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Clanis bilineata Walker (Lepidoptera: Sphingidae), a burgeoning edible insect, is experiencing rising demand in China and other regions. Despite this interest, larval production is currently constrained by the limitations of artificial production technologies, particularly the selection of optimal host plants. This study rigorously evaluated the performance of C . bilineatha larvae on four main host plants: round-leaf soybean, pointed-leaf soybean, black locust, and kudzu. Preference tests demonstrated that the larvae were most attracted to black locust (34.76 ± 4.65%), with subsequent preferences for kudzu (25.00 ± 6.12%), round-leaf soybean (23.17 ± 2.79%), and pointed-leaf soybean (14.02 ± 4.74%). No significant preference differences were noted between round-leaf soybean and either black locust or kudzu. In feeding assays, the larvae exhibited a marked preference for round-leaf soybean (37.36 ± 0.81 g, total feeding amount for larvae), followed by kudzu (37.26 ± 0.82 g), pointed-leaf soybean (35.38 ± 1.31 g), and black locust (28.53 ± 0.81 g). When the larvae were fed on round-leaf soybean, they exhibited significantly higher survival rate (39.33 ± 0.90%), body weight (9.75 ± 0.07 g), total biomass (383.43 ± 7.35 g), pupation rate (87.78 ± 1.73%), and egg production (189.80 ± 1.06 eggs/female) compared to other hosts. These findings uncovered that round-leaf soybean significantly enhances larval performance, suggesting its potential for improving C . bilineata larval production and sustainability in cultivation systems.

Background Chemical pesticides face significant challenges regarding their efficacy and environmental impact. Plant-based food attractants have emerged as a promising green alternative for pest control. However, their field application is limited by the short duration of effectiveness, necessitating improved carrier systems for sustained release. Electrospinning is a promising technology in this field, with core-shell fibers offering superior performance in efficient loading and sustained release compared to uniaxial fibers, highlighting their potential for further development. Results In this study, core-shell micro-nano fiber mats were prepared via coaxial electrospinning using multiple environmentally friendly polymers. These mats were firstly and successfully loaded with food attractants bisexually attractive to Loxostege sticticalis adults, including 1-octen-3-ol, trans-2-hexenal, linalool, and anethole, enabling sustained release and effective trapping. The components in the core-shell spinning solution were chemically compatible, and after spinning, the poly(3-hydroxybutyrate-co-4-hydroxybutyrate)/polycaprolactone (PHB/PCL) in the shell layer and polyethylene oxide (PEO) in the core layer formed core-shell fibers with clear boundaries. The mats achieved an average encapsulation efficiency of 78% for active ingredients, with a sustained release profile that delivered over 60% of the attractants within 80 days while mitigating early burst release. Electroantennogram and behavioral studies revealed that the mats retained electrophysiological activity for at least 90 days, effectively attracting male and female adult insects even after 75 days. Field trials demonstrated that the mats significantly outperformed commercial slow-release carriers, attracting a higher number of L. sticticalis adults. Additionally, the mats exhibited strong stress resistance, biodegradability, and environmental compatibility, effectively protecting active molecules while minimizing ecological impact. Conclusions The developed fiber mats provide a highly efficient, eco-friendly carrier for plant-based food attractants, offering prolonged efficacy and improved insect trapping performance. This study highlights their potential for sustainable agriculture and pest management, paving the way for greener alternatives to chemical pesticides. Graphical abstract

Background Entomopathogenic nematodes (EPNs) emerge as compatible alternatives to conventional insecticides in controlling Holotrichia parallela larvae (Coleoptera: Scarabaeidae). However, the immune responses of H. parallela against EPNs infection remain unclear. Results In present research, RNA-Seq was firstly performed. A total of 89,427 and 85,741 unigenes were achieved from the midgut of H. parallela larvae treated with Heterorhabditis beicherriana LF for 24 and 72 h, respectively; 2545 and 3156 unigenes were differentially regulated, respectively. Among those differentially expressed genes (DEGs), 74 were identified potentially related to the immune response. Notably, some immune-related genes, such as peptidoglycan recognition protein SC1 (PGRP-SC1), pro-phenoloxidase activating enzyme-I (PPAE-I) and glutathione s-transferase (GST), were induced at both treatment points. Bioinformatics analysis showed that PGRP-SC1, PPAE-I and GST were all involved in anti-parasitic immune process. Quantitative real-time PCR (qRT-PCR) results showed that the three immune-related genes were expressed in all developmental stages; PGRP-SC1 and PPAE-I had higher expressions in midgut and fat body, respectively, while GST exhibited high expression in both of them. Moreover, in vivo silencing of them resulted in increased susceptibility of H. parallela larvae to H. beicherriana LF. Conclusion These results suggest that H. parallela PGRP-SC1, PPAE-I and GST are involved in the immune responses to resist H. beicherriana LF infection. This study provides the first comprehensive transcriptome resource of H. parallela exposure to nematode challenge that will help to support further comparative studies on host-EPN interactions.

Odorant binding proteins play a crucial role in transporting semiochemicals across the sensillum lymph to olfactory receptors within the insect antennal sensilla. In this study, the general odorant binding protein 2 gene was cloned from the antennae of Loxostege sticticalis, using reverse transcription PCR and rapid amplification of cDNA ends. Recombinant LstiGOBP2 was expressed in Escherichia coli and purified by Ni ion affinity chromatography. Real-time PCR assays indicated that LstiGOBP2 mRNA is expressed mainly in adult antennae, with expression levels differing with developmental age. Ligand-binding experiments using N-phenyl-naphthylamine (1-NPN) as a fluorescent probe demonstrated that the LstiGOBP2 protein has binding affinity to a broad range of odorants. Most importantly, trans-11-tetradecen-1-yl acetate, the pheromone component of Loxostege sticticalis, and trans-2-hexenal and cis-3-hexen-1-ol, the most abundant plant volatiles in essential oils extracted from host plants, had high binding affinities to LstiGOBP2 and elicited strong electrophysiological responses from the antennae of adults.

The tobacco cutworm, Spodoptera litura F. (Lepidoptera: Noctuidae), is a polyphagous pest that causes enormous losses in tobacco production as it develops resistance to pesticides in China. Telenomus remus N. (Hymenoptera: Scelionidae) is an effective egg parasitoid for the genus Spodoptera. However, little is known about the volatile compounds that attract the parasitoid. In the present study, we investigated the response of T. remus to volatiles released from various sources, including Spodoptera litura egg masses, healthy tobacco plants, plants with manually introduced eggs, and plants with natural oviposition. The chemical profiles of the crude extracts were identified using gas chromatography–mass spectrometry (GC–MS), and the behavioral assay was performed using a Y‐tube olfactometer. Naïve and experienced T. remus showed significant attraction to crude extracts of S. litura egg masses and to oviposition‐induced plant volatiles (OIPVs). In subsequent experiments, different concentrations and combinations of the synthetic volatiles obtained from these crude extracts were tested. Notably, the binary and ternary mixtures showed significant attraction, where the binary combination of linalool (L) with tetracosane (T) and linalool (L) with cis‐3‐hexenyl acetate (C) was strongly attractive. These results underscore the potential of using synthetic volatile dispensers, particularly binary combinations with precise doses, to enhance retention of T. remus and improve the efficacy of the biological control program. The parasitoid T. remus was attracted to OIPVs and S. litura egg volatiles. At lower concentrations of 1 and 10 ng/μL the parasitoid showed the highest attraction. The binary combinations of linalool (L) with tetracosane (T) and linalool (L) with cis‐3‐hexenyl acetate (C) strongly attracted the parasitoids.

It was previously thought that the odorant binding proteins (OBPs) in the sensillum lymph might serve as carriers, which could carry lipophilic odorant molecules to olfactory receptors. In this study, two novel OBP genes of the scarab beetle Holotrichia oblita were screened using an antennal cDNA library. The full cDNA of HoblOBP3 and HoblOBP4 was cloned using reverse transcription PCR and rapid amplification of the cDNA ends. Homology modeling of both OBPs was performed using SWISS-MODEL on-line tools. Next, the two OBPs were expressed in Escherichia coli and purified using Ni ion affinity chromatography. The ligand-binding properties of HoblOBP3 and HoblOBP4 in 42 ligands respectively were measured using the fluorescence probe N-phenyl-naphthylamine (1-NPN). The results obtained from competitive binding assays demonstrated that HoblOBP4 showed a broader range of binding affinities to the test compounds, while HoblOBP3 displays more specific binding affinity. Furthermore, other OBPs and CSPs were expressed in Escherichia coli and purified using Ni ion affinity chromatography. Binding curves were measured for binary mixtures of OBPs and CSPs using 1-NPN, and the Scatchard plots exhibited \"J\"-like nonlinear correlation trends in some samples. In addition, competitive binding assays of the HoblOBP1 and HoblOBP2 mixtures and of the HoblOBP2 and HoblOBP4 mixtures with representative compounds unexpectedly demonstrated good affinity, which revealed extreme differences that were only obtained using the individual proteins. In the immunocytochemical analysis, colocalization of HoblOBP1 and HoblOBP2, and of HoblOBP2 and HoblOBP4, was detected in the sensilla basiconica and sensilla placodea, respectively. All of these results suggested that HoblOBP1 and HoblOBP2, as well as HoblOBP2 and HoblOBP4, may serve as heterodimers in the sensillum lymph.

Soybean root rot, primarily caused by Fusarium oxysporum, leads to severe root decay and substantial yield losses in Glycine max. This study screened ten entomopathogenic nematode-associated symbiotic bacteria for antagonistic activity against F. oxysporum. Among them, Xenorhabdus budapestensis XH-4 exhibited the strongest in vitro inhibition, suppressing mycelial growth by more than 73%. Antifungal activity was primarily attributed to extracellular metabolites, as both fermentation broth and cell-free culture supernatant were effective, whereas bacterial cell suspensions showed no significant inhibition. In greenhouse experiments, 40% (v/v) XH-4 reduced the disease index by 75–80%, comparable to the chemical fungicide metalaxyl–hymexazol. Genome mining revealed 20 biosynthetic gene clusters encoding diverse secondary metabolites, including fabclavine, fabclavine pyrrolizixenamide A, and putrebactin/avaroferrin, which likely underpin the strain antifungal efficacy. Additionally, XH-4 enhanced soybean antioxidant capacity and activated the phenylpropanoid pathway, indicating a dual mechanism involving direct antagonism and induced systemic resistance. These findings support the development of XH-4 as an environmentally friendly biofungicide for sustainable management of soybean root rot.

Chemosensory proteins (CSPs) play important roles in chemical communication by insects, as they recognize and transport environmental chemical signals to receptors within sensilla. In this study, we identified HoblCSP1 and HoblCSP2 from a cDNA library of Holotrichia oblita antennae, successfully expressed them in E. coli and purified them by Ni ion affinity chromatography. We then measured the ligand-binding specificities of HoblCSP1 and HoblCSP2 to 50 selected ligands in a competitive binding assay. These results demonstrated that HoblCSP1 and HoblCSP2 have similar ligand-binding spectra. Both proteins displayed the highest affinity for β-ionone, α-ionone and cinnamaldehyde, indicating that they prefer binding to odorants other than sex pheromones. Additionally, immuno-localization revealed that HoblCSP1 is highly concentrated in sensilla basiconica, while HoblCSP2 is specifically localized to sensilla placodea. In conclusion, HoblCSP1 and HoblCSP2 are responsible for binding to general odorants with slightly different specificities due to their different in vivo environments.

Holotrichia parallela is one of the agriculturally important scarab beetle pests in China. In this study, HparOBP14 was cloned, which is the most abundantly expressed among the OBP genes in the legs of female H. parallela adults. Sequence comparison and phylogenetic analysis showed that HparOBP14 has a Plus-C structure motif. The expression profile analysis revealed that HparOBP14 expression was the highest in the female antennae and then in the legs. The fluorescence competitive binding experiment of the recombinant HparOBP14 protein showed that HparOBP14 had an affinity with 6-methyl-5-heptene-2-one (plant volatile), 3-methylindole, p-cymene, methanol, formaldehyde, α-pinene, and geraniol (organic fertilizer volatile). Knockdown HparOBP14 expression decreased significantly the EAG response of the injected female adults to p-cymene, methanol, formaldehyde, α-pinene, and geraniol. Similarly, the injected female adults were significantly less attracted to geraniol and methanol. Therefore, HparOBP14 might bind organic matter volatiles during oviposition. These results are not only helpful to analyze the olfactory recognition mechanism of female adult H. parallela when choosing suitable oviposition sites, but also to provide target genes for green prevention and control of H. parallela in the future.

Objective Previous studies showed that flight muscles degenerate after migration in some aphid species; however, the underlying molecular mechanism remains virtually unknown. In this study, using the wheat aphid, Sitobion avenae , we aim to investigate aphid flight muscle degeneration and the underlying molecular mechanism. Results Sitobion avenae started to differentiate winged or wingless morphs at the second instar, the winged aphids were fully determined at the third instar, and their wings were fully developed at the fourth instar. After migration, the aphid flight muscles degenerated via programmed cell death, which is evidenced by a Terminal deoxynucleotidyl transferase dUTP-biotin nick-end labeling assay. Then, we identified a list of differentially expressed genes before and after tethered flights using differential-display reverse transcription-PCR. One of the differentially expressed genes, ubiquitin-ribosomal S27a, was confirmed using qPCR. Ubiquitin-ribosomal S27a is drastically up regulated following the aphids’ migration and before the flight muscle degeneration. Our data suggested that aphid flight muscles degenerate after migration. During flight muscle degeneration, endogenous proteins may be degraded to reallocate energy for reproduction.