The microRNA-7322-5p/p38/Hsp19 axis modulates Chilo suppressalis cell-defences against Cry1Ca: an effective target for a stacked transgenic rice approach.

Bacillus thuringiensis (Bt)-secreted crystal (Cry) toxins form oligomeric pores in host cell membranes and are a common element in generating insect-resistant transgenic crops. Although Cry toxin function has been well documented, cellular defences against pore-formation have not been as well developed. Elucidation of the processes underlying this defence, however, could contribute to the development of enhanced Bt crops. Here, we demonstrate that Cry1Ca-mediated downregulation of microRNA-7322-5p (miR-7322-5p), which binds to the 3' untranslated region of p38, negatively regulates the susceptibility of Chilo suppressalis to Cry1Ca. Moreover, Cry1Ca exposure enhanced phosphorylation of Hsp19, and hsp19 downregulation increased susceptibility to Cry1Ca. Further, Hsp19 phosphorylation occurs downstream of p38, and pull-down assays confirmed the interactions between Hsp19 and Cry1Ca, suggesting that activation of Hsp19 by the miR-7322-5p/p38/Hsp19 pathway promotes Cry1Ca sequestration. To assess the efficacy of targeting this pathway in planta, double-stranded RNA (dsRNA) targeting C. suppressalis p38 (dsp38) was introduced into a previously generated cry1Ca-expressing rice line (1CH1-2) to yield a single-copy cry1Ca/dsp38 rice line (p38-rice). Feeding on this rice line triggered a significant reduction in C. suppressalis p38 expression and the line was more resistant to C. suppressalis than 1CH1-2 in both short term (7-day) and continuous feeding bioassays as well as field trials. These findings provide new insights into invertebrate epithelium cellular defences and demonstrate a potential new pyramiding strategy for Bt crops.

Transcriptomic analysis of salivary gland and proteomic analysis of oral secretion in Helicoverpa armigera under cotton plant leaves, gossypol, and tannin stresses.

Gossypol and tannin are involved in important chemical defense processes in cotton plants. In this study, we used transcriptomics and proteomics to explore the changes in salivary gland functional genes and oral secretion (OS) proteins after feeding with artificial diet (containing gossypols and tannins) and cotton plant leaves. We found that dietary cotton plant leaves, gossypols and tannins exerted adverse impacts on the genes that regulated the functions of peptidase, GTPase, glycosyl hydrolases in the salivary glands of the Helicoverpa armigera (H. armigera). However, GST, UGT, hydrolases, and lipase genes were up-regulated to participate in the detoxification and digestive of H. armigera. The oral secretory proteins of H. armigera were significantly inhibited under the stress of gossypol and tannin, such as enzyme activity, but some proteins (such as PZC71358.1) were up-regulated and involved in immune and digestive functions. The combined analysis of transcriptomics and metabolomics showed a weak correlation, and the genes and proteins involved were mainly in digestive enzyme activities. Our work clarifies the deleterious physiological impacts of gossypols and tannins on H. armigera and reveals the mechanism by which H. armigera effectively mitigate the phytotoxic effects through detoxification and immune systems.

Isolation and functional analysis of OsAOS1 promoter for resistance to Nilaparvata lugens Stål infestation in rice.

Insect pests have a great impact on the yield and quality of crops. Insecticide applications are an effective method of pest control, however, they also have adverse effects on the environment. Using insect-inducible promoters to drive insect-resistant genes in transgenic crops is a potential sustainable pest management strategy, but insect-inducible promoters have been rarely reported. In this study, we found rice allene oxide synthase gene (AOS, LOC_Os03g12500) can be highly upregulated following brown planthopper (Nilaparvata lugens Stål, BPH) infestation. Then, we amplified the promoter of OsAOS1 and the ß- glucuronidase reporter gene was used to analyze the expression pattern of the promoter. Through a series of 5' truncated assays, three positive regulatory regions in response to BPH infestation in the promoter were identified. The transgenic plants, P1R123-min 35S and P1TR1-min 35S promoter-driven snowdrop lectin (Galanthus nivalis agglutinin, GNA) gene, demonstrated the highest expression levels of GNA and lowest BPH survival. Our work identified a BPH-inducible promoter and three positive regions within it. Transgenic rice with GNA driven by OsAOS1 promoter and positive regions exhibited an expected lethal effect on BPH. This study proved the application potential of BPH-inducible promoter and provided a novel path for the selection of insect-resistant tools in the future.

Repellency, contact toxicity, and anti-oviposition effects of three ethanol-extracted plant essential oils on Bemisia tabaci (Hemiptera: Aleyrodidae).

There are more than 1200 species of whiteflies found across the globe. Due to the high level of resistance of whitefly against synthetic insecticides, alternate pest management measures have their significance. Plant essential oils (EOs) affect insect pests in many ways, such as via stimulatory, deterrent, toxic, and hormonal effects. This study was designed to determine the repellency of EOs, toxicity, and oviposition deterrent activities of Allium ascalonicum, Cinnamomum camphora, and Mentha haplocalyx against adult whiteflies. In repellency determination experiments, a single tomato plant was treated with 10 ml of ethanol-extracted EO with 1000 ppm concentration. Results showed that C. camphora EO was the most repellent for whitefly compared to M. haplocalyx and A. ascalonicum. The oviposition deterrent experiments revealed that C. camphora has the highest oviposition deterrent effect, followed by M. haplocalyx and A. ascalonicum. A single plant treatment method was used to assess the contact toxicity of three EOs against whitefly after 12, 24, 48, and 72 h of exposure. The results revealed that C. camphora is more toxic to whitefly than M. haplocalyx and A. ascalonicum. After determining the antagonistic effects of these EOs, the oils were analyzed using gas chromatography/mass spectrometry to identify the chemical components. It can be concluded that C. camphora is the most effective oil EO in terms of toxicity, repellence, and oviposition deterrence, followed by M. haplocalyx and A. ascalonicum under greenhouse conditions. Our results introduce some new eco-friendly plant EOs to control whiteflies.

Notch is an alternative splicing gene in brown planthopper, Nilaparvata lugens.

Much research has assumed that Notch codes one protein. Then the protein will be cleaved into two parts and regenerates a heterodimers receptor to construct Notch signal pathways to regulate development in the past three decades. Here, we show that Notch in brown planthopper is a complex alternatively spliced gene has at least three transcriptional start sites, four exon skips, and 21 transcriptional endpoints that uses these to form variants and codes a series of proteins. When used dsRNAs to suppression different regions of the full-length variant NlNF resulted in a similar phenotype. Insects were molting after treatment, sensation circles on antennas near to root decayed, bristles on wings shortened, thickened or disappeared, accompanied by thickening veins and blades of fore-wing apex regions thickened. These results suggested that Notch influenced developmental of sensation circles, bristles, veins, and blades in nymph late periods. This study has deepened our understanding of Notch.

miR-34 modulates wing polyphenism in planthopper.

Polyphenism is a successful strategy adopted by organisms to adapt to environmental changes. Brown planthoppers (BPH, Nilaparvata lugens) develop two wing phenotypes, including long-winged (LW) and short-winged (SW) morphs. Though insulin receptor (InR) and juvenile hormone (JH) have been known to regulate wing polyphenism in BPH, the interaction between these regulators remains largely elusive. Here, we discovered that a conserved microRNA, miR-34, modulates a positive autoregulatory feedback loop of JH and insulin/IGF signaling (IIS) pathway to control wing polyphenism in BPH. Nlu-miR-34 is abundant in SW BPHs and suppresses NlInR1 by targeting at two binding sites in the 3'UTR of NlInR1. Overexpressing miR-34 in LW BPHs by injecting agomir-34 induces the development towards SW BPHs, whereas knocking down miR-34 in SW BPHs by injecting antagomir-34 induces more LW BPHs when another NlInR1 suppressor, NlInR2, is also suppressed simultaneously. A cis-response element of Broad Complex (Br-C) is found in the promoter region of Nlu-miR-34, suggesting that 20-hydroxyecdysone (20E) might be involved in wing polyphenism regulation. Topic application of 20E downregulates miR-34 expression but does not change wing morphs. On the other hand, JH application upregulates miR-34 expression and induces more SW BPHs. Moreover, knocking down genes in IIS pathway changes JH titers and miR-34 abundance. In all, we showed that miRNA mediates the cross talk between JH, 20E and IIS pathway by forming a positive feedback loop, uncovering a comprehensive regulation mechanism which integrates almost all known regulators controlling wing polyphenism in insects.

Risk factors and the utility of three different kinds of prediction models for postoperative fatigue after gastrointestinal tumor surgery.

BACKGROUND: Postoperative fatigue (POF) is a common complication after gastrointestinal tumor surgery, and it also brings negative effect on prognosis and life quality. However, there are no prediction models for POF, and studies of risk factors are not comprehensive. Therefore, the aim of this study is to investigate the risk factors and pick out the best prediction model for POF and to validate it. METHODS: A prospective study was conducted for patients undergoing elective gastrointestinal tumor surgery. Physiological, psychological, and socioeconomic factors were collected. Logistic regression, back-propagation artificial neural networks (BP-ANNs), and classification and regression tree (CART) were constructed and compared. RESULTS: A total of 598 patients consisting of 463 derivation sample and 135 validation sample were included. The incidence of POF in derivation sample, validation sample, and total were 58.3%, 57.0%, and 58.7%, respectively. Logistic regression results showed age, higher degree of education, lower personal monthly income, advanced cancer, hypoproteinemia, preoperative anxiety or depression, and limited social support were risk factors for POF. Receiver operating characteristic curve (ROC) and performance indices were used to test three models. BP-ANN was the best by the comparison of models, and its strong predictive performance was also validated. CONCLUSIONS: More attention should be paid on specific patients after gastrointestinal tumor surgery. BP-ANN is a powerful mathematical tool that could predict POF exactly. It may be used as a noninvasive screening tool to guide clinicians for early identification of high-risk patients and grading interventions.

Emamectin benzoate induced enzymatic and transcriptional alternation in detoxification mechanism of predatory beetle Paederus fuscipes (Coleoptera: Staphylinidae) at the sublethal concentration.

In this study, the detoxification enzyme activity and the transcriptional profile changes in the second instar through RNA-sequencing technology due to emamectin benzoate (EB) were assessed. The cytochrome P450 monooxygenases (P450) enzyme activity was not altered by EB due to the change in concentration and exposure time in all treatments. The glutathione S-transferase (GST) enzyme was not considerably varying in all treatments, while exposure time significantly changed the enzyme activity. Results showed that the esterase (Ests) activity was elevated with the increasing concentrations and exposure time. Two libraries were generated, containing 107,767,542 and 108,142,289 clean reads for the samples treated with LC30 of EB and control. These reads were grouped into 218,070 transcripts and 38,097 unigenes. A total of 2257 differentially expressed genes (DEGs) were identified from these unigenes, of which 599 up-regulated and 1658 were down-regulated. The majority of these DEGs related to pesticides resistance were identified in numerous Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, e.g., steroid hormone biosynthesis, glutathione metabolism, drug metabolism-other enzymes, chemical carcinogenesis, pathways of cancer, metabolism of xenobiotics by cytochrome P450, drug metabolism of cytochrome P450, linoleic acid metabolism, retinol metabolism, and insect hormone biosynthesis. These pathways also shared the same genes as cytochrome P450 monooxygenases (P450s), glutathione S-transferases (GSTs), Esterase (Ests), UDP-glucosyltransferases (UGTs), and ATP-binding cassettes (ABCs). A heatmap analysis also showed that regulation of genes in a pathway causes a series of gene expression regulation in subsequent pathways. Our quantitative reverse transcription-PCR (qRT-PCR) results were consistent with the DEG's data of transcriptome analysis. The comprehensive transcriptome sequence resource attained through this study evidence that the EB induces significant modification in enzyme activity and transcriptome profile of Paederus fuscipes, which may enable more significant molecular underpinnings behind the insecticide-resistance mechanism for further investigations.

Decapentaplegic function in wing vein development and wing morph transformation in brown planthopper, Nilaparvata lugens.

The decapentaplegic (dpp) gene plays a variety of roles in diverse cellular and molecular processes of the growth and development. In insects, dpp is mainly required for dorsal-ventral patterning and appendage formation. The brown planthopper (BPH) Nilaparvata lugens, a major pest of rice, possesses two distinct wing morphs described as long-winged (LW) and short-winged (SW) morphs. With our lab-maintained stable strains of LW and SW BPH, RNA interference (RNAi) was used to research the functions of N. lugens dpp (Nldpp) on wing development. Silencing of Nldpp in the SW strain led to the significant lengthening of the forewing, while Nldpp-knockdown in the LW strain resulted in distorted wings. Moreover, knockdown of Nldpp caused the complete absence of wing veins. During the development of wing-pads, the Nldpp abundance in the terga of the SW strain was significantly higher than that of the LW strain. Through controlling the direction of wing morph transformation, we found that the expression level of Nldpp increased in the NlInR1-knockdown BPH (tending to SW) and abundance of Nldpp declined after dsNlInR2 injection (tending to LW). Our results showed that Nldpp is mainly responsible for the formation and development of veins in BPH. Also, Nldpp can be regulated by NlInR1/2 and participate in the wing morph transformation.

Diversity, Tissue Localization, and Infection Pattern of Bacterial Symbionts of the White-Backed Planthopper, Sogatella furcifera (Hemiptera: Delphacidae).

The white-backed planthopper (WBPH), Sogatella furcifera (Horváth), is a destructive pest of rice. Bacterial symbionts play an important role in insect hosts, especially hemipteran hosts. This study was designed to examine the bacterial symbionts of the WBPH using 16S rDNA high-throughput sequencing. A total of 63 and 177 operational taxonomic units (OTUs) were identified in females and males of three WBPH populations, respectively. These OTUs included bacteria of 75 genera from 11 phyla, where Wolbachia, Cardinium, and Asaia were the dominant genera, accounting for over 97.99% of all the symbiotic bacteria. Fluorescence in situ hybridization detected Wolbachia, Cardinium, and Asaia in the salivary glands, guts, testes, and eggs of the WBPH, indicating the potential for both horizontal and vertical transmission. Moreover, the infection pattern of the three dominant bacterial symbionts was detected in six WBPH populations. The frequencies of Wolbachia infection of females and Cardinium infection of both sexes were over 96.7%. Wolbachia infection of males ranged between 46.7 and 63.3%, which was significantly lower than that observed for females. Asaia infection of both sexes varied substantially among the populations. These results indicate that the complex host-symbiotic bacteria interaction is influenced by host sex and geographical origin and potentially by the transmission modes of the symbionts.

The inhibitory effect of microRNA-1827 on anoikis resistance in lung adenocarcinoma A549 cells via targeting caveolin-1.

Anoikis resistance is a critical process for cancer cell metastasis in non-small cell lung cancer (NSCLC), and microRNA-1827 (miR-1827) is closely correlated with NSCLC metastasis. In this study, we aimed to evaluate the role of miR-1827 in regulating the anoikis resistance of NSCLC. The results showed that miR-1827 level was decreased in tumor tissues and cells and was correlated with tumor grade and lymph node (LN) metastasis. Overexpression of miR-1827 inhibited anchorage-independent growth and anoikis resistance in A549 cells. Bioinformatics and functional analysis identified that caveolin-1 (CAV-1) is directly targeted by miR-1827. Restoration of CAV-1 significantly attenuated miR-1827's effect on anoikis resistance in A549 cells. Our data identified a novel signaling axis of miR-1827/CAV-1 in regulating anoikis resistance, which might serve as a potential therapeutic target for metastatic NSCLC.

Safety assessment of transgenic Cry2Aa rice to a generalist predator, Paederus fuscipes Curtis (Coleoptera: Staphylinidae).

The insecticidal crystal proteins of Cry2A family from Bacillus thuringiensis (Bt) are important candidate proteins expressed in gene pyramiding Bt crops. A transgenic rice line (T2A-1) harboring a synthetic Cry2A* (Cry2Aa) gene showed effective resistance to some lepidopteran rice pests. As a generalist predator in rice ecosystems, the rove beetle (Paederus fuscipes) can prey on many rice insect pests such as planthoppers. Considering the possible exposure of Cry2Aa to P. fuscipes through tritrophic food chain, it is necessary to assess the potential risks of T2A-1 rice to this predator. In this study, a tritrophic experiment was conducted to assess the prey-mediated effects of Cry2Aa on P. fuscipes through the T2A-1 rice-Nilaparvata lugens-P. fuscipes food chain. After preying on N. lugens nymphs reared on T2A-1, no accumulated Cry2Aa could be detected in P. fuscipes adults, despite Cry2Aa being detected in N. lugens. In addition, no harmful effects were detected on the life table parameters of P. fuscipes in this tritrophic chain. Additionally, direct exposure to a high dose of purified Cry2Aa protein, representing the worst case scenario, showed no significant adverse effects on the development of P. fuscipes. These results showed that transgenic Cry2Aa rice had no harmful effects on P. fuscipes.

Assessing the effects of Cry2Aa protein on Habrobracon hebetor (Hymenoptera: Braconidae), a parasitoid of Indian meal moth, Plodia interpunctella (lepidoptera: Pyralidae).

Transgenic crops express Cry proteins exhibit high resistant to target insect pests. When we evaluate the effects of Cry proteins on the parasitoid of target insect pest via tritrophic experiments (transgenic plant-target insect pest-parasitoid) host quality of parasitoids might decrease because of insecticidal protein ingestion, this would cause host-quality mediated effects and influence the accuracy of biosafety assessment. In the current study, high dose of Cry2Aa protein was injected into the hemolymph of Plodia interpunctella by microinjection, and the hemolymph was used as the carrier to deliver Cry protein to Habrobracon hebetor, which has been previously reported as an ectoparasitoid of P. interpunctella larval, in order to avoid the "host-quality mediated effects". Results showed that injected Cry2Aa remained at high concentration and bioactive in the hemolymph of P. interpunctella parasitized by H. hebetor, the hemolymph of P. interpunctella could be used as carriers of Cry protein to H. hebetor, and high dose of Cry2Aa have no negative impacts on the development time, weight of pupa, sex ratio, adults weight (male and female), adult longevity and fecundity, and the activity of stress-related enzymes of H. hebetor. However, the hemolymph of P. interpunctella injected into Galanthus nivalis agglutinin (the positive control) showed significant negative impact on these parameters measured in the present study of H. hebetor. This indicated that Cry2Aa protein had no detrimental effects on the biological parameters of H. hebetor measured in the current study. Meanwhile, this study provides a new method for the safety evaluation of the ectoparasitoids of target pest and might be expanded to the other species of ectoparasitoids of target insects of Cry proteins in biosafety risk assessment.

Effect of sarcopenia on clinical outcomes following digestive carcinoma surgery: a meta-analysis.

BACKGROUND: The effect of sarcopenia on digestive carcinoma surgery outcomes is controversial. We aimed to assess the effect of sarcopenia defined by the European Working Group on Sarcopenia in Older People (EWGSOP) or the Asian Working Group for Sarcopenia (AWGS) on outcomes following digestive carcinoma surgery. METHODS: Eligible studies were searched from PubMed, EMBASE and other databases from inception to April 2018. We conducted a meta-analysis to estimate the risk ratios or mean differences of outcomes in the sarcopenia group versus the non-sarcopenia group. Stratified analyses and sensitivity analyses were performed. RESULTS: We included 11 cohort studies, with a sarcopenia prevalence ranging from 11.6 to 33.0%. Sarcopenia was associated with an increased risk of total complications (RR = 1.87, P < 0.00001), major complications (RR = 2.45, P = 0.002), re-admissions (RR = 2.53,P < 0.0001), infections (RR = 2.23, P = 0.09), severe infections (RR = 2.96, P = 0.04), 30-day mortality (RR = 3.36, P = 0.001), longer hospital stay (MD = 4.61, P = 0.001) and increased hospitalization expenditures (SMD = 0.25, P = 0.02). Sarcopenia differentially affected outcomes when stratified, and the results were stable. CONCLUSIONS: Sarcopenia defined by the EWGSOP or AWGS Consensus was a high-risk factor for digestive carcinoma surgery outcomes. Different tumour site and muscle mass measurements are the sources of heterogeneity. More high-quality studies are needed.

Candidate detoxification-related genes in brown planthopper, Nilaparvata lugens, in response to ß-asarone based on transcriptomic analysis.

Nilaparvata lugens(Stål) is a serious pest of rice and has evolved different levels of resistance against most chemical pesticides. ß-asarone is the main bioactive insecticidal compound of Acorus calamus L. that shows strong insecticidal activity against pests. In this study, we conducted a bioassay experiment to determine the contact toxicity of ß-asarone to N. lugens nymphs. The LD30 sublethal dose was 0.106 µg per nymph, with 95% confidence limits of 0.070-0.140 µg. We applied the LD30 concentration of ß-asarone to nymphs for 24 h or 72 h and then performed a transcriptome sequence analysis by referencing the N. lugens genome to characterize the variation. The transcriptomic analysis showed that several GO terms and KEGG pathways presented significant changes. Individually, 126 differentially expressed genes (DEGs), including 72 upregulated and 54 downregulated genes, were identified at 24 h, and 1771 DEGs, including 882 upregulated and 889 downregulated genes, were identified at 72 h. From the DEGs, we identified a total of 40 detoxification-related genes, including eighteen Cytochrome P450 monooxygenase genes (P450s), three Glutathione S-transferase genes, one Carboxylesterase gene, twelve UDP-glucosyltransferases and six ATP-binding cassette genes. We selected the eighteen P450s for subsequent verification by quantitative PCR. These findings indicated that ß-asarone presented strong contact toxicity to N. lugens nymphs and induced obvious variation of detoxification-related genes that may be involved in the response to ß-asarone.

Lethal and sublethal effects of emamectin benzoate on the rove beetle, Paederus fuscipes, a non-target predator of rice brown planthopper, Nilaparvata lugens.

The use of pesticides in rice can not only manage the pest but also influence non-target organisms. The rove beetle (Paederus fuscipes), which is an important predator of the brown plant hopper (Nilaparvata lugens) in rice ecosystems, was tested to investigate acute and chronic effects of emamectin benzoate. The results from this study show that the LC50 of emamectin benzoate to adults of P. fuscipes at 72 h was 3.07 (1.84-4.54) mg a.i. L-1, whereas the LC50 of emamectin benzoate to the second instar larvae of P. fuscipes at 72 h was 2.58 (1.95-3.19) mg a.i. L-1. Tested sublethal doses (LC10 and LC30) had significant effects on the second instar developmental time of P. fuscipes compared with that of the control. The LC30 dose had a negative influence on the pre-imaginal developmental duration and the feeding potential of treated P. fuscipes larvae. Additionally, the LC30 reduced the pre-oviposition period, the fecundity and the body weight of adults emerged from treated larvae of P. fuscipes. In the sublethal experiment with adults, the fecundity and the feeding potential were significantly reduced at the LC30 dose. These results revealed that sublethal doses of emamectin benzoate negatively affected the development and biological activities of P. fuscipes, and we conclude that more attention should be paid to the use of this chemical as part of integrated pest management strategies.

Bt rice in China - focusing the nontarget risk assessment.

Bt rice can control yield losses caused by lepidopteran pests but may also harm nontarget species and reduce important ecosystem services. A comprehensive data set on herbivores, natural enemies, and their interactions in Chinese rice fields was compiled. This together with an analysis of the Cry protein content in arthropods collected from Bt rice in China indicated which nontarget species are most exposed to the insecticidal protein and should be the focus of regulatory risk assessment.

Knockdown of TOR causing ovarian diapause in a genetically stable brachypterous strain of Nilaparvata lugens.

Brown planthopper (BPH), Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), is one of the most damaging pests of rice crops. BPH is a migratory insect with a delayed ovarian development in migrants classified as reproductive diapause. The molecular mechanism of reproductive diapause remains unclear, although we suspect it might be regulated by one or more nutrient signaling pathways. The target of rapamycin (TOR) pathway regulates cell growth in response to nutritional information, which raised a hypothesis that TOR mediates BPH reproductive diapause. We used a pure brachypterous strain (BS) and a predominantly macropterous strain (MS) to investigate the roles of NlTOR in BPH reproductive diapause. We found that NlTOR is expressed from the nymphal to adult stages, with a higher expression level of NlTOR in BS adults at 1, 2, and 4 days posteclosion than in MS at the same time points. Injection of dsNlTOR into BS nymphs resulted in the termination of BPH female ovary development and the retardation of nymph development. We infer that TOR signaling functions in BPH reproductive diapause by regulating the expression of NlFoxA and NlVitellogenin.

Integrated Omics Analysis Reveals Key Pathways in Cotton Defense against Mirid Bug (Adelphocoris suturalis Jakovlev) Feeding.

The recent dominance of Adelphocoris suturalis Jakovlev as the primary cotton field pest in Bt-cotton-cultivated areas has generated significant interest in cotton pest control research. This study addresses the limited understanding of cotton defense mechanisms triggered by A. suturalis feeding. Utilizing LC-QTOF-MS, we analyzed cotton metabolomic changes induced by A. suturalis, and identified 496 differential positive ions (374 upregulated, 122 downregulated) across 11 categories, such as terpenoids, alkaloids, phenylpropanoids, flavonoids, isoflavones, etc. Subsequent iTRAQ-LC-MS/MS analysis of the cotton proteome revealed 1569 differential proteins enriched in 35 metabolic pathways. Integrated metabolome and proteome analysis highlighted significant upregulation of 17 (89%) proteases in the α-linolenic acid (ALA) metabolism pathway, concomitant with a significant increase in 14 (88%) associated metabolites. Conversely, 19 (73%) proteases in the fructose and mannose biosynthesis pathway were downregulated, with 7 (27%) upregulated proteases corresponding to the downregulation of 8 pathway-associated metabolites. Expression analysis of key regulators in the ALA pathway, including allene oxidase synthase (AOS), phospholipase A (PLA), allene oxidative cyclase (AOC), and 12-oxophytodienoate reductase3 (OPR3), demonstrated significant responses to A. suturalis feeding. Finally, this study pioneers the exploration of molecular mechanisms in the plant-insect relationship, thereby offering insights into potential novel control strategies against this cotton pest.