【Aim】 The purpose of this study is to evaluate the effects of flonicamid on the predation ability of the 3rd instar larvae of Harmonia axyridis. 【Methods】 The residual film method was employed to observe and analyze the effects of exposure of the 3rd instar larvae of H. axyridis to 0.146 and 0.292 mg/mL of flonicamid on their predation ability on Aphis gossypii adults, including the predation amount, instantaneous attack rate, handling time and searching efficiency.【Results】 The predatory functional responses of the 3rd instar larvae of H. axyridis treated with 0.146 and 0.292 mg/mL of flonicamid to A. gossypii adults followed the type Ⅱ functional responses, which can be described by Holling’s disc equation. Compared with the control group treated with acetone, exposure of 0.146 and 0.292 mg/mL of flonicamid had negative impacts on the predation functional response parameters and searching efficiency of the 3rd instar larvae of H. axyridis， and the effects on the predation and searching ability became more significant with the increase of the concentration of flonicamid. In the control group, and treatment groups with 0.146 and 0.292 mg/mL of flonicamid, the instantaneous attack rates of the 3rd instar larvae of H. axyridis were 0.9260, 1.4451 and 2.1197, respectively, the maximum daily predation amounts were 392.62, 52.63 and 32.15 individuals, respectively, and the handling time was 0.0025, 0.0190 and 0.0311 h, respectively. 【Conclusion】 Exposure of the 3rd instar larvae of H. axyridis to flonicamid has negative impacts on their predation function on A. gossypii adults. Therefore, in order to better protect and utilize natural enemies, when implementing the prevention and control of A. gossypii using H. axyridis, consideration should be given to reducing or not using chemical pesticides.

 Invasive alien insects, as dangerous pests in newly introduced areas, present challenges such as delayed detection, difficult monitoring, rapid outbreaks and incomplete eradication. These issues have long been the emphases and difficulties in the field of biosecurity worldwide. In this article, we made an overview of the major progress in the studies on the mechanisms of population outbreak and causing disaster, monitoring and early warning technologies, and control measures for invasive alien insects in China. We also summarized and introduced the main contents of this special issue from three aspects: The researches on population dynamics monitoring, mechanisms of insect resistance, and green control technologies for pest insects. Finally, we prospected the development trends of standardization, informatization, intelligence, and greening of monitoring and control of invasive alien insects in the future, and proposed the key directions for future control and management strategies for these pests, in order to promote more efficient, integrated and sustainable control approaches through technological innovation.

【Aim】To explore the role of heat shock protein Hsp70 genes of Myzus persicae in response to high and low temperature stress. 【Methods】 RT-qPCR was used to detect the relative expression levels of eight MpHsp70 genes (MpHsp70-1, MpHsp70-2, MpHsp70A1, MpHsp70B2, MpHsp68a, MpHsp68b, MpHsc70-4 and MpHsp70) in different wingless adult tissues (head, midgut, embryo and cuticle) and wingless adults under high temperature (36 ℃) and low temperature (4 ℃) stress for different duration (0, 30, 60, 90, 120 and 150 min), respectively. RNAi was used to silence two key MpHsp70 genes (MpHsp70A1 and MpHsp68a), and the survival rate and number of nymphs produced of wingless adults were observed and calculated at 120 min under high temperature treatment (36 ℃) and 30 min under low temperature treatment (4 ℃).【Results】 The eight MpHsp70 genes were expressed in different wingless adult tissues of M. persicae, and the expression levels of MpHsp70, MpHsp70A1, MpHsp70B2, MpHsc70-4 and MpHsp68b in the cuticile were significantly higher than those in the other tissues. The expression levels of MpHsp70-1, MpHsp70-2 and MpHsp68a in the embryo of M. persicae were significantly higher than those in other tissues. High and low temperature stress had significant induction effect on the expression of MpHsp70 genes in wingless adult of M. persicae. The expression levels of MpHsp70-1, MpHsp70A1, MpHsp70-2, MpHsp68a and MpHsp68b all increased and then decreased under 4 ℃ stress, and reached the highest at 30 min under 4 ℃ stress, which were significantly higher than those of the control. Under 36 ℃ stress, the expressions levels of MpHsp70-1, MpHsp70A1, MpHsp70-2, MpHsc70-4, MpHsp68a and MpHsp68b increased first and then decreased. The expression level of MpHsp70-1 reached the highest at 60 min after 36 ℃ stress, and those of the other genes reached the highest at 120 min after 36 ℃ stress. After the silence of MpHsp68a, the survival rate and number of nymphs produced of M. persicae under high and low temperature stress were significantly decreased and those after the silence of MpHsp70A1 under high temperature stress were extremely significantly decreased as compared with those of the control group. 【Conclusion】MpHsp70 genes of M. persicae can respond to high and low temperature stress, and play an important role in the molecular mechanism of resistance to temperature stress of M. persicae.
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【Aim】The aim of this study is to clarify the level of the field-evolved resistance of Tuta absoluta to spinetoram and its potential resistance risk, in order to provide a theoretical basis for the rational use of spinetoram to control T. absoluta and slowing development of its resistance to spinetoram. 【Methods】 The leaf-dipping method was used to determine the resistance levels of 18 field populations of T. absoluta collected from five provinces (municipalities or autonomous regions) in northern China to spinetoram. To assess the resistance risk of T. absoluta to spinetoram, 10-generation consecutive selections with spinetoram were carried out in the spinetoram-susceptible strain of T. absoluta via the leaf-dipping method. After that, the realized heritability (h²) of resistance was calculated using Tabashnik’s method for threhold trait agalysis, and the resistance development rates under different selection pressures were predicted based on the data of selection. 【Results】 Among the 18 field populations of T. absoluta, three populations including the populations from Miyun and Huairou in Beijing, and Baotou in Inner Mongolia, exhibited low-level resistance to spinetoram, with the resistance ratios of 6.7, 6.0 and 7.1, respectively. On the other hand, the other 15 populations of T. absoluta were susceptible to spinetoram. After 10-generation consecutive selections with spinetoram, T. absoluta developed 8.9-fold resistance to spinetoram, with the h2 of 0.1973. It was predicted that under different selection pressures (mortality=50%, 60%, 70%, 80% and 90%), T. absoluta needed 11.56, 9.50, 7.92, 6.60 and 5.23 generations, respectively, to develop 10-fold resistance to spinetoram, and 23.12, 18.99, 1583, 13.19 and 10.47 generations, respectively, to develop 100-fold resistance to spinetoram. 【Conclusion】 Due to the risk of T. absoluta developing resistance to spinetoram, it is essential to strengthen insecticide management in the field and emphasize the rotation with alternative types of insecticides to prolong the lifecycle of this insecticide.

【Aim】 Through laboratory insecticidal effect determination and field control efficacy evaluation, the insecticides with high control efficacy against Tuta absoluta were screened to satisfy the demand for emergency control of this pest in production.【Methods】 T. absoluta larvae collected from tomato plants in the field were reared in the laboratory for more than 45 generations, the effects of 10 commonly used insecticides of seven categories including 5% emamectin benzoate water dispersible granule (WG), 60 g/L spinetoram suspension concentrate (SC) and 5% spinosad SC (antibiotics), 200 g/L chlorantraniliprole SC (bisamide), 150 g/L indoxacarb emulsifiable concentrate (EC)(oxadiazine), 10% chlorfenapyr SC (pyrroles), 240 g/L methoxyfenozide SC (hormone), 2.5% rotenone EC (botanical source), and 8 000 IU/μL Bacillus thuringiensis SC and 30 billion spores/g Beauveria bassiana wettable powder (WP)(microbial source) on the hatching rates of T. absoluta eggs and the  mortality rates of the 2nd instar larvae were determined by indoor egg-dipping method and leaf-dipping method, respectively. In July 2023, five insecticide formulations with strong insecticidal effects on T. absoluta in laboratory bioassay were sprayed to open field tomatoes in Xinjiang, Northwest China to evaluate their field control efficacy against T. absoluta. 【Results】 Laboratory bioassay results showed that the 10 pesticides had different effects on the hatching of T. absoluta eggs, the microbial insecticides 8 000 IU/μL B. thuringiensis SC and 30 billion spores/g B. bassiana WP and the botanical insecticide 2.5% rotenone EC had no significant effect on the hatching of eggs at 7 d after treatment, while the chemical insecticides 240 g/L methoxyfenozide SC, 60 g/L spinetoram SC and 5% emamectin benzoate WG exhibited significant inhibitory effects on the hatching of eggs in 5 d. The 10 pesticides had different lethal effects on the 2nd instar larvae of T. absoluta. Among the chemical pesticides, 60 g/L spinetoram SC showed the highest insecticidal activity against the 2nd instar larvae, causing 100.00% mortality rate at 1-4 d post treatment, and 10% chlorfenapyr SC and 150 g/L indoxacarb EC causing 100.00% mortality rate at 3 and 4 d post treatment, while the botanical insecticide 2.5% rotenone  EC  and the microbial insecticide  30 billion spores/g B. bassiana WP had lower lethal effects on the 2nd instar larvae during the 4-d treatment. Field experiment results revealed that the control efficacy of the tested five insecticide formulations against T. absoluta was most obvious at 7 d after application. The control efficacy of 60 g/L spinetoram SC, 200 g/L chlorantraniliprole SC and 10% chlorfenapyr SC against T. absoluta was ranked the top three, being 91.14%, 90.29% and 88.67%, respectively.【Conclusion】 Through laboratory bioassay and field control efficacy evaluation, it was found that 60 g/L spinetoram SC, 200 g/L chlorantraniliprole SC and 10% chlorfenapyr SC at their recommended dosages can be used for chemical control of T. absoluta in tomato production, which can provide guidance for the formulation of comprehensive control plans for T. absoluta and the selection of field control agents.

【Aim】 Previous study found that the overexpression of CYP6AY3v2 and CYP439A1v3 in deltamethrin-resistant strain JH-del of Laodelphax striatellus is the main mechanism of deltamethrin resistance of L. striatellus. The aim of this study is to clarify the mechanism of up-regulated expression of CYP6AY3v2 and CYP439A1v3 in L. striatellus and to reveal the regulatory signal pathways. 【Methods】 The expression levels of long wavelength-sensitive opsin (LOW) gene LWO of the G proteincoupled receptor (GPCR) family A in the 4th instar nymphs of the deltamethrin-resistant strain JHdel and the sensitive strain JHS of L. striatellus were detected by quantitative PCR. RNAi and Bupivacaine HCL were used to interfere with LWO/AC/cAMP/PKA signal pathway genes LWO, AC-2, AC-3, PKA-1, PKA-2 and PKA-3 of the 3rd instar nymphs of L. striatellus strain JH-del, and bioassay was used to detect the change in the sensitivity of L. striatellus to deltamethrin so as to verify that LWO-activated downstream AC/cAMP/PKA/CYP450s signal pathway genes to mediate deltamethrin resistance of L. striatellus by increasing its own expression level. Transgenic Drosophila combined with GAL4/UAS system and insect baculovirus expression system were used to heterogeneously express the L. striatellus LWOi n the 3-day-old female adults of Drosophila melanogaster and Sf9 cells of Spodoptera frugiperda to verify the function of LWO. 【Results】 The relative expression level of LWO  in the deltamethrin-resistant L. striatellus strain JH-del was 1.54-fold as high as that in the sensitive strain JHS. When any node of the LWO/AC/cAMP/PKA signal pathway in the 3rd instar nymph of L. striatellus strain JH-del was interfered by feeding dsRNAs of target genes, the expression levels of CYP6AY3v2 and CYP439A1v3 that metabolize deltamethrin downstream of this signal pathway were significantly decreased, and the sensitivity of the 3rd instar nymphs of strain JH-del of L. striatellus was restored as compared with that in the control group (fed with ds GFP). After heterologous expression of LWO in D. melanogaster and Sf9 cells, the resistance of D. melanogaster and Sf9 cells to deltamethrin increased significantly, and the increase of deltamethrin resistance was also mediated by LWO/AC/cAMP/PKA/CYP450s signal pathway of L. striatellus. 【Conclusion】 LWO receptor activates downstream AC/cAMP/PKA/CYP450s signal pathway by increasing its own expression level, which mediates deltamethrin resistance of L. striatellus. The results provide a theoretical basis for resistance management of L. striatellus and screening of new insecticide targets.

As the most diverse group of animals on earth, insects are closely related to human production and activities, making entomological research both theoretically significant and practically valuable. In the past decade, the rapid development of novel methods and technologies has greatly promoted the research progress of entomological research. In this article, we provided a comprehensive overview of the applications of emerging methods and technologies in such fields as entomological morphological identification, molecular mechanism and pest management, focusing on the main contents of this special issue from seven aspects: Micro-computed tomography, RNA interference, gene editing, artificial intelligence-driven intelligent recognition, nanotechnology, regulation of insect-microorganism symbiotes, and olfactory behavior regulation. We also proposed the challenges faced by the large-scale application and sustainable development of these technologies, and prospected the future development trend.

 【Aim】 This study aims to reveal the impact of the climatic features of Hainan Island, South China on the occurrence patterns of the invasive pest Brontispa longissima, so as to provide a scientific basis for developing effective control strategies. 【Methods】From 2021 to 2023, a monthly survey was conducted on Palmae plants within a 500-m range of expressways and national highways on Hainan Island. The information such as the latitude and longitude of the area of B. longissima infestations, host species, and the number of damaged plants was recorded. The spatio-temporal occurrence patterns of the occurrence area of B. longissima were analyzed by statistical methods, while the influences of ten climatic factors on the occurrence area of B. longissima were explored using the random forest model. 【Results】 The occurrence area of B. longissima had been increasing annually, showing a unimodal pattern within the annual cycle, with the peak occurring in May or June. The damage of B. longissima during the wet season (May to October) was more severe than that during the dry season (November to April of the next year). The damage caused by B. longissima was mainly concentrated in the coastal areas of Hainan Island, with the eastern coastal regions being the most severely affected. According to the climate zone division, the southeastern region had the largest occurrence area of B. longissima. The occurrence area in the northeastern region was larger than that in the southwestern region from March to August, but from September to February of the next year, the occurrence area in the southwestern region exceeded that in the northeastern region. According to the random forest feature importance ranking for the occurrence area of B. longissima and climatic factors, precipitation and relative humidity were the main climatic factors affecting the occurrence area of B. longissima, with the percentages of increase of mean square error (IncMSE%) of 28.14% and 27.39%, respectively.【Conclusion】 The occurrence area of B. longissima on Hainan Island is closely related to climatic conditions, displaying distinct spatio-temporal occurrence patterns. The wet season (May to October) is a critical period for control efforts, with damage reaching its annual peak particularly in May and June. Coastal areas of Hainan Island, especially in the eastern and southeastern climate zones, are the key regions for focus, requiring enhanced monitoring and management. In the northeastern climate zones, emphasis should be paid on the control in spring and summer (March to August), while in the southwestern region efforts should be strengthened during the autumn and winter months (September to February of the next year). This study provides a reference for scientifically formulating regional and seasonal control strategies for B. longissima.

 Tomato is one of the most important horticultural crops, and China is the largest producer of tomato in the world. In recent years, the tomato industry is facing increasingly severe pest threats including the traditional important pests (Bemisi tabaci, Frankliniella occidentalis and Helicoverpa armigera) and the newly emerged invasive pest Tuta absoluta. Elucidating the defensive mechanism of tomato especially wild tomato germplasm resource, which has significantly higher resistance to pests, can provide important genetic resources for breeding process of insect-resistant tomato varieties. Meanwhile, the key insect-resistant metabolites of tomato can also offer valuable insights into the development of new safer and more eco-friendly botanical pesticides. In this article, we summarized the interactions between tomato pests and host plants like tomato across multiple levels of insect resistance mechanisms in plants. Key topics include: (1) the recognition of saliva proteins from piercing-sucking and chewing insects by tomatoes and its impact on anti-insect immunity; (2) the signal transduction networks of insect resistance and the regulatory mechanisms of core defense-related transcription factors in tomato; (3) structural and metabolic bases of insect resistance in plants, such as trichomes, acylsugars, phenolamides, steroidal alkaloids, and volatile compounds, which respond to pest attacks and confer insect resistance through molecular and ecological pathways. Future research should leverage emerging technologies like single-cell transcriptomics and spatial transcriptomics, combined with gene editing and genetic manipulation tools, to further clarify the signaling pathways of insect resistance and the synthesis and regulation of defense compounds in tomato. These efforts will deepen our understanding of plant-insect interactions and lay a theoretical foundation for breeding high-yield, insect-resistant tomato varieties.

【Aim】 This study aims to enrich the basic information of 14-3-3ζ gene of Apis cerana cerana, so as to provide a reference and basis for its further functional study. 【Methods】 The coding sequence (CDS) of 14-3-3ζ gene was amplified by RT-PCR, followed by TA cloning and Sanger sequencing. The physicochemical properties and molecular features of 14-3-3ζ were predicted using the relevant software, and the phylogenetic analysis of 14-3-3ζ was performed. RT-qPCR was used to detect the expression levels of 14-3-3ζ gene in different developmental stages (egg, larva, pre-pupa, pupa and adult), and different tissues (antennae, midgut, fat body, hypopharyngeal gland, brain, cuticle and venom gland) of the newly emerged adult workers of Ap. cerana cerana, as well as in the guts of the 4-, 5- and 6-day-old larvae of Ap. cerana cerana after inoculating the 3-day-old larval workers with Ascosphaera apis.【Results】 The CDS of 14-3-3ζ gene of Ap. cerana cerana was successfully cloned, including 744 nucleotides and encoding 247 amino acids. 14-3-3ζ of Ap. cerana cerana had the molecular weight of about 28.0 kD, included 26 phosphorylation sites, four structural domains and one conserved motif, but had no transmembrane domains and signal peptides. The 14-3-3ζ proteins of Ap. cerana cerana, Ap. mellifera, Ap. laboriosa, Ap. florea, Ceratina calcarata, Bombus pyrosoma, B. terrestris, Megachile rotundata, Osmia lignaria and Habropoda laboriosa all contained four identical conserved motifs and one same structural domain (14-3-3_1). The 14-3-3ζ proteins of Ap. cerana cerana and Ap. mellifera clustered into a single clade on the phylogenetic tree. The expression level of 14-3-3ζ gene in Ap. cerana cerana eggs was significantly higher than those in the 3-day-old larvae, 1-day-old prepupae, 2-day-old prepupae and 4-day-old pupae. The differences in the expression level of 14-3-3ζ gene in various day-old adult workers of Ap. cerana cerana were non-significant. The expression level of 14-3-3ζ gene in the venom gland of the newly emerged adult workers of Ap. cerana cerana was the highest, significantly higher than those in the antennae, midgut, hypopharyngeal gland, brain, cuticle and fat body. Following inoculation of the 3-day-old larval workers of Ap. cerana cerana with As. apis, the expression levels of 14-3-3ζ gene in the 4-, 5- and 6-day-old larval worker guts of Ap. cerana cerana were significantly down-regulated as compared with those in the control group.【Conclusion】 Ap. cerana cerana 14-3-3ζ gene is specifically and highly expressed in the venom gland and egg of worker, and the expression of 14-3-3ζ gene in the larval guts is activated in the process of As. apis infection. 14-3-3ζ is a putative hydrophilic, non-transmembrane and intracellular protein, and highly conserved in Ap. cerana cerana and the above other ten bee species. There is the closest genetic relationship between 14-3-3ζ of Ap. cerana cerana and Ap. mellifera.
Key words: Apis cerana cerana; 14-3-3; molecular features; expression pattern; Ascospaera apis

【Aim】 The objective of this study is to investigate the oviposition deterrent and antifeedant effects of plant essential oils (EOs) on a major invasive pest, the fall armyworm, Spodoptera frugiperda, and provide a new method for the green control of this pest. 【Methods】 S. frugiperda collected from a maize field in Zhanjiang, Guangdong Province, South China and reared indoors for several generations was used. In the laboratory, the oviposition deterrent activities of EOs from 21 plants (Perilla frutescens, Cupressus funebris, Acorus calamus, Artemisia argyi, Capsicum annuum, Myristica fragrans, Zingiber officinale, Piper nigrum, Allium sativum, Cedrus deodara, Cinnamomum cassia, Mentha canadensis, Mentha spicata, Litsea cubeba, Melia azedarach, Citrus limon, Camellia sinensis, Cymbopogon citratus, Eucalyptus globules, Cinnamomum camphora and Plectranthus hadiensis) against S. frugiperda adults were investigated using the behavior selection method, while the antifeedant activities of these EOs against the 2nd instar larvae of S. frugiperda were evaluated using the leaf dish feeding method. 【Results】 At the concentration of 2.5 mL/L, Capsicum annuum EO and Melia azedarach EO had the best oviposition deterrent effects on S. frugiperda adults, with the deterrent rates of 89.13% and 88.83%, respectively. At the concentration of 5 mL/L, Capsicum annuum EO showed the best oviposition deterrent effect on S. frugiperda adults, with the deterrent rate of 100.00%. At the concentration of 10 mL/L, the EOs from Perilla frutescens, Cupressus funebris, Capsicum annuum, Myristica fragrans, Piper nigrum, Allium sativum, Cedrus deodara and Citrus limon showed obvious oviposition deterrent effects on S. frugiperda adults. Piper nigrum EO at the concentration of 2.5 mL/L had the best non-selective and selective antifeedant effects on the 2nd instar larvae of S. frugiperda, with the antifeedant rates of 98.67% and 97.37%, respectively. At the concentrations of 5 and 10 mL/L, Piper nigrum EO also exhibited the best antifeedant effects on the 2nd instar larvae of S. frugiperda, both with the antifeedant rate of 100.00%. 【Conclusion】 At relatively low concentrations, Cayenne pepper EO and Melia azedarach EO showed highly effectiveness in deterring oviposition of S. frugiperda adults, and Piper nigrum EO showed excellent antifeedant activity against the 2nd instar larvae of S. frugiperda. These plant EOs demonstrated promising application potential in the green control of S. frugiperda.

【Aim】 The aim of this study is to investigate the role of spalt major, a regulator of wing development, in wing differentiation of the pea aphid, Acyrthosiphon pisum, and to further reveal the molecular mechanism of non-genetic wing polyphenism in insects. 【Methods】 A high ratio of winged offspring was induced by density in the parthenogenetic wingless A. pisum under the laboratory conditions and the expression level of Apsal was detected by qRT-PCR. dsRNAs were synthesized, including dsApsal targeting spalt major gene (Apsal), dsApWnt2 targeting Wnt-2 (ApWnt2) and dsApdpp targeting decapentaplegic (Apdpp) in A. pisum. ApWnt2 and Apdpp are upstream of Apsal in the Wg/Wnt and Dpp pathways, respectively. Nanocarrier-mediated body wall penetration was used to interfere with Apsal, ApWnt2 and Apdpp in wingless female adults and their newly born nymphs, and then the ratio of winged morph was recorded, and the gene expression levels were analyzed by qRT-PCR. 【Results】 More than 80% of winged offspring of A. pisum could be stably induced by high density treatment on the female adults. The expression level of Apsal, a key gene in wing development, was increased in the winged offspring aphid induced by density as compared with that in the wingless solitarily bred aphid. Interfering the expression of Apsal by dsApsal resulted in a significant decrease of the ratio of winged morph compared with the control group (dseGFP). When the expression of upstream gene ApWnt2 or Apdpp was interfered, the expression level of Apsal decreased as compared with that in the control group (dseGFP). 【Conclusion】 Apsal regulates wing differentiation of A. pisum and is activated by both Wg/Wnt and Dpp signaling pathways. This study further reveals the molecular mechanism of aphid wing differentiation, providing theoretical and technical bases for RNAi-based pest management to prevent aphids from migrating to new hosts.

【Aim】The aim of this study is to investigate the biological function of ABC transporter genes in the development of resistance to indoxacarb in Spodoptera frugiperda, so as to provide a theoretical basis for the comprehensive control of this pest. 【Methods】 Indoxacarb alone and combined with ABC transporter inhibitor verapamil hydrochloride were used to treat the 3rd instar larvae of the indoxacarb-resistant population DC-22 and the indoxacarb-susceptible strain WH of S. frugiperda by the topical application method, and the median lethal concentration (LC₅₀) and the synergistic ratio of verapamil hydrochloride to indoxacarb were calculated at 24 h after treatment. The expression levels of seven ABC transporter genes (SfABCG20, SfABCC2, SfABCF4, SfABCA1, SfABCA5, SfABCG23 and SfABCG9) in the 3rd instar larvae of the indoxacarb-susceptible strain WH and four indoxacarb-resistant populations， including DC-22, CX-22, MY-22 and RH-22, were examined. The highly expressed ABC transporter gene SfABCG23 in response to indoxacarb was silenced through RNAi by injecting dsSfABCG23 into the 3rd instar larvae of DC-22 and WH. The expression level of SfABCG23 was detected by RT-qPCR at 48 h after RNAi, and the mortality was detected at 24 h after exposure to LC₃₀of indoxacarb following RNAi.【Results】Verapamil hydrochloride significantly increased the susceptibility of the indoxacarb-resistant population DC-22 to indoxacarb, with the synergistic ratio of 1.73. The expression levels of SfABCG23 in the 3rd instar larvae of the indoxacarb-resistant populations DC-22 and CX-22 were up-regulated by 2.56- and 4.05-fold, respectively, as compared with that in the indoxacarb-susceptible strain WH, and the expression level of SfABCG23 was significantly positively correlated with the resistance ratio, with the correlation coefficient of 0.941. After dsSfABCG23 injection, the gene silencing efficiency was 65.04% and 39.55%, respectively, in the indoxacarb-resistant population DC-22 and the indoxacarb-susceptible strain WH, and compared with the dsGFP-injected control group, the dsSfABCG23 injection increased the mortality of the 3rd instar larvae of the indoxacarb-resistant population DC-22 and the indoxacarb-susceptible strain WH, by 30.55% and 25.00%, respectively, at 24 h after exposure to indoxacarb. 【Conclusion】 The results of this study suggest that ABC transporter genes play an important role in regulating the development of resistance in the indoxacarb-resistant population of S. frugiperda, and the overexpression of SfABCG23 may play an important role in the development of resistance to indoxacarb in S. frugiperda.

 One of the hot topics in entomology today is the symbiotic microorganism-host interactions in insects. Symbiotic microorganisms in insects can not only provide nutrients for the growth and development of the hosts, but also synthesize many active substances that regulate the ecological adaptability, stress resistance, diversity formation, reproduction, and mating behavior of hosts. Despite of their widespread use, microbial insecticides have led to varying degrees of resistance in pests due to the toxicity of their metabolites and their long-term application in the field. The development of this resistance is closely related to the symbiotic microorganisms in pests. In this article, we reviewed the role of insect symbiotic microorganisms in the development of host resistance to adversity stresses, with focuses on the role of insect symbiotic microorganisms in assisting hosts to resist infestation by different species of biocontrol fungi/bacteria, such as Beauveria, Metarhizium, Pandora neoaphidis and Bacillus thuringiensis. In addition, we deeply clarified the defense mechanisms of symbiotic microorganisms against exogenous pathogen infestation. The present studies showed that the symbiotic microorganisms against exogenous pathogens are mainly distributed in the epidermis, antennal glands and gut of insects, and the symbiotic microorganisms such as Pantoea, Pseudomonas, Wolbachia, Citrobacter freundii and Arsenophonus are more prominent in assisting the hosts to fight the pathogen infestation, having a protective effect on various insect hosts. In addition, these symbiotic microorganisms improve the resistance of insects to pathogens mainly through three pathways: Competition for nutrients with exogenous microorganisms, secretion of antimicrobial substances and modulation of the immune system. This article provides new ideas for a comprehensive elucidation of the mutualistic interactions among pathogenic microorganisms, insects and symbiotic microorganisms, and can also serve as a valuable reference for the development of pest biological control strategies based on the regulation of symbiotic relationships.

【Aim】 The objective of this study is to elucidate the regulatory function of piR-ame-1186994 of Apis mellifera ligustica, so as to offer a scientific basis for further investigation of the underlying regulatory mechanism of piR-ame-1186994. 【Methods】 The expression and sequence authenticity of piR-ame-1186994 in the 6-day-old adult worker’s midgut, 12-day-old adult drone’s testis and 7-day-old adult queen’s ovary of A. m. ligustica were verified by Stem-loop RT-PCR and Sanger sequencing, respectively. Relevant software was utilized to predict the target mRNAs of piR-ame-1186994 followed by GO and KEGG database annotation. Regulatory networks related to developmental signaling pathways, energy metabolism pathways and cellular and humoral immune pathways were further constructed. Newly emerged adult workers were fed with mimic and mimic-NC (negative control) of piR-ame-1186994, followed by the detection of the relative expression levels of piR-ame-1186994 and its key target genes (YAP1 and PLD2) in the midguts of adult workers using RT-qPCR. 【Results】 The specific fragment of piR-ame-1186994 was amplified from the 6-day-old adult worker’s midgut, 12-day-old adult drone’s testis and 7-day-old adult queen’s ovary of A. m. ligustica. piR-ame-1186994 targeted 1 097 mRNAs, which could be annotated to 30 GO terms involved in metabolic process, binding, cell, etc., and 182 KEGG pathways including Wnt signaling pathway, endocytosis and oxytocin signaling pathway. Thirty-six and 16 target mRNAs were respectively involved in five developmental-related signaling pathways (mTOR, Wnt, Hippo, AMPK and Notch signaling pathways) and four pathways related to energy metabolism (amino sugar and nucleotide sugar metabolism, fructose and mannose metabolism, glycolysis/gluconeogenesis, and pentose phosphate pathway), respectively. Additionally, 29 and eight target mRNAs were engaged in four cellular immune pathways (lysosome, endocytosis, phagosome and ubiquitin-mediated protein degradation) and three humoral immune pathways (PI3K-Akt, MAPK and FoxO signaling pathways), respectively. In the mimic-piR group, the expression level of piR-ame-1186994 was significantly up-regulated in the 1-, 3- and 5-day-old workers’ midguts and up-regulated extremely significantly in the 2- and 4-day-old workers’ midguts, the expression level of the target gene YAP1 was extremely significantly down-regulated in the 1-, 3-, 4- and 5-day-old workers’ midguts and significantly down-regulated in the 2-day-old worker’s midgut, and the expression level of target gene PLD2 was significantly down-regulated in the 2-, 3- and 5-day-old workers’ midguts and downregulated extremely significantly in the 4-day-old worker’s midguts as compared with those in the mimic-NC group. 【Conclusion】 piR-ame-1186994 exists and expresses in the worker’s midgut, drone’s testes and queen’s ovary of A. m. ligustica. piR-ame-1186994 potentially modulates the development and immunity of worker’s midgut through targeting and negatively regulating the expression of YAP1 and PLD2.

【Aim】 Picromerus lewisi is a significant predatory natural enemy insect distributed in multiple countries of Asia, such as China, Korea and Japan, primarily used for controlling lepidopteran pests. Venom plays a crucial role in causing rapid paralysis and death of preys during hunting. The aim of this study is to understand the transcriptomic characteristics of the venom glands of P. lewisi, explore the diversity of toxins in P. lewisi, and establish a foundation for further research on the composition and function of the venom in P. lewisi.【Methods】Transcriptome sequencing was conducted on the venom glands of adults and the 5th instar nymphs of P. lewisi collected from Qianxinan Prefecture, Guizhou Province using the high-throughput Illumina NovaSeq 6000 platform. The resulting data were annotated using the NR, NT, Pfam, KOG/COG, Swiss-Prot, KEGG and GO databases. Gene expression in the venom gland samples of P. lewisi was assessed using the FPKM method, and DESeq was employed for the differential expression analysis of venom gland transcriptomes between adult and the 5th instar nymph. The differentially expressed genes (DEGs) between the adult and the 5th instar nymphal venom gland transcriptomes were screened using the criteria of |log₂(Fold change)|>1 and P<0.05, followed by GO functional enrichment analysis and KEGG pathway enrichment analysis of the identified DEGs. The 33 215 transcripts obtained from the sequenced venom gland transcriptomes of adults and the 5th instar nymphs of P. lewisi were subjected to BLAST comparisons in the UniProt database.【Results】Transcriptome sequencing of the venom glands of adults and the 5th instar nymphs of P. lewisi assembled to 22 242 unigenes with an average length of 949 bp. A total of 15 364 unigenes were annotated to the NR, NT, Pfam, Swiss-Prot, GO, KOG/COG and KEGG databases, corresponding to 10 closely related species including three species of true bugs and two species of spiders. A total of 344 DEGs were screened between the venom gland transcriptomes of adults and the 5th instar nymphs of P. lewisi, with 218 genes up-regulated and 126 genes down-regulated. A total of 443 sequences encoding 33 distinct types of toxin-related proteins were identified.【Conclusion】The transcriptome data from the venom glands of both the 5th instar nymphs and adults of P. lewisi were sequenced and obtained in this study. Differential proteins between the venom gland transcriptomes of adults and the 5th instar nymphs were screened, and sequences associated with toxin proteins were identified. This research lays a theoretical foundation for the identification of components in the venom of P. lewisi and the investigation of their biological functions.

【Aim】To explore the effect of Lactobacillus on the transcription level of antimicrobial peptide genes in Bombyx mori. 【Methods】After spraying the suspension (2×10 ⁸ CFU/mL) of Lactobacillus plantarum FLPL028, L. rhamnosus FLRH956 and L. reuteri FLRE589 to the mulberry leaves (20 μL/cm ²) to feed the 1st instar larvae of B. mori, the RNAref transcriptome sequencing of the 5th instar larvae was performed and the mortality rate before cocooning of B. mori after feeding the 5th instar larvae with the mulberry leaves sprayed with the suspension (2×10 ⁶ CFU/mL, 20 μL/cm ²) of Serratia marcescens was calculated. The numbers of viable bacteria of S. marcescens were counted at 4 h after incubation with L. plantarum FLPL028, L. rhamnosus FLRH956 and L. reuteri FLRE589, respectively. The expression levels of immune-related genes including LOC101742127, glv1,glv2, CecA, LOC101739681, CecD, Attacin1, Leb3 and Lzm (antimicrobial peptide genes), LOC692824 (lectin gene), PGRP-S1 and LOC101738493(Toll/Imd signaling pathway-related genes), and Pi3k60, MAPK and Ras2(PI3K and MAPK signaling pathway-related genes) in the 5th instar larvae of B. mori were detected by qPCR. 【Results】After the 1st instar larvae of B. mori were fed with L. plantarum FLPL028, L. rhamnosus FLRH956 and L. reuteri FLRE589, respectively, the transcription levels of most antimicrobial peptide genes, including Moricin, glv4-like and glv2, were significantly decreased in the 5th instar larvae, and that of Moricin decreased the most, as compared with those of the control group. After the 1st instar larvae of B. mori were fed with L. plantarum FLPL028, L. rhamnosus FLRH956 and L. reuteri FLRE589, respectively, the transcription levels of lectin genes such as CTL10, CTL19 and LOC101736606, and Toll/Imd signaling pathway-related genes PGRP-S2, LOC101738325 and LOC101738493 in the 5th instar larvae decreased, and those of PI3K and MAPK signaling pathway-related genes Pi3k60 and MAPK were increased by about 2.4- and 2.1-fold, respectively, as compared with those of the control group. However, the above three species of Lactobacillus had antagonistic effects on S. marcescens, and reduced the mortality rate of B. mori in the model group (only fed with S. marcescens) from 83% to less than 35%, among them L. reuteri FLRE589 had the best antagonistic effect on S. marcescens, causing only 18.1% mortality rate of the 5th instar larvae of B. mori. The basic change trends of the expression levels of LOC101742127, glv1, glv2, CecA, LOC101739681, CecD, Attacin1, Leb3, Lzm, LOC692824, LOC101738493, PGRP-S1, Pi3k60, MAPK and Ras2 were consistent with those of RNAref transcriptome sequencing results. The supernatant of the fermentation of these three species of Lactobacillus could effectively kill S. marcescens and reduce the number of viable bacteria of S. marcescens.【Conclusion】Lactobacilli inhibits the expression of antimicrobial peptide genes and Toll/Imd immune pathway-related genes in B. mori, reduces the innate immune response of B. mori, but is conducive to the harmony between Lactobacilli and B. mori. In addition, Lactobacilli can also improve the acquired immunity of B. mori by activating the PI3K and MAPK signaling pathways. This finding will help to understand the immune system of B. mori more comprehensively and provide a new strategy for the prevention and control of diseases in B. mori industry.

【Aim】 Ionotropic receptors (IRs) are crucial for insects’ olfactory, gustatory, temperature, and humidity sensing capabilities. This study aims to identify the IR gene family in Bactrocera dorsalis at the whole-genome level combined with transcriptome data, so as to provide a theoretical basis for further research on the functions of ionotropic receptor genes in B. dorsalis. 【Methods】 IR genes in the whole genome of B. dorsalis were identified by using hidden Markov models, multiple sequence alignments, gene domain analysis, and phylogenetic analysis. Chromosomal localization, full-length gene structure, protein conserved motifs, and collinearity analysis of IR gene family were examined by using TBtools. Evolutionary pressures were assessed with EasyCodeML. Hisat2 and Stringtie software was employed to quantify and analyze the expression patterns of IR genes in the peripheral nervous tissues (antenna, mouthparts, foreleg, midleg, hindleg, and genitalia) of B. dorsalis. qPCR was used for verification. 【Results】 A total of 39 candidate IR genes were identified within the whole genome of B. dorsalis. All the above candidate IR genes were located on autosomes, with 26 of them having complete full-length CDS structures supported by transcriptomic evidence. These full-length IR genes shared 2-8 identical protein conserved motifs, reflecting their structural conservation. Twenty-eight candidate IR genes were demonstrated the collinearity with those of other true flies in the genus Bactrocera, and one IR gene was found to be under strong negative selection. Seventeen candidate IR genes with collinearity were expressed in the peripheral nervous tissues of B. dorsalis, among them, 10 IR genes were exclusively expressed in the antennae, two IR genes only expressed in the mouthparts, and one IR gene solely expressed in the ovipositor. Additionally, one IR gene was expressed only in the antennae and male genitalia, one IR gene was expressed across the tissues except for the ovipositor, two IR genes were expressed across all peripheral nervous tissues, six IR genes were expressed differentially between female and male, and the remaining 11 candidate IR genes showed no expression in peripheral nervous tissues. 【Conclusion】 In this study we systematically identified the IR gene family in B. dorsalis using genomic and transcriptomic data. We analyzed the structural features, evolutionary relationships, and expression patterns of the IR gene family members, providing a theoretical foundation for further functional studies on the IR genes of B. dorsalis.

 Pollinators (including bees, butterflies, beetles, flies, moths, etc.) play an irreplaceable and important role in ecosystems, and they directly affect plant reproduction and ecological balance. With the rapid growth of global population and social development, serious problems such as ecological damage and environmental pollution have occurred and exacerbated challenges for pollinators, such as habitat loss and the use of chemical pesticides, synergistic effects of climate change and pathogen transmission, which have many negative impacts on the stability of ecosystems. Therefore, strengthening research on pollinators and exploring their physiological, morphological, behavioural and ecological characteristics as well as their coevolutionary relationship with plants not only contribute to an indepth understanding of the functional mechanisms of biodiversity and ecosystems, but also provide a fundamental scientific basis for the conservation and use of pollinator resources. This special issue of pollinators presented some latest domestic research progress of pollinators, which may promote exchanges and cooperation in the field of pollinators research and advance the development of the discipline in this field, so as to provide a scientific basis for the construction of China’s ecological civilization and the sustainable development of the environment.

【Aim】This study aims to explore the potential regulatory roles of the demethylase AmALKBH genes in Apis mellifera ligustica in the growth and developmental process and mRNA methylation, so as to provide the new theoretical basis and references for the further functional study of AmALKBHs. 【Methods】 Multiple software was used to bioinformatically analyze the five AmALKBH genes (AmALKBH1, AmALKBH4, AmALKBH6, AmALKBH7 and AmALKBH8), homologous sequence alignment was performed and phylogenetic tree of ALKBHs from different species was constructed. qRT-PCR was employed to examine the expression levels of these five AmALKBH genes in different developmental stages (egg, larva and pupa), brains of the 1-, 7- and 18-day-old adult workers, and various tissues (antennae, brain, hypopharyngeal gland, crop, midgut, ileum, rectum, fat body and venom gland) of the 1-day-old adult workers of A. m. ligustica. Additionally, high performance liquid chromatography tandem triple quadrupole mass spectrometry (HPLC-QqQ-MS/MS) was used to quantify the levels of mRNA m ⁶A methylation in samples throughout the above different developmental stages and brains of the 1-, 7- and 18-day-old adult workers of A. m. ligustica. 【Results】The five AmALKBH genes (AmALKBH1, AmALKBH4, AmALKBH6, AmALKBH7 and AmALKBH8) in A. m. ligustica encode proteins of 311, 297, 229, 228 and 585 amino acids, respectively, with conserved motifs and structural domains, being hydrophilic spherical proteins. These five proteins are different in their physicochemical properties. The five AmALKBHs of A. m. ligustica exhibited 12.92%-46.46% amino acid sequence identities with ALKBHs of Drosophila melanogaster, Homo sapiens, Mus musculus, Arabidopsis thaliana and Escherichia coli, and showed the closest genetic relationships with ALKBHs from A. cerana, Vespa velutina, and  Bombus terrestris.Along with the embryonic development, there was a progressive reduction in the expression levels of AmALKBH4 and AmALKBH7, concomitant with an elevation in the expression level of AmALKBH6. The expression level of AmALKBH8 reached its peak in the middle stage of egg development (36 h post oviposition), and then declined thereafter. The expression of AmALKBH1 was undetectable. During the larval and pupal stages, the expression levels of all the five AmALKBH genes gradually increased with the developmental progress, reaching the peak in the brown-eyed white pupae and brown-eyed light-colored pupae before decreasing slightly in the newly emerged adults. These five AmALKBH genes were highly expressed in the antennae and brain of the 1-day-old adult workers of A. m. ligustica. The expression levels of AmALKBH6, AmALKBH7 and AmALKBH8 in the brains of the 1-day-old adult workers were significantly lower than those in the brains of the 7- and 18-day-old adult workers. An obvious decrease in the level of mRNA m ⁶A methylation was observed from the early stage to middle stage of egg development, with a notable increase in the late stage of egg development (60 h post oviposition). The level of mRNA m ⁶A methylation exhibited a similar trend as the gene expression level, peaking earlier in the pink-eyed white pupae. While the mRNA m ⁶A methylation level was the highest in the brains of the 1-day-old adult workers but reduced in the brains of the 7- and 18-day-old adult workers. 【Conclusion】 This study result demonstrated the high conservation of the ALKBH gene family among species. Differences in the sequence characteristics, protein structure and expression patterns were observed among the five AmALKBH genes in A. m. ligustica, indicating their distinct functional features. The expression levels of AmALKBH genes and the levels of mRNA m ⁶A methylation dynamically changed throughout the developmental stages of A. m. ligustica and were found to be correlated, suggesting a potential regulatory role of AmALKBH genes in the growth and development of A. m. ligustica. These findings lay a solid foundation for uncovering the molecular mechanisms, by which AmALKBH genes regulate growth and development, providing a new theoretical basis for further enhancing the value of honeybees in basic research and pollination applications.

The red imported fire ant, Solenopsis invicta, is an invasive pest that poses a serious threat to agricultural and forestry industries, people’s health, public facilities and biodiversity. Its extreme aggressiveness and strong environmental adaptability make the control of this species a significant challenge. As a soil-dwelling social insect, S. invicta relies primarily on its developed chemical communication system. S. invicta uses a variety of semiochemicals as carriers to efficiently transmit information with other organisms inside and outside nests, thereby coordinating the behavior of ant colonies and completing important life activities. Therefore, understanding the characteristics of the chemical communication of S. invicta may help to timely control the spread of this species. In this article, we focused on the chemical communication system of S. invicta, summarizing the regulatory roles of various important semiochemicals inside and outside the colonies of S. invicta in the social behaviors of ant colonies, including the trail pheromone and alarm pheromone of S. invicta during foraging and dealing with dangers, cuticular hydrocarbons (CHCs)-based individual recognition inside and outside nests and necrophoric behavior, the ability of interspecies eavesdropping to other organisms and the queen pheromone for regulating the development direction of larval ants. We also reviewed and summerized the contemporary applications and problems of S. invicta semiochemicals, with the aim of providing a theoretical reference for its green prevention and control.

【Aim】The aim of this study is to explore the function of the ferritin1HCH (Fer1) gene HvFer1 in the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata, and to provide a theoretical basis for the biological control of H. vigintioctopunctata. 【Methods】The amino acid sequence of HvFer1 was subjected to blastp analysis followed by multiple alignment with Fer1 sequences from different species. The expression levels of HvFer1 in different developmental stages (egg, 1st-4th instar larvae, pupa, and female and male adults) and different tissues (head, midgut, fat body, cuticle and Malpighian tubules) of the 4th instar larvae of H. vigintioctopunctata were detected by RT-qPCR. HvFer1 was silenced by injecting 200 ng/individual of dsHvFer1 into the 3rd instar larvae of H. vigintioctopunctata. Then the larval phenotype, survival rate and duration from the 4th instar larva to pupa, as well as the expression level of dual oxidase (Duox) gene HvDuox, the level of reactive oxygen species (ROS), the content of malondialdehyde (MDA), and bacterial diversity in the midgut were observed and determined. 【Results】The coleopteran Fer1 sequences clustered into one clade, and HvFer1 was the most closely related to Fer1s of Harmonia axyridis and Coccinella septempunctata. The expression level of HvFer1 was the highest in male adults and the lowest in the 4th instar larvae of H. vigintioctopunctata. The expression level of HvFer1 in the midgut of the 4th instar larvae of H. vigintioctopunctata was significantly higher than those in the other tissues. Compared with the control group (dsGFP injection), injection of 200 ng/individual of dsHvFer1 into the 3rd instar larvae of H. vigintioctopunctata significantly prolonged the duration from the 4th instar larva to pupa, and prevented the formation of some spines and spots in pupae. The elyta of the emerged adults of H. vigintioctopunctata were abnormally developed after dsHvFer1 injection, with spots fused or deficient. The expression level of HvFer1 in the 3rd instar larvae of H. vigintioctopunctata on the 2nd day post injection of dsHvFer1 showed no significant difference from that in the control group, but the expression level of HvFer1 on the 4th day post injection of dsHvFer1 was significantly lower than that in the control group. Compared with the control group, HvFer1 silencing caused severe oxidative stress in H. vigintioctopunctata larvae, significantly up-regulated the expression level of HvDuox, ROS level and MDA content in the larval midgut, changed the structural composition of midgut bacteria, reduced the relative abundance of Enterobacter and Chryseobacterium, and also increased the relative abundance of Serratia, Pseudomonas, Acinetobacter and other intestinal bacteria.【Conclusion】 HvFer1 plays a crucial role in regulating the larval development, adult phenotype, and maintaining the larval gut immunity and microbial homeostasis of H. vigintioctopunctata. HvFer1 is expected to be a candidate target gene for controlling H. vigintioctopunctata based on RNAi technology.

【Aim】 This study focuses on the cloning and prokaryotic expression of the sex pheromone biosynthesis activating neuropeptide gene PxylPBAN in the diamondback moth, Plutella xylostella. Additionally, the effects of RNAi-mediated PxylPBAN silencing on the expression of genes involved in both the juvenile hormone pathway and the sex pheromone synthesis pathway were detected to explore the role of PxylPBAN in sex pheromone synthesis. 【Methods】 The full-length CDS of PxylPBAN  in P. xylostella was cloned using RT-PCR. PxylPBAN was expressed in a prokaryotic expression system and subsequently purified. RNAi was conducted by injecting ds PxylPBAN into female pupae of P. xylostella, followed by the RT-qPCR verification of the expression levels of PxylPBAN, juvenile hormone pathway genes PxylJHAMT, PxylnJHBP, PxylhJHBP and PxylMet, as well as the sex pheromone synthesis pathway genes PxylACC and PxylFAR6 at 24, 48 and 72 h post injection. 【Results】 The cloning process successfully generated a 582 bp full-length CDS for PxylPBAN (GenBank accession number: LOC105391112), of P. xylostella, encoding a 193 amino acid protein with the estimated molecular weight of approximately 21.85 kD. The recombinant PxylPBAN protein was produced through prokaryotic expression. RNAi result revealed a significant down-regulation in expression level of PxylPBAN following the injection of ds PxylPBAN compared to the control group injected with ds EGFP of P. xylostella, with the most pronounced decrease observed at 24 h post injection. Additionally, the expression levels of PxylJHAMT, PxylnJHBP, PxylhJHBP, PxylMet, PxylACC and PxylFAR6 of P. xylostella in the ds PxylPBAN-injected group were significantly reduced as compared to those in the control group. 【Conclusion】 This study successfully obtained the recombinant PxylPBAN protein and identified PxylPBAN as a key gene in the juvenile hormone pathway and sex pheromone synthesis pathway in P. xylostella. These findings establish a theoretical basis for understanding the regulatory relationship between PxylPBAN and juvenile hormones in sex pheromone biosynthesis in P. xylostella.

 RNA interference (RNAi) has been widely applied in the study of gene function in insects and the development of novel biological strategies for the selective control of agricultural pests due to its high conservation, strong specificity, easy operation and efficient silencing. However, the practical application of RNAi in pest control also faces challenges, among which the efficiency of RNAi is the most significant limiting factor. In this review, we summarized and discussed the three main pathways of the molecular mechanisms of RNAi, the impact of nucleases on the stability of doublestranded RNA (dsRNA) and RNAi efficiency, and the research progress in using nanoparticle materials to encapsulate and deliver dsRNA. We also provide a perspective on improving RNAi efficiency by inhibiting nuclease activity in insects and combining it with target gene dsRNA, with the aim of providing references for the biological control of pests and the development of green insecticides based on nanoparticles. One of the main factors limiting the efficiency of RNAi in insects is the stability of dsRNA. Combining dsRNA with different nanoparticles to form polymers can increase the stability of dsRNA in insects, thereby enhancing RNAi efficiency. Different nanoparticles provide various ways to deliver dsRNA in RNAi applications. Nanoparticles, with their advantages of safety, low-toxicity, low-cost, good biocompatibility and high RNAi efficiency, have become a research hotspot in both domestic and international research, and are increasingly applied in insect RNAi studies, demonstrating great potential and value in pest control and the development of green insecticides. In the future, the functional design of nanomaterials should be enhanced, low-cost production processes should be developed, and an ecological risk assessment system should be established to promote the practical application and sustainable development of RNAi technology in agriculture.

-【Aim】Tuta absoluta is a newly invaded devastating pest on tomatoes in China, and poses a significant threat to tomato production. Olfactory behavior manipulation technique is an important component of the green control techniques of T. absoluta. By exploring the effects of the components of floral scents, tomato plant volatiles and volatiles from traditional food baits (fermented sugar water and sugar vinegar solution) on the behavior of T. absoluta, this study aims to provide a reference for the development of olfactory behavior manipulation technique for this pest. 【Methods】First, the electroantennogram (EAG) response experiments were carried out to assay the EAG responses of the female and male adults of T. absoluta to 24 compounds including nine compounds of floral scents ［linalool, methyl o-anisate, ethyl hexanoate, cis-jasmone, (-)-limonene, phenylacetaldehyde, β-myrcene, methyl salicylate and ethyl benzoate］, four components of tomato plant volatiles (octyl acetate, catechol, resorcinol and hydroquinone), 11 volatile compounds of from traditional food baits (2-ethyltoluene, decanal, ethyl decanoate, 3-furaldehyde, 1, 2-diethylbenzene, ethyl heptanoate, ethyl octanoate, n-propylbenzene, ethyl nonanoate, benzaldehyde and 1, 4-diethylbenzene) at the doses of 1, 10， 100 and 1 000 μg. Then, the olfactory behavior responses were determined with Y-tube olfactometer to test the attractiveness of 12 compounds screened out by the above experiments with high EAG responses at the doses of 1, 10, 100 and 1 000 μg to the female and male adults of T. absoluta. Finally, the oviposition selection behavior was determined with cage experiments to test the oviposition attraction effects of six effective attractants screened out by the above tests on the female adults of T. absoluta. 【Results】 Twelve volatile compounds, including linalool（10, 100 and 1 000 μg）, decanal（1, 10, 100 and 1 000 μg）, methyl o-anisate（10, 100 and 1 000 μg）, octyl acetate（10, 100 and 1 000 μg）, 3-furaldehyde（1, 10, 100 and 1 000 μg）, ethyl heptanoate（10, 100 and 1 000 μg）, phenylacetaldehyde（10, 100 and 1 000 μg）, resorcinol（1, 10, 100 and 1 000 μg）, methyl salicylate（1, 10, 100 and 1 000 μg）, hydroquinone（1, 10, 100 and 1 000 μg）, ethyl benzoate（1, 10, 100 and 1 000 μg） and 1,4-diethylbenzene（100 and 1 000 μg） elicited high EAG responses in both female and male adults of T. absoluta. Results from the olfactometer bioassay indicated that six compounds including decanal(10 μg), ethyl heptanoate(10 and 1 000 μg), octyl acetate(100 μg), resorcinol(10 μg), methyl salicylate(1 000 μg), and ethyl benzoate(100 and 1 000 μg) had attraction effects on T. absoluta adults, with decanal(10 μg), ethyl heptanoate(10 and 1 000 μg), resorcinol(10 and 1 000 μg) and ethyl benzoate(100 and 1 000 μg) showing significant attraction to female or male adults, and decanal(10 μg) and ethyl benzoate(100 μg) exhibiting bisexual attraction effects at the same dose. Oviposition selection results demonstrated that decanal (10  μg), ethyl heptanoate (100 μg), octyl acetate (1 000 μg), resorcinol (10 μg), methyl salicylate (1 000 μg), and ethyl benzoate (1 000 μg) showed significant oviposition attractiveness to female adults of T. absoluta. 【Conclusion】 Six volatile compounds including decanal, ethyl heptanoate, octyl acetate, resorcinol, methyl salicylate and ethyl benzoate show significant attractive and oviposition stimulant activities to T. absoluta adults, and decanal and ethyl benzoate have the potential to be developed as bisexual attractants. These results provide references for the development of green control techniques for T. absoluta.

【Aim】This study aims to clarify the influence of multiple generations of selection with tetraniliprole on the population fitness and detoxification enzyme activities of Tuta absoluta, so as to provide a theoretical basis for the scientific and rational use of this insecticide. 【Methods】 After T. absoluta was selected with tetraniliprole for 18 generations using spraying method in the laboratory, the toxicity of tetraniliprole to the 2nd-3rd instar larvae of T. absoluta larvae in 48 h was determined by leaf-dipping method, and a tereniliprole-resistant population of T. absoluta was obtained. The fitness of the tereniliprole-resistant population of T. absoluta was evaluated by constructing the two-sex life table, and the activities of mixed-function oxidase (MFO), carboxylesterase (CarE) and glutathione S-transferase (GST) in the 2nd-3rd instar larvae were determined. 【Results】 After 18 generations of laboratory selection, T. absoluta developed 24.60-fold resistance to tetraniliprole. The larval duration, pupal duration, total preoviposition period (TPOP) and mean generation time (T) of the tetraniliprole-resistant population were all prolonged, and the 1st instar larval duration and mean generation time (T) increased significantly by 0.43 and 0.61 d, respectively, as compared to those of the tereniliprole-susceptible population. In addition, compared to the tereniliprole-susceptible population, the tereniliprole-resistant population had decreased intrinsic rate of increase (r), net reproductive rate (λ) and finite rate of increase (R₀). The tereniliprole-resistant population had a relative fitness of 0.90, indicating a survival disadvantage. The activities of GST and MFO in the 2nd-3rd instar larvae of the tereniliprole-resistant population increased significantly, being 1.25 and 1.21 times as high as those of the tereniliprole-susceptible population, respectively, while the CarE activity in the 2nd-3rd instar larvae of the tereniliprole-resistant population showed no significant change as compared with that in the tereniliprole-susceptible population. 【Conclusion】 The resistance of T. absoluta to tetraniliprole increased after selection for 18 generations in laboratory, and MFO and GST might be involved in the development of tetraniliprole resistance. The resistance of T. absoluta to tetraniliprole has a fitness cost for growth and development.

 【Aim】 The small cocoon mutant sc was discovered among the offspring of space silkworm (Bombyx mori), exhibiting slow larval development and reduced consumption of mulberry leaves. We speculate that genes related to growth and development pathways of the sc mutant may be affected by the gene mutation. Our previous research indicated that the sc mutant is controlled by a pair of recessive genes located on the 3rd linkage group of B. mori, but the responsible gene has not yet been identified. This study aims to provide insights into the identification of the responsible gene for the sc mutant and the analysis of its molecular mechanisms through comparative transcriptomic analysis between the sc mutant and the normal cocoon strain (TG) derived from space B. mori.【Methods】 The head and midgut tissues from the day-4 5th instar larvae of sc and TG were collected for transcriptome sequencing (RNA-seq), respectively. The differentially expressed genes (DEGs) were obtained by comparative transcriptome analysis. Then GO annotation and KEGG enrichment analysis of DEGs were performed. qRT-PCR was employed to validate the expression levels of randomly selected DEGs in sc and TG and investigate the expression levels of the genes of interest in sc. 【Results】 A total of 1 528 DEGs were detected in heads in the comparison group TG vs sc, with 820 DEGs showing up-regulated expression and 708 DEGs showing down-regulated expression. Similarly, 1 401 DEGs were identified in the midguts of the comparison group TG vs sc, with 683 DEGs showing up-regulated expression and 718 DEGs showing down-regulated expression. The GO analysis indicated that in biological processes, the majority of DEGs in the head and midgut were implicated in cellular process, metabolic process, biological regulation, response to stimulus, etc. In terms of molecular functions, most DEGs were associated with binding, catalytic activity, structural molecule activity, transporter activity and ATP-dependent activity. DEGs in the head and midgut were implicated in signaling pathways associated with the growth and development of B. mori, including the Hippo, Insulin and mTOR pathways. The qRT-PCR analysis revealed that the gene expression trend was consistent with the transcriptome sequencing result, and compared to TG, the sc mutant had the key genes BMSK0008105, BMSK0009907, BMSK0002689, BMSK0000286, BMSK0012340 and BMSK00083629 involved in growth and development signaling pathways with differential expression. 【Conclusion】 The differential expression of the critical genes in growth and development signaling pathways of sc and TG disturbs the key physiological processes like energy metabolism, organogenesis and cell growth, proliferation and apoptosis, thereby affecting the body development of the small cocoon mutant sc. These findings contribute to understanding the molecular mechanisms underlying the formation of the sc mutant and offer valuable experimental data for further exploration into the regulation of B. mori body size.

【Aims】 To clarify the morphological and biological characteristics of the tea weevil, Myllocerinus aurolineatus, so as to provide a scientific basis for the accurate identification and the development of green control technologies for this insect pest.【Methods】 M. aurolineatus larvae and adults were reared with field-collected soils and fresh tea branches, respectively, in the laboratory under (25±1) ℃. The main morphological characteristics of M. aurolineatus at various developmental stages were observed under light microscope. Field investigations were conducted to learn the migratory behavior of M. aurolineatus larvae in different soil layers of tea plantation, and the distribution pattern in soil upon pupation. The calling, mating and phototactic behaviors of M. aurolineatus adults were recorded and described with a combination of laboratory observation and behavior test.【Results】The freshly laid eggs of M. aurolineatus are creamy white and gradually turn light yellow. The fleshy larvae have no legs and are usually curved or shaped like the letter C. The naked pupa is milky white with an obvious mouthpart and a pair of wing buds. The mature adults are grayish-black and possess yellowish-green shiny scales and stripes on the sheath wings. And the body size of females is always slightly larger than that of males. M. aurolineatus overwinters in the larval stage in the soil layer of 20-30 cm in depth from the ground and its pupation peak occurs in mid-late April. For pupation, the mature larvae will migrate to the soil layer of 0-10 cm in depth from the ground, and the larvae and pupae in the soil layer of 0-5 cm in depth from the ground accounted for 75.83% of the population. M. aurolineatus adults usually mate from 16:00 to 4:00, and copulate in a “false male-above” position, in which the male sits on top of the female. The whole mating period lasts for 44-132 min, and the mating procedure consists of four stages including pre-copulation, copulation, insemination and post-copulatory mate guarding. In addition, both female and male adults of M. aurolineatus are strongly phototactic in the evening. 【Conclusion】In the present study, the main morphological characteristics of M. aurolineatus at various developmental stages have been clarified, and the vertical migrating behaviors of larvae in the soil of the tea plantation and the vertical distribution patterns in different soil layers during the peak period of pupation have been examined. In addition, the mating and phototactic behaviors of the adults have been preliminarily revealed. The results not only provide references for the accurate identification of M. aurolineatus, but also can give theoretical supports for the development of green control technologies.

【Aim】 Grapholita molesta is a fruit pest that poses a significant threat to fruit production globally. Sex-pheromone-mediated mating disruption technology is a highly specific, efficient, non-toxic, and ecologically beneficial control method. The control efficacy of this technology mainly depends on the dosage of sex pheromones and the density of the mating disruption products. This study aims to explore the accurate use of sex-pheromone-mediated mating disruption technology, and to provide new data support for effectively improving the green control level of G. molesta and realizing the safety of fruit production in China. 【Methods】 In this study, G. molesta adults were treated with the sex pheromone containing four components ［(Z)-8-dodecenyl acetate, (E)-8-dodecenyl acetate, (Z)-8-dodecen-l-ol, and dodecanol］ at five different dosages (0.0131, 0.131, 1.31, 13.1 and 131 mg). The electroantennogram (EAG) responses of unmated and once-mated male adults to different dosages of sex pheromone were measured. The number of mating pairs, mating duration and mating day-old age of female and male adults, as well as the change characteristics of the number of eggs laid per female, number of eggs hatched and egg hatching rate were measured in the environments with different dosages of sex pheromone using an indoor cage method. The rate of mating disruption of sex pheromone to G. molesta and the fruit-boring control efficacy were validated in the field. 【Results】 The EAG response results showed that, as the sex pheromone dosage increased, the EAG response of unmated male adults of G. molesta became stronger, reaching the maximum at the dosage of 13.1 mg. However, the sex pheromone dosage had no significant effect on the EAG response of once-mated male adults. Indoor cage test indicated that the sex pheromone at the dosages of 1.31-131 mg significantly reduced the number of mating pairs between female and male adults compared to the control (n-hexane). Although there were no significant differences in mating duration, mating day-old age, and the number of eggs laid per female between the sex pheromone treatment group and the control group, all the five dosages of sex pheromone significantly reduced the number of eggs hatched and egg hatching rate. When pregnant females were directly exposed to different dosages of sex pheromone, the number of eggs laid per female showed no significant change, but the sex pheromone at the dosages of 0.131-131 mg significantly reduced the number of hatched eggs and egg hatching rate compared to the control. In the field, the sex pheromone at the dosage of 131 mg resulted in significantly higher rate of mating disruption and fruit-boring control efficacy against G. molesta than that at the dosage of 13.1 mg. There was no significant difference in the rate of mating disruption or fruit-boring control efficacy against G. molesta between hanging 60 and 90 mating disruption products per 667 m2.【Conclusion】 Considering both control efficacy and economic cost, a sex pheromone dosage of 131 mg with 60 mating disruption products per 667 m² is suitable for mating disruption technology of G. molesta in the field. These results provide the theoretical basis of mating disruption technology to precisely control G. molesta.

 【Aim】 To identify the gustatory receptor (GR) family genes of Anopheles sinensis at the whole genome level, analyze the structure and characteristics of the GR gene family members, and predict the possible biological functions of the GR family members, so as to provide an information framework for the GR family genes of An. sinensis. 【Methods】 Using the known GR amino acid sequences of An. gambiae, Culex quinquefasciatus, Aedes aegypti, and Drosophila melanogaster downloaded from NCBI and Vectorbase databases as inquiry sequences, the GR family genes of An. sinensis were searched and identified at the whole genome level by Blast and HMM methods and named. Using bioinformatics methods, the basic characteristics (physicochemical properties, gene structure and genome localization), conservative domains and Ka/Ks ratios of the GR genes in An. sinensis were predicted. Using MEGA software and maximum likelihood (ML) method, a phylogenetic tree of proteins encoded by the GR genes of An. sinensis and An. gambiae was constructed.【Results】 A total of 54 GR genes were screened and identified in the genome of An. sinensis. According to the classification of An. gambiae and phylogenetic relationship, AsGRs of An. sinensis were divided into four subfamilies of CO₂ receptor, bitter receptor, sugar receptor and pheromone receptor, with 3, 40, 6 and 5 receptors, respectively. CO₂ receptors and sugar receptors were clustered into a single branch, pheromone receptors were clustered into a single branch, and bitter receptors were scattered throughout the phylogenetic tree. The 54 GR genes of An. sinensis were located on the chromosomes Chr.1 and Chr.2. The amino acid numbers of AsGRs are 128-3 429, with the molecular weight of 14.85-397.08 kD, the theoretical isoelectric point (pI) of 5.16-9.84, and the hydrophilicity index (GRAVY) of between -0.023-0.838. The exons of the 54 An. sinensis AsGR genes ranged from 1 to 23, with a wide range. The Ka/Ks ratios of most of the direct homologous gene pairs between An. sinensis and An. gambiae were less than 1, indicating that the GR family genes of An. sinensis were mainly affected by purification selection during the evolutionary process. 【Conclusion】 This study conducted genome-wide identification and characteristic analysis of the GR family genes of An. sinensis, enriching the genome database of An. sinensis and laying a certain foundation for subsequent functional research on the GR genes of An. sinensis.

【Aim】 To determine the toxicity of plumbagin (an important plant secondary substance from Chinese medicine Plumbago zeylanica) to the 2nd instar larvae of Spodoptera frugiperda and ascertain the sublethal effect of a sublethal concentration (LC₂₅) of plumbagin on the growth, development and detoxifying enzyme activities of the F₀ and F₁ generations of S. frugiperda. 【Methods】 The toxicity of plumbagin against the 2nd instar larvae of S. frugiperda (the 2nd day after ecdysis) was determined using insecticide incorporated artificial diet bioassay. After the 2nd instar larvae were exposed to LC₂₅ (0.343 mg/g) of plumbagin, the larval duration, pupation rate, pupal duration, female and male adult longevity, and number of eggs laid per female adult of the F₀ and F₁ generations of S. frugiperda were recorded, the age-stage, two-sex life table was constructed, and the activities of three detoxifying enzymes including carboxylesterase (CarE), glutathione S-transferase (GST) and cytochrome P450 monooxygenase (CYP450) were determined at 24 h after treatment. 【Results】 The median lethal concentration (LC₅₀) and LC₂₅ values of plumbagin against the 2nd instar larvae of S. frugiperda in 7 d were 0.607 and 0.343 mg/g, respectively. For the parental generation (F₀) of S. frugiperda, LC₂₅ of plumbagin significantly decreased the pupation rate by 25.13%, significantly shortened the female and male pupal duration by 1.29 and 1.08 d, respectively, significantly decreased the number of eggs laid per female by 47%, and significantly shortened the oviposition period and adult longevity by 1.75 and 1.19 d, respectively, compared to the vehicle control (0.1% acetone). For the offspring generation (F₁) of S. frugiperda, LC₂₅ of plumbagin only significantly shortened the mean generation time (T) by 0.91 d, compared to the vehicle control. LC₂₅ of plumbagin significantly induced the activities of the three detoxifying enzymes CarE, GST and CYP450 in the 2nd instar larvae of S. frugiperda after treatment for 24 h, which were increased to 1.28-, 1.30- and 1.42-fold as high as those in the vehicle control. 【Conclusion】 LC₂₅of plumbagin had obvious adverse effects on the growth, development and fecundity of the F₀ generation of S. frugiperda, and significantly increased the activities of the three detoxifying enzymes CarE, GST and CYP450 in the 2nd instar larvae after treatment for 24 h, which will be helpful for using the plant secondary substance plumbagin as one of potent biocontrol strategies for this pest.

【Aim】To elucidate the repellent activities of three plant-derived compounds neochamaejasmine A, thymol and azadirachtin against adult Tribolium confusum and their effects on its olfaction-related genes. 【Methods】 The relative electroantennogram (EAG) response values of T. confusum adults to 0.005, 0.05, 0.5, 5 and 50 μg/μL neochamaejasmine A, thymol and azadirachtin were measured. The olfactory behavior responses of T. confusum adults to neochamaejasmine A, thymol and azadirachtin at the concentrations of 0.5, 5 and 50 μg/μL were studied by Y-tube olfactometer. Impregnated filter paper method was used to determine the repellent rates of 20, 30, 40, 50 and 60 mg/L neochamaejasmine A, thymol and azadirachtin to T. confusum adults. Transcriptomes of the head of T. confusum adults treated with median lethal concentration (LC₅₀)(28.37 mg/L) neochamaejasmine A, LC₅₀(51.05 mg/L) thymol and LC₅₀(42.20 mg/L) azadirachtin were sequenced and comparatively analyzed using RNA-Seq. 【Results】The relative EAG response value of T. confusum adults was proportional to the concentrations of neochamaejasmine A, thymol and azadirachtin. At the maximum concentration of 50 μg/μL, neochamaejasmine A, thymol and azadirachtin resulted in the relative EAG response values of T. confusum adults of 1.61, 2.69 and 2.34 mV, respectively, and the selection rates of 86.48%, 73.89% and 78.33%, respectively. The repellent rates (74.67% and 80.82%, respectively) of 60 mg/L thymol to T. confusum adults were the highest at 2 and 4 h, and the repellent grades reached grades Ⅳ and Ⅴ, respectively. After treatment with thymol, neochamaejasmine A and azadirachtin, there were 569, 597 and 661 differentially expressed genes in the head transcriptomes of T. confusum adults, respectively, 412, 434 and 498 differentially expressed genes were annotated with GO function, respectively, and the differentially expressed genes were identified to 26, 33 and 32 KEGG pathways, respectively, compared with the control group. Thirteen, four and five olfaction-related genes with differential expression were found after treatment with thymol, neochamaejasmine A and azadirachtin, respectively. 【Conclusion】In this study, the repellent activities of the three plant-derived compounds to T. confusum adults were determined and the head transcriptome of T. confusum adults was sequenced. It is speculated that the differentially expressed genes may play an important role in T. confusum repelling the three plant-derived compounds.

【Aim】 Using CRISPR/Cas9 gene editing technology to perform single-target knockout of Spodoptera frugiperda doublesex ( Sfdsx), this study aims to explore the effect of Sfdsx on the wing development of male adults of S. frugiperda. 【Methods】CRISPR/Cas9 gene editing technology was used to knock out the female and male common region of Sfdsx in embryos of S. frugiperda and construct Sfdsx mutant. After pupation, the differences in the morphological characteristics of pupa and adult wings of the Sfdsx mutant were observed.【Results】 The Sfdsx mutant of S. frugiperda exhibited significant male-biased sex ratio (female∶male=0∶14), and the genital pores on the 8th-9th abdominal segments of its male pupae were severely twisted. The wing traits of the male adult mutant tended to be neutral in sex, in which，the renal patches in the center of the forewing were deformed, the black spots at the end of the wing disappeared, and the wing scales were arranged in disorder. The hind wings were deformed in wingspan, the arrangement of wing scales changed, and small black spots developed.【Conclusion】CRISPR/Cas9-mediated knockout of the common region of Sfdsx led to wing deformation of male adults of S. frugiperda. The results of our study provide an ideal gene target and theoretical basis to modulate the development of S. frugiperda through sterile insect technology (SIT).

【Aim】 Neoseiulus setarius, a newly identified predatory mite species discovered in the Inner Mongolia Region, North China, has great potential to control small pests, such as thrips. When predatory mites were released in regions characterized by considerable temperature variations between day and night, a sudden decrease in temperature may potentially influence their predation capacity. Rapid cold hardening(RCH)can improve the cold tolerance of ectotherms. This study aims to explore whether N. setarius has this phenotypic plasticity and its response to short-term low-temperature stress by examining the RCH response of N. setarius and its fitness before and after cold hardening. 【Methods】 The critical temperature (at which the survival rate is below 10%) for female adults of N. setarius to survive was measured. The survival rates of female adults of N. setarius after exposure to different low temperatures (0, 2.5, 5, 7.5 and 10 ℃) were counted to determine the optimal acclimation temperature and duration. The RCH responses of various developmental stages of N. setarius were also investigated under the optimal acclimation temperature and duration. Moreover, the effects of RCH on the predatory and reproductive potential of female adults of N. setarius were evaluated by using the 1st instar nymphs of Frankliniella intonsa as prey, which were aged 1-2-day-old after hatching from the eggs laid on beans within 1 d. 【Results】 The critical low temperature for survival of female adults of N. setarius was -21 ℃. Exposure to 2.5 ℃ for 2 h was found to be the optimal condition to induce RCH in female adults of N. setarius, effectively increasing their survival rate. There was only 3.33% survival rate when adult females of N. setarius were directly transferred from the room temperature of 25 ℃ to -21 ℃ for 2 h, while the survival rate of adult females was increased to 68.33% after cold hardening at 2.5 ℃ for 2 h and then exposure to -21 ℃ for 2 h. Similarly, the survival rates of other developmental stages of N. setarius significantly increased after cold hardening at 2.5 ℃ for 2 h and then exposure to -21 ℃ for 2 h. The survival rates of non-hardened eggs, larvae, protonymphs, deutonymphs and male adults of N. setarius after exposure to -21 ℃ for 2 h were 6.67%, 21.67%, 1.67%, 5.00% and 5.00%, respectively, while after cold hardening at 2.5 ℃ for 2 h, the survival rates of the corresponding developmental stages of N. setarius after exposure to -21 ℃ for 2 h were increased to 60.00%, 33.33%, 20.00%, 31.67% and 51.67%, respectively. Meanwhile, the predation and oviposition of the female adults of N. setarius on F. intonsa were not reduced after cold hardening. 【Conclusion】 All developmental stages of N. setarius have the ability to respond to RCH. Exposure to 2.5 ℃ for 2 h can improve the cold tolerance of N. setarius without reducing its predation fitness against F. intonsa, which provides a basis for the field releasing of this mite in regions with significant variations in temperature throughout the day.

 【Aim】 This study aims to explore the relationship between the changes in cytochrome c oxidase (COX) activity and gene expression of COX subunits SmCOX1 and SmCOX2 in Sitodiplosis mosellana and diapause development, so as to provide a theoretical basis for elucidating the energy metabolism mechanism of diapause of S. mosellana. 【Methods】 The SmCOX activities in S. mosellana larvae at different stages in the natural diapause process including pre-diapause, diapause, post-diapause quiescence and post-diapause development were measured using a COX assay kit. The full-length cDNA sequences of SmCOX1 and SmCOX2 were cloned using RT-PCR and RACE techniques, and analyzed by bioinformatics. qPCR technique was used to detect the expression pattern of SmCOX1 and SmCOX2 in the diapause process of S. mosellana. 【Results】 The SmCOX activity in larvae of S. mosellana in the diapause stage significantly decreased as compared to that in larvae in the pre-diapause stage, remained low level in larvae throughout the diapause stage, significantly increased in larvae in the post-diapause quiescent stage, and increased again in larvae in the post-diapause developmental stage which was significantly higher than those in larvae in the diapause and post-diapause quiescent stages. The open reading frames of SmCOX1 and SmCOX2 (GenBank accession number: PP466915 and PP466916, respectively) obtained were 1 536 and 660 bp in length, respectively, and contained more than 75% A+T base. SmCOX1 and SmCOX2 encoded the proteins of 511 and 220 amino acids with the predicted molecular weight of 57.70 and 25.76 kD, respectively. The amino acid sequence analysis indicated that SmCOX1 and SmCOX2 contained the classical REDOX copper coenzyme factors (CuA and CuB) and heme-containing metal centers heme a and heme a3 of mitochondria COX catalytic core. SmCOX1 and SmCOX2 displayed the highest amino acid sequence identity and the closest relationship to corresponding homologues from Orseolia oryzae (Cecidomyiidae). qPCR result showed that the expression levels of SmCOX1 and SmCOX2 exhibited significant difference in the diapause process of S. mosellana. The expression levels of SmCOX1 and SmCOX2 in larvae in the pre-diapause and post-diapause developmental stages were the highest, followed by those in larvae in the post-diapause quiescence stage, and those in larvae in the diapause stage were the lowest, showing the change trend basically consistent with the activity change of SmCOX. 【Conclusion】 The decrease of SmCOX activity and SmCOX1/2 expression during the diapause of S. mosellana were closely related to low oxygen consumption in diapause stage, while their increase was closely related to more energy demand in the diapause termination and post-diapause developmental stages.

【Aim】 This study aims to clone the matrix metalloproteinase-14 gene AcMMP14 in Apis cerana and to analyze its molecular characteristics and expression pattern, so as to offer the reference and basis for continuous and further functional study of AcMMP14. 【Methods】 Total RNA from the 6-day-old larval gut of A. cerana workers was extracted, and the coding sequence (CDS) of AcMMP14 was amplified by PCR. Relevant bioinformatic software was employed to predict the physicochemical property and molecular characteristics of AcMMP14, to identify structural domains and conserved motifs in MMP14s from A. cerana and other bee species, followed by phylogenetic analysis. RT-qPCR was utilized to detect the relative expression levels of AcMMP14 in egg, larva, prepupa, pupa and different day-old adult workers, and antenna, brain, cuticle, fat body, venom gland, midgut and hypopharyngeal gland of the adult workers, and the relative expression levels of AcMMP14 in the midgut of the newly emerged 1-day-old adult A. cerana workers at 1-4 d post inoculation with Nosema ceranae. 【Results】 AcMMP14 has a molecular formula of  C₃₀₆₈H₄₅₈₁N₈₀₃O₉₁₅S₂₀, a molecular weight of about 68.00 kD, a liposoluble coefficient of 64.12, a theoretical isoelectric point of 5.85, and an average hydrophilic coefficient of -0.47. AcMMP14 contains one signal peptide and 36 phosphorylation sites, and can be simultaneously located in mitochondria, nucleus and cytoplasm. Three same structural domains and five same conserved motifs were included in MMP14 proteins from A. cerana and other 10 bee species. MMP14 of A. cerana and A. dorsata had the amino acid sequence identity of 94.23% and clustered into one clade on the phylogenetic tree. AcMMP14 was differentially expressed in egg, 3-day-old larva, 1- and 2-day-old prepupae and 4-day-old pupa of A. cerana workers, and the expression level of AcMMP14 was the highest in the 4-day-old pupa and significantly higher than those in the 3-day-old larva and 2-day-old prepupa. AcMMP14 was differentially expressed in different day-old adult workers, and the expression level of AcMMP14 was the highest in the 15-day-old adult and significantly higher than those in the 1-, 2-, 6-, 12- and 17-day-old adult. AcMMP14 was differentially expressed in the seven tissues of adult workers, and the expression level of AcMMP14 was the highest in the antenna and significantly higher than those in the brain, cuticle, fat body, venom gland, midgut and hypopharyngeal gland. The expression levels of AcMMP14 in workers’ midguts of adults at 1 and 2 d post inoculation with N. ceranae were significantly down-regulated, while those in workers’ midguts of adults at 3 and 4 d post inoculation with N. ceranae were down-regulated without significant difference as compared with those of the control group. 【Conclusion】 AcMMP14 is a potential hydrophilic and secretary protein. AcMMP14 plays a putative key role at different developmental stages and in different adult tissues of A. cerana workers. AcMMP14 in the adult worker’s midgut is activated and expressed at early stage of the 1st proliferation cycle of N. ceranae.

【Aim】 This study aims to explore the sublethal effects oflambda-cyhalothrin on Conogethes  punctiferalis, so as to provide a theoretical basis for its scientific application in field management of this pest. 【Methods】 The 48-h toxicity of lambda-cyhalothrin to the 3-day-old adults of C. punctiferalis was determined using the residual film method. Sublethal concentrations (LC₁, LC₅ and LC₁₀) of lambda-cyhalothrin were applied to the 3-day-old adults of C. punctiferalis, the adult longevity, fecundity and survival rate of the F₀ generation, and the larval and pupal duration, fecundity, hatching rate, pupation rate and emergence rate of the F₁ generation were observed and recorded. The age-stage, two-sex life table was constructed for the F₁ generation. 【Results】 Within 48-h exposure, the LC₁, LC₅ and LC₁₀ values of lambda-cyhalothrin against the 3-day-old adults of C. punctiferalis were 0.233, 0.601 and 0.995 mg/L, respectively. Exposure of the 3-day-old adults of C. punctiferalis to LC₁, LC₅ and LC₁₀ of lambda-cyhalothrin resulted in reductions in the average number of eggs laid per female by 5.56%, 24.55% and 51.06%, respectively. After the 3-day-old adults of C. punctiferalis were exposed to LC₁₀ of lambda-cyhalothrin, the adult longevity of the F₀ generation was significantly shortened by 1.60 d, and the oviposition period significantly decreased by 1.29 d, as compared to those in the control group exposed to acetone. The results for the F₁ population showed that, in the treatment groups with LC₁, LC₅ and LC₁₀ of lambda-cyhalothrin, the survival rates at the egg stage decreased by 8.70%, 13.75% and 30.38%, respectively, the average number of eggs laid per female decreased by 38.05%, 30.75% and 24.84%, respectively, and the survival rates at the pupal stage decreased by 2.70%, 13.87% and 27.68%, respectively, as compared to those in the control group. In addition, in the treatment groups with LC₁, LC₅ and LC₁₀ of lambda-cyhalothrin, the peak values of the age-specific reproductive rate (f_x) curve in the F₁ generation of C. punctiferalis decreased by 39.76%, 34.68% and 39.56%, respectively, and the pre-oviposition period, the peak of the female age-specific fecundity, intrinsic rate of increase (r), finite rate of increase (λ) and net rate of increase of the F₁ generation (R₀) were all significantly reduced as compared to those in the control group. 【Conclusion】 Sublethal concentrations of lambda-cyhalothrin significantly reduced the longevity and fecundity of the F₀ generation of C. punctiferalis, and also caused a significant decline in the number of eggs laid of the F₁ generation, thereby suppressing population growth. The results of this study provide a theoretical foundation for the scientific application of lambda-cyhalothrin in the management of C. punctiferalis.

 Reactive oxygen species (ROS) is a general term for a class of oxygen-containing free radicals formed due to incomplete oxidation of oxygen molecules, or peroxides that are easy to form oxygen free radicals. When insects are invaded by pathogens, the ROS defense system mediated by dual oxidase (DUOX) will respond quickly to produce a large amount of ROS to resist the invasion of pathogenic bacteria, and then play a role in regulating the immune defense process of insects. However, high level of ROS can damage the biological macromolecules such as proteins, DNA and lipids in cells, causing damage to insect cells and affecting the normal development of insects. In order to avoid damage from excessive oxidative stress, insects have formed a complete antioxidant defense system mainly composed of antioxidant enzymes and small molecule antioxidants to prevent excessive damage from occurring. It is interesting that changes in ROS levels in cells can play completely different roles in regulating insect lifespan: for certain insects the accumulation of a large amount of ROS could lead to a shortened lifespan, while for some other insects the presence of high physiological levels of ROS could also induce diapause and prolong their lifespan. Studies on the regulatory mechanisms of ROS in insect lifespan have achieved a lot of progress in recent years. Therefore, in this article, we comprehensively reviewed the sources and influencing factors of insect ROS, the defense mechanisms mediated by ROS, and for the first time made a summary and outlook on the specific roles of ROS in regulating insect lifespan, so as to provide a reference for subsequent research on ROS-related topics.

【Aim】High-fat diet has been proven to induce various diseases such as hyperlipidemia, posing a threat to human health. This study investigated the effects of high-fat diet on the growth and development, economic traits, lipid metabolism and immune function of Bombyx mori by adding soybean oil to mulberry leaves, to evaluate the feasibility of using B. mori to construct a hyperlipidemia disease model for human health research. 【Methods】 The day-0 4th instar and day-0 5th instar larvae of B. mori were fed with high-fat diets (mulberry leaves treated with 0.5%, 1.0% and 2.0% soybean oil, respectively) until maturity. The body weight of various day-old larvae and the cocoon layer rate were measured. The triglyceride contents in the fat body and serum of the 5th instar larvae of B. mori fed with the mulberry leaves treated with 0.5% soybeal oil were detected using a triglyceride test kit. Oil red O staining was used to observe the morphological changes and average optical density values of lipid droplets. Real-time fluorescence quantitative PCR was used to analyze the transcription levels of eight immune-related genes (BmCecD2, BmCecA, BmCecB, BmCecE, BmCec-CBM2-2, BmSpz1, BmDual and BmKayak) in the 5th instar larval fat bodies. 【Results】Compared with the control group (fed with regular mulberry leaves), high-fat diet (nulberry leaves treated with 0.5%, 1.0% and 2.0% soybean oil, respectively) significantly inhibited the body weight gain of B. mori larvae and significantly reduced the cocoon layer rate. Compared with the control, a high-lipid diet (mulbery leaves treated with 0.5% soybean oil) significantly increased the triglyceride content in the fat body and serum of the 5th instar larvae of B. mori, resulted in abnormal morphology of fat body and accumulation of lipid droplets, and significantly down-regulated the expression levels of the tested eight innate immune-related genes in the fat body of the 5th instar larvae. Among them, the expression levels of BmCecE, BmDual and BmKayak were down-regulated by more than 90%.【Conclusion】The high-fat diet leads to slow growth, delayed development, decreased economic traits, disrupted lipid metabolism and weakened immune regulatory function in B. mori. Among them, the increase in triglyceride content is similar to the physiological and pathological characteristics of human hyperlipidemia. This study provides a theoretical basis for constructing disease models using insects.

【Aim】 To reveal the functions of the cytochrome P450 (CYP450）genes, HcCYP4S47 and HcCYP332A29, in Hyphantria cunea in response to chlorantraniliprole stress.【Methods】The diets containing LC₃₀ (0.06 mg/L) of chlorantraniliprole were fed to the 3rd instar larvae of the laboratory population, northern population (Tieling population from Liaoning), and southern population (Dawu population from Xiaogan, Hubei) of H. cunea via mixing the pesticide into the diets, and the surviving larvae were collected at 6, 12, 24 and 48 h after feeding treatment, respectively. The CYP450 activity in the 3rd instar larvae of H. cunea was determined. The expression levels of HcCYP4S47 and HcCYP332A29 in the 3rd instar larvae of the three geographical populations after exposure to 0.06 mg/L of chlorantraniliprole were analyzed using RT-qPCR. Additionally, HcCYP4S47 and HcCYP332A29 in the 3rd instar larvae of the three geographical populations were silenced by RNAi technology, and the expression levels of target genes were detected by RT-qPCR after dsRNA injection. The diets containing 0.06 mg/L of chlorantraniliprole were fed to the gene-silenced 3rd instar larvae of the three geographical populations, and the survival rates of H. cunea larvae were recorded at 12, 24, 36, 48, 60 and 72 h after feeding.【Results】 After exposure to 0.06 mg/L of chlorantraniliprole, the CYP450 activities in the 3rd instar larvae of the northern and southern populations of H. cunea were higher than those in the laboratory population at four treatment time points (6, 12, 24 and 48 h). Specifically, the CYP450 activity in the 3rd instar larvae of the northern population in the chlorantraniliprole treatment group exhibited the highest levels at 24 and 48 h after treatment, while that in the 3rd instar larvae of the southern population in the chlorantraniliprole treatment group peaked at 12 and 24 h after treatment. Phylogenetic analysis indicated that HcCYP4S47 and HcCYP332A29 belong to the CYP4 and CYP3 subfamilies, respectively. The expression levels of HcCYP4S47 and HcCYP332A29 were significantly higher in the 3rd instar larvae of the northern and southern populations than those in the 3rd instar larvae of the laboratory population. After exposure to 0.06 mg/L of chlorantraniliprole, the expression levels of HcCYP4S47 and HcCYP332A29 were significantly up-regulated in the 3rd instar larvae of the laboratory population as compared to those in the control group, peaking at 12 h post treatment. The expression levels of HcCYP4S47 in the 3rd instar larvae of the southern and northern populations were significantly down-regulated at 6, 12 and 48 h after chlorantraniliprole treatment, that in the northern population was significantly up-regulated and that in the southern population showed no significant change at 24 h after chlorantraniliprole treatment, as compared to that in the control group. The expression level of HcCYP332A29 in the 3rd instar larvae of the northern population in the chlorantraniliprole treatment group was significantly decreased at 6 and 12 h, but significantly up-regulated at 48 h, as compared to that in the control group. The expression level of HcCYP332A29 in the 3rd instar larvae was significantly decreased at 6 and 12 h after chlorantraniliprole treatment but significantly up-regulated at 48 h after chlorantraniliprole treatment as compared to that in the control group. In the 3rd instar larvae of the southern population, the expression level of HcCYP332A29 was significantly down-regulated at 6 h after chlorantraniliprole treatment, followed by a significant increase at 12 and 24 h. Silencing HcCYP332A29 and HcCYP4S47 increased the susceptibility of the 3rd instar larvae of H. cunea to chlorantraniliprole. 【Conclusion】 HcCYP332A29 and HcCYP4S47 play crucial roles in the response of H. cunea to chlorantraniliprole stress.