Sublethal emamectin benzoate suppresses multi-transgenerational reproduction and alters symbiotic bacteria of Thrips tabaci

Sublethal emamectin benzoate suppresses multi-transgenerational reproduction and alters symbiotic bacteria of Thrips tabaci | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Sublethal emamectin benzoate suppresses multi-transgenerational reproduction and alters symbiotic bacteria of Thrips tabaci Chunjie Xian, Miaomiao Xin, xiaoyun wang, Xiangzhen Zhu, Li Wang, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8257750/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 13 You are reading this latest preprint version Abstract Thrips tabaci (Tysanoptera: Tripidae) is a globally significant agricultural pest whose control relies heavily on insecticides. This study comprehensively assessed the toxicity and sublethal effects of emamectin benzoate (EB) against T. tabaci and explored its underlying mechanisms. EB exhibited high acute toxicity, and at the LC 20 level, it significantly reduced longevity and fecundity in the F 0 generation, with these inhibitory effects persisting into the F 1 generation. Molecular analyses revealed that the P450 gene CYP6K1 is crucial in the T. tabaci ’s response to insecticide stress, and RNAi-mediated suppression of CYP6K1 expression significantly increased susceptibility. Additionally, sublethal dose EB exposure altered the gut microbiota, marked by a decline in Acinetobacter and an expansion of Pantoea . We propose that T. tabaci mounts a coordinated defense mediated by both the host CYP6K1 gene and intestinal symbionts, and disruption of this system impairs tolerance. These findings provide multi-level insights into the effect of EB on T. tabaci , supporting the development of integrated management strategies targeting both host detoxification and microbial communities. pests insecticide toxicity sublethal effects growth and development transcriptome 16S rRNA sequencing Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Key Message Emamectin benzoate shows dual toxicity across T. tabaci generations. The P450 gene CYP6K1 modulates insecticide susceptibility. Gut microbiome shifts accompany sublethal insecticide exposure. Host‑symbiont co‑response underpins pesticide tolerance in thrips. Targeting detox genes and microbiota may improve integrated pest control. Introduction Thrips tabaci (Tysanoptera: Tripidae), well-known as cotton/onion thrips, is a polyphagous agricultural pest with extensive host range and distributed mostly in tropical, subtropical and temperate areas of the world. Combined with sucking sap from leaves especially young ones, flowers, and fruits, it can transmit viral plant diseases through leaf scraping, eventually causing substantial yield losses of various crop in the world every year(Naeem-Ullah et al., 2020 ). Up to now, chemical control using insecticides remains the primary strategy for managing thrips infestations on crops (Reitz et al., 2020 ; Sun et al., 2023 ).However, the long-term use of insecticides has led to the development of resistance to multiple chemical classes for thrips populations(Shen et al., 2023 ; Wakil et al., 2023 ) . Emamectin benzoate (EB), a semi-synthetic insecticide that belongs to a class of avermectin family, derived from the natural fermentation of a naturally occurring soil actinomycete Streptomyces avermitilis , exhibits high bioactivity against major crop pests alongside a favorable environmental profile characterized by low toxicity and minimal residues(D. S. Yang et al., 2017 ; Zhou et al., 2016 ). These properties underpin its widespread utilization as a key intervention for pest control in diverse agricultural systems, particularly in vegetable, fruit, and cotton production(Abbas et al., 2023 ; Abdel-Baky, Alhewairini, & Bakry, 2019 ; El-Saleh et al., 2025 ). The toxicity of EB lies in the specific and irreversible disruption of unique neurological targets in insects, leading to neuromuscular paralysis. These characteristics account for its high insecticidal activity, low toxicity, and environmental safety, establishing its importance in integrated pest management strategies(Z. Liu et al., 2025 ). Beyond acute lethality, the sublethal effects of EB constitute another dimension of its ecological impact. These effects arise from persistent field exposure to low concentrations, a common scenario resulting from insecticide degradation and the cryptic nature of pest feeding. Following field application, insecticides undergo gradual degradation by environmental factors such as sunlight, rain and so on. While high doses can elicit strong lethal effects on target pests, prolonged exposure to sublethal concentrations may induce a range of sublethal effects(Mougabure-Cueto, Fronza, & Nattero, 2024 ). Considerable studies have demonstrated that these effects—such as extended developmental duration, reduced longevity, and diminished fecundity—can paradoxically lead to population resurgence in pest species (Afza et al., 2023 ; Y. X. Chen et al., 2023 ), and transgenerational impacts on offspring(H. Gao et al., 2024 ; Guo et al., 2023 ).The minute body size and cryptic behavior of Thrips tabaci , which often conceals it within plant structures, significantly increase the likelihood of its exposure to sublethal doses of emamectin benzoate. Therefore, investigating the sublethal impacts of emamectin benzoate on T. tabaci is critical, as it benefits for enhancing the insecticidal efficacy of this compound and optimizing integrated pest management strategies. This study focuses on exploring the sublethal effects of emamectin benzoate on T. tabaci , and confirms its inhibitory impact on thrips reproduction and development for successive two generations eventually. Through an integrated approach incorporating bioassays, transcriptome sequencing, and 16S rRNA microbiome analysis, we systematically investigated this insecticide’s toxic action mechanisms and the corresponding detoxification metabolic responses of T. tabaci . Taken together, these results are expected to provide a critical theoretical foundation for the rational field application of emamectin benzoate and for the development of sustainable resistance management strategies against T. tabaci . Materials and methods Insects and insecticides A thelytokous population of Thrips tabaci was used in this study, originally provided by the Cotton Research Institute of the Chinese Academy of Agricultural Sciences. The T. tabaci were reared on red cabbage ( Brassica oleracea var. capitata rubra ) leaves placed in cages (29 cm × 19 cm × 13 cm) under the control condition of 25 ± 1°C, 50 ± 5% relative humidity, and a photoperiod of 16 h light : 8h dark. Emamectin benzoate (EB) insecticide (Pesticide Registration No.: PD20170668) was purchased from Shandong Feixiang Agricultural Development Co., Ltd., China. Toxicity bioassays EB toxicity to Thrips tabaci was evaluated by leaf-dip bioassay(Y. X. Chen et al., 2023 ). A stock solution was prepared by dissolving an appropriate amount of EB in distilled water containing 0.01% Triton X-100 as a surfactant. This stock solution was then serially diluted with the 0.01% Triton X-100 aqueous solution to create a series of EB concentrations (50, 25, 12.5, 6.25, and 3.125 mg/L). A 0.01% Triton X-100 aqueous solution served as the control. Cabbage leaves discs (2 cm in diameter), prepared with a hole puncher, were immersed in the test solutions or control for 15 seconds. After air-drying, the treated leaf discs were placed in plastic containers. Twenty-five newly emerged, healthy adult thrips were placed into each container. All containers were then kept in the climate chamber under the same control condition as above. Mortality was assessed after 48 hours. Individuals that showed no movement when gently prodded with a fine brush were considered dead. The experiment was repeated 3 times for each concentration. Sublethal effects of EB on T. tabaci The sublethal effects were evaluated using the leaf-dipping method with the LC 20 concentration of EB, prepared by serial dilution of the stock solution with distilled water containing 0.1% (v/v) Triton X-100. Fresh cabbage leaves were immersed in the LC 20 solution for 15 seconds, air-dried. Then 60 newly emerged, healthy adult thrips (designated as the F 0 generation) were transferred to those treated leaves, with one individual per leaf. A control group was established following the same protocol using leaves treated only with the 0.1% Triton X-100 solution. After 48 hours of exposure, the surviving F 0 adults were transferred to fresh, untreated cabbage leaves in separate container and maintained until their natural death, respectively. The longevity of these F 0 adults was recorded daily. As eggs of T. tabaci laid within cabbage leaves could not be directly observed, the number of hatched nymphs was used as a proxy for estimating fecundity(Gu et al., 2023 ; X. Yang, Sun, Chi, Kang, & Zheng, 2020 ). Nymphs were counted only after confirming that no further eclosion occurred on the leaves. To obtain the F 1 generation, newly hatched nymphs produced by the LC 20 -treated F 0 adults were transferred to separate petri dish containing a fresh cabbage leaf without EB exposure, respectively. Approximately 60 nymphs were set up and leaves were replaced daily to ensure a sufficient food supply. The developmental stage, survival, and mortality were recorded daily. Upon pupation and subsequent adult emergence, daily fecundity of F 1 generation thrips was recorded until death. The F 2 generation was obtained from eggs laid by the F 1 adults, and the same observational procedures as for the F 1 generation were repeated to record their biological data. The life table parameters of F 1 and F 2 generation thrips in the control group was obtained according to the same procedure as in LC 20 -treated group. RNA extraction and transcriptome sequencing Newly emerged adult thrips were fed for 48 hours on cabbage leaf discs treated with the LC 20 concentration of EB as described above. The surviving individuals were collected, immediately frozen in liquid nitrogen, and stored at -80℃ until RNA extraction. Three biological replicates with approximately 40 individuals in each repeat were included for the LC 20 treated group and 0.1% Triton X-100 solution treated group, respectively. Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions, and the integrity was verified through 1% agarose gel electrophoresis. RNA concentration and purity were assessed using a NanoDrop spectrophotometer. Only high-quality RNA samples were used for subsequent library construction and transcriptome sequencing. Specifically, mRNA was enriched from extracted RNA using oligo(dT) beads for library construction, and then sequenced on an Illumina NovaSeq 6000 platform to generate paired-end reads. Raw reads were quality-controlled and aligned to the T. tabaci reference genome (Y. Gao et al., 2024 )(). Gene expression levels were quantified using RSEM and expressed as FPKM(Smith-Unna, Boursnell, Patro, Hibberd, & Kelly, 2016 ). Differentially expressed genes (DEGs) were identified with a threshold of |log 2 FC| ≥ 1 and an adjusted p-value < 0.05. Functional enrichment analysis of KEGG pathways was performed, with terms having an adjusted p-value ≤ 0.05 considered as significant criterion. DNA extraction and 16S rRNA sequencing Sample collection was performed as described in section 2.4. For each treatment, 30 surface-sterilized adult T. tabaci were pooled as one biological replicate, with five replicates per group. Total genomic DNA from the thrips microbiota was extracted and purified using the TIANGEN DNA Purification Kit (Tiangen Biotech, Beijing, China). following the manufacturer’s protocol. The quality of the extracted DNA was checked via 1% agarose gel electrophoresis, and the NanoDrop 2000C spectrophotometer was used to measure the concentration and purity of obtained DNA. The full-length 16S rRNA gene was amplified from genomic DNA using barcoded primers 27F (5’-AGRGTTYGATYMTGGCTCAG-3’) and 1492R (5’-RGYTACCTTGTTACGACTT-3’). After library preparation with the SMRTbell prep kit 3.0, sequencing was performed on the Pacbio Sequel IIe platform to generate HiFi reads. The resulting circular consensus sequencing (CCS) reads were processed using the QIIME2(Bolyen et al., 2019 ) pipeline with the DADA2(Callahan et al., 2016 ) plugin for denoising and Amplicon Sequence Variant (ASV) calling. Sequences annotated as chloroplasts or mitochondria were removed, and all samples were rarefied to 6,000 sequences per sample for downstream analysis. Taxonomic assignment of ASVs was performed using a consensus approach. Alpha diversity (Chao1, Ace, Shannon, Simpson indices) and beta diversity (Principal Component Analysis based on Bray-Curtis distance) were calculated to assess microbial community structure. Statistical analysis Adult mortality data of T. tabaci were compiled in Microsoft Excel 2019. Toxicity parameters (LC 20 , LC 50 , LC 95 , etc.) were derived by probit analysis using IBM SPSS Statistics 27. The age-stage, two-sex life table data for the F 0 –F 2 generations were analyzed with the TWOSEX-MSChart program, applying 100,000 bootstrap replications for estimating life table parameters under LC 20 stress. Data visualization was performed with GraphPad Prism 8. Statistical significance ( P < 0.05) was assessed by an independent samples t-test, and qRT-PCR data were normalized via the 2 –ΔΔCt method(Livak & Schmittgen, 2001 ). Results Sublethal effects of EB on F 0 of T. tabaci The indoor toxicity of EB against T. tabaci was evaluated using the leaf-dipping method (Table 1 ), and the LC 50 , LC 20 , and LC 95 concentration of EB for T. tabaci was determined as 9.631 mg/L, 4.014 mg/L, and 53.259 mg/L, respectively. The corresponding 95% confidence intervals were 6.839–13.053 mg/L, 2.191–5.811 mg/L, and 33.704-119.101 mg/L. The correlation coefficients (R²) for the toxicity regression lines all exceeded 0.993, indicating a high reliability of the bioassay in this study. Based on these results, the LC 20 concentration was selected as the sublethal dose for subsequent experiments to investigate the effects of EB on the growth, development, and reproductive capacity of T. tabaci across three successive generations. Table 1 Toxicity of emamectin benzoate to T. tabaci Insecticide n Slope ± SE* LC20(mg/L) (95%CL) † LC50(mg/L) (95%CL) † LC95(mg/L) (95%CL) † R2 Emamectin benzoate 450 2.215 ± 0.36 4.014 (2.191–5.811) 9.631 (6.839–13.053) 53.259 (33.704-119.101) 0.993 *, Standard error. †, 95% confidence interval. Exposure to the sublethal concentration (LC 20 ) of EB significantly impaired the reproduction and development of the parental generation (F 0 ) of T. tabaci (Table 2 ). Compared to the control group, the treated adults exhibited a substantial reduction in adult longevity (from 19.73 d to 8.57 d), total fecundity (from 94.99 eggs to 17.05 eggs), and oviposition period (from 18.90 d to 7.91 d). Table 2 Effects of emamectin benzoate on F 0 generation of T. tabaci at LC 20 concentration Parameters Control Emamectin benzoate Adult Longevity (days) 19.45 ± 0.94a 8.57 ± 0.29b Fecundity (eggs/female) 94.99 ± 5.25a 17.05 ± 0.74b Oviposition period (days) 18.90 ± 1.01a 7.91 ± 0.32b Gross Reproductive Rate GRR (offspring per individual) 106.25 ± 5.16a 19.17 ± 0.72b Net reproductive rate R 0 (offspring per individual) 94.99 ± 5.25a 17.04 ± 0.86b Intrinsic rate of increase r (days − 1 ) 0.2042 ± 1.79a 0.1472 ± 2.47b Finite rate of increase λ (days − 1 ) 1.2226 ± 2.19a 1.1585 ± 2.86b Mean generation time T (days) 22.30 ± 0.19a 19.26 ± 5.61b Note: Values are presented as mean ± SE. Means within a row followed by different lowercase letters are significantly different between two groups (independent samples t -test, p < 0.05). The sublethal concentration (LC 20 ) of EB also significantly influenced the population dynamics of the parental (F 0 ) generation of T. tabaci (Table 2 ). Compared to the control, the LC 20 EB treated T. tabaci exhibited a sharp decline in both the gross reproductive rate (GRR) and the net reproductive rate (R 0 ), which were significantly reduced by 82.0% (from 106.25 to 19.17) and 82.1% (from 94.99 to 17.04), respectively. Furthermore, the intrinsic rate of increase (r) decreased from 0.2042 d⁻¹ to 0.1472 d⁻¹, and the finite rate of increase (λ) dropped from 1.2226 d⁻¹ to 1.1585 d⁻¹ in the LC 20 EB treated population, indicating a severely constrained expansion potential. Concurrently, the mean generation time (T) was significantly shortened by approximately 3.04 days, suggesting an accelerated life cycle under insecticide stress, which occurred at the expense of reproductive fitness. The age-specific survival rate (l x ) of the EB-treated group began to decline on day 21, obviously earlier than that of the control group (Fig. 1 A). The age-specific fecundity (m x ) curve for the F 0 generation under LC 20 EB exposure was consistently lower than that of the control and peaked at the 16th day with a value of 4.23, which was both significantly lower and earlier than the peak value of 6.55 at the 24th day in the control (Fig. 1 B). The age-stage-specific life expectancy (e xj ) results indicated that the life expectancy of T. tabaci was 28 days in the EB treated group, significantly shorter than 40 days compared to the control (Fig. 1 C), demonstrating an obvious adverse impact on the expected lifespan of T. tabaci due to stress of LC 20 EB. Furthermore, the age-stage-specific reproductive value (v xj ) peaked at 34.66 on day 18 in the control group, whereas the LC 20 EB-treated group reached a lower peak value of 12.22 on day 16 (Fig. 1 D). Transgenerational effects of sublethal EB on T. tabaci Multigenerational influence of sublethal EB stress on T. tabaci was also assessed through the life table parameters analysis (Table 3 ). For the 1st generation (F1) thrips, their total longevity was not significantly affected, however, several key developmental and reproductive parameters were markedly altered due to their mothers’ exposure to LC 20 EB. For example, the adult longevity was significantly shortened by 27.8% (from 19.41 to 14.02 days), and the oviposition period decreased by 26.9% (from 18.65 to 13.64 days). Concurrently, compared to the control, the egg stage duration was significantly reduced from 10.91 to 7.69 days. In contrast, the durations of the first instar nymphal and pupal stages were both significantly prolonged. More critically, fecundity was drastically reduced from 99.41 to 34.02 eggs per female, namely falling by 65.8%. Consequently, significant detrimental effects were also observed in population growth parameters: the gross reproductive rate (GRR) decreased from 114.43 to 38.10 (-66.7%), the net reproductive rate (R 0 ) fell from 99.41 to 34.02 (-65.8%), the intrinsic rate of increase (r) dropped from 0.2017 to 0.1223 d − 1 , and the finite rate of increase (λ) declined from 1.2223 to 1.1301 d − 1 . Besides, the mean generation time (T) was significantly extended from 22.80 to 28.83 days. Unexpectedly, the sublethal effects of EB on T. tabaci largely subsided in the F 2 generation. Only adult longevity was significantly affected which decreased from 19.50 to 16.67 days. Life table parameters related to developmental, reproductive, and population growth were approximate between LC 20 EB treated group and control group, indicating a recovery of T. tabaci from the transgenerational sublethal effects. Table 3 Life table parameters of F 1 -F 2 generation of T. tabaci after treatment with sublethal concentrations of emamectin benzoate in F 0 generation Parameters F1 generation F2 generation Control Emamectin benzoate Control Emamectin benzoate Egg stage (days) 10.91 ± 0.12a 7.69 ± 0.28b 7.58 ± 0.17a 7.40 ± 0.20a First instar nymph stage (days) 3.18 ± 0.23b 4.89 ± 4.78a 4.40 ± 0.19a 4.76 ± 0.09a Second instar nymph stage (days) 2.47 ± 0.17a 2.75 ± 6.52a 2.75 ± 0.12a 2.79 ± 7.12a Pupa stage (days) 1.71 ± 0.14b 3.14 ± 5.18a 2.15 ± 0.13a 2.00 ± 8.68a Adult Longevity (days) 19.41 ± 1.13a 14.02 ± 0.28b 19.50 ± 0.99a 16.67 ± 1.02b Longevity (days) 37.81 ± 1.27a 35.35 ± 1.23a 38.00 ± 0.94a 35.76 ± 0.95a Fecundity (eggs/female) 99.41 ± 7.18a 34.02 ± 0.71b 87.70 ± 6.27a 81.89 ± 3.54a Oviposition period (days) 18.65 ± 1.07a 13.64 ± 0.25b 18.40 ± 0.97a 16.06 ± 1.06a Gross Reproductive Rate GRR (offspring per individual) 114.43 ± 6.80a 38.10 ± 0.67b 108.65 ± 9.95a 89.72 ± 3.39a Net reproductive rate R0 (offspring per individual) 99.41 ± 7.18a 34.02 ± 0.71b 87.70 ± 6.29a 76.48 ± 5.56a Intrinsic rate of increase r (days -1 ) 0.2017 ± 3.45a 0.1223 ± 8.11b 0.1775 ± 3.68a 0.1751 ± 3.23a Finite rate of increase λ (days -1 ) 1.2223 ± 4.22a 1.1301 ± 9.16b 1.1942 ± 4.40a 1.1914 ± 3.84a Mean generation time T (days) 22.80 ± 0.35b 28.83 ± 0.19a 25.21 ± 0.57a 24.75 ± 0.26a Note: Values are presented as mean ± SE. Means within a row of F1 or F2 generation followed by different lowercase letters are significantly different between two groups (independent samples *t*-test, *p* < 0.05). Four age-specific life parameters of T. tabaci were affected in F 1 generation due to their mothers’ (F 0 ) exposure to LC 20 EB (Fig. 2 ). Although the F 1 generation exposed to LC 20 EB exhibited a later decline in age-specific survival rate (l x ) and maintained higher overall survival rates, it consistently exhibited reduced fecundity (m x ) curves relative to the control group over its entire life course. Specifically, the m x peaked 3.95 at the 23th day in the LC 20 EB treated group, which was substantially lower than 7.38 in the control on day 22 (Fig. 2 C). Besides, the age-stage-specific life expectancy (e xj ) curve showed the total longevity of the F 1 generation in the LC 20 treatment group was 37.70 d with no significant difference from that in the control of 37.81 d (Fig. 2 C). In contrast, the age-speciffc reproductive value (v xj ) peak in the LC 20 EB group was both markedly delayed, occurring on day 36 compared to day 20 in the control, and significantly reduced in magnitude, reaching only 22.34 d − 1 versus 33.15 d − 1 in the control (Fig. 2 D). In the F 2 generation, age-specific survival rate (l x ), the life expectancy (e xj ) and fecundity (m x ) curves did not differ significantly between the LC 20 treatment and the control groups with both exhibiting a similar gradual decline (Fig. 2 , E-G). However, a delay and reduction in the reproductive peak was observed: the v xj peak in the LC 20 treatment group occurred on day 23 with a value of 27.43, which was later and lower than that (31.93 on day 20) of the control (Fig. 2 H). Effects of sublethal EB on the gene expression of T. tabaci Transcriptomic analysis of six biological samples from T. tabaci —LC 20 EB-treated group and a control group—yielded a total of 38.7 Gb of clean data. Each sample produced no less than 6.14 Gb of clean data, with a Q30 base percentage of 95.99%, meeting the core quality control threshold of Q30 > 95%. These results confirm the high quality and reliability of the sequencing data for subsequent analyses. Principal component analysis (PCA) revealed a clear separation between the insecticide-treated and control groups (Fig. 3 A), indicating that LC 20 EB stress induced substantial gene response for T. tabaci . Correlation analysis of genes expression profiles among samples further demonstrated high intra-group reproducibility, with correlation coefficients all above 0.961 within groups, in contrast to lower inter-group correlations (Fig. 3 B). Based on these, the impact of sublethal EB on gene expression in T. tabaci was assessed furthermore, and a total of 1066 differentially expressed genes (DEGs) were, among which 421 were up-regulated and 645 were down-regulated (Fig. 3 C). KEGG pathway enrichment analysis revealed the overall trends with DEGs in T. tabaci under LC 20 EB stress. The most pronounced responses were observed in carbohydrate metabolism pathways, such as starch and sucrose metabolism and fructose and mannose metabolism, suggesting that sublethal EB may trigger a reprogramming of energy metabolism in T. tabaci to meet potential energy demands. Concurrently, pathways closely associated with stress response and cellular regulation, including the MAPK and ErbB signaling pathways, also showed moderate enrichment, indicating potential disruptions in neural transmission and cellular homeostasis. Furthermore, alterations in pathways such as regulation of the actin cytoskeleton and protein processing in the endoplasmic reticulum reflect the multifaceted potential effects of sublethal doses of EB on cellular structure and function. Together, these findings provide preliminary insights and directions for further validation toward a systematic understanding of the molecular adaptation mechanisms of T. tabaci under sublethal EB stress (Fig. 3 D). Influence of sublethal EB treatment on bacterial communities of T. tabaci Sequencing of 10 samples yielded a total of 343,160 single-end reads. Following quality control, denoising, and chimera removal, 126,701 high-quality sequences were retained, resulting in 501 amplicon sequence variants (ASVs) (Table S2). Alpha diversity analysis, as measured by the Shannon index, revealed a slight reduction in microbial diversity in the EB-treated group compared to the control; however, the difference was not statistically significant (P = 0.2963; adjusted P = 0.5925) (Fig. 4 A), indicating that LC 20 EB exposure did not substantially alter the overall microbiota diversity of T. tabaci . Principal coordinate analysis (PCoA) based on genus-level community composition showed a modest separation between the control and EB-treated groups (Fig. 4 B), suggesting a potential shift in microbial structure of T. tabaci in response to EB treatment. Nevertheless, this trend did not reach statistical significance, which may be attributable to the limited sample size or considerable inter-individual variation. At the genus level (Fig. 4 C), the core microbiota in control T. tabaci consisted of Serratia (25.98%), Acinetobacter (44.03%), Pantoea (23.24%), Rosenbergiella (4.96%), and Erwinia (0.22%). After LC 20 EB treatment, the relative abundances of Serratia (29.49%), Pantoea (39.47%), Rosenbergiella (9.24%), and Erwinia (1.29%) increased to varying degrees, with the most pronounced rise observed in Pantoea . In contrast, the abundance of Acinetobacter decreased to 17.26%. At the species level (Fig. 4 E), the relative abundance of Pantoea ananatis significantly increased from 22.68% in the control group to 39.46% in the EB-treated group, indicating its possible roles in enhancing tolerance of T. tabaci to EB stress. Similarly, species within the genus Serratia , including S. rubidaea and S. marcescens , increased by 1.21% and 4.47%, respectively, also suggesting their potential involvement in the host’s stress response. Conversely, the relative abundance of Acinetobacter species (e.g., A. lactucae and unclassified Acinetobacter ) decreased in the treated group, with A. lactucae showing the most dramatic decline—from 26.54% to 0.25%. This shift may contribute to altered gut microenvironment homeostasis in T. tabaci . Collectively, these findings indicate that sublethal EB stress significantly restructured the core microbiota of T. tabaci . KEGG functional prediction based on PICRUSt2 analysis of 16S rRNA gene sequences indicated that LC 20 EB treatment altered the functional potential of the microbial community in T. tabaci (Fig. 4 C). Compared to the control group, the treated group exhibited differences in the abundance of several Level 2 metabolic pathways. Pathways closely related to fundamental material and energy metabolism—such as carbohydrate metabolism, amino acid metabolism, and lipid metabolism—were generally downregulated. In contrast, pathways involved in xenobiotic biodegradation and metabolism, membrane transport, and signal transduction, which are associated with environmental stress response and specific physiological processes, were upregulated in the LC 20 EBtreated group. Identification of key detoxification genes involving the adaptation of T. tabaci to sublethal EB stress Given its marked upregulation (log 2 FC = 1.04, P < 3.6e − 17 ) in response to sublethal EB exposure, we hypothesized that CYP6K1 plays a critical role in the detoxification process and may contribute to tolerance in T. tabaci . To functionally validate this hypothesis, we employed RNA interference to knock down its expression. RNA interference experiments demonstrated that feeding adult T. tabaci with dsRNA for 48 hours via a double-membrane method significantly reduced the expression of the CYP6K1 gene by 51.32% compared to the control group (P < 0.05; Fig. 5 ). After a 48-hour feeding period with synthesized dsCYP6K1 , adult thrips were subjected to a bioassay using the sublethal concentration LC 20 emamectin benzoate to determine changes in insecticide susceptibility. The results showed that the mortality rate of RNAi-treated thrips following LC 20 emamectin benzoate exposure reached 78.19%, representing a significant increase of 58.88% compared to the control mortality of 19.31% (P < 0.001). This dramatic increase in susceptibility confirms that CYP6K1 is a key mediator of sublethal EB tolerance. Discussion In this study, LC 50 of EB to T. tabaci was identified as 9.631 mg/L (Table 1 ), suggesting the high toxicity to this disgusting pest. Similarly, a synthesis of considerable previous bioassay data confirms potent and broad-spectrum insecticidal activity of EB. Against major lepidopteran pests, EB exhibits high toxicity, with reported LC 50 values of 0.019 mg/L for Spodoptera frugiperda (Fiaboe, Fening, Gbewonyo, & Deshmukh, 2023 ), 0.631 mg/L for S. litura (Devi, Mahajan, Saini, & Kaur, 2024 ), 0.464 mg/L for Helicoverpa zea (López, Latheef, & Hoffmann, 2010 ), 0.173 mg/L for Plutella xylostella (K. X. Liu, Guo, Zhang, & Xue, 2022 ), and 0.010 mg/L for S. littoralis (El-Saleh et al., 2025 ). Notably, the efficacy of EB extends beyond Lepidoptera to include diverse pests of fruit crops (e.g., Drosophila suzukii with the LC 50 of 0.021 mg/L(H. Gao et al., 2024 )), soybeans (e.g., Riptortus pedestris with the LC 50 of 7.681 mg/L(Guo et al., 2023 )), citrus (e.g., Panonychus citri with the LC 50 of 0.350 mg/L(Khan et al., 2021 )), and oil palm (e.g., Rhynchophorus ferrugineus with the LC 50 of 0.144 mg/L(Rezk et al., 2024 )), underscoring its wide applicability. Besides, comprehensive analysis of EB’s toxicity to several thrips species showed that the LC 50 of EB against Frankliniella occidentalis (7.997 mg/L) was determined to be comparable to our result, (Shen et al., 2023 ), however, its efficacy fluctuated with LC 50 from 27.731 to 34.536 mg/L when resisting against T. palmi from different populations, thereby deserves more attention in using this appropriately for precise thrips control. The suitability of an insecticide for integrated pest management (IPM) depends not only on its efficacy against target pests but also on its selectivity toward non-target organisms, particularly natural enemies. Available data indicate that emamectin benzoate exhibits relatively low toxicity to some certain predators such as Chrysoperla sinica (Shan et al., 2020 ), yet also poses a high risk to numerous beneficial arthropods with low LC 50 from 7.41(4.88–11.26) mg/L, including Cotesia marginiventris (Hou et al., 2024 ), honey bees (Abdu-Allah & Pittendrigh, 2018 ), Trichogramma japonicum Ashmead (Zhu et al., 2025 ) and poses a moderate threat to the rove beetle ( Paederus fuscipes (Khan, Nawaz, Hua, Cai, & Zhao, 2018 )). Consequently, a comprehensive evaluation of the safety and ecological risks of EB is also needed, including its effects on key thrips natural enemies such as predatory bugs, mites, and parasitoids, to guide scientifical usage of this pesticide. Sublethal dose insecticides causing by gradual degradation after field spraying usually produce unintended effects on target pest or environment. In this study, exposure to the sublethal concentration (LC 20 ) of EB significantly suppressed the F 0 generation of T. tabaci , especially resulting in reduced adult longevity and fecundity (Table 2 , Fig. 1 ). These findings align with previously documented sublethal effects in other insect species. For instance, sublethal doses of emamectin benzoate induced severe physiological and reproductive inhibition in Riptortus pedestris (Guo et al., 2023 ) and significantly decreased egg-laying in Mamestra brassicae (Moustafa, Kákai, Awad, & Fónagy, 2016 ).The sublethal effects also persisted into the F 1 generation of T. tabaci , where both developmental and reproductive parameters were significantly impaired compared to the control (Table 3 , Fig. 2 ). This transgenerational suppression is consistent with observations by Rezk et al.(Rezk et al., 2024 ), who also reported significant inhibiting of offspring population parameters following parental exposure to serial sublethal (LC 10 , LC 25 , LC 50 ) EB in Rhynchophorus ferrugineus . Similarly, sublethal concentrations of EB was found can induce long-term population suppression in Plutella xylostella , although a transient stimulation of oviposition may occur(K. X. Liu et al., 2022 ). Likewise, they significantly impair the development and reproduction of S. frugiperda , leading to comprehensive declines in population growth parameters such as the intrinsic rate of increase (C. Y. Chen, Tang, Zhao, Zhang, & Zhang, 2025 ). EB at sublethal doses also disrupt normal development and severely inhibit reproduction of Helicoverpa armigera , thereby substantially diminishing population growth potential through these "hidden" effects (Dong LiXia, Rui ChangHui, Ren Long, & Tan XiaoWei, 2011). However, a recent study by Shun-fan Wu et al (Gao et al., 2025 ) reported that lethal doses of EB accelerated ovarian development and increased mature egg load in female Nilaparvata lugens , potentially leading to pest resurgence. Collectively, these studies indicate that emamectin benzoate not noly exhibits high toxicity for effective controlling against a wide range of pests, but also can exert sustained suppression under residual sublethal concentrations, however it may also trigger population outbreaks in a few species. Furthermore, this study also revealed that LC 20 EB exposure also significantly altered the microbial community structure of T. tabaci , characterized primarily by a marked increase in the relative abundance of Pantoea and a sharp decline in Acinetobacter (Fig. 4 ). This finding offers new insights into the insecticidal mechanism of EB from a microbiological perspective. Previous research has established that Acinetobacter plays a critical role in insect responses to environmental stress; for example, it assists the Camellia weevil in adapting to its host plant by degrading tea saponins(Song, Shu, & Zhang, 2025 ), and Acinetobacter baumannii enhances the resistance of Nilaparvata lugens to the entomopathogenic fungus Metarhizium anisopliae (Tang et al., 2024 ). These findings suggest that Acinetobacter may aid insects in maintaining homeostasis and coping with external stressors through multiple mechanisms. Therefore, the observed decline of Acinetobacter following sublethal EB exposure likely compromised the physiological resilience of T. tabaci , increasing its susceptibility to the insecticide. Concurrently, Pantoea also known as a dominant bacterial genus in P. xylostella , has been implicated as a potential target for pest control, as reducing its density in P. xylostella effectively achieved population suppression of host (Li, Jin, Li, Cheng, & Jin, 2017 ). The microbial dysbiosis observed here, marked by the replacement of Acinetobacter by Pantoea as the dominant genus, may disrupt host gut function and, combined with the loss of Acinetobacter , collectively exacerbate the physiological stress imposed by EB on T. tabaci . Overall, this study provides a new perspective on the interaction between emamectin benzoate and the microbiota of T. tabaci , although the precise causal mechanisms, including the potential role of Acinetobacter in direct insecticide detoxification, require further validation. RNA interference experiments provided direct evidence elucidating the central role of CYP6K1 in the response of T. tabaci to emamectin benzoate stress. Silencing this gene resulted in a significant increase in thrips mortality under LC 20 EB exposure, confirming that CYP6K1 is one of the key determinants mediating tolerance of T. tabaci to EB. More importantly, this study is the first to reveal that the stress effects of emamectin benzoate on T. tabaci may operate simultaneously across two dimensions: host genes and symbiotic microbiota. Our results indicate that under emamectin benzoate stress, T. tabaci not only upregulates CYP6K1 gene expression in an attempt to metabolize and detoxify the insecticide but also undergoes drastic changes in gut microbiota structure, particularly a significant reduction in the abundance of Acinetobacter , which may contribute to stress resistance. Based on these findings, we hypothesize that in response to sublethal doses of emamectin benzoate, T. tabaci activates a coordinated defense network involving both host P450 detoxification genes and specific intestinal symbiotic bacteria. Disruption of any key component in this defense system, whether a host gene or a member of the symbiotic microbiota, may lead to the collapse of the thrips’ tolerance and result in higher mortality. Conclusion In summary, this study delineates the response of T. tabaci to emamectin benzoate stress at both the transcriptional and microbiome levels, and functionally validates CYP6K1 as one of core tolerance gene. Future studies should further decipher the interactions between host detoxification mechanisms and specific gut microbiota functions. Specially, a key focus would be to determine whether Acinetobacter can directly degrade emamectin benzoate or indirectly enhances tolerance by modulating host immunity. Such investigations are crucial for building a theoretical foundation for developing integrated control strategies against Thrips tabaci . Declarations Ethical approval We declare that all applicable national and provincial guidelines for the care and use of animals were followed. Competing interests The authors declare no competing interests. Funding This research was funded by the Central Public-interest Scientific Institution Basal Research Fund (No.1610162023010), National Key R&D Program of China (Grant No. 2022YFD1400300), Biological Breeding-Major Projects (2023ZD04062), the China Agriculture Research System, and Agricultural Science and Technology Innovation Program (ASTIP) (CAAS-ZDRW202412). Author Contribution Jichao Ji, Junyu Luo, and Jinjie Cui conceived and supervised the project. Miaomiao Xin and Chunjie Xian: Data curation, Formal analysis, Writing–original draft, Writing – review & editing. Xiaoyun Wang: Data curation, Formal analysis. Miaomiao Xin, Li Wang, Xiangzhen Zhu performed the experiments; Kaixin Zhang, Dongyang Li reviewed the original draft; Jichao Ji reviewed, edited and polished the manuscript. All authors read and approved the final manuscript. Acknowledgements We thank the editors and reviewers for their constructive comments on our work. The studies were conducted in the laboratories of the Institute of Cotton Research, Chinese Academy of Agricultural Sciences. Availability of data and materials All data and materials needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary material. Additional data related to this paper may be requested from the authors. References Abbas, A., Zhao, C. R., Arshad, M., Han, X., Iftikhar, A., Hafeez, F., . . . Ullah, F. (2023). Sublethal effects of spinetoram and emamectin benzoate on key demographic parameters of fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae) under laboratory conditions. Environ Sci Pollut Res Int, 30 (34), 82990-83003. http://doi:10.1007/s11356-023-28183-8 Abdel-Baky, N. F., Alhewairini, S. S., & Bakry, M. M. S. (2019). EMAMECTIN-BENZOATE AGAINST Tuta absoluta MEYRICK AND Spodoptera littoralis BOISDUVAL LARVAE. Pakistan Journal of Agricultural Sciences, 56 (3), 801-808. http://doi:10.21162/pakjas/19.8082 Abdu-Allah, G. A. M., & Pittendrigh, B. R. (2018). Lethal and sub-lethal effects of select macrocyclic lactones insecticides on forager worker honey bees under laboratory experimental conditions. Ecotoxicology, 27 (1), 81-88. http://doi:10.1007/s10646-017-1872-6 Afza, R., Afzal, A., Riaz, M. A., Majeed, M. Z., Idrees, A., Qadir, Z. A., . . . Li, J. (2023). Sublethal and transgenerational effects of synthetic insecticides on the biological parameters and functional response of Coccinella septempunctata (Coleoptera: Coccinellidae) under laboratory conditions. Frontiers in Physiology, 14 , 1088712-1088725. http://doi:10.3389/fphys.2023.1088712 Bolyen, E., Rideout, J. R., Dillon, M. R., Bokulich, N., Abnet, C. C., Al-Ghalith, G. A., . . . Caporaso, J. G. (2019). Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nature Biotechnology, 37 (8), 852-857. http://doi:10.1038/s41587-019-0209-9 Callahan, B. J., McMurdie, P. J., Rosen, M. J., Han, A. W., Johnson, A. J. A., & Holmes, S. P. (2016). DADA2: High-resolution sample inference from Illumina amplicon data. Nature Methods, 13 (7), 581-583. http://doi:10.1038/nmeth.3869 Chen, C. Y., Tang, Y. T., Zhao, Y. X., Zhang, X. F., & Zhang, K. (2025). Life table study of sublethal concentrations of emamectin benzoate against Spodoptera frugiperda (Lepidoptera, Noctuidae). Journal of Insect Science, 25 (1), 7. http://doi:10.1093/jisesa/ieaf014 Chen, Y. X., Tian, H. J., Lin, S., Yu, Y., Xie, L. C., Li, H., . . . Wei, H. (2023). Sublethal effects of emamectin benzoate on development, reproduction, and vitellogenin and vitellogenin receptor gene expression in Thrips hawaiiensis (Thysanoptera: Thripidae). J Insect Sci, 23 (3). http://doi:10.1093/jisesa/iead035 Devi, M., Mahajan, A., Saini, H. S., & Kaur, S. (2024). The impact of lethal and sub-lethal exposure of emamectin benzoate on populations of Spodoptera litura (Lepidoptera: Noctuidae) under laboratory conditions. Toxicon, 250 , 108121-108129. http://doi:10.1016/j.toxicon.2024.108121 Dong LiXia, D. L., Rui ChangHui, R. C., Ren Long, R. L., & Tan XiaoWei, T. X. (2011). Effect of sublethal dose of emamectin benzoate on growth and development of Helicoverpa armigera (Hubner). Acta Phytophylacica Sinica, 38 (6), 539-544. Retrieved from ://CABI:20123048930 http://www.wanfangdata.com.cn El-Saleh, M. A., Aioub, A. A., El-Sheikh, E. A., Desuky, W. M. H., Alkeridis, L. A., Al-Shuraym, L. A., . . . Hamed, I. A. (2025). Comparative Toxicological Effects of Insecticides and Their Mixtures on Spodoptera littoralis (Lepidoptera: Noctuidae). Insects, 16 (8). http://doi:10.3390/insects16080821 Fiaboe, K. R., Fening, K. O., Gbewonyo, W. S. K., & Deshmukh, S. (2023). Bionomic responses of Spodoptera frugiperda (J. E. Smith) to lethal and sublethal concentrations of selected insecticides. PLoS One, 18 (11), e0290390-e0290410. http://doi:10.1371/journal.pone.0290390 Gao, H., Wang, Y., Chen, P., Zhang, A., Zhou, X., & Zhuang, Q. (2024). Toxicity of Eight Insecticides on Drosophila suzukii and Its Pupal Parasitoid Trichopria drosophilae. Insects, 15 (11), 910-924. http://doi:10.3390/insects15110910 Gao, Y., Ji, J., Xu, C., Wang, L., Zhang, K., Li, D., . . . Luo, J. (2024). Chromosome-level genome assembly of cotton thrips Thrips tabaci (Thysanoptera: Thripidae). Sci Data, 11 (1), 1003-1015. http://doi:10.1038/s41597-024-03737-8 Gao, Y., Su, S. C., Xing, J. Y., Liu, Z. Y., Nässel, D. R., Bass, C., . . . Wu, S. F. (2025). Pesticide-induced resurgence in brown planthoppers is mediated by action on a suite of genes that promote juvenile hormone biosynthesis and female fecundity. Elife, 12 . http://doi:10.7554/eLife.91774 Gu, Z., Zhang, T., Long, S., Li, S., Wang, C., Chen, Q., . . . Cao, Y. (2023). Responses of Thrips hawaiiensis and Thrips flavus populations to elevated CO2 concentrations. Journal of Economic Entomology, 116 (2), 416-425. http://doi:10.1093/jee/toad026 Guo, J., An, J., Chang, H., Li, Y., Dang, Z., Wu, C., & Gao, Z. (2023). The Lethal and Sublethal Effects of Lambda-Cyhalothrin and Emamectin Benzoate on the Soybean Pest Riptortus pedestris (Fabricius). Toxics, 11 (12), 971-988. http://doi:10.3390/toxics11120971 Hou, Y. Y., Zang, Z. Y., Lü, W. J., Xu, W., Desneux, N., & Zang, L. S. (2024). Transgenerational hormesis and sublethal effects of five key insecticides for controlling Spodoptera frugiperda on its endoparasitoid Cotesia marginiventris. Pest Manag Sci, 80 (4), 1681-1691. http://doi:10.1002/ps.7899 Khan, M. M., Ali, M. W., Hafeez, M., Fan, Z. Y., Ali, S., & Qiu, B. L. (2021). Lethal and sublethal effects of emamectin benzoate on life-table and physiological parameters of citrus red mite, Panonychus citri. Exp Appl Acarol, 85 (2-4), 173-190. http://doi:10.1007/s10493-021-00667-7 Khan, M. M., Nawaz, M., Hua, H., Cai, W., & Zhao, J. (2018). Lethal and sublethal effects of emamectin benzoate on the rove beetle, Paederus fuscipes, a non-target predator of rice brown planthopper, Nilaparvata lugens. Ecotoxicol Environ Saf, 165 , 19-24. http://doi:10.1016/j.ecoenv.2018.08.047 Li, W. H., Jin, D. C., Li, F. L., Cheng, Y., & Jin, J. X. (2017). Metabolic phenomics of bacterium Pantoea sp. from larval gut of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae). Symbiosis, 72 (2), 135-142. http://doi:10.1007/s13199-016-0453-4 Liu, K. X., Guo, Y., Zhang, C. X., & Xue, C. B. (2022). Sublethal effects and reproductive hormesis of emamectin benzoate on Plutella xylostella. Front Physiol, 13 , 1025959. http://doi:10.3389/fphys.2022.1025959 Liu, Z., Lyu, B., Lu, H., Tang, J. H., Zhang, Q. K., & Jiao, B. (2025). The toxicological mechanism of emamectin benzoate in Spodoptera frugiperda via regulating gut microbiota. Entomologia Generalis, 45 (1), 253-263. http://doi:10.1127/entomologia/2024/2913 Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25 (4), 402-408. http://doi:10.1006/meth.2001.1262 López, J. D., Jr., Latheef, M. A., & Hoffmann, W. C. (2010). Effect of emamectin benzoate on mortality, proboscis extension, gustation and reproduction of the corn earworm, Helicoverpa zea. J Insect Sci, 10 (89), 1-16. http://doi:10.1673/031.010.8901 Mougabure-Cueto, G., Fronza, G., & Nattero, J. (2024). What happens when the insecticide does not kill? A review of sublethal toxicology and insecticide resistance in triatomines. Medical and Veterinary Entomology . http://doi:10.1111/mve.12753 Moustafa, M. A. M., Kákai, A., Awad, M., & Fónagy, A. (2016). Sublethal effects of spinosad and emamectin benzoate on larval development and reproductive activities of the cabbage moth, Mamestra brassicae L. (Lepidoptera: Noctuidae). Crop Protection, 90 , 197-204. http://doi:10.1016/j.cropro.2016.09.004 Naeem-Ullah, U., Ramzan, M., Bokhari, S. H. M., Saleem, A., Qayyum, M. A., Iqbal, N., . . . Saeed, S. (2020). Insect Pests of Cotton Crop and Management Under Climate Change Scenarios. In Environment, Climate, Plant and Vegetation Growth (pp. 367-396). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-030-49732-3_15 Reitz, S. R., Gao, Y., Kirk, W. D. J., Hoddle, M. S., Leiss, K. A., & Funderburk, J. E. (2020). Invasion Biology, Ecology, and Management of Western Flower Thrips. Annu Rev Entomol, 65 , 17-37. http://doi:10.1146/annurev-ento-011019-024947 Rezk, A. A., Naqqash, M. N., Sattar, M. N., Mehmood, K., Elshafie, H., & Al-Khayri, J. M. (2024). Sublethal effect of emamectin benzoate on age-stage, two-sex life table and population projection of red palm weevil, Rhynchophorus ferrugineus. Sci Rep, 14 (1), 22565. http://doi:10.1038/s41598-024-70042-0 Shan, Y. X., Zhu, Y., Li, J. J., Wang, N. M., Yu, Q. T., & Xue, C. B. (2020). Acute lethal and sublethal effects of four insecticides on the lacewing (Chrysoperla sinica Tjeder). Chemosphere, 250 , 126321. http://doi:10.1016/j.chemosphere.2020.126321 Shen, X. J., Chen, J. C., Cao, L. J., Ma, Z. Z., Sun, L. N., Gao, Y. F., . . . Wei, S. J. (2023). Interspecific and intraspecific variation in susceptibility of two co-occurring pest thrips, Frankliniella occidentalis and Thrips palmi, to nine insecticides. Pest Manag Sci, 79 (9), 3218-3226. http://doi:10.1002/ps.7502 Smith-Unna, R., Boursnell, C., Patro, R., Hibberd, J. M., & Kelly, S. (2016). TransRate: reference-free quality assessment of de novo transcriptome assemblies. Genome Research, 26 (8), 1134-1144. http://doi:10.1101/gr.196469.115 Song, F., Shu, J. P., & Zhang, S. K. (2025). Gut bacterium Acinetobacter sp. assists Camellia weevil with host plant adaptation by degrading tea saponin via the benzoate pathway. Microbiome, 13 (1), 139-165. http://doi:10.1186/s40168-025-02131-9 Sun, Y., Hu, C., Chen, G., Li, X., Liu, J., Xu, Z., . . . Zhang, X. (2023). Insecticide-mediated changes in the population and toxicity of the thrips species, Frankliniella occidentalis (Pergande) and Thrips flavus (Schrank) (Thysanoptera: Thripidae). Journal of Economic Entomology, 117 (1), 293-301. http://doi:10.1093/jee/toad226 Tang, C., Hu, X., Tang, J. F., Wang, L., Liu, X. W., Peng, Y. F., . . . Xie, J. Q. (2024). The symbiont Acinetobacter baumannii enhances the insect host resistance to entomopathogenic fungus Metarhizium anisopliae. Communications Biology, 7 (1), 1184. http://doi:10.1038/s42003-024-06779-1 Wakil, W., Gulzar, S., Wu, S., Rasool, K. G., Husain, M., Aldawood, A. S., & Toews, M. D. (2023). Development of Insecticide Resistance in Field Populations of Onion Thrips, Thrips tabaci (Thysanoptera: Thripidae). Insects, 14 (4), 376. http://doi:10.3390/insects14040376 Yang, D. S., Cui, B., Wang, C. X., Zhao, X., Zeng, Z. H., Wang, Y., . . . Cui, H. X. (2017). Preparation and Characterization of Emamectin Benzoate Solid Nanodispersion. Journal of Nanomaterials, 15 (7), 495-511. http://doi:10.1155/2017/6560780 Yang, X., Sun, L., Chi, H., Kang, G., & Zheng, C. (2020). Demography of Thrips palmi (Thysanoptera: Thripidae) Reared on Brassica oleracea (Brassicales: Brassicaceae) and Phaseolus vulgaris (Fabales: Fabaceae) With Discussion on the Application of the Bootstrap Technique in Life Table Research. J Econ Entomol, 113 (5), 2390-2398. http://doi:10.1093/jee/toaa171 Zhou, L., Luo, F. J., Zhang, X. Z., Jiang, Y. P., Lou, Z. Y., & Chen, Z. M. (2016). Dissipation, transfer and safety evaluation of emamectin benzoate in tea. Food Chemistry, 202 , 199-204. http://doi:10.1016/j.foodchem.2015.11.069 Zhu, W., Guo, Q., Chen, M., Wang, J., Zhang, Y., & Ma, R. (2025). Comprehensive assessment of the acute lethal, risk level, and sub-lethal effects of four insecticides on Trichogramma ostriniae. PLoS One, 20 (6), e0325733. http://doi:10.1371/journal.pone.0325733 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 06 Feb, 2026 Reviews received at journal 30 Jan, 2026 Reviews received at journal 28 Jan, 2026 Reviews received at journal 22 Jan, 2026 Reviews received at journal 16 Jan, 2026 Reviewers agreed at journal 13 Jan, 2026 Reviewers agreed at journal 13 Jan, 2026 Reviewers agreed at journal 12 Jan, 2026 Reviewers agreed at journal 12 Jan, 2026 Reviewers invited by journal 12 Jan, 2026 Editor assigned by journal 03 Dec, 2025 Submission checks completed at journal 03 Dec, 2025 First submitted to journal 02 Dec, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8257750","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":573916283,"identity":"f18c2303-6522-497d-8b87-a0c69da8dd04","order_by":0,"name":"Chunjie Xian","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Chunjie","middleName":"","lastName":"Xian","suffix":""},{"id":573916284,"identity":"cf700eff-772d-46a9-84f1-6d56f0765ff5","order_by":1,"name":"Miaomiao Xin","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Miaomiao","middleName":"","lastName":"Xin","suffix":""},{"id":573916285,"identity":"b540a866-8532-4f14-a6f3-21be28e340ca","order_by":2,"name":"xiaoyun wang","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"xiaoyun","middleName":"","lastName":"wang","suffix":""},{"id":573916290,"identity":"8c52f049-8cc6-4f28-9349-dc37d2d91ac2","order_by":3,"name":"Xiangzhen Zhu","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Xiangzhen","middleName":"","lastName":"Zhu","suffix":""},{"id":573916293,"identity":"faa6afe1-ca27-4db2-97e8-47086f484dc1","order_by":4,"name":"Li Wang","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Li","middleName":"","lastName":"Wang","suffix":""},{"id":573916303,"identity":"8a97f0fc-4ebb-48ec-8b52-cfb8017c4200","order_by":5,"name":"Dongyang Li","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Dongyang","middleName":"","lastName":"Li","suffix":""},{"id":573916304,"identity":"950c10b0-643c-4752-aca7-0d2481aa8892","order_by":6,"name":"Kaixin Zhang","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Kaixin","middleName":"","lastName":"Zhang","suffix":""},{"id":573916305,"identity":"b437268d-3764-440c-ac28-78990b6dabe0","order_by":7,"name":"Xueke Gao","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Xueke","middleName":"","lastName":"Gao","suffix":""},{"id":573916309,"identity":"809cc591-9722-4d0e-9b7c-8f416857fad9","order_by":8,"name":"Jichao Ji","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAtUlEQVRIiWNgGAWjYBACAxDBw8DGwM/MfPgBaVok29nSDEjRAmSc51GQIEqLuXTvMYm3bXzyxod5gPprbKIJarGccy5Ncs4ZNsNth3kPPGA4lpbbQNBhN3LMpHkq2BLMDvMlGDA2HCZWiwFbgnEzj4EECVqAthgwk6DF2BLklxmHgYGcQKRfDG+8bTsmz99/+PCDDzU2hLUAAQswOo5BmAlEKAcB5g8MDDVEqh0Fo2AUjIIRCQAgEDoorJLalgAAAABJRU5ErkJggg==","orcid":"","institution":"","correspondingAuthor":true,"prefix":"","firstName":"Jichao","middleName":"","lastName":"Ji","suffix":""},{"id":573916315,"identity":"ebc5894e-7086-4526-a88d-f5a93c91a77d","order_by":9,"name":"Junyu Luo","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Junyu","middleName":"","lastName":"Luo","suffix":""},{"id":573916316,"identity":"abb24e28-48d1-45c0-adf3-aa45b1584f83","order_by":10,"name":"Jinjie Cui","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Jinjie","middleName":"","lastName":"Cui","suffix":""}],"badges":[],"createdAt":"2025-12-02 08:23:08","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8257750/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8257750/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":100259936,"identity":"d7e5e20d-a602-42b3-bf16-8137592a7679","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1091628,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.docx","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/6b68a9ae2694e39a76f0fda5.docx"},{"id":100370899,"identity":"25db4289-25a2-4169-9a24-a760d52ec70a","added_by":"auto","created_at":"2026-01-16 08:08:57","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":10702,"visible":true,"origin":"","legend":"","description":"","filename":"4472938ae94245a99aae29a328d0e635.json","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/f40469ae51a0e71302fc943f.json"},{"id":100259941,"identity":"5c6a0a15-3a0c-463c-a38a-4e76da19d858","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"xml","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":155088,"visible":true,"origin":"","legend":"","description":"","filename":"4472938ae94245a99aae29a328d0e6351enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/03d39328c6513ea4307814e5.xml"},{"id":100259933,"identity":"c1330a82-cbc1-43c6-8afa-225215747cf0","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"jpeg","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":209361,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/92ba520f9b53d9f3863da34e.jpeg"},{"id":100372451,"identity":"7bf64346-8555-4dbf-8591-fc7f865b5cdf","added_by":"auto","created_at":"2026-01-16 08:12:26","extension":"jpeg","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":351339,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/ee1732e869f34b388af03afd.jpeg"},{"id":100371750,"identity":"1f70169c-8a07-42c4-944c-0d3de31ce848","added_by":"auto","created_at":"2026-01-16 08:10:48","extension":"jpeg","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":235613,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/6da080817e337baedeb79421.jpeg"},{"id":100372409,"identity":"1180d9d6-5115-484c-b442-8c51f533b5d6","added_by":"auto","created_at":"2026-01-16 08:12:16","extension":"jpeg","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":238500,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/a604ad73af40fb904a8c571a.jpeg"},{"id":100371882,"identity":"41ffd4e9-c132-4192-9479-99adda5f7707","added_by":"auto","created_at":"2026-01-16 08:11:08","extension":"jpeg","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":63634,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/fc780d21342ab82235d14060.jpeg"},{"id":100371577,"identity":"1870e5d2-e8e5-4111-a8d4-81086f474a10","added_by":"auto","created_at":"2026-01-16 08:10:34","extension":"png","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":77327,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/03db17740df75577f7fa2d1b.png"},{"id":100372484,"identity":"1ce56f5d-c559-4b39-9589-e24dc137ebbc","added_by":"auto","created_at":"2026-01-16 08:12:29","extension":"png","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":123979,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/95215eee39ea1d0e46647c71.png"},{"id":100372275,"identity":"45a23aa4-0d61-4c1b-85b7-fcaa0bfff3ae","added_by":"auto","created_at":"2026-01-16 08:11:54","extension":"png","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":59818,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/d25aa8b4d5fc932e4c76f265.png"},{"id":100259942,"identity":"26848a01-516f-43d7-a614-0906ea1eb404","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"png","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":61756,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/5d96d86faa7a7ba149ab08ef.png"},{"id":100259944,"identity":"5c1edd42-c915-4152-8d72-ed69c986be9c","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"png","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":14531,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/2e8b4f0c64717fe59fda9bb9.png"},{"id":100259946,"identity":"a9647d3b-35be-42fa-bf58-25ff0b905a23","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"xml","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":153319,"visible":true,"origin":"","legend":"","description":"","filename":"4472938ae94245a99aae29a328d0e6351structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/8a50f5d4b3e3e6856ec4dd7d.xml"},{"id":100259947,"identity":"e48a587e-4ba2-4d92-8d91-0e666dfbbfc4","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"html","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":167112,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/ef28eb79221cd508fbd2fc4d.html"},{"id":100259929,"identity":"bda5f08a-57b4-47ac-939a-6859d8589af5","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":78078,"visible":true,"origin":"","legend":"\u003cp\u003eAge-specific survival rate (l\u003csub\u003ex\u003c/sub\u003e), age-specific fecundity (m\u003csub\u003ex\u003c/sub\u003e), age-specific life expectancy (e\u003csub\u003ex\u003c/sub\u003e), and reproductive values of specific age-stage (Vxj) of the parental \u003cem\u003eT. tabaci \u003c/em\u003e(F\u003csub\u003e0\u003c/sub\u003e generation) exposed to LC\u003csub\u003e20\u003c/sub\u003e EB.\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/8f5d3d0f59ec2cf4a65d89ab.jpg"},{"id":100371996,"identity":"841df0c3-c543-48a5-b377-598d8a2107de","added_by":"auto","created_at":"2026-01-16 08:11:20","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":85458,"visible":true,"origin":"","legend":"\u003cp\u003eAge-specific survival rate (l\u003csub\u003ex\u003c/sub\u003e), age-specific fecundity (m\u003csub\u003ex\u003c/sub\u003e), age-specific life expectancy (e\u003csub\u003ex\u003c/sub\u003e), and reproductive values of specific age-stage (V\u003csub\u003exj\u003c/sub\u003e) of the F\u003csub\u003e1\u003c/sub\u003e-F\u003csub\u003e2\u003c/sub\u003e \u003cem\u003eT. tabaci\u003c/em\u003e after F\u003csub\u003e0\u003c/sub\u003e generation\u003cem\u003e \u003c/em\u003eexposed to LC\u003csub\u003e20\u003c/sub\u003e EB.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/bf4eb50e74f90149b487646c.jpg"},{"id":100259931,"identity":"8198c5dc-6315-4c2e-9045-4587070f33bb","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":81924,"visible":true,"origin":"","legend":"\u003cp\u003eImpact of LC\u003csub\u003e20\u003c/sub\u003e EB exposure on the \u003cem\u003eT. tabaci\u003c/em\u003e at gene expression level. (A) Principal Component Analysis, PCA, (B) Inter-sample correlation analysis, (C) Volcano plot of DEGs, (D) KEGG pathway enrichment analysis of DEGs.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/55a9023a3246fa68b3de090a.jpg"},{"id":100259932,"identity":"a4c8eceb-ec22-4f50-b539-eb2a105953b4","added_by":"auto","created_at":"2026-01-14 16:50:40","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":86647,"visible":true,"origin":"","legend":"\u003cp\u003eMicrobiota analysis of \u003cem\u003eT. tabaci\u003c/em\u003e following exposure to LC\u003csub\u003e20\u003c/sub\u003e EB stress via 16S rRNA sequencing. (A) Shannon index of alpha diversity, (B) Principal coordinate analysis (PCoA) of microbial communities at the genus level, Microbiota composition of \u003cem\u003eT. tabaci\u003c/em\u003e at the genus (C) and species (D) level, (E) Predictive functional profiling of the microbiota.\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/78ece5b8e64cd7f64ad9ea8c.jpg"},{"id":100371893,"identity":"a97690d3-4e03-469d-97ae-33e9cc601399","added_by":"auto","created_at":"2026-01-16 08:11:09","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":24939,"visible":true,"origin":"","legend":"\u003cp\u003eA: Relative expression level of \u003cem\u003edsGFP\u003c/em\u003e and \u003cem\u003edsCYP6K1\u003c/em\u003e after RNAi; B: Mortality of \u003cem\u003eT. tabaci\u003c/em\u003eafter RNAi the expression of \u003cem\u003eCYP6K1\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eNote: Bar graphs represent mean ± standard error; asterisks indicate significance levels (**P \u0026lt; 0.05, ***P \u0026lt; 0.001).\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/3154ac854623ef5e6addab9a.jpg"},{"id":100383617,"identity":"3d46601f-da9e-48ec-be08-9654b30c6609","added_by":"auto","created_at":"2026-01-16 10:47:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1403169,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8257750/v1/e0cacab1-7a7a-44b5-968d-f87ae776eeb0.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Sublethal emamectin benzoate suppresses multi-transgenerational reproduction and alters symbiotic bacteria of Thrips tabaci","fulltext":[{"header":"Key Message","content":"\u003cul start=\"50\"\u003e\n \u003cli\u003eEmamectin benzoate shows dual toxicity across \u003cem\u003eT. tabaci\u003c/em\u003e generations.\u003c/li\u003e\n \u003cli\u003eThe P450 gene\u003cem\u003e\u0026nbsp;CYP6K1\u003c/em\u003e modulates insecticide susceptibility.\u003c/li\u003e\n \u003cli\u003eGut microbiome shifts accompany sublethal insecticide exposure.\u003c/li\u003e\n \u003cli\u003eHost‑symbiont co‑response underpins pesticide tolerance in thrips.\u003c/li\u003e\n \u003cli\u003eTargeting detox genes and microbiota may improve integrated pest control.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eThrips tabaci\u003c/em\u003e (Tysanoptera: Tripidae), well-known as cotton/onion thrips, is a polyphagous agricultural pest with extensive host range and distributed mostly in tropical, subtropical and temperate areas of the world. Combined with sucking sap from leaves especially young ones, flowers, and fruits, it can transmit viral plant diseases through leaf scraping, eventually causing substantial yield losses of various crop in the world every year(Naeem-Ullah et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Up to now, chemical control using insecticides remains the primary strategy for managing thrips infestations on crops (Reitz et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Sun et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).However, the long-term use of insecticides has led to the development of resistance to multiple chemical classes for thrips populations(Shen et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Wakil et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) .\u003c/p\u003e \u003cp\u003eEmamectin benzoate (EB), a semi-synthetic insecticide that belongs to a class of avermectin family, derived from the natural fermentation of a naturally occurring soil actinomycete \u003cem\u003eStreptomyces avermitilis\u003c/em\u003e, exhibits high bioactivity against major crop pests alongside a favorable environmental profile characterized by low toxicity and minimal residues(D. S. Yang et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). These properties underpin its widespread utilization as a key intervention for pest control in diverse agricultural systems, particularly in vegetable, fruit, and cotton production(Abbas et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Abdel-Baky, Alhewairini, \u0026amp; Bakry, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; El-Saleh et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). The toxicity of EB lies in the specific and irreversible disruption of unique neurological targets in insects, leading to neuromuscular paralysis. These characteristics account for its high insecticidal activity, low toxicity, and environmental safety, establishing its importance in integrated pest management strategies(Z. Liu et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Beyond acute lethality, the sublethal effects of EB constitute another dimension of its ecological impact. These effects arise from persistent field exposure to low concentrations, a common scenario resulting from insecticide degradation and the cryptic nature of pest feeding.\u003c/p\u003e \u003cp\u003eFollowing field application, insecticides undergo gradual degradation by environmental factors such as sunlight, rain and so on. While high doses can elicit strong lethal effects on target pests, prolonged exposure to sublethal concentrations may induce a range of sublethal effects(Mougabure-Cueto, Fronza, \u0026amp; Nattero, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Considerable studies have demonstrated that these effects\u0026mdash;such as extended developmental duration, reduced longevity, and diminished fecundity\u0026mdash;can paradoxically lead to population resurgence in pest species (Afza et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Y. X. Chen et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), and transgenerational impacts on offspring(H. Gao et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Guo et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).The minute body size and cryptic behavior of \u003cem\u003eThrips tabaci\u003c/em\u003e, which often conceals it within plant structures, significantly increase the likelihood of its exposure to sublethal doses of emamectin benzoate. Therefore, investigating the sublethal impacts of emamectin benzoate on \u003cem\u003eT. tabaci\u003c/em\u003e is critical, as it benefits for enhancing the insecticidal efficacy of this compound and optimizing integrated pest management strategies.\u003c/p\u003e \u003cp\u003eThis study focuses on exploring the sublethal effects of emamectin benzoate on \u003cem\u003eT. tabaci\u003c/em\u003e, and confirms its inhibitory impact on thrips reproduction and development for successive two generations eventually. Through an integrated approach incorporating bioassays, transcriptome sequencing, and 16S rRNA microbiome analysis, we systematically investigated this insecticide\u0026rsquo;s toxic action mechanisms and the corresponding detoxification metabolic responses of \u003cem\u003eT. tabaci\u003c/em\u003e. Taken together, these results are expected to provide a critical theoretical foundation for the rational field application of emamectin benzoate and for the development of sustainable resistance management strategies against \u003cem\u003eT. tabaci\u003c/em\u003e.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eInsects and insecticides\u003c/h2\u003e \u003cp\u003eA thelytokous population of \u003cem\u003eThrips tabaci\u003c/em\u003e was used in this study, originally provided by the Cotton Research Institute of the Chinese Academy of Agricultural Sciences. The \u003cem\u003eT. tabaci\u003c/em\u003e were reared on red cabbage (\u003cem\u003eBrassica oleracea var. capitata rubra\u003c/em\u003e) leaves placed in cages (29 cm × 19 cm × 13 cm) under the control condition of 25 ± 1°C, 50 ± 5% relative humidity, and a photoperiod of 16 h light : 8h dark. Emamectin benzoate (EB) insecticide (Pesticide Registration No.: PD20170668) was purchased from Shandong Feixiang Agricultural Development Co., Ltd., China.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eToxicity bioassays\u003c/h3\u003e\n\u003cp\u003eEB toxicity to \u003cem\u003eThrips tabaci\u003c/em\u003e was evaluated by leaf-dip bioassay(Y. X. Chen et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). A stock solution was prepared by dissolving an appropriate amount of EB in distilled water containing 0.01% Triton X-100 as a surfactant. This stock solution was then serially diluted with the 0.01% Triton X-100 aqueous solution to create a series of EB concentrations (50, 25, 12.5, 6.25, and 3.125 mg/L). A 0.01% Triton X-100 aqueous solution served as the control. Cabbage leaves discs (2 cm in diameter), prepared with a hole puncher, were immersed in the test solutions or control for 15 seconds. After air-drying, the treated leaf discs were placed in plastic containers. Twenty-five newly emerged, healthy adult thrips were placed into each container. All containers were then kept in the climate chamber under the same control condition as above. Mortality was assessed after 48 hours. Individuals that showed no movement when gently prodded with a fine brush were considered dead. The experiment was repeated 3 times for each concentration.\u003c/p\u003e \u003cp\u003e \u003cb\u003eSublethal effects of EB on\u003c/b\u003e \u003cb\u003eT. tabaci\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe sublethal effects were evaluated using the leaf-dipping method with the LC\u003csub\u003e20\u003c/sub\u003e concentration of EB, prepared by serial dilution of the stock solution with distilled water containing 0.1% (v/v) Triton X-100. Fresh cabbage leaves were immersed in the LC\u003csub\u003e20\u003c/sub\u003e solution for 15 seconds, air-dried. Then 60 newly emerged, healthy adult thrips (designated as the F\u003csub\u003e0\u003c/sub\u003e generation) were transferred to those treated leaves, with one individual per leaf. A control group was established following the same protocol using leaves treated only with the 0.1% Triton X-100 solution. After 48 hours of exposure, the surviving F\u003csub\u003e0\u003c/sub\u003e adults were transferred to fresh, untreated cabbage leaves in separate container and maintained until their natural death, respectively. The longevity of these F\u003csub\u003e0\u003c/sub\u003e adults was recorded daily. As eggs of \u003cem\u003eT. tabaci\u003c/em\u003e laid within cabbage leaves could not be directly observed, the number of hatched nymphs was used as a proxy for estimating fecundity(Gu et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; X. Yang, Sun, Chi, Kang, \u0026amp; Zheng, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Nymphs were counted only after confirming that no further eclosion occurred on the leaves.\u003c/p\u003e \u003cp\u003eTo obtain the F\u003csub\u003e1\u003c/sub\u003e generation, newly hatched nymphs produced by the LC\u003csub\u003e20\u003c/sub\u003e-treated F\u003csub\u003e0\u003c/sub\u003e adults were transferred to separate petri dish containing a fresh cabbage leaf without EB exposure, respectively. Approximately 60 nymphs were set up and leaves were replaced daily to ensure a sufficient food supply. The developmental stage, survival, and mortality were recorded daily. Upon pupation and subsequent adult emergence, daily fecundity of F\u003csub\u003e1\u003c/sub\u003e generation thrips was recorded until death. The F\u003csub\u003e2\u003c/sub\u003e generation was obtained from eggs laid by the F\u003csub\u003e1\u003c/sub\u003e adults, and the same observational procedures as for the F\u003csub\u003e1\u003c/sub\u003e generation were repeated to record their biological data. The life table parameters of F\u003csub\u003e1\u003c/sub\u003e and F\u003csub\u003e2\u003c/sub\u003e generation thrips in the control group was obtained according to the same procedure as in LC\u003csub\u003e20\u003c/sub\u003e-treated group.\u003c/p\u003e\n\u003ch3\u003eRNA extraction and transcriptome sequencing\u003c/h3\u003e\n\u003cp\u003eNewly emerged adult thrips were fed for 48 hours on cabbage leaf discs treated with the LC\u003csub\u003e20\u003c/sub\u003e concentration of EB as described above. The surviving individuals were collected, immediately frozen in liquid nitrogen, and stored at -80℃ until RNA extraction. Three biological replicates with approximately 40 individuals in each repeat were included for the LC\u003csub\u003e20\u003c/sub\u003e treated group and 0.1% Triton X-100 solution treated group, respectively. Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions, and the integrity was verified through 1% agarose gel electrophoresis. RNA concentration and purity were assessed using a NanoDrop spectrophotometer. Only high-quality RNA samples were used for subsequent library construction and transcriptome sequencing. Specifically, mRNA was enriched from extracted RNA using oligo(dT) beads for library construction, and then sequenced on an Illumina NovaSeq 6000 platform to generate paired-end reads. Raw reads were quality-controlled and aligned to the \u003cem\u003eT. tabaci\u003c/em\u003e reference genome (Y. Gao et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)(). Gene expression levels were quantified using RSEM and expressed as FPKM(Smith-Unna, Boursnell, Patro, Hibberd, \u0026amp; Kelly, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Differentially expressed genes (DEGs) were identified with a threshold of |log\u003csub\u003e2\u003c/sub\u003eFC| ≥ 1 and an adjusted p-value \u0026lt; 0.05. Functional enrichment analysis of KEGG pathways was performed, with terms having an adjusted p-value ≤ 0.05 considered as significant criterion.\u003c/p\u003e\n\u003ch3\u003eDNA extraction and 16S rRNA sequencing\u003c/h3\u003e\n\u003cp\u003eSample collection was performed as described in section 2.4. For each treatment, 30 surface-sterilized adult \u003cem\u003eT. tabaci\u003c/em\u003e were pooled as one biological replicate, with five replicates per group. Total genomic DNA from the thrips microbiota was extracted and purified using the TIANGEN DNA Purification Kit (Tiangen Biotech, Beijing, China). following the manufacturer’s protocol. The quality of the extracted DNA was checked via 1% agarose gel electrophoresis, and the NanoDrop 2000C spectrophotometer was used to measure the concentration and purity of obtained DNA. The full-length 16S rRNA gene was amplified from genomic DNA using barcoded primers 27F (5’-AGRGTTYGATYMTGGCTCAG-3’) and 1492R (5’-RGYTACCTTGTTACGACTT-3’). After library preparation with the SMRTbell prep kit 3.0, sequencing was performed on the Pacbio Sequel IIe platform to generate HiFi reads. The resulting circular consensus sequencing (CCS) reads were processed using the QIIME2(Bolyen et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) pipeline with the DADA2(Callahan et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) plugin for denoising and Amplicon Sequence Variant (ASV) calling. Sequences annotated as chloroplasts or mitochondria were removed, and all samples were rarefied to 6,000 sequences per sample for downstream analysis. Taxonomic assignment of ASVs was performed using a consensus approach. Alpha diversity (Chao1, Ace, Shannon, Simpson indices) and beta diversity (Principal Component Analysis based on Bray-Curtis distance) were calculated to assess microbial community structure.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eAdult mortality data of \u003cem\u003eT. tabaci\u003c/em\u003e were compiled in Microsoft Excel 2019. Toxicity parameters (LC\u003csub\u003e20\u003c/sub\u003e, LC\u003csub\u003e50\u003c/sub\u003e, LC\u003csub\u003e95\u003c/sub\u003e, etc.) were derived by probit analysis using IBM SPSS Statistics 27. The age-stage, two-sex life table data for the F\u003csub\u003e0\u003c/sub\u003e–F\u003csub\u003e2\u003c/sub\u003e generations were analyzed with the TWOSEX-MSChart program, applying 100,000 bootstrap replications for estimating life table parameters under LC\u003csub\u003e20\u003c/sub\u003e stress. Data visualization was performed with GraphPad Prism 8. Statistical significance (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05) was assessed by an independent samples t-test, and qRT-PCR data were normalized via the 2\u003csup\u003e–ΔΔCt\u003c/sup\u003e method(Livak \u0026amp; Schmittgen, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e "},{"header":"Results","content":"\u003cp\u003e \u003cb\u003eSublethal effects of EB on F\u003c/b\u003e \u003csub\u003e \u003cb\u003e0\u003c/b\u003e \u003c/sub\u003e \u003cb\u003eof\u003c/b\u003e \u003cb\u003eT. tabaci\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe indoor toxicity of EB against \u003cem\u003eT. tabaci\u003c/em\u003e was evaluated using the leaf-dipping method (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), and the LC\u003csub\u003e50\u003c/sub\u003e, LC\u003csub\u003e20\u003c/sub\u003e, and LC\u003csub\u003e95\u003c/sub\u003e concentration of EB for \u003cem\u003eT. tabaci\u003c/em\u003e was determined as 9.631 mg/L, 4.014 mg/L, and 53.259 mg/L, respectively. The corresponding 95% confidence intervals were 6.839–13.053 mg/L, 2.191–5.811 mg/L, and 33.704-119.101 mg/L. The correlation coefficients (R²) for the toxicity regression lines all exceeded 0.993, indicating a high reliability of the bioassay in this study. Based on these results, the LC\u003csub\u003e20\u003c/sub\u003e concentration was selected as the sublethal dose for subsequent experiments to investigate the effects of EB on the growth, development, and reproductive capacity of \u003cem\u003eT. tabaci\u003c/em\u003e across three successive generations.\u003c/p\u003e\u003cdiv class=\"gridtable\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eToxicity of emamectin benzoate to T. tabaci\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInsecticide\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSlope ± SE*\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLC20(mg/L)\u003c/p\u003e \u003cp\u003e(95%CL) †\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLC50(mg/L)\u003c/p\u003e \u003cp\u003e(95%CL) †\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLC95(mg/L)\u003c/p\u003e \u003cp\u003e(95%CL) †\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eR2\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEmamectin\u003c/p\u003e \u003cp\u003ebenzoate\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e450\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.215 ± 0.36\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.014\u003c/p\u003e \u003cp\u003e(2.191–5.811)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.631\u003c/p\u003e \u003cp\u003e(6.839–13.053)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e53.259\u003c/p\u003e \u003cp\u003e(33.704-119.101)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.993\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003e*, Standard error. †, 95% confidence interval.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003eExposure to the sublethal concentration (LC\u003csub\u003e20\u003c/sub\u003e) of EB significantly impaired the reproduction and development of the parental generation (F\u003csub\u003e0\u003c/sub\u003e) of \u003cem\u003eT. tabaci\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Compared to the control group, the treated adults exhibited a substantial reduction in adult longevity (from 19.73 d to 8.57 d), total fecundity (from 94.99 eggs to 17.05 eggs), and oviposition period (from 18.90 d to 7.91 d).\u003c/p\u003e\u003cp\u003e \u003c/p\u003e\u003cdiv class=\"gridtable\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffects of emamectin benzoate on F\u003csub\u003e0\u003c/sub\u003e generation of \u003cem\u003eT. tabaci\u003c/em\u003e at LC\u003csub\u003e20\u003c/sub\u003e concentration\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEmamectin benzoate\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAdult Longevity (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19.45 ± 0.94a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.57 ± 0.29b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFecundity (eggs/female)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e94.99 ± 5.25a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.05 ± 0.74b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOviposition period (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.90 ± 1.01a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.91 ± 0.32b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGross Reproductive Rate \u003cem\u003eGRR\u003c/em\u003e (offspring per individual)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e106.25 ± 5.16a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.17 ± 0.72b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNet reproductive rate \u003cem\u003eR\u003c/em\u003e\u003csub\u003e\u003cem\u003e0\u003c/em\u003e\u003c/sub\u003e (offspring per individual)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e94.99 ± 5.25a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.04 ± 0.86b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntrinsic rate of increase \u003cem\u003er\u003c/em\u003e (days\u003csup\u003e− 1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.2042 ± 1.79a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.1472 ± 2.47b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFinite rate of increase λ (days\u003csup\u003e− 1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.2226 ± 2.19a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.1585 ± 2.86b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMean generation time T (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e22.30 ± 0.19a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.26 ± 5.61b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003eNote: Values are presented as mean ± SE. Means within a row followed by different lowercase letters are significantly different between two groups (independent samples \u003cem\u003et\u003c/em\u003e-test, p \u0026lt; 0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003eThe sublethal concentration (LC\u003csub\u003e20\u003c/sub\u003e) of EB also significantly influenced the population dynamics of the parental (F\u003csub\u003e0\u003c/sub\u003e) generation of \u003cem\u003eT. tabaci\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Compared to the control, the LC\u003csub\u003e20\u003c/sub\u003e EB treated \u003cem\u003eT. tabaci\u003c/em\u003e exhibited a sharp decline in both the gross reproductive rate (GRR) and the net reproductive rate (R\u003csub\u003e0\u003c/sub\u003e), which were significantly reduced by 82.0% (from 106.25 to 19.17) and 82.1% (from 94.99 to 17.04), respectively. Furthermore, the intrinsic rate of increase (r) decreased from 0.2042 d⁻¹ to 0.1472 d⁻¹, and the finite rate of increase (λ) dropped from 1.2226 d⁻¹ to 1.1585 d⁻¹ in the LC\u003csub\u003e20\u003c/sub\u003e EB treated population, indicating a severely constrained expansion potential. Concurrently, the mean generation time (T) was significantly shortened by approximately 3.04 days, suggesting an accelerated life cycle under insecticide stress, which occurred at the expense of reproductive fitness.\u003c/p\u003e\u003cp\u003eThe age-specific survival rate (l\u003csub\u003ex\u003c/sub\u003e) of the EB-treated group began to decline on day 21, obviously earlier than that of the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). The age-specific fecundity (m\u003csub\u003ex\u003c/sub\u003e) curve for the F\u003csub\u003e0\u003c/sub\u003e generation under LC\u003csub\u003e20\u003c/sub\u003e EB exposure was consistently lower than that of the control and peaked at the 16th day with a value of 4.23, which was both significantly lower and earlier than the peak value of 6.55 at the 24th day in the control (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). The age-stage-specific life expectancy (e\u003csub\u003exj\u003c/sub\u003e) results indicated that the life expectancy of \u003cem\u003eT. tabaci\u003c/em\u003e was 28 days in the EB treated group, significantly shorter than 40 days compared to the control (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC), demonstrating an obvious adverse impact on the expected lifespan of \u003cem\u003eT. tabaci\u003c/em\u003e due to stress of LC\u003csub\u003e20\u003c/sub\u003e EB. Furthermore, the age-stage-specific reproductive value (v\u003csub\u003exj\u003c/sub\u003e) peaked at 34.66 on day 18 in the control group, whereas the LC\u003csub\u003e20\u003c/sub\u003e EB-treated group reached a lower peak value of 12.22 on day 16 (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eD).\u003c/p\u003e\u003cp\u003e \u003cb\u003eTransgenerational effects of sublethal EB on\u003c/b\u003e \u003cb\u003eT. tabaci\u003c/b\u003e\u003c/p\u003e\u003cp\u003eMultigenerational influence of sublethal EB stress on \u003cem\u003eT. tabaci\u003c/em\u003e was also assessed through the life table parameters analysis (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). For the 1st generation (F1) thrips, their total longevity was not significantly affected, however, several key developmental and reproductive parameters were markedly altered due to their mothers’ exposure to LC\u003csub\u003e20\u003c/sub\u003e EB. For example, the adult longevity was significantly shortened by 27.8% (from 19.41 to 14.02 days), and the oviposition period decreased by 26.9% (from 18.65 to 13.64 days). Concurrently, compared to the control, the egg stage duration was significantly reduced from 10.91 to 7.69 days. In contrast, the durations of the first instar nymphal and pupal stages were both significantly prolonged. More critically, fecundity was drastically reduced from 99.41 to 34.02 eggs per female, namely falling by 65.8%. Consequently, significant detrimental effects were also observed in population growth parameters: the gross reproductive rate (GRR) decreased from 114.43 to 38.10 (-66.7%), the net reproductive rate (R\u003csub\u003e0\u003c/sub\u003e) fell from 99.41 to 34.02 (-65.8%), the intrinsic rate of increase (r) dropped from 0.2017 to 0.1223 d\u003csup\u003e− 1\u003c/sup\u003e, and the finite rate of increase (λ) declined from 1.2223 to 1.1301 d\u003csup\u003e− 1\u003c/sup\u003e. Besides, the mean generation time (T) was significantly extended from 22.80 to 28.83 days.\u003c/p\u003e\u003cp\u003eUnexpectedly, the sublethal effects of EB on \u003cem\u003eT. tabaci\u003c/em\u003e largely subsided in the F\u003csub\u003e2\u003c/sub\u003e generation. Only adult longevity was significantly affected which decreased from 19.50 to 16.67 days. Life table parameters related to developmental, reproductive, and population growth were approximate between LC\u003csub\u003e20\u003c/sub\u003e EB treated group and control group, indicating a recovery of \u003cem\u003eT. tabaci\u003c/em\u003e from the transgenerational sublethal effects.\u003c/p\u003e\u003cdiv class=\"gridtable\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eLife table parameters of F\u003csub\u003e1\u003c/sub\u003e-F\u003csub\u003e2\u003c/sub\u003e generation of \u003cem\u003eT. tabaci\u003c/em\u003e after treatment with sublethal concentrations of emamectin benzoate in F\u003csub\u003e0\u003c/sub\u003e generation\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eF1 generation\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c6\" namest=\"c4\"\u003e \u003cp\u003eF2 generation\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eEmamectin\u003c/p\u003e \u003cp\u003ebenzoate\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEmamectin\u003c/p\u003e \u003cp\u003ebenzoate\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEgg stage (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.91 ± 0.12a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.69 ± 0.28b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e7.58 ± 0.17a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.40 ± 0.20a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFirst instar nymph stage (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.18 ± 0.23b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.89 ± 4.78a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e4.40 ± 0.19a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.76 ± 0.09a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSecond instar nymph stage (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.47 ± 0.17a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.75 ± 6.52a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e2.75 ± 0.12a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.79 ± 7.12a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePupa stage (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.71 ± 0.14b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.14 ± 5.18a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e2.15 ± 0.13a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.00 ± 8.68a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAdult Longevity (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19.41 ± 1.13a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.02 ± 0.28b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e19.50 ± 0.99a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16.67 ± 1.02b\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLongevity (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e37.81 ± 1.27a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35.35 ± 1.23a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e38.00 ± 0.94a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e35.76 ± 0.95a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFecundity (eggs/female)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e99.41 ± 7.18a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.02 ± 0.71b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e87.70 ± 6.27a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e81.89 ± 3.54a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOviposition period (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.65 ± 1.07a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.64 ± 0.25b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e18.40 ± 0.97a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16.06 ± 1.06a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGross Reproductive Rate \u003cem\u003eGRR\u003c/em\u003e (offspring per individual)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e114.43 ± 6.80a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e38.10 ± 0.67b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e108.65 ± 9.95a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e89.72 ± 3.39a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNet reproductive rate \u003cem\u003eR0\u003c/em\u003e (offspring per individual)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e99.41 ± 7.18a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.02 ± 0.71b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e87.70 ± 6.29a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e76.48 ± 5.56a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIntrinsic rate of increase \u003cem\u003er\u003c/em\u003e (days\u003csup\u003e-1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.2017 ± 3.45a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.1223 ± 8.11b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.1775 ± 3.68a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.1751 ± 3.23a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFinite rate of increase λ (days\u003csup\u003e-1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.2223 ± 4.22a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.1301 ± 9.16b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e1.1942 ± 4.40a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.1914 ± 3.84a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMean generation time T (days)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e22.80 ± 0.35b\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e28.83 ± 0.19a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e25.21 ± 0.57a\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e24.75 ± 0.26a\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"6\"\u003eNote: Values are presented as mean ± SE. Means within a row of F1 or F2 generation followed by different lowercase letters are significantly different between two groups (independent samples *t*-test, *p* \u0026lt; 0.05).\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eFour age-specific life parameters of \u003cem\u003eT. tabaci\u003c/em\u003e were affected in F\u003csub\u003e1\u003c/sub\u003e generation due to their mothers’ (F\u003csub\u003e0\u003c/sub\u003e) exposure to LC\u003csub\u003e20\u003c/sub\u003e EB (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Although the F\u003csub\u003e1\u003c/sub\u003e generation exposed to LC\u003csub\u003e20\u003c/sub\u003e EB exhibited a later decline in age-specific survival rate (l\u003csub\u003ex\u003c/sub\u003e) and maintained higher overall survival rates, it consistently exhibited reduced fecundity (m\u003csub\u003ex\u003c/sub\u003e) curves relative to the control group over its entire life course. Specifically, the m\u003csub\u003ex\u003c/sub\u003e peaked 3.95 at the 23th day in the LC\u003csub\u003e20\u003c/sub\u003e EB treated group, which was substantially lower than 7.38 in the control on day 22 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC). Besides, the age-stage-specific life expectancy (e\u003csub\u003exj\u003c/sub\u003e) curve showed the total longevity of the F\u003csub\u003e1\u003c/sub\u003e generation in the LC\u003csub\u003e20\u003c/sub\u003e treatment group was 37.70 d with no significant difference from that in the control of 37.81 d (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC). In contrast, the age-speciffc reproductive value (v\u003csub\u003exj\u003c/sub\u003e) peak in the LC\u003csub\u003e20\u003c/sub\u003e EB group was both markedly delayed, occurring on day 36 compared to day 20 in the control, and significantly reduced in magnitude, reaching only 22.34 d\u003csup\u003e− 1\u003c/sup\u003e versus 33.15 d\u003csup\u003e− 1\u003c/sup\u003e in the control (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD).\u003c/p\u003e\u003cp\u003eIn the F\u003csub\u003e2\u003c/sub\u003e generation, age-specific survival rate (l\u003csub\u003ex\u003c/sub\u003e), the life expectancy (e\u003csub\u003exj\u003c/sub\u003e) and fecundity (m\u003csub\u003ex\u003c/sub\u003e) curves did not differ significantly between the LC\u003csub\u003e20\u003c/sub\u003e treatment and the control groups with both exhibiting a similar gradual decline (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, E-G). However, a delay and reduction in the reproductive peak was observed: the v\u003csub\u003exj\u003c/sub\u003e peak in the LC\u003csub\u003e20\u003c/sub\u003e treatment group occurred on day 23 with a value of 27.43, which was later and lower than that (31.93 on day 20) of the control (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eH).\u003c/p\u003e\u003cp\u003e \u003cb\u003eEffects of sublethal EB on the gene expression of\u003c/b\u003e \u003cb\u003eT. tabaci\u003c/b\u003e\u003c/p\u003e\u003cp\u003eTranscriptomic analysis of six biological samples from \u003cem\u003eT. tabaci\u003c/em\u003e—LC\u003csub\u003e20\u003c/sub\u003e EB-treated group and a control group—yielded a total of 38.7 Gb of clean data. Each sample produced no less than 6.14 Gb of clean data, with a Q30 base percentage of 95.99%, meeting the core quality control threshold of Q30 \u0026gt; 95%. These results confirm the high quality and reliability of the sequencing data for subsequent analyses.\u003c/p\u003e\u003cp\u003ePrincipal component analysis (PCA) revealed a clear separation between the insecticide-treated and control groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA), indicating that LC\u003csub\u003e20\u003c/sub\u003e EB stress induced substantial gene response for \u003cem\u003eT. tabaci\u003c/em\u003e. Correlation analysis of genes expression profiles among samples further demonstrated high intra-group reproducibility, with correlation coefficients all above 0.961 within groups, in contrast to lower inter-group correlations (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). Based on these, the impact of sublethal EB on gene expression in \u003cem\u003eT. tabaci\u003c/em\u003e was assessed furthermore, and a total of 1066 differentially expressed genes (DEGs) were, among which 421 were up-regulated and 645 were down-regulated (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC).\u003c/p\u003e\u003cp\u003eKEGG pathway enrichment analysis revealed the overall trends with DEGs in \u003cem\u003eT. tabaci\u003c/em\u003e under LC\u003csub\u003e20\u003c/sub\u003e EB stress. The most pronounced responses were observed in carbohydrate metabolism pathways, such as starch and sucrose metabolism and fructose and mannose metabolism, suggesting that sublethal EB may trigger a reprogramming of energy metabolism in \u003cem\u003eT. tabaci\u003c/em\u003e to meet potential energy demands. Concurrently, pathways closely associated with stress response and cellular regulation, including the MAPK and ErbB signaling pathways, also showed moderate enrichment, indicating potential disruptions in neural transmission and cellular homeostasis. Furthermore, alterations in pathways such as regulation of the actin cytoskeleton and protein processing in the endoplasmic reticulum reflect the multifaceted potential effects of sublethal doses of EB on cellular structure and function. Together, these findings provide preliminary insights and directions for further validation toward a systematic understanding of the molecular adaptation mechanisms of \u003cem\u003eT. tabaci\u003c/em\u003e under sublethal EB stress (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD).\u003c/p\u003e\u003cp\u003e \u003cb\u003eInfluence of sublethal EB treatment on bacterial communities of\u003c/b\u003e \u003cb\u003eT. tabaci\u003c/b\u003e\u003c/p\u003e\u003cp\u003eSequencing of 10 samples yielded a total of 343,160 single-end reads. Following quality control, denoising, and chimera removal, 126,701 high-quality sequences were retained, resulting in 501 amplicon sequence variants (ASVs) (Table S2). Alpha diversity analysis, as measured by the Shannon index, revealed a slight reduction in microbial diversity in the EB-treated group compared to the control; however, the difference was not statistically significant (P = 0.2963; adjusted P = 0.5925) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA), indicating that LC\u003csub\u003e20\u003c/sub\u003e EB exposure did not substantially alter the overall microbiota diversity of \u003cem\u003eT. tabaci\u003c/em\u003e. Principal coordinate analysis (PCoA) based on genus-level community composition showed a modest separation between the control and EB-treated groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB), suggesting a potential shift in microbial structure of \u003cem\u003eT. tabaci\u003c/em\u003e in response to EB treatment. Nevertheless, this trend did not reach statistical significance, which may be attributable to the limited sample size or considerable inter-individual variation.\u003c/p\u003e\u003cp\u003eAt the genus level (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC), the core microbiota in control \u003cem\u003eT. tabaci\u003c/em\u003e consisted of \u003cem\u003eSerratia\u003c/em\u003e (25.98%), \u003cem\u003eAcinetobacter\u003c/em\u003e (44.03%), \u003cem\u003ePantoea\u003c/em\u003e (23.24%), \u003cem\u003eRosenbergiella\u003c/em\u003e (4.96%), and \u003cem\u003eErwinia\u003c/em\u003e (0.22%). After LC\u003csub\u003e20\u003c/sub\u003e EB treatment, the relative abundances of \u003cem\u003eSerratia\u003c/em\u003e (29.49%), \u003cem\u003ePantoea\u003c/em\u003e (39.47%), \u003cem\u003eRosenbergiella\u003c/em\u003e (9.24%), and \u003cem\u003eErwinia\u003c/em\u003e (1.29%) increased to varying degrees, with the most pronounced rise observed in \u003cem\u003ePantoea\u003c/em\u003e. In contrast, the abundance of \u003cem\u003eAcinetobacter\u003c/em\u003e decreased to 17.26%.\u003c/p\u003e\u003cp\u003eAt the species level (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE), the relative abundance of \u003cem\u003ePantoea ananatis\u003c/em\u003e significantly increased from 22.68% in the control group to 39.46% in the EB-treated group, indicating its possible roles in enhancing tolerance of \u003cem\u003eT. tabaci\u003c/em\u003e to EB stress. Similarly, species within the genus \u003cem\u003eSerratia\u003c/em\u003e, including \u003cem\u003eS. rubidaea\u003c/em\u003e and \u003cem\u003eS. marcescens\u003c/em\u003e, increased by 1.21% and 4.47%, respectively, also suggesting their potential involvement in the host’s stress response. Conversely, the relative abundance of \u003cem\u003eAcinetobacter\u003c/em\u003e species (e.g., \u003cem\u003eA. lactucae\u003c/em\u003e and unclassified \u003cem\u003eAcinetobacter\u003c/em\u003e) decreased in the treated group, with \u003cem\u003eA. lactucae\u003c/em\u003e showing the most dramatic decline—from 26.54% to 0.25%. This shift may contribute to altered gut microenvironment homeostasis in \u003cem\u003eT. tabaci\u003c/em\u003e. Collectively, these findings indicate that sublethal EB stress significantly restructured the core microbiota of \u003cem\u003eT. tabaci\u003c/em\u003e.\u003c/p\u003e\u003cp\u003eKEGG functional prediction based on PICRUSt2 analysis of 16S rRNA gene sequences indicated that LC\u003csub\u003e20\u003c/sub\u003e EB treatment altered the functional potential of the microbial community in \u003cem\u003eT. tabaci\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC). Compared to the control group, the treated group exhibited differences in the abundance of several Level 2 metabolic pathways. Pathways closely related to fundamental material and energy metabolism—such as carbohydrate metabolism, amino acid metabolism, and lipid metabolism—were generally downregulated. In contrast, pathways involved in xenobiotic biodegradation and metabolism, membrane transport, and signal transduction, which are associated with environmental stress response and specific physiological processes, were upregulated in the LC\u003csub\u003e20\u003c/sub\u003e EBtreated group.\u003c/p\u003e\u003cp\u003e \u003cb\u003eIdentification of key detoxification genes involving the adaptation of\u003c/b\u003e \u003cb\u003eT. tabaci\u003c/b\u003e \u003cb\u003eto sublethal EB stress\u003c/b\u003e\u003c/p\u003e\u003cp\u003eGiven its marked upregulation (log\u003csub\u003e2\u003c/sub\u003eFC = 1.04, P \u0026lt; 3.6e\u003csup\u003e− 17\u003c/sup\u003e) in response to sublethal EB exposure, we hypothesized that \u003cem\u003eCYP6K1\u003c/em\u003e plays a critical role in the detoxification process and may contribute to tolerance in \u003cem\u003eT. tabaci\u003c/em\u003e. To functionally validate this hypothesis, we employed RNA interference to knock down its expression. RNA interference experiments demonstrated that feeding adult \u003cem\u003eT. tabaci\u003c/em\u003e with dsRNA for 48 hours via a double-membrane method significantly reduced the expression of the \u003cem\u003eCYP6K1\u003c/em\u003e gene by 51.32% compared to the control group (P \u0026lt; 0.05; Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). After a 48-hour feeding period with synthesized \u003cem\u003edsCYP6K1\u003c/em\u003e, adult thrips were subjected to a bioassay using the sublethal concentration LC\u003csub\u003e20\u003c/sub\u003e emamectin benzoate to determine changes in insecticide susceptibility. The results showed that the mortality rate of RNAi-treated thrips following LC\u003csub\u003e20\u003c/sub\u003e emamectin benzoate exposure reached 78.19%, representing a significant increase of 58.88% compared to the control mortality of 19.31% (P \u0026lt; 0.001). This dramatic increase in susceptibility confirms that \u003cem\u003eCYP6K1\u003c/em\u003e is a key mediator of sublethal EB tolerance.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this study, LC\u003csub\u003e50\u003c/sub\u003e of EB to \u003cem\u003eT. tabaci\u003c/em\u003e was identified as 9.631 mg/L (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), suggesting the high toxicity to this disgusting pest. Similarly, a synthesis of considerable previous bioassay data confirms potent and broad-spectrum insecticidal activity of EB. Against major lepidopteran pests, EB exhibits high toxicity, with reported LC\u003csub\u003e50\u003c/sub\u003e values of 0.019 mg/L for \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e (Fiaboe, Fening, Gbewonyo, \u0026amp; Deshmukh, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), 0.631 mg/L for \u003cem\u003eS. litura\u003c/em\u003e (Devi, Mahajan, Saini, \u0026amp; Kaur, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), 0.464 mg/L for \u003cem\u003eHelicoverpa zea\u003c/em\u003e (L\u0026oacute;pez, Latheef, \u0026amp; Hoffmann, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), 0.173 mg/L for \u003cem\u003ePlutella xylostella\u003c/em\u003e (K. X. Liu, Guo, Zhang, \u0026amp; Xue, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), and 0.010 mg/L for \u003cem\u003eS. littoralis\u003c/em\u003e(El-Saleh et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Notably, the efficacy of EB extends beyond Lepidoptera to include diverse pests of fruit crops (e.g., \u003cem\u003eDrosophila suzukii\u003c/em\u003e with the LC\u003csub\u003e50\u003c/sub\u003e of 0.021 mg/L(H. Gao et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)), soybeans (e.g., \u003cem\u003eRiptortus pedestris\u003c/em\u003e with the LC\u003csub\u003e50\u003c/sub\u003e of 7.681 mg/L(Guo et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e)), citrus (e.g., \u003cem\u003ePanonychus citri\u003c/em\u003e with the LC\u003csub\u003e50\u003c/sub\u003e of 0.350 mg/L(Khan et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2021\u003c/span\u003e)), and oil palm (e.g., \u003cem\u003eRhynchophorus ferrugineus\u003c/em\u003e with the LC\u003csub\u003e50\u003c/sub\u003e of 0.144 mg/L(Rezk et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)), underscoring its wide applicability. Besides, comprehensive analysis of EB\u0026rsquo;s toxicity to several thrips species showed that the LC\u003csub\u003e50\u003c/sub\u003e of EB against \u003cem\u003eFrankliniella occidentalis\u003c/em\u003e (7.997 mg/L) was determined to be comparable to our result, (Shen et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), however, its efficacy fluctuated with LC\u003csub\u003e50\u003c/sub\u003e from 27.731 to 34.536 mg/L when resisting against \u003cem\u003eT. palmi\u003c/em\u003e from different populations, thereby deserves more attention in using this appropriately for precise thrips control.\u003c/p\u003e \u003cp\u003eThe suitability of an insecticide for integrated pest management (IPM) depends not only on its efficacy against target pests but also on its selectivity toward non-target organisms, particularly natural enemies. Available data indicate that emamectin benzoate exhibits relatively low toxicity to some certain predators such as \u003cem\u003eChrysoperla sinica\u003c/em\u003e (Shan et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), yet also poses a high risk to numerous beneficial arthropods with low LC\u003csub\u003e50\u003c/sub\u003e from 7.41(4.88\u0026ndash;11.26) mg/L, including \u003cem\u003eCotesia marginiventris\u003c/em\u003e (Hou et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), honey bees (Abdu-Allah \u0026amp; Pittendrigh, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), \u003cem\u003eTrichogramma japonicum Ashmead\u003c/em\u003e (Zhu et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) and poses a moderate threat to the rove beetle (\u003cem\u003ePaederus fuscipes\u003c/em\u003e(Khan, Nawaz, Hua, Cai, \u0026amp; Zhao, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2018\u003c/span\u003e)). Consequently, a comprehensive evaluation of the safety and ecological risks of EB is also needed, including its effects on key thrips natural enemies such as predatory bugs, mites, and parasitoids, to guide scientifical usage of this pesticide.\u003c/p\u003e \u003cp\u003eSublethal dose insecticides causing by gradual degradation after field spraying usually produce unintended effects on target pest or environment. In this study, exposure to the sublethal concentration (LC\u003csub\u003e20\u003c/sub\u003e) of EB significantly suppressed the F\u003csub\u003e0\u003c/sub\u003e generation of \u003cem\u003eT. tabaci\u003c/em\u003e, especially resulting in reduced adult longevity and fecundity (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). These findings align with previously documented sublethal effects in other insect species. For instance, sublethal doses of emamectin benzoate induced severe physiological and reproductive inhibition in \u003cem\u003eRiptortus pedestris\u003c/em\u003e(Guo et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) and significantly decreased egg-laying in \u003cem\u003eMamestra brassicae\u003c/em\u003e(Moustafa, K\u0026aacute;kai, Awad, \u0026amp; F\u0026oacute;nagy, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).The sublethal effects also persisted into the F\u003csub\u003e1\u003c/sub\u003e generation of \u003cem\u003eT. tabaci\u003c/em\u003e, where both developmental and reproductive parameters were significantly impaired compared to the control (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This transgenerational suppression is consistent with observations by Rezk et al.(Rezk et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), who also reported significant inhibiting of offspring population parameters following parental exposure to serial sublethal (LC\u003csub\u003e10\u003c/sub\u003e, LC\u003csub\u003e25\u003c/sub\u003e, LC\u003csub\u003e50\u003c/sub\u003e) EB in \u003cem\u003eRhynchophorus ferrugineus\u003c/em\u003e. Similarly, sublethal concentrations of EB was found can induce long-term population suppression in \u003cem\u003ePlutella xylostella\u003c/em\u003e, although a transient stimulation of oviposition may occur(K. X. Liu et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Likewise, they significantly impair the development and reproduction of \u003cem\u003eS. frugiperda\u003c/em\u003e, leading to comprehensive declines in population growth parameters such as the intrinsic rate of increase (C. Y. Chen, Tang, Zhao, Zhang, \u0026amp; Zhang, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). EB at sublethal doses also disrupt normal development and severely inhibit reproduction of \u003cem\u003eHelicoverpa armigera\u003c/em\u003e, thereby substantially diminishing population growth potential through these \"hidden\" effects (Dong LiXia, Rui ChangHui, Ren Long, \u0026amp; Tan XiaoWei, 2011). However, a recent study by Shun-fan Wu \u003cem\u003eet al\u003c/em\u003e (Gao et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) reported that lethal doses of EB accelerated ovarian development and increased mature egg load in female \u003cem\u003eNilaparvata lugens\u003c/em\u003e, potentially leading to pest resurgence. Collectively, these studies indicate that emamectin benzoate not noly exhibits high toxicity for effective controlling against a wide range of pests, but also can exert sustained suppression under residual sublethal concentrations, however it may also trigger population outbreaks in a few species.\u003c/p\u003e \u003cp\u003eFurthermore, this study also revealed that LC\u003csub\u003e20\u003c/sub\u003e EB exposure also significantly altered the microbial community structure of \u003cem\u003eT. tabaci\u003c/em\u003e, characterized primarily by a marked increase in the relative abundance of \u003cem\u003ePantoea\u003c/em\u003e and a sharp decline in \u003cem\u003eAcinetobacter\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). This finding offers new insights into the insecticidal mechanism of EB from a microbiological perspective. Previous research has established that \u003cem\u003eAcinetobacter\u003c/em\u003e plays a critical role in insect responses to environmental stress; for example, it assists the \u003cem\u003eCamellia weevil\u003c/em\u003e in adapting to its host plant by degrading tea saponins(Song, Shu, \u0026amp; Zhang, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), and \u003cem\u003eAcinetobacter baumannii\u003c/em\u003e enhances the resistance of \u003cem\u003eNilaparvata lugens\u003c/em\u003e to the entomopathogenic fungus \u003cem\u003eMetarhizium anisopliae\u003c/em\u003e (Tang et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). These findings suggest that \u003cem\u003eAcinetobacter\u003c/em\u003e may aid insects in maintaining homeostasis and coping with external stressors through multiple mechanisms. Therefore, the observed decline of \u003cem\u003eAcinetobacter\u003c/em\u003e following sublethal EB exposure likely compromised the physiological resilience of \u003cem\u003eT. tabaci\u003c/em\u003e, increasing its susceptibility to the insecticide. Concurrently, \u003cem\u003ePantoea\u003c/em\u003e also known as a dominant bacterial genus in \u003cem\u003eP. xylostella\u003c/em\u003e, has been implicated as a potential target for pest control, as reducing its density in \u003cem\u003eP. xylostella\u003c/em\u003e effectively achieved population suppression of host (Li, Jin, Li, Cheng, \u0026amp; Jin, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The microbial dysbiosis observed here, marked by the replacement of \u003cem\u003eAcinetobacter\u003c/em\u003e by \u003cem\u003ePantoea\u003c/em\u003e as the dominant genus, may disrupt host gut function and, combined with the loss of \u003cem\u003eAcinetobacter\u003c/em\u003e, collectively exacerbate the physiological stress imposed by EB on \u003cem\u003eT. tabaci\u003c/em\u003e. Overall, this study provides a new perspective on the interaction between emamectin benzoate and the microbiota of \u003cem\u003eT. tabaci\u003c/em\u003e, although the precise causal mechanisms, including the potential role of \u003cem\u003eAcinetobacter\u003c/em\u003e in direct insecticide detoxification, require further validation.\u003c/p\u003e \u003cp\u003eRNA interference experiments provided direct evidence elucidating the central role of \u003cem\u003eCYP6K1\u003c/em\u003e in the response of \u003cem\u003eT. tabaci\u003c/em\u003e to emamectin benzoate stress. Silencing this gene resulted in a significant increase in thrips mortality under LC\u003csub\u003e20\u003c/sub\u003e EB exposure, confirming that \u003cem\u003eCYP6K1\u003c/em\u003e is one of the key determinants mediating tolerance of \u003cem\u003eT. tabaci\u003c/em\u003e to EB. More importantly, this study is the first to reveal that the stress effects of emamectin benzoate on \u003cem\u003eT. tabaci\u003c/em\u003e may operate simultaneously across two dimensions: host genes and symbiotic microbiota. Our results indicate that under emamectin benzoate stress, \u003cem\u003eT. tabaci\u003c/em\u003e not only upregulates \u003cem\u003eCYP6K1\u003c/em\u003e gene expression in an attempt to metabolize and detoxify the insecticide but also undergoes drastic changes in gut microbiota structure, particularly a significant reduction in the abundance of \u003cem\u003eAcinetobacter\u003c/em\u003e, which may contribute to stress resistance. Based on these findings, we hypothesize that in response to sublethal doses of emamectin benzoate, \u003cem\u003eT. tabaci\u003c/em\u003e activates a coordinated defense network involving both host P450 detoxification genes and specific intestinal symbiotic bacteria. Disruption of any key component in this defense system, whether a host gene or a member of the symbiotic microbiota, may lead to the collapse of the thrips\u0026rsquo; tolerance and result in higher mortality.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, this study delineates the response of \u003cem\u003eT. tabaci\u003c/em\u003e to emamectin benzoate stress at both the transcriptional and microbiome levels, and functionally validates \u003cem\u003eCYP6K1\u003c/em\u003e as one of core tolerance gene. Future studies should further decipher the interactions between host detoxification mechanisms and specific gut microbiota functions. Specially, a key focus would be to determine whether \u003cem\u003eAcinetobacter\u003c/em\u003e can directly degrade emamectin benzoate or indirectly enhances tolerance by modulating host immunity. Such investigations are crucial for building a theoretical foundation for developing integrated control strategies against \u003cem\u003eThrips tabaci\u003c/em\u003e.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eEthical approval\u003c/h2\u003e \u003cp\u003eWe declare that all applicable national and provincial guidelines for the care and use of animals were followed.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eCompeting interests\u003c/strong\u003e \u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis research was funded by the Central Public-interest Scientific Institution Basal Research Fund (No.1610162023010), National Key R\u0026amp;D Program of China (Grant No. 2022YFD1400300), Biological Breeding-Major Projects (2023ZD04062), the China Agriculture Research System, and Agricultural Science and Technology Innovation Program (ASTIP) (CAAS-ZDRW202412).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eJichao Ji, Junyu Luo, and Jinjie Cui conceived and supervised the project. Miaomiao Xin and Chunjie Xian: Data curation, Formal analysis, Writing\u0026ndash;original draft, Writing \u0026ndash; review \u0026amp; editing. Xiaoyun Wang: Data curation, Formal analysis. Miaomiao Xin, Li Wang, Xiangzhen Zhu performed the experiments; Kaixin Zhang, Dongyang Li reviewed the original draft; Jichao Ji reviewed, edited and polished the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eWe thank the editors and reviewers for their constructive comments on our work. The studies were conducted in the laboratories of the Institute of Cotton Research, Chinese Academy of Agricultural Sciences.\u003c/p\u003e\u003ch2\u003eAvailability of data and materials\u003c/h2\u003e \u003cp\u003eAll data and materials needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary material. Additional data related to this paper may be requested from the authors.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbbas, A., Zhao, C. R., Arshad, M., Han, X., Iftikhar, A., Hafeez, F., . . . Ullah, F. (2023). Sublethal effects of spinetoram and emamectin benzoate on key demographic parameters of fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae) under laboratory conditions. \u003cem\u003eEnviron Sci Pollut Res Int, 30\u003c/em\u003e(34), 82990-83003. http://doi:10.1007/s11356-023-28183-8\u003c/li\u003e\n\u003cli\u003eAbdel-Baky, N. F., Alhewairini, S. S., \u0026amp; Bakry, M. M. S. (2019). EMAMECTIN-BENZOATE AGAINST Tuta absoluta MEYRICK AND Spodoptera littoralis BOISDUVAL LARVAE. \u003cem\u003ePakistan Journal of Agricultural Sciences, 56\u003c/em\u003e(3), 801-808. http://doi:10.21162/pakjas/19.8082\u003c/li\u003e\n\u003cli\u003eAbdu-Allah, G. A. M., \u0026amp; Pittendrigh, B. R. (2018). Lethal and sub-lethal effects of select macrocyclic lactones insecticides on forager worker honey bees under laboratory experimental conditions. \u003cem\u003eEcotoxicology, 27\u003c/em\u003e(1), 81-88. http://doi:10.1007/s10646-017-1872-6\u003c/li\u003e\n\u003cli\u003eAfza, R., Afzal, A., Riaz, M. A., Majeed, M. Z., Idrees, A., Qadir, Z. A., . . . Li, J. (2023). Sublethal and transgenerational effects of synthetic insecticides on the biological parameters and functional response of Coccinella septempunctata (Coleoptera: Coccinellidae) under laboratory conditions. \u003cem\u003eFrontiers in Physiology, 14\u003c/em\u003e, 1088712-1088725. http://doi:10.3389/fphys.2023.1088712\u003c/li\u003e\n\u003cli\u003eBolyen, E., Rideout, J. R., Dillon, M. R., Bokulich, N., Abnet, C. C., Al-Ghalith, G. A., . . . Caporaso, J. G. (2019). Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. \u003cem\u003eNature Biotechnology, 37\u003c/em\u003e(8), 852-857. http://doi:10.1038/s41587-019-0209-9\u003c/li\u003e\n\u003cli\u003eCallahan, B. J., McMurdie, P. J., Rosen, M. J., Han, A. W., Johnson, A. J. A., \u0026amp; Holmes, S. P. (2016). DADA2: High-resolution sample inference from Illumina amplicon data. \u003cem\u003eNature Methods, 13\u003c/em\u003e(7), 581-583. http://doi:10.1038/nmeth.3869\u003c/li\u003e\n\u003cli\u003eChen, C. Y., Tang, Y. T., Zhao, Y. X., Zhang, X. F., \u0026amp; Zhang, K. (2025). Life table study of sublethal concentrations of emamectin benzoate against Spodoptera frugiperda (Lepidoptera, Noctuidae). \u003cem\u003eJournal of Insect Science, 25\u003c/em\u003e(1), 7. http://doi:10.1093/jisesa/ieaf014\u003c/li\u003e\n\u003cli\u003eChen, Y. X., Tian, H. J., Lin, S., Yu, Y., Xie, L. C., Li, H., . . . Wei, H. (2023). Sublethal effects of emamectin benzoate on development, reproduction, and vitellogenin and vitellogenin receptor gene expression in Thrips hawaiiensis (Thysanoptera: Thripidae). \u003cem\u003eJ Insect Sci, 23\u003c/em\u003e(3). http://doi:10.1093/jisesa/iead035\u003c/li\u003e\n\u003cli\u003eDevi, M., Mahajan, A., Saini, H. S., \u0026amp; Kaur, S. (2024). The impact of lethal and sub-lethal exposure of emamectin benzoate on populations of Spodoptera litura (Lepidoptera: Noctuidae) under laboratory conditions. \u003cem\u003eToxicon, 250\u003c/em\u003e, 108121-108129. http://doi:10.1016/j.toxicon.2024.108121\u003c/li\u003e\n\u003cli\u003eDong LiXia, D. L., Rui ChangHui, R. C., Ren Long, R. L., \u0026amp; Tan XiaoWei, T. X. (2011). Effect of sublethal dose of emamectin benzoate on growth and development of Helicoverpa armigera (Hubner). \u003cem\u003eActa Phytophylacica Sinica, 38\u003c/em\u003e(6), 539-544. Retrieved from \u0026lt;Go to ISI\u0026gt;://CABI:20123048930 http://www.wanfangdata.com.cn\u003c/li\u003e\n\u003cli\u003eEl-Saleh, M. A., Aioub, A. A., El-Sheikh, E. A., Desuky, W. M. H., Alkeridis, L. A., Al-Shuraym, L. A., . . . Hamed, I. A. (2025). Comparative Toxicological Effects of Insecticides and Their Mixtures on Spodoptera littoralis (Lepidoptera: Noctuidae). \u003cem\u003eInsects, 16\u003c/em\u003e(8). http://doi:10.3390/insects16080821\u003c/li\u003e\n\u003cli\u003eFiaboe, K. R., Fening, K. O., Gbewonyo, W. S. K., \u0026amp; Deshmukh, S. (2023). Bionomic responses of Spodoptera frugiperda (J. E. Smith) to lethal and sublethal concentrations of selected insecticides. \u003cem\u003ePLoS One, 18\u003c/em\u003e(11), e0290390-e0290410. http://doi:10.1371/journal.pone.0290390\u003c/li\u003e\n\u003cli\u003eGao, H., Wang, Y., Chen, P., Zhang, A., Zhou, X., \u0026amp; Zhuang, Q. (2024). Toxicity of Eight Insecticides on Drosophila suzukii and Its Pupal Parasitoid Trichopria drosophilae. \u003cem\u003eInsects, 15\u003c/em\u003e(11), 910-924. http://doi:10.3390/insects15110910\u003c/li\u003e\n\u003cli\u003eGao, Y., Ji, J., Xu, C., Wang, L., Zhang, K., Li, D., . . . Luo, J. (2024). Chromosome-level genome assembly of cotton thrips Thrips tabaci (Thysanoptera: Thripidae). \u003cem\u003eSci Data, 11\u003c/em\u003e(1), 1003-1015. http://doi:10.1038/s41597-024-03737-8\u003c/li\u003e\n\u003cli\u003eGao, Y., Su, S. C., Xing, J. Y., Liu, Z. Y., N\u0026auml;ssel, D. R., Bass, C., . . . Wu, S. F. (2025). Pesticide-induced resurgence in brown planthoppers is mediated by action on a suite of genes that promote juvenile hormone biosynthesis and female fecundity. \u003cem\u003eElife, 12\u003c/em\u003e. http://doi:10.7554/eLife.91774\u003c/li\u003e\n\u003cli\u003eGu, Z., Zhang, T., Long, S., Li, S., Wang, C., Chen, Q., . . . Cao, Y. (2023). Responses of Thrips hawaiiensis and Thrips flavus populations to elevated CO2 concentrations. \u003cem\u003eJournal of Economic Entomology, 116\u003c/em\u003e(2), 416-425. http://doi:10.1093/jee/toad026\u003c/li\u003e\n\u003cli\u003eGuo, J., An, J., Chang, H., Li, Y., Dang, Z., Wu, C., \u0026amp; Gao, Z. (2023). The Lethal and Sublethal Effects of Lambda-Cyhalothrin and Emamectin Benzoate on the Soybean Pest Riptortus pedestris (Fabricius). \u003cem\u003eToxics, 11\u003c/em\u003e(12), 971-988. http://doi:10.3390/toxics11120971\u003c/li\u003e\n\u003cli\u003eHou, Y. Y., Zang, Z. Y., L\u0026uuml;, W. J., Xu, W., Desneux, N., \u0026amp; Zang, L. S. (2024). Transgenerational hormesis and sublethal effects of five key insecticides for controlling Spodoptera frugiperda on its endoparasitoid Cotesia marginiventris. \u003cem\u003ePest Manag Sci, 80\u003c/em\u003e(4), 1681-1691. http://doi:10.1002/ps.7899\u003c/li\u003e\n\u003cli\u003eKhan, M. M., Ali, M. W., Hafeez, M., Fan, Z. Y., Ali, S., \u0026amp; Qiu, B. L. (2021). Lethal and sublethal effects of emamectin benzoate on life-table and physiological parameters of citrus red mite, Panonychus citri. \u003cem\u003eExp Appl Acarol, 85\u003c/em\u003e(2-4), 173-190. http://doi:10.1007/s10493-021-00667-7\u003c/li\u003e\n\u003cli\u003eKhan, M. M., Nawaz, M., Hua, H., Cai, W., \u0026amp; Zhao, J. (2018). Lethal and sublethal effects of emamectin benzoate on the rove beetle, Paederus fuscipes, a non-target predator of rice brown planthopper, Nilaparvata lugens. \u003cem\u003eEcotoxicol Environ Saf, 165\u003c/em\u003e, 19-24. http://doi:10.1016/j.ecoenv.2018.08.047\u003c/li\u003e\n\u003cli\u003eLi, W. H., Jin, D. C., Li, F. L., Cheng, Y., \u0026amp; Jin, J. X. (2017). Metabolic phenomics of bacterium Pantoea sp. from larval gut of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae). \u003cem\u003eSymbiosis, 72\u003c/em\u003e(2), 135-142. http://doi:10.1007/s13199-016-0453-4\u003c/li\u003e\n\u003cli\u003eLiu, K. X., Guo, Y., Zhang, C. X., \u0026amp; Xue, C. B. (2022). Sublethal effects and reproductive hormesis of emamectin benzoate on Plutella xylostella. \u003cem\u003eFront Physiol, 13\u003c/em\u003e, 1025959. http://doi:10.3389/fphys.2022.1025959\u003c/li\u003e\n\u003cli\u003eLiu, Z., Lyu, B., Lu, H., Tang, J. H., Zhang, Q. K., \u0026amp; Jiao, B. (2025). The toxicological mechanism of emamectin benzoate in Spodoptera frugiperda via regulating gut microbiota. \u003cem\u003eEntomologia Generalis, 45\u003c/em\u003e(1), 253-263. http://doi:10.1127/entomologia/2024/2913\u003c/li\u003e\n\u003cli\u003eLivak, K. J., \u0026amp; Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-\u0026Delta;\u0026Delta;CT method. \u003cem\u003eMethods, 25\u003c/em\u003e(4), 402-408. http://doi:10.1006/meth.2001.1262\u003c/li\u003e\n\u003cli\u003eL\u0026oacute;pez, J. D., Jr., Latheef, M. A., \u0026amp; Hoffmann, W. C. (2010). Effect of emamectin benzoate on mortality, proboscis extension, gustation and reproduction of the corn earworm, Helicoverpa zea. \u003cem\u003eJ Insect Sci, 10\u003c/em\u003e(89), 1-16. http://doi:10.1673/031.010.8901\u003c/li\u003e\n\u003cli\u003eMougabure-Cueto, G., Fronza, G., \u0026amp; Nattero, J. (2024). What happens when the insecticide does not kill? A review of sublethal toxicology and insecticide resistance in triatomines. \u003cem\u003eMedical and Veterinary Entomology\u003c/em\u003e. http://doi:10.1111/mve.12753\u003c/li\u003e\n\u003cli\u003eMoustafa, M. A. M., K\u0026aacute;kai, A., Awad, M., \u0026amp; F\u0026oacute;nagy, A. (2016). Sublethal effects of spinosad and emamectin benzoate on larval development and reproductive activities of the cabbage moth, Mamestra brassicae L. (Lepidoptera: Noctuidae). \u003cem\u003eCrop Protection, 90\u003c/em\u003e, 197-204. http://doi:10.1016/j.cropro.2016.09.004\u003c/li\u003e\n\u003cli\u003eNaeem-Ullah, U., Ramzan, M., Bokhari, S. H. M., Saleem, A., Qayyum, M. A., Iqbal, N., . . . Saeed, S. (2020). Insect Pests of Cotton Crop and Management Under Climate Change Scenarios. In \u003cem\u003eEnvironment, Climate, Plant and Vegetation Growth\u003c/em\u003e (pp. 367-396). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-030-49732-3_15\u003c/li\u003e\n\u003cli\u003eReitz, S. R., Gao, Y., Kirk, W. D. J., Hoddle, M. S., Leiss, K. A., \u0026amp; Funderburk, J. E. (2020). Invasion Biology, Ecology, and Management of Western Flower Thrips. \u003cem\u003eAnnu Rev Entomol, 65\u003c/em\u003e, 17-37. http://doi:10.1146/annurev-ento-011019-024947\u003c/li\u003e\n\u003cli\u003eRezk, A. A., Naqqash, M. N., Sattar, M. N., Mehmood, K., Elshafie, H., \u0026amp; Al-Khayri, J. M. (2024). Sublethal effect of emamectin benzoate on age-stage, two-sex life table and population projection of red palm weevil, Rhynchophorus ferrugineus. \u003cem\u003eSci Rep, 14\u003c/em\u003e(1), 22565. http://doi:10.1038/s41598-024-70042-0\u003c/li\u003e\n\u003cli\u003eShan, Y. X., Zhu, Y., Li, J. J., Wang, N. M., Yu, Q. T., \u0026amp; Xue, C. B. (2020). Acute lethal and sublethal effects of four insecticides on the lacewing (Chrysoperla sinica Tjeder). \u003cem\u003eChemosphere, 250\u003c/em\u003e, 126321. http://doi:10.1016/j.chemosphere.2020.126321\u003c/li\u003e\n\u003cli\u003eShen, X. J., Chen, J. C., Cao, L. J., Ma, Z. Z., Sun, L. N., Gao, Y. F., . . . Wei, S. J. (2023). Interspecific and intraspecific variation in susceptibility of two co-occurring pest thrips, Frankliniella occidentalis and Thrips palmi, to nine insecticides. \u003cem\u003ePest Manag Sci, 79\u003c/em\u003e(9), 3218-3226. http://doi:10.1002/ps.7502\u003c/li\u003e\n\u003cli\u003eSmith-Unna, R., Boursnell, C., Patro, R., Hibberd, J. M., \u0026amp; Kelly, S. (2016). TransRate: reference-free quality assessment of de novo transcriptome assemblies. \u003cem\u003eGenome Research, 26\u003c/em\u003e(8), 1134-1144. http://doi:10.1101/gr.196469.115\u003c/li\u003e\n\u003cli\u003eSong, F., Shu, J. P., \u0026amp; Zhang, S. K. (2025). Gut bacterium Acinetobacter sp. assists Camellia weevil with host plant adaptation by degrading tea saponin via the benzoate pathway. \u003cem\u003eMicrobiome, 13\u003c/em\u003e(1), 139-165. http://doi:10.1186/s40168-025-02131-9\u003c/li\u003e\n\u003cli\u003eSun, Y., Hu, C., Chen, G., Li, X., Liu, J., Xu, Z., . . . Zhang, X. (2023). Insecticide-mediated changes in the population and toxicity of the thrips species, Frankliniella occidentalis (Pergande) and Thrips flavus (Schrank) (Thysanoptera: Thripidae). \u003cem\u003eJournal of Economic Entomology, 117\u003c/em\u003e(1), 293-301. http://doi:10.1093/jee/toad226\u003c/li\u003e\n\u003cli\u003eTang, C., Hu, X., Tang, J. F., Wang, L., Liu, X. W., Peng, Y. F., . . . Xie, J. Q. (2024). The symbiont Acinetobacter baumannii enhances the insect host resistance to entomopathogenic fungus Metarhizium anisopliae. \u003cem\u003eCommunications Biology, 7\u003c/em\u003e(1), 1184. http://doi:10.1038/s42003-024-06779-1\u003c/li\u003e\n\u003cli\u003eWakil, W., Gulzar, S., Wu, S., Rasool, K. G., Husain, M., Aldawood, A. S., \u0026amp; Toews, M. D. (2023). Development of Insecticide Resistance in Field Populations of Onion Thrips, Thrips tabaci (Thysanoptera: Thripidae). \u003cem\u003eInsects, 14\u003c/em\u003e(4), 376. http://doi:10.3390/insects14040376\u003c/li\u003e\n\u003cli\u003eYang, D. S., Cui, B., Wang, C. X., Zhao, X., Zeng, Z. H., Wang, Y., . . . Cui, H. X. (2017). Preparation and Characterization of Emamectin Benzoate Solid Nanodispersion. \u003cem\u003eJournal of Nanomaterials, 15\u003c/em\u003e(7), 495-511. http://doi:10.1155/2017/6560780\u003c/li\u003e\n\u003cli\u003eYang, X., Sun, L., Chi, H., Kang, G., \u0026amp; Zheng, C. (2020). Demography of Thrips palmi (Thysanoptera: Thripidae) Reared on Brassica oleracea (Brassicales: Brassicaceae) and Phaseolus vulgaris (Fabales: Fabaceae) With Discussion on the Application of the Bootstrap Technique in Life Table Research. \u003cem\u003eJ Econ Entomol, 113\u003c/em\u003e(5), 2390-2398. http://doi:10.1093/jee/toaa171\u003c/li\u003e\n\u003cli\u003eZhou, L., Luo, F. J., Zhang, X. Z., Jiang, Y. P., Lou, Z. Y., \u0026amp; Chen, Z. M. (2016). Dissipation, transfer and safety evaluation of emamectin benzoate in tea. \u003cem\u003eFood Chemistry, 202\u003c/em\u003e, 199-204. http://doi:10.1016/j.foodchem.2015.11.069\u003c/li\u003e\n\u003cli\u003eZhu, W., Guo, Q., Chen, M., Wang, J., Zhang, Y., \u0026amp; Ma, R. (2025). Comprehensive assessment of the acute lethal, risk level, and sub-lethal effects of four insecticides on Trichogramma ostriniae. \u003cem\u003ePLoS One, 20\u003c/em\u003e(6), e0325733. http://doi:10.1371/journal.pone.0325733\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-pest-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pest","sideBox":"Learn more about [Journal of Pest Science](https://www.springer.com/journal/10340)","snPcode":"10340","submissionUrl":"https://submission.nature.com/new-submission/10340/3","title":"Journal of Pest Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"pests, insecticide toxicity, sublethal effects, growth and development, transcriptome, 16S rRNA sequencing","lastPublishedDoi":"10.21203/rs.3.rs-8257750/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8257750/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cem\u003eThrips tabaci\u003c/em\u003e (Tysanoptera: Tripidae) is a globally significant agricultural pest whose control relies heavily on insecticides. This study comprehensively assessed the toxicity and sublethal effects of emamectin benzoate (EB) against \u003cem\u003eT. tabaci\u003c/em\u003e and explored its underlying mechanisms. EB exhibited high acute toxicity, and at the LC\u003csub\u003e20\u003c/sub\u003e level, it significantly reduced longevity and fecundity in the F\u003csub\u003e0\u003c/sub\u003e generation, with these inhibitory effects persisting into the F\u003csub\u003e1\u003c/sub\u003e generation. Molecular analyses revealed that the P450 gene \u003cem\u003eCYP6K1\u003c/em\u003e is crucial in the \u003cem\u003eT. tabaci\u003c/em\u003e\u0026rsquo;s response to insecticide stress, and RNAi-mediated suppression of \u003cem\u003eCYP6K1\u003c/em\u003e expression significantly increased susceptibility. Additionally, sublethal dose EB exposure altered the gut microbiota, marked by a decline in \u003cem\u003eAcinetobacter\u003c/em\u003e and an expansion of \u003cem\u003ePantoea\u003c/em\u003e. We propose that \u003cem\u003eT. tabaci\u003c/em\u003e mounts a coordinated defense mediated by both the host \u003cem\u003eCYP6K1\u003c/em\u003e gene and intestinal symbionts, and disruption of this system impairs tolerance. These findings provide multi-level insights into the effect of EB on \u003cem\u003eT. tabaci\u003c/em\u003e, supporting the development of integrated management strategies targeting both host detoxification and microbial communities.\u003c/p\u003e","manuscriptTitle":"Sublethal emamectin benzoate suppresses multi-transgenerational reproduction and alters symbiotic bacteria of Thrips tabaci","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-14 16:50:35","doi":"10.21203/rs.3.rs-8257750/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-06T14:22:29+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-30T07:56:50+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-28T09:33:26+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-22T12:07:09+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-16T18:02:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"131117377877492037709470459854139451382","date":"2026-01-13T12:20:52+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"92006354007693908485690762206119402813","date":"2026-01-13T08:28:31+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"162325690267915839851267997213868969994","date":"2026-01-13T04:04:35+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"311385935516715826272259578717992441465","date":"2026-01-12T15:16:06+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-12T15:10:03+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-12-03T08:08:11+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-12-03T08:07:38+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Pest Science","date":"2025-12-02T08:04:54+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-pest-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pest","sideBox":"Learn more about [Journal of Pest Science](https://www.springer.com/journal/10340)","snPcode":"10340","submissionUrl":"https://submission.nature.com/new-submission/10340/3","title":"Journal of Pest Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"78a58926-26ba-4711-84c4-0cf6a68cd377","owner":[],"postedDate":"January 14th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-03-26T13:56:35+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-14 16:50:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8257750","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8257750","identity":"rs-8257750","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}