eusociality. We found that in worker fat body, more JH-regulated genes were up- rather than down-regulated, and enriched for metabolic and biosynthetic pathways. This transcriptomic pattern is consistent with earlier evidence that JH is the major gonadotropin in bumble bees. In the brain, more JH-regulated genes were down- rather than up-regulated and enriched for protein turnover pathways. Brain ribosomal protein gene expression shows a similar trend of downregulation in dominant workers, which naturally have high JH titers. In other species, similar downregulation of protein turnover is found in aging brains or under stress, associated with compromised long-term memory and health. These findings suggest a previously unknown gonadotropin-mediated tradeoff. Analysis of published data reveals no such downregulation of protein turnover pathways in the brain of honey bee workers, which exhibit more complex eusociality and in which JH is not a gonadotropin but rather regulates division of labor. These results suggest that the evolution of complex eusociality in honey bees was associated with modifications in hormonal signalling supporting extended and extremely high fertility while reducing the ancient costs of high gonadotropin titers to the brain.

Reproductive workers insufficiently signal their reproductive ability in a paper wasp

Abstract Why workers forfeit direct reproduction is a crucial question in eusocial evolution. Worker reproduction provides an excellent opportunity to understand the mechanism of kin conflict resolution between the queen and workers. We evaluated behavioral and physiological differences among females in the paper wasp Polistes chinensis antennalis to examine why some workers reproduce under queenright conditions. Reproductive workers were old and foraged less early in the season; their cuticular hydrocarbon (CHC) profiles overlapped with those of queens but were significantly different. The distinct CHC profile of the eggs of the queen likely represented a cue for policing against those by workers. Juvenile hormone (JH) and dopamine seemed to be associated with gonadotropic function, and the JH level of reproductive workers was similar to that of the queen. The high JH level of reproductive workers likely facilitated their reproduction even under queenright conditions. Gene expression levels of the queen and reproductive workers differed only in vitellogenin. These results suggest that worker reproduction is facilitated by an increase in JH level; however, CHC is not a fertility-linked signal, but a queen-linked signal; consequently, reproductive workers without a queen-linked signal might be allowed to stay within the colony.

Epigenetic Regulator CoREST Controls Social Behavior in Ants.

Ants acquire distinct morphological and behavioral phenotypes arising from a common genome, underscoring the importance of epigenetic regulation. In Camponotus floridanus, "Major" workers defend the colony, but can be epigenetically reprogrammed to forage for food analogously to "Minor" workers. Here, we utilize reprogramming to investigate natural behavioral specification. Reprogramming of Majors upregulates Minor-biased genes and downregulates Major-biased genes, engaging molecular pathways fundamental to foraging behavior. We discover the neuronal corepressor for element-1-silencing transcription factor (CoREST) is upregulated upon reprogramming and required for the epigenetic switch to foraging. Genome-wide profiling during reprogramming reveals CoREST represses expression of enzymes that degrade juvenile hormone (JH), a hormone elevated upon reprogramming. High CoREST, low JH-degrader expression, and high JH levels are mirrored in natural Minors, revealing parallel mechanisms of natural and reprogrammed foraging. These results unveil chromatin regulation via CoREST as central to programming of ant social behavior, with potential far-reaching implications for behavioral epigenetics.

Diapause hormone directly stimulates the prothoracic glands of diapause larvae under juvenile hormone regulation in the bamboo borer, Omphisa fuscidentalis Hampson.

Larval diapause in many lepidopteran insects is induced and maintained by high juvenile hormone (JH). In the case of the bamboo borer, Omphisa fuscidentalis, the effect of JH is the opposite: The application of juvenile hormone analog (JHA: S-methoprene) terminates larval diapause, unlike in other insect species. Here, we analyzed the expression of JH-receptor Met, DH-PBAN, and Kr-h1 in the subesophageal ganglion (SG) from October to April using semi-quantitative polymerase chain reaction (PCR). The results show that OfMet and OfDH-PBAN messenger RNA in the SG are mainly expressed during the larval diapause stage, while OfKr-h1 increases during the pupal stage. Using tissue culture techniques and an enzyme-linked immunosorbent assay (ELISA), diapause hormone (DH) was found to induce ecdysteroidogenesis in the culture medium of the prothoracic gland (PG) after incubation for 30 min with 25 ng and 50 ng of DH. Thus, DH is a novel stimulator for the PG. We identified a DHR homolog in the bamboo borer and confirmed that it is expressed in the PG. In addition, for in vitro experiments, DH increased the expression levels of OfDHR, OfEcR-A, and ecdysone-inducible genes in the PG. These results demonstrate that DH can function as a prothoracicotropic factor, and this function of DH might be through of DHR expressed on PG cells. Consequently, DH is one of the key factors in larval diapause break which is triggered by JH in the bamboo borer, O. fuscidentalis.

Juvenile hormone and parental care in subsocial insects: implications for the role of juvenile hormone in the evolution of sociality

Juvenile hormone (JH) has both gonadotropic and non-gonadotropic roles in eusocial insects. There is controversy over whether the non-gonadotropic role is novel, related specifically to social evolution, or is a second conserved role. Study of subsocial insects suggests that when JH is non-gonadotropic, the specific associations of high JH, low vitellogenin, suppressed ovarian development and elevated metabolism may parallel those in workers of eusocial insects. This suggests that a more fundamental understanding of JH in insect biology is required.

Differential gene expression underlying ovarian phenotype determination in honey bee, Apis mellifera L., caste development

Adult honey bee queens and workers drastically differ in ovary size. This adult ovary phenotype difference becomes established during the final larval instar, when massive programmed cell death leads to the degeneration of 95–99% of the ovariole anlagen in workers. The higher juvenile hormone (JH) levels in queen larvae protect the ovaries against such degeneration. To gain insights into the molecular architecture underlying this divergence critical for adult caste fate and worker sterility, we performed a microarray analysis on fourth and early fifth instar queen and worker ovaries. For the fourth instar we found nine differentially expressed genes (DEGs) with log2FC > 1.0, but this number increased to 56 in early fifth-instar ovaries. We selected 15 DEGs for quantitative PCR (RT-qPCR) analysis. Nine differed significantly by the variables caste and/or development. Interestingly, genes with enzyme functions were higher expressed in workers, while those related to transcription and signaling had higher transcript levels in queens. For the RT-qPCR confirmed genes we analyzed their response to JH. This revealed a significant up-regulation for two genes, a short chain dehydrogenase reductase (sdr) and a heat shock protein 90 (hsp90). Five other genes, including hsp60 and hexamerin 70b (hex70b), were significantly down-regulated by JH. The sdr gene had previously come up as differentially expressed in other transcriptome analyses on honey bee larvae and heat shock proteins are frequently involved in insect hormone responses, this making them interesting candidates for further functional assays.

Sexual difference in juvenile-hormone titer in workers leads to sex-biased soldier differentiation in termites

In termites, the soldier caste, with its specialized defensive morphology, is one of the most important characteristics for sociality. Most of the basal termite species have both male and female soldiers, and the soldier sex ratio is almost equal or only slightly biased. However, in the apical lineages (especially family Termitidae), there are many species that have soldiers with strongly biased sex ratio. Generally in termites, since high juvenile hormone (JH) titer is required for soldier differentiation from a worker via a presoldier stage, it was hypothesized that the biased soldier-sex ratio was caused by differences in JH sensitivity and/or JH titer between male and female workers. Therefore, we focused on the presoldier differentiation and the worker JH titer in species with only male soldiers (Nasutitermes takasagoensis) and with both male and female soldiers (Reticulitermes speratus) in natural conditions. In the former species, there are four types of workers; male minor, male medium, female medium and female major workers, and presoldiers differentiate from male minor workers. First, we tried to artificially induce presoldiers from male and female workers. In N. takasagoensis, the presoldier differentiation rate and mortality was significantly higher in male minor workers. Morphological analyses showed that both male and female induced presoldiers possessed normal soldier-specific morphologies. It was suggested that female workers, from which soldiers do not differentiate under natural conditions, also maintained the physiological and developmental potential for soldier differentiation. In R. speratus, however, no differences were observed in solder differentiation rate and mortality between male and female workers. Second, the JH titers of each sex/type of workers were quantified by high performance liquid chromatography–mass spectrometry in two different seasons (April and December). The results showed that, in N. takasagoensis, JH titer in male minor workers was consistently higher than those in other worker types. In R. speratus, in contrast, there were no significant differences in JH titers between male and female workers. These results suggested that, in N. takasagoensis, male minor workers maintain JH titers at a high level throughout a year, and this may cause the male-biased presoldier differentiation.

Expressions of juvenile hormone biosynthetic genes during presoldier differentiation in the incipient colony of Zootermopsis nevadensis (Isoptera: Archotermopsidae)

Soldier differentiations require high juvenile hormone (JH) titers in workers and two molts via a presoldier stage. However, it is still unclear when the endogenous JH titer increases in workers, because identifying an individual before it undergoes the presoldier molt is quite difficult in natural conditions. In an incipient colony of Zootermopsis nevadensis Hagen, the oldest 3rd instar larva (Νo. 1 larva) always differentiates into a presoldier and the second 3rd instar larva (Νo. 2 larva) molts into a 4th instar larva. Consequently, by focusing on the ontogeny of the Νo. 1 larva, the timing of the increase in the JH titer can be determined. Here, we compared the expression levels of JH biosynthetic genes in the heads of Νo. 1 and Νo. 2 larvae. Most genes involved in JH biosynthesis were found in a genome database, and two candidate JH epoxidase (CYP15A1) genes were identified. Expression levels of JH acid methyltransferase (JHAMT) and one candidate CYP15A1, probably involved in the final steps of the JH biosynthesis, were specifically high in Νo. 1 larvae at 2–3 and 4–5 days after the appearance, respectively. Expression patterns of these genes may be used for the prediction of endogenous JH titer changes during presoldier differentiation.

Knockdown of the juvenile hormone receptor gene inhibits soldier-specific morphogenesis in the damp-wood termite Zootermopsis nevadensis (Isoptera: Archotermopsidae)

The Methoprene-tolerant (Met) protein has been established as a juvenile hormone (JH) receptor. Knockdown of the Met gene caused precocious metamorphosis and suppression of ovarian development. However, the function of Met in caste development of social insects is unclear. In termites, JH acts as a central factor for caste development, especially for soldier differentiation, which involves two molts from workers via a presoldier stage. Increased JH titer in workers is needed for the presoldier molt, and the high JH titer is maintained throughout the presoldier period. Although presoldiers have the fundamental morphological features of soldiers, the nature of the cuticle is completely different from that of soldiers. We expected that JH signals via Met are involved in soldier-specific morphogenesis of the head and mandibles during soldier differentiation, especially in the presoldier period, in natural conditions. To test this hypothesis, we focused on soldier differentiation in an incipient colony of the damp-wood termite Zootermopsis nevadensis. Met homolog (ZnMet) expression in heads increased just after the presoldier molt. This high expression was reduced by ZnMet double stranded (dsRNA) injection before the presoldier molt. Although this treatment did not cause any morphological changes in presoldiers, it caused strong effects on soldiers, their mandibles being significantly shorter and head capsules smaller than those of control soldiers. Injection of ZnMet dsRNA throughout the presoldier stage did not affect the formation of soldier morphology, including cuticle formation. These results suggested that the rapid increase in ZnMet expression and subsequent activation of JH signaling just after the presoldier molt are needed for the formation of soldier-specific weapons. Therefore, besides its established role in insect metamorphosis, the JH receptor signaling also underlies soldier development in termites.

High juvenile hormone titre and abdominal activation of JH signalling may induce reproduction of termite neotenics.

Termite castes are a key example of polyphenism, in which reproductive division of labour is clearly seen in colonies. The reproductive castes in termites include primary and neotenic reproductives; primary reproductives found a new colony whereas neotenics succeed them in the reproductive role when the primary reproductives die or become senescent. Neotenics usually differentiate from nymphs or workers by developing functional gonads while retaining juvenile characteristics; however, the developmental mechanism during neotenic differentiation remains poorly understood. Juvenile hormone (JH) mediates a number of aspects of developmental regulation in caste differentiation in termites. In the present study we quantified JH titres in neotenic reproductives of Reticulitermes speratus, and compared these with other developmental stages. In addition, expression changes in JH signalling gene homologues (Methoprene-tolerant [Met], Krüppel-homolog1, Broad-Complex) in the head, thorax and abdomen were investigated during neotenic differentiation. Finally, we examined the function of Met in reproduction of neotenics by RNA interference (RNAi). Our results showed that the JH titres of neotenics were significantly higher than those of nymphs and workers. JH signalling genes were highly expressed in neotenic abdomens, compared with those in workers and nymphs. Met RNAi resulted in the inhibition of vitellogenin gene expression in newly moulted neotenics. These results suggest that the fertility of neotenics might be controlled by a large increase of JH titres and body-part-specific activation of JH signalling pathways.

Juvenile hormone, metabolic rate, body mass and longevity costs in parenting burying beetles

Levels of juvenile hormone (JH) are elevated during parental care in burying beetles (Nicrophorus) at a time when ovarian activity is suppressed, suggesting that JH plays an alternative role to its better known gonadotropic function. Because parental activity in burying beetles is time-intensive, it might be expected to be energetically stressful and to impart longevity costs. We predicted that the active (feeding) stage of care would be associated with elevated JH and greater energy expenditure (higher metabolic rate, lower body mass), and that this stress would result in shorter life span. Parents experimentally manipulated into providing more care for young had increased levels of JH. For females, there was a significant longevity cost associated with parental care but not with mating or egg production. During carcass preparation, females and males that would later regurgitate to larvae showed an initial increase in body mass followed by a significant decrease in mass during the intense period of parental provisioning of young (during and just after the JH peak). Males that provided little care (on small carcasses with a female partner) showed no such anticipatory weight gain. An indirect measure of metabolic rate (VCO2) was nearly twice as high in caregiving females compared to nonbreeding females. These results suggest that the energy demands and/or high JH levels during care extract a significant cost on longevity. We propose that JH has evolved to play a novel role in parental care in burying beetles associated with extreme energy demands during feeding of offspring.

Juvenile Hormone Biosynthesis Gene Expression in the corpora allata of Honey Bee (Apis mellifera L.) Female Castes

Juvenile hormone (JH) controls key events in the honey bee life cycle, viz. caste development and age polyethism. We quantified transcript abundance of 24 genes involved in the JH biosynthetic pathway in the corpora allata-corpora cardiaca (CA-CC) complex. The expression of six of these genes showing relatively high transcript abundance was contrasted with CA size, hemolymph JH titer, as well as JH degradation rates and JH esterase (jhe) transcript levels. Gene expression did not match the contrasting JH titers in queen and worker fourth instar larvae, but jhe transcript abundance and JH degradation rates were significantly lower in queen larvae. Consequently, transcriptional control of JHE is of importance in regulating larval JH titers and caste development. In contrast, the same analyses applied to adult worker bees allowed us inferring that the high JH levels in foragers are due to increased JH synthesis. Upon RNAi-mediated silencing of the methyl farnesoate epoxidase gene (mfe) encoding the enzyme that catalyzes methyl farnesoate-to-JH conversion, the JH titer was decreased, thus corroborating that JH titer regulation in adult honey bees depends on this final JH biosynthesis step. The molecular pathway differences underlying JH titer regulation in larval caste development versus adult age polyethism lead us to propose that mfe and jhe genes be assayed when addressing questions on the role(s) of JH in social evolution.

Presoldier differentiation of Australian termite species induced by juvenile hormone analogues

Termite soldiers have species-specific defensive morphologies. High juvenile hormone (JH) titre is required for workers to moult into presoldiers (intermediates between workers and soldiers). Artificial inductions of presoldiers using JH or JH analogues (JHAs) have been conducted in many species, but levels of intergeneric and species-specific JH sensitivity are not clear. Here, we investigated mortality and presoldier induction rates after the same JHA treatments in three Australian termites: the rhinotermitids Coptotermes lacteus and Schedorhinotermes intermedius, and the early branching termite species Mastotermes darwiniensis. Induced presoldiers, especially in S. intermedius (soldiers with labral brush-like weapons), were used for the observation of outer morphologies using scanning electron microscopy. In C. lacteus, large numbers of presoldiers were induced by all JHAs examined, and the mortality rate of workers was quite high. On the other hand, presoldier induction rates were very low in S. intermedius and zero in M. darwiniensis. However, mortality rates of these two species were quite different. In M. darwiniensis, the numbers of dead individuals in almost all JHA-treated dishes were not significantly different from those of an acetone control. These results suggest that JHA sensitivity might be quite different among the three species, although further analyses using multiple colonies are needed to confirm this possibility. Scanning electron microscopy observation in S. intermedius showed that soldier-specific formations of frontal pores in the heads, including a median furrow running from the frontal pore to the apical part of the labrum, and many campaniform sensillae and short bristles at the end of the labrum were all initiated at the presoldier stage.