An insect chemosensory protein facilitates locust avoidance to fungal pathogens via recognition of fungal volatiles.

Locusts (Locusta migratoria) are one of the most destructive insect pests worldwide. Entomopathogenic fungi can infect and kill locusts, with Metarhizium acridum having evolved as a specialized acridid pathogen. However, locusts have evolved countermeasures to limit or avoid microbial pathogens, although the underlying molecular mechanisms behind these defenses remain obscure. Here, we demonstrate that L. migratoria exhibit avoidance behaviors towards M. acridum contaminated food via recognition of fungal volatiles, with locust perception of the volatile mediated by the LmigCSP60 chemosensory protein. RNAi-knockdown of LmigCSP60 lowered locust M. acridum avoidance behavior and increased infection and mortality. The fungal volatile, 2-phenylethanol (PEA), was identified to participate in locust behavioral avoidance. RNAi-knockdown of LmigCSP60 reduced antennal electrophysiological responses to PEA and impaired locust avoidance to the compound. Purified LmigCSP60 was able to bind a set of fungal volatiles including PEA. Furthermore, reduction of PEA emission by M. acridum via construction of a targeted gene knockout mutant of the alcohol dehydrogenase gene (ΔMaAdh strain) that contributes to PEA production reduced locust avoidance behavior towards the pathogen. These findings identify an olfactory circuit used by locusts to detect and avoid potential microbial pathogens before they are capable of initiating infection and highlight behavioral and olfactory adaptations affecting the co-evolution of host-pathogen interactions.

Insecticidal potential of a Consolida ajacis extract and its major compound (ethyl linoleate) against the diamondback moth, Plutella xylostella.

The diamondback moth (Plutella xylostella) is one of the most destructive lepidopteran pests of cruciferous vegetables. However, DBM has developed resistance to current chemical and biological insecticides used for its control, indicating the necessity for finding new insecticides against it. Bio-insecticides derived from plant extracts are eco-friendly alternatives to synthetic pesticides. The aims of this study were to evaluate the insecticidal activity of Consolida ajacis seed extracts against DBM, the underlying mechanism of the control effect of promising extracts, and the identification of the main insecticidal compounds of these extracts. The results showed that ethyl acetate extract of C. ajacis seed exhibited strong contact toxicity (LC50: 5.05 mg/mL), ingestion toxicity, antifeedant, and oviposition deterrent activities against DBM, among the extracts evaluated. At 72 h, glutathiase, acetylcholinesterase, carboxylesterase, peroxidase, and superoxide dismutase activities were inhibited, but catalase activity was activated. The main compound identified from the extract was ethyl linoleate, which had the most significant insecticidal activity on the diamondback moths. This study's findings provide a better understanding of the insecticidal activity of ethyl acetate extract obtained from C. ajacis and its main component (ethyl linoleate). This will help in the development of new insecticides to control DBM.

Comparative analysis of gut microbiota and immune genes linked with the immune system of wild and captive Spodoptera frugiperda (Lepidoptera: Noctuidae).

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is one of the most highly polyphagous invasive pests causing serious damage to maize crops in China. However, little is known about the gut immune responses to the environment, particularly along the migration routes in Jianghuai, China, throughout the autumn and winter. In this study, high-throughput sequencing and real-time quantitative PCR (RT-qPCR) were employed to examine the variations in immune genes and gut microbiome communities between captive and wild fall armyworm populations. Results showed that the diversity and community of the gut's microbes were higher in wild populations, and the average weighted UniFrac distance between bacterial taxa varied. A wide variety of immune genes were more abundant in the wild populations than in others. Results indicated that diets and different survival conditions impacted the gut microbiota and immune system of S. frugiperda, which was crucial for environmental adaptation. These differences in gut microbiota and immune responses between wild and captive Fall armyworms are critical for comprehending the symbiotic relationship between microbes, immune genes, and hosts. They also highlight the need for increased focus on developing more effective and environmentally friendly pest control methods.

Comparative transcriptome analysis reveals key candidate genes mediating ovarian development in Spodoptera frugiperda fed on two host plants.

The fall armyworm (FAW), Spodoptera frugiperda, is a highly polyphagous lepidopteran pest, with its growth and adaptation affected by different host plants. However, little is known about the effects of host plants on ovarian development in this species. Thus, we evaluated the effects of feeding on corn (Zea mays L.) and goosegrass (Eleusine indica), on the ovarian development of S. frugiperda. Using various stages of S. frugiperda, we also evaluated the larval and pupal weights, number of eggs, and differentiation of ovarioles over time. Results showed that females fed on goosegrass had shorter ovarioles and laid less eggs than those fed on corn. Transcriptome analysis identified 3,213 genes involved in ovarian development in the fall armyworm. Of these, 881 genes were differentially expressed when fed on corn and goosegrass. The analysis also indicated that the hormone biosynthetic pathways may be involved in the reproductive system. In relation to the reproductive function, nine juvenile hormone (JH) biosynthetic genes, four 20-hydroxyecdysone (20E) biosynthetic genes, and four ovary-relevant functional genes were identified. The time course of the expression profiles of these hormone- and ovary development-related genes was measured by quantitative real-time PCR (qRT-PCR). In total, six of them showed a decreasing trend in the ovary of the FAW fed on goosegrass, while two genes showed an increasing trend. Our results showed that significant changes in the reproductive activity/ovary development in the FAW occurred in response to different diets. These results serve as bases for evaluating how optimal host plants and feeding preference affect ovarian development in the FAW.

The complete mitochondrial genome sequence and phylogenetic position of Xylota coquilletti (Soo-Hyun Jeong, Ho-Yeon Han 2019) (Diptera: Syrphidae: Eristalinae: Xylotini).

The complete mitochondrial genome sequence of the Xylota coquilletti (Diptera: Syrphidae: Eristalinae: Xylotini) was sequenced and reported for the first time. The whole genome was 15,920 bp in length with the 37 classical eukaryotic mitochondrial genes and a control region. The nucleotide composition was included by 40.5% A, 39.6% T, 11.7% C, and 8.2% G, meaning that A + T (80.1%) was much greater than C + G (19.9%). It consisted of 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), 13 protein-coding genes (PCGs), and a control region (CR). Phylogenetic analyses were performed using 13 PCGs and it was found that Xylota coquilletti was sister to Ferdinadea cupera. All this information could complement the mitochondrial data for a new tribe of Eristalinae.

Identification and Functional Analysis of Glutathione S-Transferases from Sitophilus zeamais in Olfactory Organ.

Odorant-degrading enzymes (ODEs) play an important role in rapidly degrading and inactivating odorant molecules that have completed information transmission, as well as in maintaining the stability and sensitivity of insect olfactory sensing systems. Glutathione S-transferases (GSTs), as a group of ODEs, supposedly bear the ability to catalyze the conjugation of glutathione (GSH) and xenobiotic odorant molecules in the degrading process. However, there are few reports regarding the role of the GST genes of Sitophilus zeamais in the degrading process. Thus, we characterized 13 full-length genes encoding GST sequences from S. zeamais, of which only SzeaGSTd1 contained a high abundance in the antennae. Ligand-binding assays implied that SzeaGSTd1 was able to catalyze the conjugation of GSH with 2, 4-dinitrochlorobenzene (CDNB). We investigated whether recombinant SzeaGSTd1 bears the ability to degrade the volatile molecules of the host; among the host volatiles, and found capryl alcohol to be a suitable substrate for SzeaGSTd1. These results strongly suggest that SzeaGSTd1 probably plays a role in auxiliary host location by degrading the host volatiles of capryl alcohol and exhibits a potential biological function in the olfactory sensing system of S. zeamais. Knowledge of the potential functions of SzeaGSTd1 will provide new ideas for biological control strategies for S. zeamais.

Differential responses of the antennal proteome of male and female migratory locusts to infection by a fungal pathogen.

The narrow host range entomopathogenic fungus, Metarhizium acridum, is an environmentally friendly acridid specific pathogen used for locust control. The locust is capable of responding within hours of infection, however, little is known concerning how the locust detects the pathogen. Here, we have identified 3213 proteins in the infected antennal proteome of the migratory locust, Locusta migratoria. iTRAQ comparative analyses of antennal proteomes identified 194 differentially abundant proteins (DAPs) between uninfected and infected males, 218 DAPs between uninfected and infected females, and 240 DAPs between infected males and infected females. In relation to olfaction, a total of 29 chemosensory proteins (CSPs), 9 odorant binding proteins (OBPs), 31 odorant receptors (ORs), and 8 ionotropic receptors (IRs) were differentially abundant after M. acridum infection, with a subset of 12 proteins found in both infected male and female antennae not present in uninfected individuals. The time course of the gene expression profiles of olfaction related DAPs were investigated by quantitative real-time PCR (qRT-PCR). Our data indicate significant changes in the antennal proteomes of male and female locusts in response to a microbial pathogen, highlighting the potential participation of olfactory processes in pathogen detection and response. BIOLOGICAL SIGNIFICANCE: The ability of an organism to detect microbial pathogens is essential for mounting a response to mitigate the spread of the infection. Using iTRAQ-based proteomic analyses changes in the protein repertoire of the antennae of male and female locusts in response to infection by a host-specific pathogen were determined. These data show proteomic alterations that are also sex-specific, identifying members of olfactory pathways that are modified in response to infection. Our data identify antennal and related olfactory proteins that are candidates for mediating host detection of pathogens, and that may contribute to subsequent behavioral and/or immune responses of the host to the infection challenge.

Sex-specific variation in the antennal proteome of the migratory locust.

Olfaction plays a critical role in mediating environmental chemical signals to affect behavior. Locusts are voracious feeders that can cause substantial agricultural and environmental damage. Here, we have identified 3473 proteins in the antennal proteome of the migratory locust, Locusta migratoria. Comparative analysis of sex variation in the antennal proteomes of male and female locusts using iTRAQ proteomics identified 260 differentially abundant proteins (DAPs). In relation to olfaction, 29 chemosensory proteins (CSPs), 9 odorant binding proteins (OBPs), 31 odorant receptors (ORs), and 8 ionotropic receptors (IRs) were identified, in the male and female antennal proteomes. Two new previously undescribed CSP proteins were identified in antennae and the expression of their respective gene coding sequences verified. In addition, sex-biased proteins were found significantly enriched in melanogenesis and riboflavin metabolism in males and in ribosome and metabolic pathways in females. The sex- and tissue-specific gene expression profiles of 29 CSPs, 31 ORs and 8 IRs were investigated by quantitative real-time PCR (qRT-PCR), revealing distinct pattern of expression or these genes in different locust tissues including the antennae, legs, wing, and palps. Our data identify sex-biased antennal proteome profiles and indicate expression of putative olfaction genes in tissues beyond the antennae. BIOLOGICAL SIGNIFICANCE: Little is known concerning sexual dimorphism in olfaction related processes at the level of the antennal proteome of insects. Using iTRAQ-based proteomic analyses the protein repertoire of the antennae of male and female locusts were determined. These data show sex-specific differences, identifying a subset of proteins, including two novel CSPs, that likely help define male versus female olfactory responses that contributes to our understanding of chemoreception and downstream behavioral consequences. The characterization of antennal and related olfactory proteins identifies potential biomarkers that can contribute to the development of control strategies against locusts and orthologous insect pests.

Nuclear hormone receptor BmFTZ-F1 is involved in regulating the fibroin heavy chain gene in the silkworm, Bombyx mori.

BACKGROUND: The synthesis of silk protein is controlled by hormones. The expression of the nuclear hormone Bmftz-f1 in the posterior silk gland (PSG) is induced by 20-hydroxyecdysone in vivo and in vitro. However, whether Bmftz-f1 regulates silk protein expression is unknown. METHODS: In our study, western blotting and quantitative polymerase chain reactions were conducted to detect the expression of FTZ-F1 in the PSG. Electrophoretic mobility shift, chromatin immunoprecipitation, far-western blotting, bimolecular fluorescence complementation, and dual luciferase reporter assays were performed to investigate the effect of FTZ-F1 on the fibH promoter. RESULTS: (1) The expression of the hormone receptor BmFTZ-F1 was opposite to that of fibH. It was highly expressed in the PSG during the fourth molting stage and the beginning of the fifth instar, and then its expression decreased gradually until it disappeared at the end of the fifth instar and the wandering stage. (2) We identified a FTZ-F1 response element 390bp upstream of the transcription initiation site of the fibH promoter. (3) BmFTZ-F1 interacted with the basic helix-loop-helix transcription factor Bmdimm. (4) BmFTZ-F1 down-regulated fibH promoter activity and counteracted the effect of Bmdimm on fibH expression. CONCLUSIONS: Integrating these results, we conclude that BmFTZ-F1 regulates the transcription of fibH by binding to the FTZ-F1 response element in the fibH promoter and counteracts the effect of Bmdimm on fibH expression. GENERAL SIGNIFICANCE: These findings provide new insights into the mechanism of regulation of the silk protein gene.

Identification of MBF2 family genes in Bombyx mori and their expression in different tissues and stages and in response to Bacillus bombysepticus infection and starvation.

The Multiprotein bridge factor 2 (MBF2) gene was first identified as a co-activator involved in BmFTZ-F1-mediated activation of the Fushi tarazu gene. Herein, nine homologous genes of MBF2 gene are identified. Evolutionary analysis showed that this gene family is insect-specific and that the family members are closely related to response to pathogens (REPAT) genes. Tissue distribution analysis revealed that these genes could be expressed in a tissue-specific manner. Developmental profiles analysis showed that the MBF2 gene family members were highly expressed in the different stages. Analysis of the expression patterns of nine MBF2 family genes showed that Bacillus bombysepticus treatment induced the up-regulation of several MBF2 family genes, including MBF2-4, -7, -9, -8. Furthermore, we found the MBF2 family genes were modulated by starvation and the expression of these genes recovered upon re-feeding, except for MBF2-5, -9. These findings suggested roles for these proteins in insect defense against pathogens and nutrient metabolism, which has an important guiding significance for designing pest control strategies.