G-Protein Coupled Receptor Gpr-1 Is Important for the Growth and Nutritional Metabolism of an Invasive Bark Beetle Symbiont Fungi Leptographium procerum.

Leptographium procerum has been demonstrated to play important roles in the invasive success of red turpentine beetle (RTB), one of the most destructive invasive pests in China. Our previous studies found that bacterial volatile ammonia plays an important role in the maintenance of the RTB-L. procerum invasive complex. In this study, we found a GPCR gene Gpr-1 that was a response to ammonia but not involved in the ammonia-induced carbohydrate metabolism. Deletion of Gpr-1 significantly inhibited the growth and pathogenicity but thickened the cell wall of L. procerum, resulting in more resistance to cell wall-perturbing agents. Further analyses suggested that Gpr-1 deletion caused growth defects that might be due to the dysregulation of the amino acid and lipid metabolisms. The thicker cell wall in the ΔGpr-1 mutant was induced through the cell wall remodeling process. Our results indicated that Gpr-1 is essential for the growth of L. procerum by regulating the nutritional metabolism, which can be further explored for potential applications in the management of RTB.

Increased environmental microbial diversity reduces the disease risk of a mosquitocidal pathogen.

IMPORTANCE: The host-specific microbiotas of animals can both reduce and increase disease risks from pathogens. In contrast, how environmental microbial communities affect pathogens is largely unexplored. Aquatic habitats are of interest because water enables environmental microbes to readily interact with animal pathogens. Here, we focused on mosquitoes, which are important disease vectors as terrestrial adults but are strictly aquatic as larvae. We identified a pathogen of mosquito larvae from the field as a strain of Chromobacterium haemolyticum. Comparative genomic analyses and functional assays indicate this strain and other Chromobacterium are mosquitocidal but are also opportunistic pathogens of other animals. We also identify a critical role for diversity of the environmental microbiota in disease risk. Our study characterizes both the virulence mechanisms of a pathogen and the role of the environmental microbiota in disease risk to an aquatic animal of significant importance to human health.

Exosomal lncRNA LINC02191 Promotes Laryngeal Squamous cell Carcinoma Progression by Targeting miR-204-5p/RAB22A Axis and Regulating PI3K/Akt/mTOR Pathway.

Recent research has explored the potential use of serum-derived biomarkers in cancer screening, and mounting evidence has illustrated the pivotal roles of long noncoding RNAs (lncRNAs) in regulating laryngeal squamous cell carcinoma (LSCC) progression. LINC02191 is a newly identified lncRNA and no studies have investigated its role in malignant tumors. This study aims to explore the functions and mechanisms of lncRNA LINC02191 in LSCC. LINC02191 was knocked down in LSCC cells using shRNAs for loss-of-function experiments. RT-qPCR revealed that LINC02191 was upregulated in LSCC patients' serum exosomes, tissues and cells. Western blotting and RT-qPCR were implemented for detecting molecular protein and RNA levels. Colony formation, CCK-8, wound healing and Transwell assays were employed for examining LSCC cell malignant behaviors in vitro. A tumor-bearing mouse model (n = 4/group) was established for examining LINC02191 role in vivo. The results showed that LINC02191 silencing hindered LSCC cell proliferation, invasiveness, migration as well as EMT in vitro and impeded tumorigenesis in xenograft mouse model. Luciferase reporter assay was utilized for verifying the interaction between LINC02191, miR-204-5p and RAB22A. Pearson correlation analysis was employed to evaluate their expression correlation in LSCC tissue specimens (N = 30). Mechanistically, LINC02191 upregulated RAB22A by binding to miR-204-5p, and knocking down LINC02191 inhibited PI3K/Akt/mTOR signaling transduction in LSCC cells and tumor-bearing mice. Moreover, RAB22A overexpression reversed LINC02191 depletion-triggered suppression of LSCC cell aggressiveness and inactivation of PI3K/Akt/mTOR signaling. In conclusion, LINC02191 aggravates LSCC by targeting miR-204-5p/RAB22A/PI3K/Akt/mTOR signaling pathway, which indicates that LINC02191 may serve as a promising target for LSCC treatment.

Quantitative transcriptomic and proteomic analyses reveal the potential maintenance mechanism of female adult reproductive diapause in Chrysoperla nipponensis.

BACKGROUND: The green lacewing Chrysoperla nipponensis is an important natural enemy of many insect pests and exhibits reproductive diapause to overwinter. Our previous studies showed that adult C. nipponensis enters reproductive diapause under a short-day photoperiod. However, the molecular mechanism underlying diapause maintenance in C. nipponensis is still unknown. RESULTS: The total lipid and triglyceride content showed the reservation and degradation of energy during diapause in C. nipponensis. Thus, we performed combined transcriptomic and proteomic analyses of female reproductive diapause in C. nipponensis at three ecophysiological phases (initiation, maintenance and termination). A total of 64 388 unigenes and 5532 proteins were identified from the transcriptome and proteome. In-depth dissection of the gene-expression dynamics revealed that differentially expressed genes and proteins were predominately involved in the lipid and carbohydrate metabolic pathways, in particular fatty acid metabolism, metabolic pathways and the citrate cycle. Among of these genes, TIM, CLK, JHAMT2, PMK, HMGS, HMGR, FKBP39, Kr-h1, Phm, ECR, IR1, ILP3, ILP4, mTOR, ACC, LSD1 and LSD2 were differentially expressed in diapause and non-diapause female adults of C. nipponensis. The expression patterns of these genes were consistent with the occurrence of vitellogenesis and expression of either Vg or VgR. CONCLUSION: Our findings indicated that diapause adult C. nipponensis accumulate energy resources to overwinter. Transcriptomic and proteomic analyses suggested candidate key genes involved in the maintenance of C. nipponensis during adult reproductive diapause. Taken together, these results provide in-depth knowledge to understand the maintenance mechanism of C. nipponensis during adult reproductive diapause. © 2023 Society of Chemical Industry.

Symbiotic microbes aid host adaptation by metabolizing a deterrent host pine carbohydrate d-pinitol in a beetle-fungus invasive complex.

The red turpentine beetle (RTB) is one of the most destructive invasive pests in China and solely consumes pine phloem containing high amounts of d-pinitol. Previous studies reported that d-pinitol exhibits deterrent effects on insects. However, it remains unknown how insects overcome d-pinitol during their host plant adaptation. We found that d-pinitol had an antagonistic effect on RTB, which mainly relied on gallery microbes to degrade d-pinitol to enhance host adaptation with mutualistic Leptographium procerum and two symbiotic bacteria, Erwinia and Serratia, responsible for this degradation. Genomic, transcriptomic, and functional investigations revealed that all three microbes can metabolize d-pinitol via different branches of the inositol pathway. Our results collectively highlight the contributions of symbiotic microbes in RTB's adaptation to living on pine, thereby facilitating outbreaks of RTB in China. These findings further enrich our knowledge of symbiotic invasions and contribute to the further understanding of plant-insect interactions.

Genome-Wide Identification and Expression Profiling of Candidate Sex Pheromone Biosynthesis Genes in the Fall Armyworm (Spodoptera frugiperda).

Spodoptera frugiperda is an agricultural pest causing substantial damage and losses to commercial crops. Sex pheromones are critical for successful mating in Lepidoptera and have been used for monitoring and control of many pest species. The sex pheromone of S. frugiperda is known, but the genes involved in its biosynthesis have not been identified. We systematically studied 99 candidate sex pheromone genes in the genome of S. frugiperda including 1 acetyl-CoA carboxylase (ACC), 11 fatty acid synthases (FASs), 17 desaturases (DESs), 4 fatty acid transport proteins (FATPs), 29 fatty acyl-CoA reductases (FARs), 17 acetyl-CoA acetyltransferases (ACTs), 5 acyl-CoA dehydrogenase (ACDs), 3 enoyl-CoA hydratases (ECHs), 3 hydroxyacyl-CoA dehydrogenases (HCDs), 6 ethyl-CoA thiolases (KCTs), and 3 acyl-CoA-binding proteins (ACBPs). Based on the comparative transcriptome results, we found 22 candidate sex pheromone biosynthesis genes predominately expressed in pheromone glands (PGs) than abdomens without PGs including SfruFAS4, SfruFATP3, SfruACD5, SfruKCT3, SfruDES2, SfruDES5, SfruDES11, SfruDES13, SfruFAR1, SfruFAR2, SfruFAR3, SfruFAR6, SfruFAR7, SfruFAR8, SfruFAR9, SfruFAR10, SfruFAR11, SfruFAR14, SfruFAR16, SfruFAR29, SfruACT6, and SfruACT10. A combination of phylogenetic and tissue-specific transcriptomic analyses indicated that SfruDES5, SfruDES11, SfruFAR2, SfruFAR3, and SfruFAR9 may be key genes involved in the sex pheromone synthesis of S. frugiperda. Our results could provide a theoretical basis for understanding the molecular mechanisms of sex pheromone biosynthesis in S. frugiperda, and also provide new targets for developing novel pest control methods based on disrupting sexual communication.

Facultative Endosymbiont Serratia symbiotica Inhibits the Apterization of Pea Aphid To Enhance Its Spread.

Aphids display wing polyphenism, and the mother can produce a wingless morph for reproduction and a winged morph for dispersal. It is believed that the wingless morph is an adaptive status under favorable conditions and is determined prenatally. In this study, we have found that winged nymphs of the pea aphid, Acyrthosiphon pisum, can change from winged to wingless during normal development. Our results showed that winged nymphs could become the wingless morph by apterization in response to changes from stressful to favorable conditions. The acquired wingless aphids had higher fecundity than the winged morph. However, this process of regression from winged to wingless morph was inhibited by Serratia symbiotica. The existence of the symbiont did not affect the body mass and fecundity of adult aphids, but it increased the body weight of nymphs and temporally increased the quantity of a primary symbiont, Buchnera aphidicola. Our results showed that despite temporal improvement of living conditions causing the induction of apterization of winged nymphs, the inhibition effect of S. symbiotica on this process was activated simultaneously. This finding, for the first time, reveals that the wingless morph can be changed postnatally, which explains a novel regulating mechanism of wing polyphenism driven by external abiotic stimuli and internal biotic regulation together in aphids. IMPORTANCE Wing polyphenism is an important adaptative response to environmental changes for aphids. Endosymbionts are widespread in aphids and also confer the ability to withstand unfavorable conditions. However, little is known about whether endosymbionts are involved in the wing polyphenism. In this study, we report a new finding that winged nymphs of the pea aphid could turn into adults without wings or wing-related structures through apterization when winged nymphs escaped from stressful to favorable environments. Further analysis revealed that the facultative symbiont S. symbiotica could prevent the temporal determination of the host in wing suppression by inhibiting apterization, to enhance its spread. Our findings provide a novel angle to understanding the wing polyphenism regulation of aphids.

The Influence of Host Aphids on the Performance of Aphelinus asychis.

The aphid parasitoid Aphelinus asychis Walker is an important biological control agent against many aphid species. In this study, we examined whether the rearing host aphid species (the pea aphid, Acyrthosiphon pisum and the grain aphid, Sitobion avenae) affect the performance of A. asychis. We found that A. pisum-reared A. asychis showed a significantly larger body size (body length and hind tibia length) and shorter developmental time than S. avenae-reared A. asychis. There was no difference in the sex ratio between them. The longevity of A. pisum-reared A. asychis was also significantly longer than that of S. aveane-reared A. asychis. Furthermore, A. pisum-reared A. asychis presented stronger parasitic capacity and starvation resistance than S. aveane-reared A. asychi. In addition, host aphid alteration experiments showed that A. asychis only takes two generations to adapt to its new host. Taken together, these results revealed that A. pisum is a better alternative host aphid for mass-rearing and releasing of A. asychis. The body size plasticity of A. asychis is also discussed.

Cellular immune responses of the yellow peach moth, Conogethes punctiferalis (Lepidoptera: Crambidae), to the entomopathogenic fungus, Beauveria bassiana (Hypocreales: Cordycipitaceae).

The yellow peach moth (YPM), Conogethes punctiferalis, is a destructive insect pest of maize in eastern China and adapts to diverse environments, especially against pathogens. In insects, innate immunity comprising both humoral and cellular defense responses, is the primary defense against invading microbial pathogens. In this study, we identified five types of circulating hemocytes from the hemolymph of YPM larvae and analyzed their alterations and functions in immune responses to the infection of Beauveria bassiana, an entomopathogenic fungus infesting many lepidopteran species. The identified hemocytes included prohemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids. Significant decreases of total and differential hemocyte counts were recorded over time in larvae, after they were injected with B. bassiana conidia. Additionally, hemocyte-mediated phagocytosis and nodulation were initiated in the hemolymph of larvae from the B. bassiana conidia challenge. The introduction of DEAE-Sepharose Fast Flow beads stained with Congo red also induced a strong encapsulation response in the larval hemolymph. Our observations unravel the occurrence of phagocytosis, nodulation and encapsulation in the hemocoel of YPM larvae to fight against the fungal infection, and offer the first insight into the YPM immune system.

Identification and Characterization of Antioxidant Enzyme Genes in Parasitoid Aphelinus asychis (Hymenoptera: Aphelinidae) and Expression Profiling Analysis under Temperature Stress.

It is well known that high temperature, a typically negative environmental factor, reduces the parasitism of a parasitoid. Generally, high temperature causes the rapid overproduction of reactive oxygen species (ROS) in organisms, and antioxidative enzymes participate in the process of resisting environmental stress by eliminating excess ROS. In the present study, we identify two superoxide dismutase (SOD), one catalase (CAT), six peroxidases (POD), and five glutathione-S-transferase (GST) genes; and the survival rate and antioxidative enzyme patterns under short-term high temperature exposure of the parasitoid wasp, A.asychis, are examined. Survival results of A.asychis reveal that females show higher thermal tolerance than males. Under short-term high-temperature exposure, in females, the expression levels of most antioxidant enzyme genes decreased first and then increased to a peak at 41 °C, while only the expression of AasyGST4 showed a continuous increase. In males, the expression patterns of most antioxidant enzyme genes fluctuated and reached a maximum at 41 °C. Moreover, the expression levels of the majority of antioxidant enzyme genes were higher in females than in males. In addition, at temperatures of and above 35 °C, the activities of these four antioxidant enzymes were induced. The results show that the antioxidant enzymes confer thermo-tolerance to A. asychis against lethal thermal stress. Our observations enrich the understanding of the response mechanism to high-temperature assaults of A. asychis.

Characterization of Antennal Chemosensilla and Associated Chemosensory Genes in the Orange Spiny Whitefly, Aleurocanthus spiniferus (Quaintanca).

The insect chemosensory system plays an important role in many aspects of insects' behaviors necessary for their survival. Despite the complexity of this system, an increasing number of studies have begun to understand its structure and function in different insect species. Nonetheless, the chemosensory system in the orange spiny whitefly Aleurocanthus spiniferus, as one of the most destructive insect pests of citrus in tropical Asia, has not been investigated yet. In this study, the sensillum types, morphologies and distributions of the male and female antennae of A. spiniferus were characterized using scanning electron microscopy. In both sexes, six different sensilla types were observed: trichodea sensilla, chaetica sensilla, microtrichia sensilla, coeloconic sensilla, basiconic sensilla, and finger-like sensilla. Moreover, we identified a total of 48 chemosensory genes, including 5 odorant-binding proteins (OBPs), 12 chemosensory proteins (CSPs), 3 sensory neuron membrane proteins (SNMPs), 6 odorant receptors (ORs), 8 gustatory receptors (GRs), and 14 ionotropic receptors (IRs) using transcriptome data analysis. Tissue-specific transcriptome analysis of these genes showed predominantly expression in the head (including antennae), whereas CSPs were broadly expressed in both head (including the antennae) and body tissue of adult A. spiniferus. In addition, the expression profiling of selected chemosensory genes at different developmental stages was examined by quantitative real time-PCR which was mapped to the transcriptome. We found that the majority of these genes were highly expressed in adults, while AspiORco, AspiGR1, AspiGR2, and AspiIR4 genes were only detected in the pupal stage. Together, this study provides a basis for future chemosensory and genomic studies in A. spiniferus and closely related species. Furthermore, this study not only provides insights for further research on the molecular mechanisms of A. spiniferus-plant interactions but also provides extensive potential targets for pest control.

DeepLabV3+/Efficientnet Hybrid Network-Based Scene Area Judgment for the Mars Unmanned Vehicle System.

Due to the complexity and danger of Mars's environment, traditional Mars unmanned ground vehicles cannot efficiently perform Mars exploration missions. To solve this problem, the DeepLabV3+/Efficientnet hybrid network is proposed and applied to the scene area judgment for the Mars unmanned vehicle system. Firstly, DeepLabV3+ is used to extract the feature information of the Mars image due to its high accuracy. Then, the feature information is used as the input for Efficientnet, and the categories of scene areas are obtained, including safe area, report area, and dangerous area. Finally, according to three categories, the Mars unmanned vehicle system performs three operations: pass, report, and send. Experimental results show the effectiveness of the DeepLabV3+/Efficientnet hybrid network in the scene area judgment. Compared with the Efficientnet network, the accuracy of the DeepLabV3+/Efficientnet hybrid network is improved by approximately 18% and reaches 99.84%, which ensures the safety of the exploration mission for the Mars unmanned vehicle system.

Genome-Wide Identification of Neuropeptides and Their Receptors in an Aphid Endoparasitoid Wasp, Aphidius gifuensi.

In insects, neuropeptides and their receptors not only play a critical role in insect physiology and behavior but also are the potential targets for novel pesticide discoveries. Aphidius gifuensis is one of the most important and widespread aphid parasitoids, and has been successfully used to control aphid. In the present work, we systematically identified neuropeptides and their receptors from the genome and head transcriptome of A. gifuensis. A total of 35 neuropeptide precursors and 49 corresponding receptors were identified. The phylogenetic analyses demonstrated that 35 of these receptors belong to family-A, four belong to family-B, two belong to leucine-rich repeat-containing GPCRs, four belong to receptor guanylyl cyclases, and four belong to receptor tyrosine kinases. Oral ingestion of imidacloprid significantly up-regulated five neuropeptide precursors and four receptors whereas three neuropeptide precursors and eight receptors were significantly down-regulated, which indicated that these neuropeptides and their receptors are potential targets of some commercial insecticides. The RT-qPCR results showed that dopamine receptor 1, dopamine receptor 2, octopamine receptor, allatostatin-A receptor, neuropeptides capa receptor, SIFamide receptor, FMRFamide receptor, tyramine receptor and short neuropeptide F predominantly were expressed in the head whilst the expression of ion transport peptide showed widespread distribution in various tissues. The high expression levels of these genes suggest their important roles in the central nervous system. Taken together, our study provides fundamental information that may further our understanding of neuropeptidergic signaling systems in the regulation of the physiology and behavior of solitary wasps. Furthermore, this information could also aid in the design and discovery of specific and environment-friendly insecticides.

A new pest management strategy: transforming a non-host plant into a dead-end trap crop for the diamondback moth Plutella xylostella L.

BACKGROUND: The use of trap crops can reduce the egg production of female Plutella xylostella in cruciferous vegetables and is an effective method for controlling this pest. To date, most of the trap plants that have been studied are cruciferous plants containing high concentrations of glucosinolates, which are more attractive to P. xylostella female adults. However, the application of these trap plants also has some limitations. Studies have shown that aqueous extracts of cruciferous plants can attract P. xylostella to lay eggs. In this study, we utilized the extract of Chinese kale to treat a non-host plant, the faba bean, and evaluated the possibility of using it as a dead-end trap plant for P. xylostella control. RESULTS: Plutella xylostella females laid significantly more eggs on faba beans that had been sprayed with the extract of Chinese kale rather than on Chinese kale itself. The first instar larvae of P. xylostella failed to survive on faba beans. Notably, the faba beans with the Chinese kale extract had the strongest attraction effect on P. xylostella females when placed 3 m away from the Chinese kale. Moreover, this attraction effect of faba beans on P. xylostella for oviposition lasted for up to 15 days. CONCLUSION: Faba bean plants sprayed with the aqueous extract of Chinese kale represent a potential dead-end trap plant for P. xylostella adults and their oviposition while being invariably deadly for their offspring. The present study provides a new proof of concept of using a non-cruciferous trap plant for P. xylostella management.

Isolation and characterization of Pseudomonas cedrina infecting Plutella xylostella (Lepidoptera: Plutellidae).

The diamondback moth, Plutella xylostella, is one of the most destructive pests worldwide and its management relies exclusively on frequent application of chemical insecticides. Resistance to common insecticides is now widespread, and novel classes of insecticides are needed. Entomopathogenic bacteria and their related products play an important role in the management of this pest. In the present work, one bacterial strain was separated from infected pupae of P. xylostella collected from field and its pathogenicity was evaluated. On the basis of the 16S ribosomal RNA sequencing, BLASTN, and phylogenetic analysis, this bacterial isolate was identified as Pseudomonas cedrina. Oral administration of P. cedrina at levels above 10,000 CFU/ml gave significant mortality to P. xylostella larvae. The pathogenicity was also observed by reduced longevity and fecundity in adult females. However, when live bacterial cells were removed, the cultured broth lost any pathogenicity. In response to the bacterial infection, P. xylostella expressed antimicrobial and stress-associated genes. A mixture treatment of P. cedrina and Bacillus thuringiensis showed an additive effect on larval mortality of P. xylostella. These results indicated that P. cedrina is an opportunistic entomopathogen without secretion of toxins. Furthermore, the additive effect of P. cedrina and B. thuringiensis provide a new insight to develop new strategy for controlling P. xylostella.

[Classification and reconstruction of acetabular defects in revision total hip arthroplasty].

As a mature technology gradually, the total hip arthroplasty has been known and conducted by more and more orthopedic surgeons. However, it also increased the number of hip revisions. The reasons for revision of hip mainly include: unstable, infection, aseptic loosening prosthesis, prosthesis peripheral fracture, liner wear and so on, and the acetabulum bone defect and the choice of acetabular prosthesis have been perplexing surgeons. In this paper, we summarise classification and reconstruction of acetabulum bone defect in hip revision oprations, compare the advantages and disadvantages of all kinds of classification, on this basis can have more than one bone grafting mode and the choice of the acetabulum prosthesis for reconstruction of acetabulum, then expound the ways of material sources, bone grafting modes and complex acetabulum bone defect reconstruction as well as the choice of the ways of fixed prosthesis, providing the basis for clinical doctors to deal with all kinds of acetabulum bone defect and the choice of the acetabulum prosthesis, guiding patient specific therapy more precisely.

Infection of Powdery Mildew Reduces the Fitness of Grain Aphids (Sitobion avenae) Through Restricted Nutrition and Induced Defense Response in Wheat.

In natural ecological systems, plants are often simultaneously attacked by both insects and pathogens, which can affect each other's performance and the interactions can be extended to higher trophic levels, such as parasitoids. The English grain aphid (Sitobion avenae) and powdery mildew (Blumeria graminis f. sp. tritici) are two common antagonists that pose a serious threat to wheat production. Numerous studies have investigated the effect of a single factor (insect or pathogen) on wheat production. However, investigation on the interactions among insect pests, pathogens, and parasitoids within the wheat crop system are rare. Furthermore, the influence of the fungicide, propiconazole, has been found to imitate the natural ecosystem. Therefore, this study investigated the effects of B. graminis on the biological performance of grain aphids and the orientation behavior of its endoparasitic wasp Aphidius gifuensis in the wheat system. Our findings indicated that B. graminis infection suppressed the feeding behavior, adult and nymph weight, and fecundity and prolonged the developmental time of S. avenae. We found that wheat host plants had decreased proportions of essential amino acids and higher content of sucrose following aggravated B. graminis infection. The contents of Pro and Gln increased in the wheat plant tissues after B. graminis infection. In addition, B. graminis infection elicited immune responses in wheat: increase in the expression of defense genes, content of total phenolic compounds, and activity of three related antioxidant enzymes. Moreover, co-infection of B. graminis and S. avenae increased the attraction to A. gifuensis compare to that after infestation with aphids alone. In conclusion, our results indicated that B. graminis infection adversely affected the performance of S. avenae in wheat through restricted nutrition and induced defense response. Furthermore, the preference of parasitoids in such an interactive environment might provide an important basis for pest management control.

Volatile ß-Ocimene Can Regulate Developmental Performance of Peach Aphid Myzus persicae Through Activation of Defense Responses in Chinese Cabbage Brassica pekinensis.

In nature, plants have evolved sophisticated defense mechanisms against the attack of pathogens and insect herbivores. Plant volatile-mediated plant-to-plant communication has been assessed in multitrophic systems in different plant species and different pest species. ß-ocimene is recognized as an herbivore-induced plant volatile that play an important role in the chemical communication between plants and pests. However, it is still unclear whether ß-ocimene can active the defense mechanism of Chinese cabbage Brassica pekinensis against the peach aphid Myzus persicae. In this study, we found that treatment of Chinese cabbage with ß-ocimene inhibited the growth of M. persicae in terms of weight gain and reproduction. Moreover, ß-ocimene treatment negatively influenced the feeding behavior of M. persicae by shortening the total feeding period and phloem ingestion and increasing the frequency of stylet puncture. When given a choice, winged aphids preferred to settle on healthy Chinese cabbage compared with ß-ocimene-treated plants. In addition, performance of the parasitoid Aphidius gifuensis in terms of Y-tube olfaction and landings was better on ß-ocimene-treated Chinese cabbage than on healthy plants. Furthermore, ß-ocimene significantly increased the expression levels of salicylic acid and jasmonic acid marker genes and the accumulation of glucosinolates. Surprisingly, the transcriptional levels of detoxifying enzymes (CYP6CY3, CYP4, and GST) in aphids reared on ß-ocimene-treated Chinese cabbage were significantly higher than those of aphids reared on healthy plants. In summary, our results indicated that ß-ocimene can activate the defense response of Chinese cabbage against M. persicae, and that M. persicae can also adjust its detoxifying enzymes machinery to counter the host plant defense reaction.

Effect of Sublethal Doses of Imidacloprid on the Biological Performance of Aphid Endoparasitoid Aphidius gifuensis (Hymenoptera: Aphidiidae) and Influence on Its Related Gene Expression.

The integrated pest management (IPM) strategy was developed and used in combination with pesticides and beneficial biological control agents. To further develop IPM efficiency, it is important to evaluate the side effects of pesticides on biological control agents. Aphidius gifuensis is one of the most important aphid natural enemies and has been successfully used to control Myzys persicae and other aphid species. Imidacloprid (IMD) is a popular pesticide used worldwide and is highly toxic to non-target arthropods. Here, we investigated the short-term sublethal toxicity of IMD in Aphidius gifuensis and its impact on the biological performance and gene expression of this parasitoid. We found that sublethal IMD doses had a significant negative effect on the life history traits of female A. gifuensis, including shortening the lifespan and lowering parasitic capacity. Moreover, exposure to sublethal IMD also adversely affected the response of A. gifuensis to aphid-infested plant volatiles. Based on the transcriptome analysis, we found that the exposure to sublethal IMD doses significantly affected expression of genes involved in the central nervous system, energy metabolism, olfactory, and detoxification system of A. gifuensis. RT-qPCR also revealed that short term expose to sublethal IMD doses significantly induced the gene expression of genes related to the central nervous system (nAChRa7, nAChRa9, TbH, OAR1, NFR, TYR, and DAR1), olfactory system (OR28 and IR8a1), and detoxification system (CYP49p3, CYP6a2, and POD), while it suppressed the expression of genes involved in the central nervous system (nAChRa4 and nAChRb1), olfactory system (Orco1, IR8a2, and GR1), and detoxification system (GST2). Furthermore, exposure to sublethal doses of IMD also significantly increased the activities of CarEs and POD, whereas we observed no influence on the activities of CAT, GST, and SOD. Our results indicate that sublethal IMD doses might adversely affect the biological performance of A. gifuensis by altering gene expression related to the function of olfactory, nervous, energy metabolism, and detoxification systems. Thus, how the use of pesticides directly affect insect population should be considered when used in conjunction with natural pest parasitoids in IPM strategies.

The Potential Coordination of the Heat-Shock Proteins and Antioxidant Enzyme Genes of Aphidius gifuensis in Response to Thermal Stress.

Aphidius gifuensis is one of the most important aphid natural enemies and has been successfully used to control Myzys persicae and other aphid species. High temperature in summer is one of the key barriers for the application of A. gifuensis in the field and greenhouse. In this work, we investigated the biological performance of A. gifuensis and the response of heat-shock proteins and antioxidant enzymes under high temperature. The results showed that A. gifuensis could not survive at 40°C and female exhibited a higher survival in 35°C. Furthermore, the short term exposure to high temperature negatively affected the performance of A. gifuensis especially parasitism efficiency. Under short-term heating, the expression of AgifsHSP, Agifl(2)efl, AgifHSP70, AgifHSP70-4 and AgifHSP90 showed an increased trend, whereas AgifHSP10 initially increased and then decreased. In 35°C, the expressions of Agifl(2)efl, AgifHSP70-4 and AgifHSP90 in female were higher than those in male, whereas the expression of AgifHSP70 exhibited an opposite trend. Besides the HSPs, we also quantified the expression levels of 11 antioxidant enzyme genes: AgifPOD, AgifSOD1, AgifSOD2, AgifSOD3, AgifCAT1, AgifCAT2, AgifGST1, AgifGST2, AgifGST3, AgifGST4 and AgifGST5. We found that the sex-specific expression of AgifSOD2, AgifSOD3, AgifPOD, AgifGST1 and AgifGST3 were highly consistent with sex-specific heat shock survival rates at 35°C. Furthermore, when the temperature was above 30°C, the activities of GST, SOD, CAT and POD were significantly increased; however, there was no significant difference of the CAT activity between the male and female at 35°C. Collectively, all of these results suggested that the protection of thermal damage is coordinated by HSPs and antioxidant enzymes in A. gifuensis. Based on the heat tolerance abilities of many aphid natural enemies, we also discussed an integrated application strategy of many aphid enemies in summer.