Horizontal transfer of mitochondrial DNA into the nuclear genome generates nuclear mitochondrial sequences (NUMTs), which serve as molecular fossils reflecting long-term mitochondrial-nuclear interactions and genome evolution. However, the biological mechanisms governing NUMT integration, retention, and evolutionary fate remain incompletely understood in domesticated animals. Here, using the latest pig reference genome assembly (Sscrofa11.1), we present a comprehensive genome-wide characterization of NUMTs in pigs and provide new insights into their genomic distribution and evolutionary constraints. We identified 513 high-confidence NUMTs, of which 460 were chromosomally mapped, accounting for 0.0106% of the nuclear genome. Beyond increased detection, our analyses reveal that pig NUMTs exhibit non-random origins, preferentially integrate into genomic regions under weak selective constraint, and are frequently associated with repetitive elements, consistent with a DNA repair-mediated insertion mechanism. NUMTs predominantly occur as short, fragmented sequences and show signatures of long-term neutral evolution, while insertions disrupting coding sequences are strongly selected against. Synteny-based analyses further identified clustered NUMT regions and duplicated NUMTs, suggesting secondary genomic duplication events following initial integration. Comparative analysis with the earlier Sscrofa10.2 assembly demonstrates that improved genome quality substantially enhances NUMT detection, particularly in repetitive and GC-rich regions, clarifying previously ambiguous sequence-context associations. Together, this high-quality pig NUMT map provides a robust foundation for future functional, evolutionary, and population-level investigations and contributes to the conservation and utilization of pig genetic resources.

Abstract Loin muscle weight is an important indicator of carcass yield in meat rabbit production, but the host genetic and intestinal microbial factors associated with its variation remain poorly understood. Because the cecum is the primary site of hindgut fermentation in rabbits, we integrated whole-genome resequencing, cecal transcriptome profiling, and cecal and rectal 16S rRNA sequencing data from 321 Kangda meat rabbits, with rectal microbiome data used as a downstream comparative reference. Compared with the rectum, the cecum contained a richer microbial community, with more ASVs and genera and significantly higher microbial diversity, whereas predicted metabolic functions were largely conserved between the two segments. Loin muscle weight showed moderate SNP-based heritability (h² = 0.39), and the cecal microbiome explained a smaller but detectable proportion of phenotypic variation (m² = 0.13). Multi-strategy microbial screening identified 19 candidate cecal genera associated with loin muscle weight, with Methanosphaera showing the strongest negative association. Host GWAS prioritized candidate loci near MEX3C and TCF4 on chromosome 10, and integration of cecal cis-eQTL and GWAS summary statistics further prioritized GJB3 as a candidate gene associated with loin muscle weight. Consistent with the cecum-centered model, host genetic relatedness was weakly but significantly correlated with cecal microbial similarity, whereas no such global association was observed for the rectal microbiome. Microbial GWAS and SMR analyses further prioritized a cecal MRAP2–Methanobrevibacter association as the main host-regulated microbial signal, while rectal analyses identified distinct segment-specific signals, including SULF1–Roseburia. These findings suggest that host genetic variation may be linked to loin muscle deposition partly through cecal gene expression and fermentation-related cecal microbial taxa, with the rectal microbiome providing comparative evidence for hindgut segment specificity. This study provides candidate host and microbial targets for future functional validation and microbiome-informed nutritional strategies to improve carcass traits in meat rabbits.

Conventional immunoassay development relies on multi-discrete steps involving bioreceptor production, conjugation, and detection that omits matrix cleanup, all of which collectively introduce variability and undermine reliability. Herein, we present an all-in-one system that integrates nanobody (Nb) harvesting, oriented immobilization, matrix cleanup, target enrichment and quantification in a single-vessel workflow, thereby markedly enhancing the reliability and practicality of immunoassays. The system is initiated by the metal chelation, which enables direct harvesting of Nb from bacterial lysate and seamlessly progresses to oxidative locking of the Nb in an oriented manner onto functionalized magnetic beads (MBs). Under saturation loading, the prepared Nb-MBs exhibited a coefficient of variation of only 4.56% in the half-maximal effective concentration (EC50) across six batches. In a well-streamlined two-step immunoassay procedure, the Nb-MBs enabled the full process from matrix cleanup to accurate aflatoxin B1 quantification to be completed in 45 min. This design not only eliminates batch-to-batch inconsistency but also resolves the long-standing trade-off between tedious matrix cleanup and assay speed. Furthermore, the Nb-MBs demonstrated excellent storage stability, anti-interference capability, and reusability, and could also serve as a cleanup material for HPLC analysis. For the first time, this work seamlessly integrates the bioreceptor production and oriented immobilization within the detection process, offering a more reliable paradigm for the next-generation immunoassays.

Background: Machine learning (ML) holds great promise for genomic breeding value prediction in livestock and poultry, yet its application in layer breeding remains limited. Methods: In this study, we used whole-genome resequencing data from 834 Wenshui Luhua Green-Shelled (WLGS) laying hens to predict genomic breeding values for eight egg production and egg quality traits using multilayer perceptron (MLP), random forest (RF), and genomic best linear unbiased prediction (GBLUP). Model performance was evaluated via 10-fold cross-validation, and the effects of data type and single nucleotide polymorphism (SNP) density were examined. Results: Heritability analysis indicated moderate heritability for egg number (EN) at 0.327. Egg weight-related traits (EW-30W, EW-40W, and EHD-40W) exhibited high heritability (0.570-0.631), while eggshell strength (ESS-40W) and thickness (EST-40W) showed moderate heritability at 0.228 and 0.220, respectively. Model comparisons revealed that RF performed best for egg shape index (ESI-30W, 0.395) and most egg quality traits, whereas GBLUP yielded optimal results for egg weight traits, achieving prediction accuracies of 0.392 for EW-30W and 0.432 for EW-40W. Whole-genome resequencing data consistently outperformed 50K chip data across all models, with GBLUP improving EW-40W prediction accuracy by 24.9%. SNP density analysis further showed that GBLUP remained stable under low-density conditions, while MLP and RF progressively improved with increasing density, with RF demonstrating the most pronounced advantage at high densities. Conclusions: In summary, the GBLUP model is suitable for traits with high heritability and low-density marker scenarios, while the RF model demonstrates significant predictive advantages for egg production and specific egg quality traits under high-density conditions. This study provides scientific basis for model selection in the genomic selection program for laying hens.

Anthrax is a highly dangerous zoonotic disease that poses a significant threat to public safety. Accurate and convenient monitoring of anthrax biomarker, 2,6-dipicolinic acid (DPA), is particularly meaningful for preventing security incident outbreaks and treatment of anthrax. Herein, a dual-emissive ratiometric fluorescent and visual sensing platform has been elaborately constructed for facile, efficient and portable detection of DPA. The newly designed dual-ligand metal-organic framework was synthesized through one-pot solvothermal process that Eu3+ coordinated with 2,5-dihydroxyterephthalic acid (DHTA) and 2,4,6-tris(4-carboxyphenyl)-1,3,5-triazine (TATB) to form rodlike europium metal-organic framework (EuDHTA). This obtained EuDHTA displayed excellent luminescent properties and satisfactory response behavior toward DPA. In the presence of target DPA, the fluorescence from DHTA at 536 nm was remarkably enhanced while the characteristic fluorescence for Eu3+ decreased dramatically that was attributed to DPA-induced structural alteration of EuDHTA. Furthermore, a hydrogel-integrated EuDHTA visual system for instrument-free analyzing DPA in real water samples and Bacillus subtilis spores was realized via a smartphone equipped with color recognition application, which revealed great possibilities of the developed sensor for on-site reliable monitoring anthrax.

Pathogens, such as nucleopolyhedroviruses (NPVs), are promising biological control agents for lepidopteran pests to protect crops. However, a major drawback, their field application is limited by slower virulence compared with chemical insecticides. Modulating host pathways to enhance viral infectivity offers a potential strategy for improving NPV-based biocontrol. In this study, the expression level of NADH:ubiquinone oxidoreductase core subunit S8 (Ndufs8) was upregulated in the insect midgut following Spodoptera frugiperda NPV (SfNPV) infection. Silencing Ndufs8 impaired mitochondrial function, increased oxidative stress, and activated autophagy, but had no significant adverse effects on larval development and survival. The Ndufs8-autophagy cascade subsequently promoted SfNPV replication in insects. Co-feeding larvae with SfNPV and nanocarrier-delivered double-stranded RNA targeting Ndufs8 (dsNdufs8) accelerated insect death compared with the virus alone, demonstrating the enhanced virulence. Furthermore, bacterially expressed short hairpin RNA against Ndufs8 (shNdufs8), followed with nanocarrier delivery, achieved effective gene silencing and increased insect mortality comparable to synthetic dsNdufs8, supporting the potential field application for scalable RNA delivery. Our findings elucidate the function of a host responsive gene in the virus-host interaction. The combined use of dsNdufs8/shNdufs8 with SfNPV highlights a practical and scalable strategy to integrate RNA interference with pathogens to improve the biocontrol efficacy against insect pests.

The rapid and convenient synthesis of ultrabright fluorescent polymer dots is attractive for enhancing lateral flow immunoassays (LFA) sensing, but remains extremely challenging due to the high-temperature and high-pressure conditions required for their conventional carbonization. Here, we report a universal, facile, and mild photochemical strategy for preparing silanized polymer dots (SPDs) using a photosensitive fluorescein derivative and silane molecules, which can be completed under ambient conditions within 30 min. Benefiting from the crosslink-enhanced emission and stability (CEES) effect, the as-prepared SPDs exhibit exceptional photoluminescence properties, achieving a high photoluminescence quantum yield (PLQY) of 96.76% (63.2-fold enhancement over the precursor) along with substantially improved photostability and storage stability. To leverage the superior fluorescence performance of SPDs, we constructed an LFA biosensor that couples them with catalytic hairpin assembly (CHA), enabling convenient and ultrasensitive detection of miRNA-21 (miR-21). The constructed fluorescent LFA biosensor exhibited an excellent detection limit of 0.072 pM and demonstrated superior reliability for miR-21 sensing in clinical serum samples. This work opens new avenues for synthesizing ultra-bright fluorescent SPDs for biosensing and broadens the prospects for their use in clinical diagnostics.

Heat stress limits dairy production. The temperature-humidity index (THI), combining temperature and relative humidity, is widely used to assess heat stress. However, in Chinese Holstein cattle, the phenotypic responses of milk traits and genotype-environment interaction mechanisms under different THI conditions are understudied. Based on 63,334 records from 7240 cows (milk yield, fat percentage, protein percentage), matched with meteorological data and 113,297 SNPs, we employed a random-effects GWAS to examine SNP effects across a continuous THI gradient, comparing results with conventional, temperature-, and humidity-interaction GWAS. As THI increased, all traits declined with distinct patterns. Random regression GWAS identified 149 significant SNP × THI interactions (5 for MY, 86 for FP, 58 for PP), distributed across BTA5, BTA6, BTA14, and BTA20. Candidate gene annotation identified 52 candidate genes near significant SNPs, of which 50 core candidate genes were supported in both temperature and humidity GWAS. The most robustly supported core candidate genes include DGAT1, CPSF1, ABCG2, MGST1, VPS28, PPP1R16A, ZNF250, GRID2, KCNC2, and LOC787350—of which DGAT1, ABCG2, and MGST1 have been functionally validated in milk production traits, whereas others represent novel candidates requiring further investigation. Temperature and THI-GWAS showed high consistency, while humidity-GWAS detected both overlapping and specific signals. Incorporating THI as a continuous environmental gradient identifies environment-dependent regulatory signals not captured by conventional GWAS, providing candidate genes that may contribute to future breeding strategies after further validation.

Abstract Background The significant temperature variations across northern and southern China have driven the adaptive evolution of Chinese native cattle breeds, allowing them to thrive in diverse and extreme bioclimate environments. Understanding how these breeds have adapted to varying temperatures is essential for identifying genetic factors that contribute to their survival in such conditions. Results In this study, using whole-genome sequence data of 336 individuals (with an average sequencing depth of 30.12 ×) from 21 cattle breeds, including 8 breeds from cold regions, 3 from warm regions, and 10 from hot regions, clear genetic differentiation among the three groups of breeds was revealed. Using whole-genome SNP, InDel, and SV data, a series of selective genomic regions, genes, and variants/SVs associated with cold or hot temperature adaptability were identified. Key genes, including KLB, HSPA4, ECSCR, DNAJC18 and SLC9A1 are speculated to be responsible for cold/hot adaptability based on the extreme difference in allele frequency of the selective variants/SVs harbored by these genes, their known biological functions, protein–protein interaction network, findings from previous studies on their relation to environmental adaptation, and their tissue specificities. Conclusions By integrating SNP, InDel, and SV data, this study provides a comprehensive genetic framework for understanding selective environmental adaptation. These findings enhance our understanding of the mechanisms underlying temperature adaptation in cattle and offer a molecular foundation for the development of new breeds.

The WRKY transcription factors play an important role in regulating plant response to drought stress. The results show that SlWRKY30 acts as a negative regulator of drought tolerance, and that overexpressing SlWRKY30 (SlWRKY30OE) promotes vegetative growth. Moreover, the expression of SlWRKY30 was induced by drought stress, and SlWRKY30OE plants exhibited more severe wilting than WT under drought conditions. SlWRKY30 overexpression led to high relative electrolyte leakage, MDA content, and ROS levels in tomatoes under drought stress. In SlWRKY30OE plants, the activities of SOD, POD, CAT and APX were lower than those of WT plants; further, the expression levels of SlFe-SOD, SlCAT1 and SlcAPX were also lower than in WT plants under drought stress. Further analysis of RNA sequencing indicated that the transcription level of the encoding photosystem I and II genes was almost down-regulated, while the degree of SlWRKY30OE plants downregulation was stronger than in WT plants. Consistently, the Pn and Fv/Fm were lower in SlWRKY30OE plants during drought stress. These results indicate that SlWRKY30 reduces drought resistance in tomato by weakening antioxidant capacity, promoting ROS accumulation and membrane damage, and impairing photosynthetic performance under drought stress.

Aflatoxin B-1 (AFB(1)), the most dangerous mycotoxins found in grains, is a powerful carcinogen. Consequently, creating immunoassay techniques which are rapid, robust, sensitive, and user-friendly for its detection is crucial to ensuring food safety. In this study, a nanobody (variable domain of heavy chain of heavy chain antibody, VHH) against AFB(1) was prepared and purified successfully based on recombinant plasmid pET22b(+)-VHH-AFB(1) obtained in previous research work. A novel nanobody-based nanoarchitectonics of indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) for the ultra-sensitive detection of AFB(1) was constructed under the optimal experimental conditions. The icCLEIA method in the study showed high sensitivity to AFB(1) with a 50 % inhibitory concentration (IC50) of 0.63 ng/mL, and a limit of detection (LOD) of 0.066 ng/mL, and a linear range (IC20 similar to IC80) of 0.15-2.90 ng/mL. The icCLEIA method based on the purified nanobody had litter cross-reactivity (CR% < 0.1) with other structural analogues, which indicated that the method had high specificity for AFB(1.) The average recoveries of AFB(1) in spiked millet samples ranged from 85.6 % to 103.4 % and the coefficient variation (CV) values were less than 3.14 %. In the experiment of recovery test, a strong linear correlation (R-2 = 0.9991) was observed by comparing the results from the nanobody-based icCLEIA with the UPLC-MS/MS reference method. The above results indicated that the nanobody-based nanoarchitectonics of icCLEIA is a reliable and effective alternative for quantifying AFB(1) in various grains.

Leaf growth is a major determinant of plant architecture and productivity. TWISTED DWARF1 (TWD1/FKBP42) is a conserved immunophilin involved in plant growth and development, but its function in tomato remains unclear. Here, we identified and characterized SlTWD1, the tomato homolog of TWD1. SlTWD1 is highly conserved among land plants and functionally complemented the developmental defects of an Arabidopsis thaliana cr-twd1 mutant. SlTWD1 was broadly expressed in vegetative and reproductive tissues. CRISPR/Cas9-mediated disruption of SlTWD1 caused severe dwarfism and reduced leaf and fruit size, whereas overexpression of SlTWD1 or a C-terminally truncated variant moderately promoted leaf growth. Cellular analyses showed that loss of SlTWD1 markedly reduced epidermal cell size, while overexpression increased cell size, indicating that SlTWD1 promotes leaf growth primarily through cell expansion. Expression changes in auxin- and cell cycle-related genes were also associated with SlTWD1 perturbation. These findings indicate that SlTWD1 functions as an evolutionarily conserved regulator of plant growth that contributes to leaf and fruit size control in tomato.

Managing late-instar lepidopteran pests remains a critical challenge due to their robust physiological and physical barriers, which severely compromise the efficacy of insecticides. Lufenuron, an insect growth regulator, controls lepidopteran pests by inhibiting chitin biosynthesis, but its efficacy against late-instar larvae needs improvement. Here, we developed a dual-barrier disruption strategy by integrating nano-lufenuron with RNA interference targeting the chitin synthase genes PxCHS1 and PxCHS2 in the diamondback moth, Plutella xylostella. The nanocarrier star polycation (SPc)-mediated lufenuron enhanced both stomach and contact toxicity to the diamondback moth larvae, and induced high expression of PxCHS1 and PxCHS2. PxCHS1 and PxCHS2 were predominantly expressed in the integument and midgut peritrophic matrix, respectively. SPc-mediated RNA interference of PxCHS1 caused cuticle thinning and enhanced the contact toxicity of nano-lufenuron, whereas silencing of PxCHS2 disrupted peritrophic matrix integrity and increased stomach toxicity. Furthermore, dsPxCHS1 and dsPxCHS2 self-assembled with the lufenuron/SPc nanocomplex, facilitating coordinated penetration and uptake across both cuticular and intestinal barriers. Co-delivery of LUF/SPc/dsPxCHS1 + dsPxCHS2 significantly increased mortality and reduced adult emergence in late-instar larvae. Collectively, this study establishes a barrier-targeted nanopesticide framework for overcoming the physical barriers and provides a novel dual-barrier-targeted nanopesticide strategy for efficient control of late-instar diamondback moth larvae, with potential reference for other lepidopteran pests.

Genomic prediction holds significant potential for advancing precision medicine in humans, as well as accelerating genetic improvement in animals and plants. For multi-trait prediction, the conventional multi-trait models are primarily based on global genetic correlations between traits. With the development of local genetic correlation (LGC) estimation methods, it is now possible to analyze LGCs confined to specific genomic regions and it is expected that incorporating LGCs into multi-trait prediction model would enhance the prediction ability. Here, we proposed three models to address this issue and evaluated their performances using simulated data and three real datasets from human, cow, and pig populations. Our results demonstrate that LGCs are heterogeneous across the genome and incorporating LGCs in multi-trait prediction would increase the prediction accuracy by an average of 12.76% ± 2.07% compared to conventional multi-trait genomic prediction method (MTGBLUP) in the real datasets. Our findings highlight the importance of considering LGCs in improving multi-trait genomic prediction. Three local genetic correlation (LGC) genomic prediction methods are proposed to incorporate LGCs into multi-trait genomic prediction and could universally improve prediction accuracy compared to conventional multi-trait prediction methods.

Competitive immunoassays for small-molecule haptens such as aflatoxin B-1 (AFB(1)) are often limited by insufficient sensitivity and narrow dynamic ranges. This study seeks to address these issues through the genetic engineering of nanobodies and the development of novel immunoassay formats using AFB(1) as a model. An innovative elution-free biopanning strategy was developed for the in vitro evolution of an AFB(1)-specific nanobody, which proved highly suitable for screening high-performance anti-hapten nanobodies. Using this strategy, several mutants with improved analytical performance were obtained, including one that showed a 3.5-fold reduction in the half-maximal inhibitory concentration (IC50) compared with the parent nanobody. Affinity kinetics of five representative mutants demonstrated a positive correlation between the IC50 values and their affinity for the artificial antigen. By leveraging these mutants, a novel sandwich-competitive immunoassay (SCI) format was constructed using two nanobodies along with a special artificial antigen. The SCI demonstrated a markedly broadened detection range (IC10-IC90) with excellent linearity (R-2 > 0.98), outperforming conventional competitive assays, which showed compromised linearity outside the IC20-IC80 range (R-2 < 0.92). The broader detection range of SCI can be attributed to the two-stage competitive binding process of the analyte to the sandwich immunocomplex. Spike-recovery experiments confirmed the accuracy and applicability of the SCI in actual sample detection. This study offers a strategy that revitalizes the in vitro evolution of nanobodies targeting haptens and advances immunoassay development for such molecules.

Eggshells not only protect the contents of the egg from external damage but are also a key factor influencing consumer choice, second only to price. In the later stages of egg production, the incidence of pimpled eggs significantly increases, severely affecting the hatchability and food safety of the eggs. This study compares the differences in the uterine proteomes and metabolomes of hens producing pimpled eggs and those producing normal eggs, aiming to identify the proteins and metabolites that may play a crucial role in the formation of pimpled eggs. A total of 242 differentially expressed proteins (DEPs) were identified in uterine tissue, of which 116 were upregulated and 126 were downregulated. Enrichment analysis revealed that the DEPs were enriched in pathways related to ion transport, energy metabolism, and immune responses. The study found that in the normal eggs (NE) group, HCO₃⁻ was predominantly transported via SLC4A1, although other transport pathways may also play a role. In contrast, in the pimpled eggs (PE) group, bicarbonate ions (HCO₃⁻) was primarily transported through SLC4A4. Additionally, a total of 44 differentially metabolites (DMs) were identified in the uterus, with 5′-Adenylic acid (ATP) being significantly downregulated in the PE group. The ions and matrix proteins required for eggshell formation are transported from uterine cells to the uterine fluid against a concentration gradient, a process that consumes a substantial amount of energy. The decrease in ATP concentration in the PE group may be a significant factor influencing the formation of pimpled eggs. Subsequently, we found that the DEPs and DMs were jointly enriched in several signaling pathways, including the FoxO signaling pathway related to energy metabolism, nicotinate and nicotinamide metabolism, and tryptophan metabolism associated with immune response. Notably, the DMs involved in these signaling pathways were all downregulated in the PE group. Our research findings indicate that SLC4A1, SLC4A2, and ATP2B4 (DEPs), along with 5′-adenylic acid and trigonelline (DMs), influence the formation of eggshells through mechanisms related to energy metabolism, ion transport, and immune response. These DEPs and DMs may serve as potential biomarkers for the genetic improvement of eggshell quality.

Cuticular proteins are essential for cuticle formation, molting, and survival in insects. However, functional analysis of cuticular proteins in the melon aphid has been limited. In this study, we identified an endocuticle structural glycoprotein (ESG) AgSgAbd-2-like in the melon aphid Aphis gossypii, which is a member of the RR-1 subfamily of the CPR (cuticular protein containing the conserved Rebers-Riddiford motif) chitin-binding proteins. When double-stranded RNA is delivered epidermally, AgSgAbd-2-like is knocked down, resulting in molting defects and mortality. The expression of AgSgAbd-2-like is comparatively low prior to molting and increases following molting. Ecdysone signaling consistently suppresses AgSgAbd-2-like. Histologically, the endocuticle and whole cuticle are thinner in AgSgAbd-2-like RNA interference (RNAi) aphids, which is a leading cause of molting defects and mortality. Furthermore, knockdown of any other homolog of ESGs, including AgSgAbd-4, AgSgAbd-4-like, AgSgAbd-8-like, and AgSgAbd-9-like, results in molting defects and death, like that by AgSgAbd-2-like RNAi. These results indicate that the melon aphid ESGs are conserved in cuticle formation and could be potential targets for RNAi-based pest management.

Locusts have been a major global agricultural pest that poses a serious threat to crop and livestock production. Entomopathogenic fungi (EPF) provide an eco-friendly control method; however, their efficacy usually takes slow and is unstable. To achieve an enhancement of the biocontrol efficacy of Beauveria bassiana (B. bassiana) against locusts, we developed a new strategy by which B. bassiana and nanocarrier-mediated dsRNA are co-applied across the locust cuticle. The nanocarrier star polycation (SPc) effectively delivers Lmidgf4 dsRNA (dsLmidgf4) into the locust, which targets Locusta migratoria imaginal disc growth factor 4 (Lmidgf4). SPc protects dsLmidgf4 from degradation by the hemolymph and enables efficient gene silencing. Furthermore, SPc has no adverse effects on B. bassiana spore germination and growth. Lmidgf4 interference leads to a thinner layer of endocuticle, thus facilitates infection of B. bassiana, and finally reduces the median lethal time of locusts infected with B. bassiana. In conclusion, the combination of B. bassiana and dsRNA/SPc complex overcomes the slow action of fungi, providing a novel strategy for field control of locusts.

Entomopathogenic fungi are eco-friendly biological method to control various agricultural pests. However, it takes time during the practical application of Beauveria bassiana in thrip management. This study investigates the role of Megalurothrips usitatus Dorsal switch protein 1 (MuDSP1), which is an immune regulatory gene during fungus infection. MuDSP1 is highly expressed at the pupal and adult stages in M. usitatus. Following the infection with B. bassiana, the expression of MuDSP1 is upregulated, as well as Phospholipase A2, lysozyme, and apolipophorin in the immune pathway. The later three genes were repressed by the interference of MuDSP1, confirming the crucial role of MuDSP1 in the immune response. Interference of MuDSP1 has no effects on the mortality of bean flower thrips, but the combination of MuDSP1 dsRNA and B. bassiana infection significantly increases the mortality caused by B. bassiana. This could be due to the attenuated immune response induced by MuDSP1 dsRNA. In conclusion, this study reveals the significant role of MuDSP1 in immune response to fungi. Simultaneous application of MuDSP1 dsRNA and B. bassiana not only provides a new strategy for high-efficient biological management but also offers an alternative approach to sustainable agricultural pest control.