Transcriptomics Reveals the Killing Mechanism by Which Entomopathogenic Fungi Manipulate the RNA Expression Profiles of Termites and Provides Inspiration for Green Pest Management.

As chemical pesticides have caused serious environmental pollution, fungus-based biological control has become a developing alternative to chemical control. Here, we aimed to determine the molecular mechanism underlying how Metarhizium anisopliae facilitated invasive infection. We found that the fungus increased its virulence by downregulating glutathione S-transferase (GST) and superoxide dismutase (SOD) throughout termite bodies. Among 13 fungus-induced microRNAs throughout termite bodies, miR-7885-5p and miR-252b upregulation significantly downregulated several mRNAs in response to toxic substances to increase the fungal virulence [e.g., phosphoenolpyruvate carboxykinase (GTP) and heat shock protein homologue SSE1]. In addition, nanodelivered small interfering RNA of GST and SOD and miR-7885-5p and miR-252b mimics increased the virulence of the fungus. These findings provide new insights into the killing mechanism of entomopathogens and their utilization of the host miRNA machinery to reduce host defenses, laying the groundwork to enhance virulence of biocontrol agents for green pest management.

Blue light stimulates light stress and phototactic behavior when received in the brain of Diaphorina citri.

Blue light with a wavelength of 400-470 nm is the composition of the visible light. However, in recent years, blue light contributed the most significance to light pollution due to the artificial light at night. Previously, we have demonstrated that the Asian citrus psyllid (ACP), Diaphorina citri, an important pest in citrus production, has significant positive phototaxis with a light-emitting diode light of 400 nm. In this study, ACP with positive phototactic behavior to 400 nm light (PH) and non-phototactic behavior to 400 nm light (NP) were collected, individually. Transcriptome dynamics of head tissues of PH and NP groups were captured by using RNA-sequencing technology, respectively. Forty-three to 46 million clean reads with high-quality values were obtained, and 1773 differential expressed genes (DEGs) were detected. Compared with the NP group, there were 841 up-regulated DEGs and 932 down-regulated DEGs in the PH group. Eight pathways were significantly enriched in the PH group in the KEGG database, while 43 up-regulated pathways and 25 down-regulated pathways were significantly enriched in the PH group in the GO database. The DGE approach was reliable validated by real time quantitative PCR. Results indicated that the blue light acted as an abiotic stress causing physiological and biochemical responses such as oxidative stress, protein denaturation, inflammation and tumor development in ACPs. Additionally, the light was absorbed by photoreceptors of ACPs, and converted into electrical signal to regulate neuromodulation. This study provides basic information for understanding the molecular mechanisms of ACP in response to blue light and provides a reference for further studies to elucidate phototactic behavior.

Tick (Acari: Ixodoidea) fauna and zoogeographic division of Jiangxi Province, China.

Tick fauna and zoogeographic distribution of Jiangxi Province remain largely unknown due to the lack of data on distribution, occurrence, and host associations of ticks. Considering this, we collected 1,817 individual samples from natural hosts, humans, and vegetation in 18 counties/districts throughout Jiangxi Province, China, from 2015 to 2021. These 1,817 individuals were found to 13 tick species, 4 genera, and 1 family. In addition, the tick sample data from 8 sampling localities (counties and districts) reported in previous studies were also included in our data. A total of 4,021 individuals, including our sample collection and the previously reported data, were assigned to at least 18 species, 6 genera, and 2 families. One newly recorded species Dermacentor sp. (near D. steini Schulze) was found; three misidentified species (Ixodes acuminatus, Haemaphysalis spinigera, and Haemaphysalis verticalis) reported previously were deleted; and one misidentified species Dermacentor auratus Supino was revised as Dermacentor steini Schulze. In addition, we divided the tick fauna in Jiangxi Province into 5 zoogeographic areas and assigned the 18 tick species collected from 26 localities to these 5 zoogeographic areas. To summarize, our findings provide valuable information on the distribution, tick-host associations, and zoogeographic division of ticks in Jiangxi Province, China. Their molecular characterizations, phylogenetic relationships, and tick-borne pathogens that they may transmit should be further explored.

Advances in insect phototaxis and application to pest management: a review.

Many insects, especially nocturnal insects, exhibit positive phototaxis to artificial lights. Light traps are currently used to monitor and manage insect pest populations, and play a crucial role in physical pest control. Efficient use of light traps to attract target insect pests is an important topic in the application of integrated pest management (IPM). Phototactic responses of insects vary among species, light characteristics and the physiological status of the insects. In addition, light can cause several biological responses, including biochemical, physiological, molecular and fitness changes in insects. In this review, we discuss several hypotheses on insect phototaxis, factors affecting insect phototaxis, insect-sensitive wavelengths, biological responses of insects to light, and countermeasures for conserving beneficial insects and increasing the effect of trapping. In addition, we provide information on the different sensitivities to wavelengths causing positive phototactic behavior in > 70 insect pest and beneficial insect species. The use of advanced light traps equipped with superior light sources, such as light-emitting diodes (LEDs), will make physical pest control in IPM more efficient. © 2019 Society of Chemical Industry.

The Mechanisms of Social Immunity Against Fungal Infections in Eusocial Insects.

Entomopathogenic fungus as well as their toxins is a natural threat surrounding social insect colonies. To defend against them, social insects have evolved a series of unique disease defenses at the colony level, which consists of behavioral and physiological adaptations. These colony-level defenses can reduce the infection and poisoning risk and improve the survival of societal members, and is known as social immunity. In this review, we discuss how social immunity enables the insect colony to avoid, resist and tolerate fungal pathogens. To understand the molecular basis of social immunity, we highlight several genetic elements and biochemical factors that drive the colony-level defense, which needs further verification. We discuss the chemosensory genes in regulating social behaviors, the antifungal secretions such as some insect venoms in external defense and the immune priming in internal defense. To conclude, we show the possible driving force of the fungal toxins for the evolution of social immunity. Throughout the review, we propose several questions involved in social immunity extended from some phenomena that have been reported. We hope our review about social 'host-fungal pathogen' interactions will help us further understand the mechanism of social immunity in eusocial insects.

Molecular characterization and juvenile hormone-regulated transcription of the vitellogenin receptor in the cabbage beetle Colaphellus bowringi.

The vitellogenin receptor (VgR) is highly expressed in the ovaries where it is responsible for vitellogenin (Vg) deposition during oogenesis in insects. Therefore, identifying the VgR of insect pests, and understanding the mechanism regulating its expression, could lead to the development of pest management strategies based on disrupting reproduction. We cloned and identified VgR in the cabbage beetle, Colaphellus bowringi, which is a serious pest of cruciferous vegetables in Asia. The regulation of VgR transcription by juvenile hormone (JH) was also investigated. The results show that C. bowringi VgR cDNA contains an open reading frame of 5310 bp encoding 1769 amino acid residues. Protein domain prediction indicates that C. bowringi VgR belongs to the LDLR gene superfamily, having the same group of structural domains that has been well characterized in other insects. VgR mRNA was highly expressed in the ovaries of reproductive female cabbage beetles. Knockdown of VgR reduced yolk deposition in the ovaries, increased the accumulation of Vg proteins in the hemolymph and decreased the transcription of Vg1 and Vg2 in the fat body. RNA interference and hormone challenge experiments showed that JH induced VgR transcription via the JH intracellular receptor methoprene-tolerant (Met) and the JH-responsive transcription factor Krüppel homolog 1 (Kr-h1). Our results suggest that there is a feedback loop between VgR transcription in the ovaries and Vg transcription in the fat body. JH acting through Met-Kr-h1 pathway induces the transcription of the VgR that is essential for Vg uptake and reproductive development. These findings not only reveal the potential JH signaling mechanism regulating VgR transcription, but may also contribute to the development of pest control strategies based on disrupting endocrine-regulated reproduction.

Green light and light stress in moth: influence on antioxidant enzymes in the oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae).

Recently, light traps equipped with light-emitting diodes (LEDs) have been widely applied for controlling nocturnal pest moths. The oriental armyworm, known as Mythimna separata Walker, is an important insect pest in eastern Asia. The present study aimed to evaluate an influence of green light irradiation on antioxidant enzymes and light stress in M. separata adults. We determined total antioxidant capacity (T-AOC) and an activity of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and glutathione-S-transferase (GST) in the moth bodies according to an exposure time (0 to 180 min) of a green LED light. By our results, we suggested that the green (520 nm) LED light irradiation could induce a weak oxidative stress in M. separata adult moths, and the moths under the green light could restore the damage caused by this light stress. Additionally, we proposed that the adaptive ability to the light stress varied between the moths of different sexes. Our results may provide a theoretical and scientific basis for elucidating a reason of the phototactic behavior of nocturnal moths, including M. separata adults.

Juvenile hormone regulates the differential expression of putative juvenile hormone esterases via methoprene-tolerant in non-diapause-destined and diapause-destined adult female beetle.

Juvenile hormone (JH) plays an essential role in regulating molting, metamorphosis, reproduction, and diapause (dormancy), in many insects and crustaceans. JH esterases (JHEs) can control JH titer by regulating JH degradation. Although the biochemistry and structure of JHEs have been well studied, regulation of their expression remains unclear. We identified three putative JHEs (JHE1, JHE2, JHE3) in the cabbage beetle Colaphellus bowringi, and investigated the regulation of their expression by JH signaling in non-diapause-destined (NDD, reproductive) and diapause-destined (DD) female adults. Sequence and phylogenetic tree analyses indicate that the three putative JHEs shared conserved motifs with the JHEs of other insects and one crustacean, and were similar to Coleopteran, Dipteran, Orthopteran, Hymenopteran, and Decapodan JHEs. They were, however, less closely related to Hemipteran and Lepidopteran JHEs. JHEs were more highly expressed in NDD female adults than in DD female adults. JH analog induction in DD female adults significantly upregulated the expression of JHE1 and JHE2, but had no effect on the expression of JHE3. Knockdown of the JH candidate receptor methoprene-tolerant (Met) in NDD female adults downregulated the expression of all three JHEs. These results suggest that JHE expression is positively correlated with JH signaling, and that Met may be involved in the JH-mediated differential expression of JHE in DD and NDD adult female C. bowringi.

Describing the Diapause-Preparatory Proteome of the Beetle Colaphellus bowringi and Identifying Candidates Affecting Lipid Accumulation Using Isobaric Tags for Mass Spectrometry-Based Proteome Quantification (iTRAQ).

Prior to entering diapause, insects must prepare themselves physiologically to withstand the stresses of arresting their development for a lengthy period. While studies describing the biochemical and cellular milieu of the maintenance phase of diapause are accumulating, few studies have taken an "omics" approach to describing molecular events during the diapause preparatory phase. We used isobaric tags and mass spectrometry (iTRAQ) to quantitatively compare the expression profiles of proteins identified during the onset of diapause preparation phase in the heads of adult female cabbage beetles, Colaphellus bowringi. A total of 3,175 proteins were identified, 297 of which were differentially expressed between diapause-destined and non-diapause-destined female adults and could therefore be involved in diapause preparation in this species. Comparison of identified proteins with protein function databases shows that many of these differentially expressed proteins enhanced in diapause destined beetles are involved in energy production and conversion, carbohydrate metabolism and transport, and lipid metabolism. Further hand annotation of differentially abundant peptides nominates several associated with stress hardiness, including HSPs and antioxidants, as well as neural development. In contrast, non-diapause destined beetles show substantial increases in cuticle proteins, suggesting additional post-emergence growth. Using RNA interference to silence a fatty acid-binding protein (FABP) that was highly abundant in the head of diapause-destined females prevented the accumulation of lipids in the fat body, a common product of diapause preparation in this species and others. Surprisingly, RNAi against the FABP also affected the transcript abundance of several heat shock proteins. These results suggest that the identified differentially expressed proteins that play vital roles in lipid metabolism may also contribute somehow to enhanced hardiness to environmental stress that is characteristic of diapause.

Sublethal Effects of Thiamethoxam on the Demographic Parameters of Myzus persicae (Hemiptera: Aphididae).

The green peach aphid, Myzus persicae Sulzer (Hemiptera: Aphididae), is an important sap-sucking pest of many crops, including Chinese cabbage, Brassinca oleracea L. The neonicotinoid insecticide thiamethoxam has been used as an effective insecticide to control M. persicae in cabbage fields. In this study, we assessed the effects of sublethal concentrations of thiamethoxam on demographic parameters of M. persicae. In leaf-dip bioassays, thiamethoxam showed a relatively high toxicity against M. persicae with an LC50 of 6.80 mg liter-1. The duration of the preadult stage was not significantly affected in the sublethal bioassay. Additionally, the longevity and adult preoviposition period were not significantly affected by sublethal thiamethoxam. However, sublethal thiamethoxam significantly increased fecundity (LC10) and prolonged the total preoviposition period (LC40). Consequently, the finite rate of increase (λ) and the intrinsic rate of increase (rm) of aphids exposed to the LC40 were significantly lower than those of control aphids, whereas the net reproductive rate (R0) was higher, and the generation time (T) and the population doubling time (DT) were longer in the treated group. Based on these results, hormesis was induced by sublethal thiamethoxam in M. persicae, with the population growth of M. persicae negatively affected at higher sublethal concentrations of thiamethoxam. Therefore, our study indicated that the possible effects of thiamethoxam on aphids require further study to develop optimized integrated pest management strategies.

Difference in diel mating time contributes to assortative mating between host plant-associated populations of Chilo suppressalis.

Behavioral isolation in animals can be mediated by inherent mating preferences and assortative traits, such as divergence in the diel timing of mating activity. Although divergence in the diel mating time could, in principle, promote the reproductive isolation of sympatric, conspecific populations, there is currently no unequivocal evidence of this. We conducted different mate-choice experiments to investigate the contribution of differences in diel mating activity to the reproductive isolation of the rice and water-oat populations of Chilo suppressalis. The results show that inter-population difference in diel mating activity contributes to assortative mating in these populations. In the rice population, most mating activity occurred during the first half of the scotophase, whereas in the water-oat population virtually all mating activity was confined to the second half of the scotophase. However, when the photoperiod of individuals from the water-oat population was altered to more closely align their mating activity with that of the rice population, mate choice was random. We conclude that inter-population differences in diel mating time contribute to assortative mating, and thereby the partial reproductive isolation, of these host-associated populations of C. suppressalis.

Influence of UV-A radiation on oxidative stress and antioxidant enzymes in Mythimna separata (Lepidoptera: Noctuidae).

Abiotic stress factors, including ultraviolet (UV) radiation, significantly affect insect life. UV-A radiation (320-400 nm) has been widely used for insect control since it increases the production of ROS and causes oxidative cell damage. In the present study, we evaluated the effects of UV-A irradiation on an important pest in China, the ear-cutting caterpillar, Mythimna separata (Lepidoptera: Noctuidae). We exposed 3-day-old M. separata adults to UV-A radiation for different periods of time (0, 30, 60, 90, and 120 min) and evaluated the resulting total antioxidant capacity and the activity of the antioxidant enzymes superoxide dismutase, catalase, peroxidase, and glutathione-S-transferase. The total antioxidant capacity significantly increased after exposure to UV-A radiation for 60 min but decreased after 90 and 120 min of exposure, compared with the control. The antioxidant activity of glutathione-S-transferase, superoxide dismutase, catalase, and peroxidase increased after 60-min exposure, and it was decreased at the longest exposure period 120 min. The longest exposure time period relatively activates the xenobiotic detoxifying enzymes like glutathione-S-transferase, superoxide dismutase, catalase, and peroxidase enzymes. The longest duration of UV-A radiation may cooperate with pesticide detoxification mechanism in insects, making them more susceptible to insecticides. Our results demonstrated that UV irradiation causes oxidative stress, affects the activity of antioxidant enzymes, and disturbs the physiology of M. separata adults.

Ultrastructure of sensilla of antennae and ovipositor of Sitotroga cerealella (Lepidoptera: Gelechiidae), and location of female sex pheromone gland.

The Angoumois grain moth, Sitotroga cerealella, is a serious pest of stored grains worldwide. Presently, the best effective control against the moth is to disrupt the sexual communication between sexes. Sexual communication in moths includes two processes in which females produce and release pheromones from the sex pheromone gland and males detect and respond to them with a relatively sophisticated olfactory system in their antennae. To better understand these processes, we studied the ultrastructure of antennal and ovipositor sensilla of S. cerealella and determined the location of the female sex pheromone gland. Seven types of antennal sensilla were identified on both sexes: sensilla trichodea, sensilla chaetica, sensilla coeloconica, sensilla styloconica, sensilla auricillica, sensilla squamiformia and BÓ§hm bristles. Of these sensilla, the sensilla trichodea were significantly more abundant on male antennae than on those of females, suggesting that these sensilla may detect the sex pheromones. On the ovipositor, only sensilla chaetica of various lengths were found. The sexual gland was an eversible sac of glandular epithelium that was situated dorsally in the intersegmental membrane between the 8th and 9th abdominal segments. These results will lead to a better understanding of mate finding with sex pheromones for this worldwide pest species.

Fatty acid synthase 2 contributes to diapause preparation in a beetle by regulating lipid accumulation and stress tolerance genes expression.

Diapause, also known as dormancy, is a state of arrested development that allows insects to survive unfavorable environmental conditions. Diapause-destined insects store large amounts of fat when preparing for diapause. However, the extent to which these accumulated fat reserves influence diapause remains unclear. To address this question, we investigated the function of fatty acid synthase (FAS), which plays a central role in lipid synthesis, in stress tolerance, the duration of diapause preparation, and whether insects enter diapause or not. In diapause-destined adult female cabbage beetles, Colaphellus bowringi, FAS2 was more highly expressed than FAS1 at the peak stage of diapause preparation. FAS2 knockdown suppressed lipid accumulation and subsequently affected stress tolerance genes expression and water content. However, silencing FAS2 had no significant effects on the duration of diapause preparation or the incidence of diapause. FAS2 transcription was suppressed by juvenile hormone (JH) and the JH receptor methoprene-tolerant (Met). These results suggest that the absence of JH-Met induces FAS2 expression, thereby promoting lipid storage in diapause-destined female beetles. These results demonstrate that fat reserves regulate stress tolerance genes expression and water content, but have no significant effect on the duration of diapause preparation or the incidence of diapause.

Differential expression of circadian clock genes in two strains of beetles reveals candidates related to photoperiodic induction of summer diapause.

Diapause (also known as dormancy) is a state of arrested development induced by photoperiod or temperature that allows insects to survive adverse environmental conditions. By regulating diapause induction, the circadian clock is involved in short-day-induced winter diapause but whether this is also the case in long-day (LD)-induced summer diapause remains unknown. The cabbage beetle Colaphellus bowringi could enter summer diapause under LD conditions. However, a non-photoperiodic-diapause (NPD) strain of this species, which was developed in our laboratory by artificial selection, could not enter diapause under LD photoperiod. Therefore, we identified circadian clock genes in this species and measured differences in their expression between a high diapause (HD) strain and the NPD strain to investigate the potential relationship between circadian clock genes and summer diapause induction in C. bowringi. We successfully cloned eight circadian clock genes and obtained intact ORFs of four; cryptochrome2, double-time, shaggy and vrille. Phylogenetic trees and sequence alignment analyses indicated that these circadian clock genes were conserved across insect taxa. The quantitative real-time PCR indicated that clock, cycle, period, timeless, cryptochrome2, and vrille were differentially expressed between HD and NPD strains reared under LD photoperiod during the diapause induction phase. These findings suggest the potential relationship between circadian clock genes and LD-regulated summer diapause induction in C. bowringi.

Association between gut microbiota and diapause preparation in the cabbage beetle: a new perspective for studying insect diapause.

Gut microbiota mediate the nutritional metabolism and play important roles in human obesity. Diapausing insects accumulate large fat reserves and develop obese phenotypes in order to survive unfavorable conditions. However, the possibility of an association between gut microbiota and insect diapause has not been investigated. We used the Illumina MiSeq platform to compare gut bacterial community composition in nondiapause- (i.e. reproductive) and diapause-destined female cabbage beetles, Colaphellus bowringi, a serious pest of vegetables in Asia. Based on variation in the V3-V4 hypervariable region of 16S ribosomal RNA gene, we identified 99 operational taxonomic units and 17 core microbiota at the genus level. The relative abundance of the bacterial community differed between reproductive and diapause-destined female adults. Gut microbiota associated with human obesity, including Bacteroidetes, Firmicutes, and Proteobacteria, showed a good correlation with diapause. This association between gut microbiota and diapause in the cabbage beetle may open a new avenue for studying insect diapause, as well as developing a natural insect obesity model with which to explore the mechanisms responsible for human obesity.

Identification and testing of oviposition attractant chemical compounds for Musca domestica.

Oviposition attractants for the house fly Musca domestica have been investigated using electrophysiological tests, behavioural assays and field tests. Volatiles were collected via head space absorption method from fermented wheat bran, fresh wheat bran, rearing substrate residue and house fly maggots. A Y-tube olfactometer assay showed that the odor of fermented wheat bran was a significant attractant for female house flies. Bioactive compounds from fermented wheat bran for house fly females were identified by electrophysiology and mass spectrophotometry and confirmed with standard chemicals. Four electrophysiologically active compounds including ethyl palmitate, ethyl linoleate, methyl linoleate, and linoleic acid were found at a proportion of 10:24:6:0.2. Functional imaging in the female antennal lobes revealed an overlapped active pattern for all chemicals. Further multiple-choice behavioural bioassays showed that these chemicals, as well as a mixture that mimicked the naturally occurring combination, increased the attractiveness of non-preferred rearing substrates of cotton and maize powder. Finally, a field demonstration test revealed that, by adding this mimic blend into a rearing substrate used to attract and breed house flies in West Africa, egg numbers laid by females were increased. These chemicals could be utilized to improve house fly production systems or considered for lure traps.

Do differences in life-history traits and the timing of peak mating activity between host-associated populations of Chilo suppressalis have a genetic basis?

The development of host races, genetically distinct populations of the same species with different hosts, is considered to be the initial stage of ecological speciation. Ecological and biological differences consistent with host race formation have been reported between water-oat and rice-associated populations of Chilo suppressalis. In order to confirm whether these differences have a genetic basis, we conducted experiments to determine the extent to which various life-history traits and the time of peak mating activity of these populations were influenced by the species of host plant larvae were raised on. Individuals from each population were reared for three consecutive generations on either water-oat fruit pulp or rice seedlings. Descendants of both populations had higher larval survival rates, shorter larval developmental periods, higher pupal weight, and longer adult forewings, when reared on water-oats than when reared on rice. The time of peak of mating activity differed between the descendants of each population, irrespective of whether they were raised on water-oats or rice. These results indicate that although some life-history traits of host-associated populations of C. suppressalis are influenced by the host plant larvae are raised on, time of peak mating activity is not. Because it is a stable, objective, phenotypic trait, further research on difference in the time of peak mating activity between host-associated populations of C. suppressalis should be conducted to clarify the mechanism responsible for host race formation in this species.

Effect of UV-A radiation as an environmental stress on the development, longevity, and reproduction of the oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae).

The ultraviolet light (UV-A) range of 320-400 nm is widely used as light trap for insect pests. Present investigation was aimed to determine the effect of UV light-A radiation on development, adult longevity, reproduction, and development of F1 generation of Mythimna separata. Our results revealed that the mortality of the second instar larvae was higher than the third and fourth instar larvae after UV-A radiation. As the time of UV-A irradiation for pupae prolonged, the rate of adult emergence reduced. Along with the extension of radiation time decreased the longevity of adult females and males. However, the radiation exposure of 1 and 4 h/day increased fecundity of female adults, and a significant difference was observed in a 1 h/day group. The oviposition rates of female adults in all the treatments were significantly higher than the control. In addition, UV-A radiation treatments resulted in declined cumulative survival of F1 immature stages (eggs, larvae, and pupae). After exposure time of 4 and 7 h/day, the developmental periods of F1 larvae increased significantly, but no significant effects on F1 pupal period were recorded.

Juvenile hormone facilitates the antagonism between adult reproduction and diapause through the methoprene-tolerant gene in the female Colaphellus bowringi.

In insects, the process whereby juvenile hormone (JH) regulates short-day (SD)-induced reproductive diapause has been previously investigated. However, we still do not understand the mechanism by which JH regulates long-day (LD)-induced reproductive diapause. In this study, we use a cabbage beetle, Colaphellus bowringi, which is a serious pest of cruciferous vegetables in Asia capable of entering reproductive diapause under LD conditions, as a model to test whether JH regulates female reproductive diapause similar to the mechanism of SD-induced diapause. Our results showed that the JH analog (JHA) methoprene significantly induced ovarian development but inhibited lipid accumulation of diapause-destined adults. Meanwhile, the transcripts of the vitellogenin (Vg) genes were upregulated, whereas the expression of the fat synthesis and stress tolerance genes were downregulated. RNA interference of the JH candidate receptor gene methoprene-tolerant (Met) blocked JH-induced ovarian development and Vg transcription, suggesting a positive regulatory function for JH-Met signaling in reproduction. Furthermore, under reproduction-inducing conditions, Met depletion promoted a diapause-like phenotype, including arrested ovarian development and increased lipid storage, and stimulated the expression of diapause-related genes involved in lipid synthesis and stress tolerance, suggesting JH-Met signaling plays an important role in the inhibition of diapause. Accordingly, our data indicate that JH acts through Met to facilitate development of the reproductive system by upregulating Vg expression while inhibiting diapause by suppressing lipid synthesis and stress tolerance in the cabbage beetle. Combined with previous studies in SD-induced reproductive diapause, we conclude that JH may regulate female reproductive diapause using a conserved Met-dependent pathway, regardless of the length of the photoperiod inducing diapause in insects.