Ascoviruses, as obligate insect pathogens that exclusively infect lepidopteran hosts, are highly reliant on the precise modulation of key host physiological processes, including metabolism, developmental signaling, and immune homeostasis, to facilitate successful infection, subsequent viral replication and transmission. The corazonin signaling pathway serves as a key regulatory pathway in insect development and molting. Herein, we found that ascoviruses modulate the Corazonin signaling pathway to affect the host molting process, thereby influencing viral replication. Therefore, understanding the molecular mechanisms between ascovirus and the corazonin signaling pathway is of particular importance. In this study, we found that corazonin (HaCrz) and the receptor (HaCrzR) were essential for promoting the molting process in response to HvAV infection. Silencing of HaCrz or HaCrzR via RNAi extended larval instar stages, elevated host mortality, reduced transcript levels of downstream molting-related genes (HaTH and HaDDC), and substantially enhanced viral replication. In contrast, exogenous injection of synthetic mature HaCrz peptide, which interacts structurally with HaCrzR as validated by molecular docking, accelerated host molting and improved antiviral responses. These findings not only revealed the mechanism by which viruses manipulate the host molting process but more importantly, demonstrated that viruses create favorable conditions for viral replication and transmission by interfering with host developmental behaviors.

Spodoptera frugiperda (fall armyworm) is a globally distributed and highly destructive migratory pest, while Serratia marcescens is a widespread entomopathogenic bacterium capable of infecting diverse insects. In this study, oral infection with S. marcescens strain SM001 exhibited potent virulence, with an LC50 of 7.40 × 106 CFU/mL at 168 h post-infection (hpi). Logistic regression modeling demonstrated that the bacterium enters an exponential proliferation phase within the hemolymph approximately 48-54 h after infection. Infection significantly upregulated Duox and Nos, induced severe oxidative stress, and activated an apoptotic cascade involving Cyt c, p53, and caspase, culminating in extensive midgut epithelial cell apoptosis. Mitigating oxidative stress via targeted Vitamin C supplementation or Duox knockdown alleviated midgut tissue damage and delayed bacterial dissemination. However, exogenous Vitamin C treatment decreased larval mortality, whereas Duox knockdown failed to improve overall survival, revealing a critical physiological trade-off between ROS-mediated pathogen clearance and host immunopathology. Overall, SM001 disrupts midgut redox homeostasis, drives excessive H2O2 accumulation, and breaches the intestinal barrier to facilitate hemocoel translocation, ultimately leading to lethal systemic septicemia. These findings highlight the pivotal role of host-mediated immunopathology in bacterial pathogenesis and provide a robust theoretical foundation for developing advanced microbial control strategies against S. frugiperda.

The emergence of pesticide resistance in Helicoverpa armigera and other pests represents a challenge, necessitating continued development of innovative insecticides. Ascoviruses are a potential insecticide for H. armigera. These double-stranded DNA viruses cause cell death and lethal disease in infected larvae. However, this disease is chronic and prolongs the lifespan of the infected larvae by several weeks. The slow killing speed limits the success of these pathogens in the insecticidal market. Here, we demonstrate that the dsRNA silencing of H. armigera neuropeptide Adipokinetic Hormones ( HaAKH ) accelerates the killing speed of Heliothis virescens ascovirus 3h (HvAV-3h) against the third-instar larvae of H. armigera. The LT50 was reduced to 3.96 days in larvae with silenced HaAKH1 and 4.7 days in those with silenced HaAKH3. Moreover, the histopathological examinations revealed the destruction of the host's fat body and epidermal tissue shrinkage after HaAKH silencing during HvAV-3h infection. Examining detoxification and antioxidant enzyme activity in HvAV-3h infected larvae showed reduced detoxification mechanisms after HaAKH gene silencing. Furthermore, the silencing of HaAKH resulted in an overall reduction in the fold changes of proline dehydrogenase. In conclusion, this study demonstrates that the ascovirus killing speed can be accelerated by interfering with the host neuropeptide-related gene expression. Moreover, the silencing of H. armigera HaAKH1 and HaAKH3 decreased the antiviral immunity against HvAV-3h.

BACKGROUND:Monitoring arthropod pests and their natural enemies provides essential information for pest control decisions in agricultural production. Traditional monitoring methods, such as trapping and visual surveys, have practical limitations because of their time-consuming preprocessing steps and the need for knowledgeable taxonomists. We tested the environmental DNA (eDNA) metabarcoding approach for monitoring cowpea (Vigna unguiculata) arthropod pests and their natural enemies across seven climatic sub-regions of China, comparing it with traditional sample collection and morphological identification methods. RESULTS:We showed that eDNA metabarcoding reveals a higher species richness of cowpea arthropods than the traditional visual method. The eDNA method detected 163 species, of which 72, 33 and 57 were pests, natural enemies and other insects, respectively, whereas the traditional method only detected 70 species. There were significant differences in alpha- and beta-diversity for arthropods on cowpea in different climatic sub-regions. We also found that longitude, altitude and geographical distance are associated with species diversity, but latitude, mean air temperature and temperature range are not. CONCLUSION:This study represents the first use of eDNA metabarcoding to investigate cowpea arthropod communities, providing a novel method for pest monitoring. © 2025 Society of Chemical Industry.

Coelophora saucia is an important natural enemy insect in agricultural production. We determined the complete mitochondrial genome of the C. saucia by high-throughput sequencing, and the mitogenome was 18,068 bp in length, with a GC content of 20.9%, encodes 2 ribosomal RNA genes, 22 transporter RNA genes, 13 protein-coding genes and with a non-coding control region. A phylogenetic tree was constructed using the maximum likelihood (ML) method, and the results indicated that the C. saucia was most closely related to Propylea japonica and Propylea quattuordecimpunctata.

Aegerolysins are lipid-binding proteins associated with multiple functions, including membrane pore-formation, insecticidal toxicity and defence against predators. Whilst distributed over the kingdoms of the Tree of Life, ascoviruses are the only representative viruses that encode an aegerolysin-like protein. Ascoviruses are entomopathogenic and possess a large dsDNA genome. The present study aimed to functionally characterize the aegerolysin-like protein of Heliothis virescens ascovirus 3h (HvAV-3h), encoded by ORF85, and to explore its potential roles in the interaction between the ascovirus and its host. Our results demonstrate the importance of this species-specific protein to HvAV-3h replication in host cells. In vivo, silencing of this gene for 12-72 h significantly increased the expression of some innate immunity-associated genes, including Toll (114-fold), IMD (44.7-fold) and Hopscotch (22.9-fold). In parallel, we detected significant gradual increases in MyD88 and Relish and decreases in PIAS. Moreover, histopathological analyses of infected larval tissues indicated reduced tissue damage after 72 h of ORF85 gene silencing. The prokaryotic expression of the HvAV-3h aegerolysin, followed by feeding to third-instar Spodoptera exigua larvae for 24 or 48 h led to significant reductions in larval weight. Moreover, the in vitro treatment demonstrated a bactericidal action against Lysinibacillus xylanilyticus, a bacterial resident of some insect guts. Overall, our findings suggest that the protein encoded by ORF85 is associated with the pathogenicity of HvAV-3h and its ability to replicate in host cells. Additionally, aegerolysin may inhibit or kill specific bacterial species in the host microbiome during infection, potentially modulating the host immune response.

Ascoviruses, a family of large, double-stranded circular DNA viruses, exhibit high host specificity and pathogenicity, suggesting their significant potential in biocontrol. A hallmark characteristic of ascovirus infection in larvae is reduced feeding and retarded growth. However, the mechanisms by which ascoviruses regulate these effects remain largely unknown. Given their crucial role in regulating larval feeding, insect neuropeptides have attracted our attention in the context of ascovirus infection. During HvAV-3h infection in S. litura, the expression levels of neuropeptide F (NPF), including NPF1 and NPF2, which are integral to feeding regulation, were significantly reduced. HvAV-3h infection impaired NPF regulation in larvae, leading to reduced food intake and larval weight gain across different physiological states. Concurrently, significant up-regulation of the NPF receptor (NPFR) was observed in the head tissue. The observed dysregulation of the NPF/NPFR signaling pathway was associated with elevated juvenile hormone (JH) titers. In contrast, the expression levels of short neuropeptide F (sNPF) and the molting hormone ecdysone remained unchanged. Moreover, histopathological analysis of the midgut revealed no epithelial cell damage. Furthermore, RNA interference of NPF1 or NPF2 significantly increased the expression of NPFR and juvenile hormone acid O-methyltransferase (JHAMT), and tended to further reduce food intake and weight gain, which consequently increased the mortality during HvAV-3h infection. HvAV-3h infection disrupts the NPF/NPFR signaling pathway in S. litura larvae, subsequently elevating JH titers, ultimately leading to reduced food intake and larval weight gain. This study enhances our understanding of the interaction between HvAV-3h and its host, and provides a theoretical basis for the development of innovative pest management strategies.

This study reports the first complete mitochondrial genome of Ischiodon scutellaris. The mitochondrial genome is 15,815 bp in length, with an A + T content of 80.7%. The genome contains 13 protein-coding genes (PCGs), 22 tRNAs, 2 rRNAs, and a control region. All PCGs initiate with ATN (G/C/T), except COX1and ND1(TTG). Three PCGs (COX1, ATP6, ND5) terminate with an incomplete T stop codon. Phylogenetic analysis based on 13 PCG sequences using maximum likelihood revealed that I. scutellaris forms a sister clade with Scaeva affinis and clusters closely with multiple Eupeodes species. This study advances understanding of syrphid mitochondrial genomics and evolutionary relationships.

Background Synthetic biology is a young but rapidly growing field that allows for assembling long DNA fragments, including complete chromosomes. A key approach for long-DNA assembly is the Transformation Associated Recombination (TAR), which relies on efficient homologous recombination in yeast cells. Recent reports indicate that the TAR method efficiently assembles some human and animal viruses characterized by their large DNA genome size. The application of the TAR method to synthesize long DNA fragments derived from insect viruses is scarce. Therefore, this study aimed to explore the TAR approach for the construction of a long DNA fragment (>44.6 Kb) from the insecticidal Heliothesis virescens ascovirus 3h (HvAV-3h) dsDNA genome to assess the suitability of this approach in genome-wide engineering studies in this family of viruses. Results The long DNA fragment assembly process involved three stages: first, we amplified 15 segments of about 2.9-3.2 Kb each via PCR. Next, we recombined these segments through three parallel TAR cycles, producing medium-sized fragments of about 15 Kb. Finally, we assembled these fragments in a single TAR cycle to form a long DNA fragment of about 44.6 kb. We identified some positive clones by colony PCR or restriction digestion pattern. To assess the quality of the assembled DNA fragment, we conducted next-generation sequencing (NGS). A comparative analysis of Sanger sequencing for medium-sized fragments and NGS data from the synthesized long-DNA fragment demonstrated a nearly matched mutation profile, suggesting that the identified mutations and deletions were present at initial synthesis. Both datasets aligned with the reference HvAV-3h strain, revealing three specific nucleotide mutations and three unique mutation regions. Conclusions Overall, the in vivo TAR assembly method efficiently assembled a long DNA fragment derived from the ascovirus genome as a template. The process is cost-effective and can be scaled up to synthesize the entire genome for gene functional studies.

Abstract Cowpea (Vigna unguiculata (L.) Walp.), as an economical crop, is one of the important pillar industries of rural revitalization strategy in China. However, cowpea planting in China is often infested and damaged by many insects during growth, especially in Hainan region with a warm and wet tropical climate. Traditional monitoring methods with technical limitation could only detect a few common significant agricultural pests, how many kinds of species associated with cowpea is unknown. Here, we employed environmental DNA (eDNA) metabarcoding to characterize cowpea associated animal community-level diversity among six planting areas in Hainan. In all, 62 species were detected, of which 99.05% was Arthropoda, suggesting that Arthropods are the main groups interacting with cowpea. Moreover, we also detected 28 pests on cowpea, predominantly belonging to Thysanoptera, Lepidoptera, Diptera and Hemiptera, of which 20 pests were first reported and need more extra attention. Furthermore, clustering results indicated that there is a certain diversity of cowpea associated animals in different regions of Hainan, but the species composition was similar in the large planting areas due to the indiscriminate use of pesticides, which need further develop scientific pesticide applications to ensure adequate species diversity. This study represents the first molecular approach to investigate the cowpea associated animal communities and provides basic information for further scientific pesticide applications.

A new species, Cephonodes sanshaensis Deng & Huang, sp. nov. is described from Xisha islands near Sansha City, China. Photographs of the adults and their genitalia are provided. The new species is similar to C. hylas (Linnaeus, 1771) and C. picus (Cramer, 1777) but can be easily distinguished by characters in the male genitalia: the right lobe of the uncus is hook-shaped with distinctly acute apex, the left valva is long and narrow with a truncate apex, and the right valva is broad and knife-shaped. Molecular analysis based on cytochrome c oxidase I gene barcode sequences is used to infer the phylogenetic position of the new species within the genus Cephonodes. An updated key and checklist to the worldwide species of the genus Cephonodes are also provided. (c) 2023 National Science Museum of Korea (NSMK) and Korea National Arboretum (KNA), Publishing Services by Elsevier. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/).

Genomes, alignments and tree files of the study "Phylogenomic exploration of taxon sampling difference yields new insights into the deep relationships of butterflies and moths (Lepidoptera)".

A robust and stable phylogenetic framework is a fundamental goal of evolutionary biology. As the third largest insect order in the world following Coleoptera and Diptera, Lepidoptera (butterflies and moths) play a central role in almost every terrestrial ecosystem as indicators of environmental change and serve as important models for biologists exploring questions related to ecology and evolutionary biology. However, for such a charismatic insect group, the higher-level phylogenetic relationships among its superfamilies are still poorly resolved. Compared to earlier phylogenomic studies, we increased taxon sampling among Lepidoptera (37 superfamilies and 68 families containing 263 taxa) and acquired a series of large amino-acid datasets from 69,680 to 400,330 for phylogenomic reconstructions. Using these datasets, we explored the effect of different taxon sampling with significant increases in the number of included genes on tree topology by considering a series of systematic errors using maximum-likelihood (ML) and Bayesian inference (BI) methods. Moreover, we also tested the effectiveness in topology robustness among the three ML-based models. The results showed that taxon sampling is an important determinant in tree robustness of accurate lepidopteran phylogenetic estimation. Long-branch attraction (LBA) caused by site-wise heterogeneity is a significant source of bias giving rise to unstable positions of ditrysian groups in phylogenomic reconstruction. Phylogenetic inference showed the most comprehensive framework to reveal the relationships among lepidopteran superfamilies, and presented some newly relationships with strong supports (Papilionoidea was sister to Gelechioidea and Immoidea was sister to Galacticoidea, respectively), but limited by taxon sampling, the relationships within the species-rich and relatively rapid radiation Ditrysia and especially Apoditrysia remain poorly resolved, which need to increase taxon sampling for further phylogenomic reconstruction. The present study demonstrates that taxon sampling is an important determinant for an accurate lepidopteran tree of life and provides some essential insights for future lepidopteran phylogenomic studies.

4莲潜根线虫病在我国的发生及研究概况 4.1 多样潜根线虫(Hirschmanniella diversa)在我国研究历史和现状 关于潜根线虫属线虫为害莲的问题,我国研究者几乎与日本同时开始重视.从1991年开始,刘安国等[53,54]发现莲腐败病的发生与地下部的线虫有关,在莲根和莲鞭上分离出潜根线虫(H.spp.),且以根部居多;线虫为害导致的伤口,利于腐败病病原菌侵染,进而诱发莲腐败病.

The small, relict-like moth family Endromidae is well-established within the superfamily Bombycoidea, but relationships within the family have remained vague for the last decade, primarily due to very limited taxon sampling. This resulted in the explicit removal of all internal suprageneric classification by Zwick et al. (2011) when they synonymized Mirinidae and the bombycid subfamilies Oberthueriinae and Prismostictinae with Endromidae. Nucleotide and amino acid data sets of the 13 mitochondrial, protein-coding genes from representatives of 13 of the 16 accepted endromid genera were used to estimate phylogenetic relationships based on maximum likelihood and Bayesian inference methods. The results strongly support Endromidae as a monophyletic group and enable the establishment and diagnosis of four subfamilies (Endrominae, Mirininae stat. rev. , Oberthueriinae stat. rev. and Prismostictinae stat. rev. ). Within subfamily Oberthueriinae, we establish three tribes: Oberthueriini stat. rev. , Andracini tribe nov. and Mustiliini tribe nov. We provide morphological diagnoses and a genus-level checklist for the three tribes. Promustilia yajiangensis Wang, X. & Zolotuhin, 2015 is transferred to Mustilizans as M. yajiangensis comb. nov. to establish reciprocal monophyly of the two genera, and Andraca gongshanensis is transferred to Pseudandraca as P. gongshanensis comb. nov. We also synonymize Andraca (Chrypathemola) syn. nov. with Andraca (Andraca), as the latter is deeply nested within the former.

Gene families, which are parts of a genome's information storage hierarchy, play a significant role in the development and diversity of multicellular organisms. Several studies have focused on the characteristics of gene families, such as function, homology, or phenotype. However, statistical and correlation analyses on the distribution of gene family members in the genome have yet to be conducted. Here, a novel framework incorporating gene family analysis and genome selection based on NMF-ReliefF is reported. Specifically, the proposed method starts by obtaining gene families from the TreeFam database and determining the number of gene families within the feature matrix. Then, NMF-ReliefF is used to select features from the gene feature matrix, which is a new feature selection algorithm that overcomes the inefficiencies of traditional methods. Finally, a support vector machine is utilized to classify the acquired features. The results show that the framework achieved an accuracy of 89.1% and an AUC of 0.919 on the insect genome test set. We also employed four microarray gene data sets to evaluate the performance of the NMF-ReliefF algorithm. The outcomes show that the proposed method may strike a delicate balance between robustness and discrimination. Additionally, the proposed method's categorization is superior to state-of-the-art feature selection approaches.

The Genus Betapsestis was established by Matsumura in 1921, based on B. takeuchii from Honshu in Japan. It belongs to the subfamily Thyatirinae in the family Drepanidae, which is distributed in the Palaearctic and Oriental regions (Laszlo et al. 2007; Matsumura 1921). Only two species, B. umbrosa (Wileman, 1911) and B. brevis (Leech, 1900), have been known to date. Among them, B. brevis is endemic to China (Shaanxi, Sichuan, and Yunan), and B. umbrosa is endemic to Japan (Hokkaido, Honshu, Shikoku, and Kyushu), respectively (Laszlo et al. 2007; Matsumura 1921; Zhao 2004; Zhuang 2017). In this study, a new species of Betapsestis, B. shennongjiaensis Lai, Huang & Han sp. nov., from Shennongjia National Park in Hubei Province, China, is described as new to sciences.