Acetyl coenzyme A carboxylase modulates lipogenesis and sugar homeostasis in Blattella germanica.

Lipid and sugar homeostasis is critical for insect development and survival. In this study, we characterized an acetyl coenzyme A carboxylase gene in Blattella germanica (BgACC) that is involved in both lipogenesis and sugar homeostasis. We found that BgACC was dominantly expressed in the fat body and integument, and was significantly upregulated after molting. Knockdown of BgACC in 5th-instar nymphs did not affect their normal molting to the next nymphal stage, but it caused a lethal phenotype during adult emergence. BgACC-RNA interference (RNAi) significantly downregulated total free fatty acid (FFA) and triacylglycerol (TAG) levels, and also caused a significant decrease of cuticular hydrocarbons (CHCs). Repression of BgACC in adult females affected the development of oocytes and resulted in sterile females, but BgACC-RNAi did not affect the reproductive ability of males. Interestingly, knockdown of BgACC also changed the expression of insulin-like peptide genes (BgILPs), which mimicked a physiological state of high sugar uptake. In addition, BgACC was upregulated when B. germanica were fed on a high sucrose diet, and repression of BgACC upregulated the expression of the glycogen synthase gene (BgGlyS). Moreover, BgACC-RNAi increased the circulating sugar levels and glycogen storage, and a longevity assay suggested that BgACC was important for the survival of B. germanica under conditions of high sucrose uptake. Our results confirm that BgACC is involved in multiple lipid biogenesis and sugar homeostasis processes, which further modulates insect reproduction and sugar tolerance. This study benefits our understanding of the crosstalk between lipid and sugar metabolism.

Diverse functions of the ecdysone receptor (EcR) in the panoistic ovary of the German cockroach.

Ecdysone regulates essential processes in insect life. Perhaps the most well-known of these are related to metamorphosis. However, ecdysone is also required to regulate the proliferation and differentiation of germ cells in the ovary. The role of ecdysone in insect oogenesis has been studied in depth in holometabolan species with meroistic ovaries, such as Drosophila melanogaster, while in hemimetabolan species with panoistic ovaries their functions are still poorly understood. In the present work, we studied the role of ecdysone in the ovary of the last nymphal instar of the cockroach Blattella germanica by using RNA interference to reduce the levels of the ecdysone receptor (EcR), and thereby deplete the expression of ecdysteroidogenic genes in the prothoracic gland. However, the expression of ecdysteroidogenic genes was upregulated in the ovary, resulting in cell overproliferation in the germarium, which appeared swollen. By analysing the expression of genes that respond to ecdysone, we found that when the source of 20E is the nymphal ovary, EcR appears to repress 20E-associated genes bypassing early genes signalling.

Expression profiles of an inactive aspartic protease (Bla g 2 allergen) in different tissues and developmental stages of the German cockroach (Blattella germanica).

Tergal glands are found in many insect species and contain constituents such as pheromones, sugars, proteins, and so forth. Preliminary studies have revealed that tergal gland secretions in the German cockroach (Blattella germanica L.) contain the human allergen Bla g 2 (B. germanica allergen 2), an inactive aspartic protease. Although Bla g 2 protein expression has been detected previously in various German cockroach body parts, including male tergal glands, studies that link protein expression in various life stages and tissues with mRNA and protein abundance have not been conducted. Therefore, the goal of this study was to measure the relative abundances of Bla g 2 protein and mRNA in different tissues and life stages of B. germanica using immunoblotting, quantitative PCR, and liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based quantitative profiling. We found that Bla g 2 protein was detected in every sampled tissue, including the male tergal glands. Protein abundance was relatively high in adult males and their tergal glands in comparison to nymphs and virgin females. Similarly, Bla g 2 mRNA transcript levels were also comparatively higher in male tergal glands and adult males. In conclusion, this study provides new information on the relative abundance and distribution of Bla g 2 allergen, a medically significant protein, in different tissues and developmental stages of the German cockroach and lays the foundation for future studies that aim to determine the function of this protein in B. germanica development.

Melanin synthesis genes BgTH and BgDdc affect body color and cuticle permeability in Blattella germanica.

Melanin is involved in cuticle pigmentation and sclerotization of insects, which is critical for maintaining structural integrity and functional completeness of insect cuticle. The 2 key enzymes of tyrosine hydroxylase (TH) and dopa decarboxylase (DDC) predicted in melanin biosynthesis are usually conserved in insects. However, it is unclear whether their function is related to epidermal permeability. In this study, we identified and cloned the gene sequences of BgTH and BgDdc from Blattella germanica, and revealed that they both showed a high expression at the molting, and BgTH was abundant in the head and integument while BgDdc was expressed highest in the fat body. Using RNA interference (RNAi), we found that knockdown of BgTH caused molting obstacles in some cockroaches, with the survivors showing pale color and softer integuments, while knockdown of BgDdc was viable and generated an abnormal light brown body color. Desiccation assay showed that the dsBgTH-injected adults died earlier than control groups under a dry atmosphere, but dsBgDdc-injected cockroaches did not. In contrast, when dsRNA-treated cockroaches were reared under a high humidity condition, almost no cockroaches died in all treatments. Furthermore, with eosin Y staining assay, we found that BgTH-RNAi resulted in a higher cuticular permeability, and BgDdc-RNAi also caused slight dye penetration. These results demonstrate that BgTH and BgDdc function in body pigmentation and affect the waterproofing ability of the cuticle, and the reduction of cuticular permeability may be achieved through cuticle melanization.

Genome-wide identification and expression pattern analysis of novel chemosensory genes in the German cockroach Blattella germanica.

The German cockroach Blattella germanica is an important urban insect pest worldwide. In many insects, chemosensation is essential for guiding their behaviors for survival. Although a large number of chemosensory-related genes have been identified in B. germanica, little information on tissue-specific and developmental expression patterns has not been uncovered yet. In this study, we performed transcriptome analysis of different B. germanica tissues to reveal novel chemosensory proteins (CSPs) and sensory neuron membrane proteins (SNMPs). In addition, a phylogenetic tree and gender-specific expression of multiple chemosensory gene families have been analyzed. We identified three CSPs genes (BgerCSP11, BgerCSP12, and BgerCSP13) and five SNMP genes in B. germanica. Tissue-specific expression profiling showed that CSP1, 8, and 9 exhibited significant expression levels in both adult and 5th instar nymph antennae. The results have paved the way for further functional study of the chemosensory mechanism in B. germanica and provided potential insecticide targets.

Binding of de novo synthesized radiolabeled juvenile hormone (JH III) by JH receptors from the Cuban subterranean termite Prorhinotermes simplex and the German cockroach Blattella germanica.

Juvenile hormone (JH) controls insect reproduction and development through an intracellular receptor complex comprising two bHLH-PAS proteins, the JH-binding Methoprene-tolerant (Met) and its partner Taiman (Tai). Many hemimetabolous insects including cockroaches strictly depend on JH for stimulation of vitellogenesis. In termites, the eusocial hemimetabolans, JH also regulates the development of caste polyphenism. Studies addressing the agonist ligand binding to recombinant JH receptors currently include three species belonging to two holometabolous insect orders, but none that would represent any of the hemimetabolous orders. Here, we examined JH receptors in two representatives of Blattodea, the cockroach Blattella germanica and the termite Prorhinotermes simplex. To test the JH-binding capacity of Met proteins from these species, we performed chemical synthesis and tritium labeling of the natural blattodean JH homolog, JH III. Our improved protocol increased the yield and specific activity of [10-3H]JH III relative to formerly available preparations. Met proteins from both species specifically bound [3H]JH III with high affinity, whereas Met variants mutated at a critical position within the ligand-binding domain were incapable of such binding. Furthermore, JH III and the synthetic JH mimic fenoxycarb stimulated dimerization between Met and Tai components of the respective JH receptors of both species. These data present primary evidence for agonist binding by JH receptors in any hemimetabolous species and provide a molecular basis for JH action in cockroaches and termites.

Modulation of fatty acid elongation in cockroaches sustains sexually dimorphic hydrocarbons and female attractiveness.

Insect cuticular hydrocarbons (CHCs) serve as important intersexual signaling chemicals and generally show variation between the sexes, but little is known about the generation of sexually dimorphic hydrocarbons (SDHCs) in insects. In this study, we report the molecular mechanism and biological significance that underlie the generation of SDHC in the German cockroach Blattella germanica. Sexually mature females possess more C29 CHCs, especially the contact sex pheromone precursor 3,11-DimeC29. RNA interference (RNAi) screen against the fatty acid elongase family members combined with heterologous expression of the genes in yeast revealed that both BgElo12 and BgElo24 were involved in hydrocarbon (HC) production, but BgElo24 is of wide catalytic activities and is able to provide substrates for BgElo12, and only the female-enriched BgElo12 is responsible for sustaining female-specific HC profile. Repressing BgElo12 masculinized the female CHC profile, decreased contact sex pheromone level, and consequently reduced the sexual attractiveness of female cockroaches. Moreover, the asymmetric expression of BgElo12 between the sexes is modulated by sex differentiation cascade. Specifically, male-specific BgDsx represses the transcription of BgElo12 in males, while BgTra is able to remove this effect in females. Our study reveals a novel molecular mechanism responsible for the formation of SDHCs and also provide evidences on shaping of the SDHCs by sexual selection, as females use them to generate high levels of contact sex pheromone.

siRNA enrichment in Argonaute 2-depleted Blattella germanica.

BACKGROUND: RNA interference (RNAi) is a cellular mechanism used to fight various threats, including transposons, aberrant RNAs, and some types of viruses. This mechanism relies on the detection of dsRNA molecules, which through a pathway involving Dicer-2 (Dcr-2) and Argonaute 2 (AGO2), produces small interfering RNAs (siRNAs) that bind to the complementary RNAs triggering their degradation. METHODS: Using the cockroach Blattella germanica as a model, we examined AGO2 activity by depleting its mRNA using RNAi and analyzing the phenotypes produced. RESULTS: Depleting AGO2 expression had no remarkable effect on nymphal development or reproduction. dsRNA treatment triggered an immediate and transitory increase in AGO2 expression, independently of Dcr-2 action. In addition, we analyzed the siRNAs generated after injecting a heterologous dsRNA in control and AGO2-depleted animals. The results revealed that obtained siRNAs mapped non-uniformly along the dsRNA sequence. In AGO2-depleted animals, the proportion of 22 nucleotide reads was higher and accumulations of reads appeared in areas less well-represented in the controls. We also detected a preference for cytosine as the first nucleotide in controls that was significantly attenuated in AGO2-depleted individuals. CONCLUSIONS/GENERAL SIGNIFICANCE: The siRNAs produced from a dsRNA mapped heterogeneously along the length of the dsRNA and this arrangement depends on the dsRNA sequence. AGO2 exerts its role as nuclease on the siRNA duplexes independently of its action on the corresponding mRNA. This study sheds light on an extremely useful process for reverse genetics in laboratories, in addition to the design of more effective, specific, and eco-friendly pest-control strategies.

E93-depleted adult insects preserve the prothoracic gland and molt again.

Insect metamorphosis originated around the middle Devonian, associated with the innovation of the final molt; this occurs after histolysis of the prothoracic gland (PG; which produces the molting hormone) in the first days of adulthood. We previously hypothesized that transcription factor E93 is crucial in the emergence of metamorphosis, because it triggers metamorphosis in extant insects. This work on the cockroach Blattella germanica reveals that E93 also plays a crucial role in the histolysis of PG, which fits the above hypothesis. Previous studies have shown that the transcription factor FTZ-F1 is essential for PG histolysis. We have found that FTZ-F1 depletion towards the end of the final nymphal instar downregulates the expression of E93, whereas E93-depleted nymphs molt to adults that retain a functional PG. Interestingly, these adults are able to molt again, which is exceptional in insects. The study of insects able to molt again in the adult stage may reveal clues about how nymphal epidermal cells definitively become adult cells, and whether it is possible to reverse this process.

Expression, Affinity, and Functional Characterization of the Specific Binding of Two Putative Pheromone-Binding Proteins in the Omnivorous German Cockroach Blattella germanica.

The German cockroach Blattella germanica (L.) is an important pest in medical, veterinary, and public health. Studies on the olfaction mechanism of hemimetabolous insects have rarely been reported, especially in cockroaches. Pheromone-binding proteins (PBPs) play a vital role in insect sex pheromone recognition, which solubilize and carry the hydrophobic pheromonal compounds through the antennal lymph to receptors. In this study, two potential PBPs (BgerOBP26 and BgerOBP40) were identified on the basis of their biased expression in male antennae using tissue transcriptome data and verified by the quantitative real-time polymerase chain reaction approach. We then expressed and purified the two identified odorant-binding proteins (OBPs) using the Escherichia coli expression system and affinity purification. In vitro binding studies showed that the two OBPs display stronger binding affinities to the female volatile sex pheromone blattellaquinone than to its analogues and contact sex pheromone components. Finally, three-dimensional modeling of the two OBPs and dock conformation with sex pheromone molecules showed BgerOBP26 has a larger odorant cavity and more conservative active amino acid residues than BgerOBP40. These results illuminated the binding characteristics of potential PBPs of B. germanica, which could lay the groundwork for improved understanding of many aspects of the chemical ecology of B. germanica. Moreover, this information complements the understanding of the olfactory molecular mechanism in cockroaches and provides potential gene targets for B. germanica control.

Eyes absent in the cockroach panoistic ovaries regulates proliferation and differentiation through ecdysone signalling.

Eyes absent (Eya), is a protein structurally conserved from hydrozoans to humans, for which two basic roles have been reported: it can act as a transcription cofactor and as a protein tyrosine phosphatase. Eya was discovered in the fly Drosophila melanogaster in relation to its function in eye development, and the same function was later reported in other insects. Eya is also involved in insect oogenesis, although studies in this sense are limited to D. melanogaster, which has meroistic ovaries, and where eya mutations abolish gonad formation. In the present work we studied the function of eya in the panoistic ovary of the cockroach Blattella germanica. We show that eya is essential for correct development of panoistic ovaries. In B. germanica, eya acts at different level and in a distinct way in the germarium and the vitellarium. In the germarium, eya contributes to maintain the correct number of somatic and germinal cells by regulating the expression of steroidogenic genes in the ovary. In the vitellarium, eya facilitates follicle cells proliferation and contributes to regulate the cell program, in the context of basal ovarian follicle maturation. Thus, eya-depleted females of B. germanica arrest the growth and maturation of basal ovarian follicles and become sterile.

Alteration of insulin and nutrition signal gene expression or depletion of Met reduce both lifespan and reproduction in the German cockroach.

In insects, nutrition and hormones coordinately regulate lifespan and reproduction, which might affect each other. We here investigated how nutrition, insulin, and juvenile hormone (JH) signal genes affect lifespan and reproduction in the German cockroach, Blattella germanica, a serious urban pest throughout the world. Starvation as well as altering insulin and nutrition signal genes by RNA interference (RNAi) knockdown of the expression of either positive or negative components in the two pathways simultaneously reduced lifespan and ootheca number of the mated female cockroaches. Meanwhile, depletion of the JH receptor Methoprene-tolerant (Met), but not kruppel homolog 1 (Kr-h1) that predominately transduces JH signaling to prevent metamorphosis, significantly reduced the two parameters. Moreover, suppressing the expression of several reproduction-related genes, including doublesex (Dsx), vitellogenin (Vg), and the Vg receptor (VgR), had little yet various effects on lifespan; nevertheless, it is likely that there are some reproduction-independent mating factors reducing lifespan. In conclusion, through blocking lifespan and reproduction in a simultaneous manner, the alteration of insulin and nutrient signal gene expression or the depletion of Met might provide a great potential for controlling the German cockroach.

The expression of cockroach insulin-like peptides is differentially regulated by physiological conditions and affected by compensatory regulation.

In insects, the insulin receptor (InR) pathway is involved in regulating key physiological processes, including juvenile hormone (JH) synthesis, vitellogenin production, and oocyte growth. This raises the question about which ligand (or ligands) binds to InR to trigger the above effects. We have cloned seven insulin-like peptides (BgILP1 to 7) from female Blattella germanica cockroaches and found that the brain expresses BgILP1 to 6, the fat body BgILP7, and the ovary BgILP2. Starvation induces the reduction of BgILP3, 5, and 6 mRNA levels in the brain, and the various BgILPs are differentially expressed during the gonadotrophic cycle. In addition, by knocking down the BgILPs we were able to identify compensatory regulation at transcriptional level between the different BgILPs, although none of the BgILP knockdown assays, including the knockdown of the seven BgILPs, produced the same phenotypes that we achieved by depleting InR. Taken together, the results indicate that B. germanica ILPs are differentially expressed in tissues and in response to physiological conditions, and that they are affected by compensatory regulation.

Myoglianin triggers the premetamorphosis stage in hemimetabolan insects.

Insect metamorphosis is triggered by a decrease in juvenile hormone (JH) in the final juvenile instar. What induces this decrease is therefore a relevant question. Working with the cockroach Blattella germanica, we found that myoglianin (Myo), a ligand in the TGF-ß signaling pathway, is highly expressed in the corpora allata (CA, the JH-producing glands) and the prothoracic gland [(PG), which produce ecdysone] during the penultimate (fifth) nymphal instar (N5). In the CA, high Myo levels during N5 repress the expression of juvenile hormone acid methyl transferase, a JH biosynthesis gene. In the PG, decreasing JH levels trigger gland degeneration, regulated by the factors Krüppel homolog 1, FTZ-F1, E93, and inhibitor of apoptosis-1. Also in the PG, a peak of myo expression in N5 indirectly stimulates the expression of ecdysone biosynthesis genes, such as neverland, enhancing the production of the metamorphic ecdysone pulse in N6. The Myo expression peak in N5 also represses cell proliferation, which can enhance ecdysone production. The data indicate that Myo triggers the premetamorphic nymphal instar in B. germanica and possibly in other hemimetabolan insects.-Kamsoi, O., Belles, X. Myoglianin triggers the premetamorphosis stage in hemimetabolan insects.

Diversity of piRNA expression patterns during the ontogeny of the German cockroach.

The Piwi-interacting RNA (piRNA) system is an evolutionarily conserved mechanism involved in the control of transposable elements and maintenance of genomic stability, especially in germ line cells and in early embryo stages. However, relevant particularities, both in mechanism and function, exist across species among metazoans and even within the insect class. As a member of the scarcely studied hemimetabolan group, Blattella germanica can be a suitable reference model to study insect evolution. We present the results of a stringent process of identification and study of expressed piRNAs for B. germanica across 11 developmental stages, ranging from unfertilized egg to nymphs and adult female. Our results confirm the dual origin of piRNA in this species, with a majority of them being generated from the primary pathway, and a smaller but highly expressed set of sequences participating in the secondary ("ping-pong") reamplification pathway. An intriguing partial complementarity in expression is observed between the piRNA of the two biogenesis pathways, with those generated in the secondary pathway being quite restricted to early embryo stages. In addition, many piRNAs are exclusively expressed in late embryo and nymphal stages. These observations point at piRNA functions beyond the role of transposon control in early embryogenesis. Our work supports the view of a more complex scenario, with different sets of piRNAs acting in different times and having a range of functions wider than previously thought.

Expansions of key protein families in the German cockroach highlight the molecular basis of its remarkable success as a global indoor pest.

The German cockroach, Blattella germanica, is a worldwide pest that infests buildings, including homes, restaurants, and hospitals, often living in unsanitary conditions. As a disease vector and producer of allergens, this species has major health and economic impacts on humans. Factors contributing to the success of the German cockroach include its resistance to a broad range of insecticides, immunity to many pathogens, and its ability, as an extreme generalist omnivore, to survive on most food sources. The recently published genome shows that B. germanica has an exceptionally high number of protein coding genes. In this study, we investigate the functions of the 93 significantly expanded gene families with the aim to better understand the success of B. germanica as a major pest despite such inhospitable conditions. We find major expansions in gene families with functions related to the detoxification of insecticides and allelochemicals, defense against pathogens, digestion, sensory perception, and gene regulation. These expansions might have allowed B. germanica to develop multiple resistance mechanisms to insecticides and pathogens, and enabled a broad, flexible diet, thus explaining its success in unsanitary conditions and under recurrent chemical control. The findings and resources presented here provide insights for better understanding molecular mechanisms that will facilitate more effective cockroach control.

Expression of juvenile hormone acid O-methyltransferase and juvenile hormone synthesis in Blattella germanica.

Juvenile hormone (JH), a sesquiterpenoid synthetized by the insect corpora allata (CA), plays critical roles in metamorphosis and reproduction. Penultimate or last step of JH synthesis is catalyzed by juvenile hormone acid O-methyltransferase (JHAMT). Here we report the cloning and expression analysis of the JHAMT orthologue in the cockroach, Blattella germanica (L.) (BgJHAMT). BgJHAMT is mainly expressed in CA, with only expression traces in ovary. Three different isoforms, differing in the 3'-UTR sequence, were identified. Isoform A shows between 35 and 65 times higher expression than B and C in CA from penultimate nymphal instar and adult females. RNAi-triggered knock down of BgJHAMT produces a dramatic reduction of JH synthesis, concomitant with a decrease of fat body vitellogenin expression and basal follicle length. BgJHAMT mRNA levels in CA of females along the gonadotrophic cycle parallel, with a slight advancement, JH synthesis profile. BgJHAMT mRNA levels were reduced in starved females and in females in which we reduced nutritional signaling by knocking down insulin receptor and target of rapamycin (TOR). Results show that conditions that modify JH synthesis in adult B. germanica females show parallel changes of BgJHAMT mRNA levels and that the JH-specific branch of the JH synthesis pathway is regulated in the same way as the mevalonate branch. Furthermore, we demonstrate that nutrition and its signaling through the insulin receptor and TOR pathways are essential for activating BgJHAMT expression, which suggests that this enzyme can be a checkpoint for the regulation of JH production in relation to nutritional status.

RNA interference and functional characterization of a tergal gland alpha amylase in the German cockroach, Blattella germanica L.

German cockroach males possess tergal glands that secrete a combination of oligosaccharides, lipids and proteins. Four major proteins occur in the secretion, with one being the 63 kDa alpha-amylase Blattella germanica Tergal Gland protein-1 (BGTG-1). Denaturing and starch gel electrophoresis coupled with peptide sequencing verified amylase activity for the BGTG-1 protein. BGTG-1 gene expression profiles were determined by using quantitative real-time PCR to compare messenger RNA abundance among isolated tissues of males, females and gravid females. Differences in BGTG-1 gene expression occurred among male tissues, with tergal gland tissue showing the highest expression. Tissues of nongravid and gravid females had significantly lower expression in comparison with male tergal glands (gravid females lowest). RNA interference (RNAi) was used to silence BGTG-1 gene expression by injecting BGTG-1 homologous double-stranded RNA (dsRNA) into male cockroaches. Groups injected with BGTG-1 dsRNA showed ∼90% lower BGTG-1 gene and protein expression compared to controls, which correlated with lower amylase activity in colorimetric assays. However, behavioural assays comparing precopulatory behaviour and mating success between RNAi and control males did not reveal differences. These results connect amylase gene expression and activity in tergal gland tissue but suggest other factors, such as other tergal gland components, may contribute more strongly to mating success.

Persistence of a sugar-rejecting cockroach genotype under various dietary regimes.

Glucose-aversion is a heritable trait that evolved in a number of German cockroach (Blattella germanica L.) populations in response to strong selection with glucose-containing insecticide baits. However, in the absence of glucose-containing bait, glucose-averse (GA) cockroaches have lower performance than wild-type (WT) cockroaches in several fitness-determining traits. We allocated 48 caged populations initiated with homozygous GA and WT adults to four dietary treatments consisting of either pure rodent chow, rodent chow mixed to yield a content of either 20% glucose or 20% fructose, or a treatment consisting of choice between the 20% glucose- and the 20% fructose-containing food. After 6 months we found significantly higher frequency of WT individuals in populations restricted to the 20% glucose food, and after 12 months all dietary treatments contained significantly more WT individuals than expected. In accompanying experiments, we found lower survival and longer development time of GA nymphs restricted to glucose-containing food. We furthermore found evidence for assortative mating of females with males from their own genotype, with significant differences within WT cockroaches. Our study shows experimental evidence that within heterogeneous populations, WT German cockroaches will over time prevail in abundance over GA individuals, even when glucose is not a dietary component.

Juvenile hormone biosynthesis in adult Blattella germanica requires nuclear receptors Seven-up and FTZ-F1.

In insects, the transition from juvenile development to the adult stage is controlled by juvenile hormone (JH) synthesized from the corpora allata (CA) glands. Whereas a JH-free period during the last juvenile instar triggers metamorphosis and the end of the growth period, the reappearance of this hormone after the imaginal molt marks the onset of reproductive adulthood. Despite the importance of such transition, the regulatory mechanism that controls it remains mostly unknown. Here, using the hemimetabolous insect Blattella germanica, we show that nuclear hormone receptors Seven-up-B (BgSvp-B) and Fushi tarazu-factor 1 (BgFTZ-F1) have essential roles in the tissue- and stage-specific activation of adult CA JH-biosynthetic activity. Both factors are highly expressed in adult CA cells. Moreover, RNAi-knockdown of either BgSvp-B or BgFTZ-F1 results in adult animals with a complete block in two critical JH-dependent reproductive processes, vitellogenesis and oogenesis. We show that this reproductive blockage is the result of a dramatic impairment of JH biosynthesis, due to the CA-specific reduction in the expression of two key JH biosynthetic enzymes, 3-hydroxy-3-methylglutaryl coenzyme A synthase-1 (BgHMG-S1) and HMG-reductase (BgHMG-R). Our findings provide insights into the regulatory mechanisms underlying the specific changes in the CA gland necessary for the proper transition to adulthood.