Insects

Editorial

Jump to: Research, Review

5 pages, 156 KB

Open AccessEditorial

RNAi in Insect Physiology: Unlocking Mechanisms and Pioneering Sustainable Pest Control

by Jisheng Liu and Luc Swevers

Insects 2026, 17(3), 333; https://doi.org/10.3390/insects17030333 - 19 Mar 2026

Viewed by 659

Abstract

The advent of RNA interference (RNAi) in 1998, marked by the discovery of gene silencing triggered by double-stranded RNA (dsRNA) in nematodes, opened a transformative chapter in molecular biology [...] Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

Research

Jump to: Editorial, Review

17 pages, 2098 KB

Open AccessArticle

Functional Characterization of Akt, a Serine/Threonine Kinase, in Tuta absoluta: A Key Regulator of Molting, Hormones, and Reproduction with Potential for RNAi Pest Control

by Chaoshan Liang, Jiahui Song, Kangkang Xu, Fanghao Wan, Guy Smagghe and Wenjia Yang

Insects 2026, 17(2), 183; https://doi.org/10.3390/insects17020183 - 8 Feb 2026

Viewed by 734

Abstract

Insect insulin signaling plays a central role in regulating development, metamorphosis, and reproduction, yet its mechanistic functions in the tomato leafminer, Tuta absoluta, a globally significant pest, remain poorly understood. This study aimed to elucidate the role of the serine/threonine kinase Akt [...] Read more.

Insect insulin signaling plays a central role in regulating development, metamorphosis, and reproduction, yet its mechanistic functions in the tomato leafminer, Tuta absoluta, a globally significant pest, remain poorly understood. This study aimed to elucidate the role of the serine/threonine kinase Akt (TaAkt) in coordinating metamorphosis and female reproductive processes. The TaAkt gene was cloned and characterized, and its spatiotemporal expression was analyzed across various developmental stages and tissues. RNA interference (RNAi) was employed to knock down TaAkt in late pupae and newly emerged females, followed by assessment of pupal-adult eclosion, chitin metabolism, 20-hydroxyecdysone (20E) titer, ovarian development, juvenile hormone (JH) levels, vitellogenin synthesis, and fecundity. Knockdown of TaAkt significantly reduced 20E titers and downregulated the expression of ecdysone biosynthesis and signaling genes, leading to pupal mortality, defective molting, and reduced chitin content. In adult females, TaAkt silencing impaired ovarian growth, decreased JH levels, suppressed vitellogenin production, and reduced egg number and hatching rates. These findings demonstrate that TaAkt exerts pleiotropic control over both metamorphic and reproductive processes in T. absoluta. The study identifies TaAkt as a promising molecular target for RNAi-based pest management strategies, offering a potential approach to simultaneously suppress survival and reproductive capacity in this economically important pest. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

16 pages, 3342 KB

Open AccessArticle

Identification and Functional Characterization of Chitinase Genes During Larva–Pupa–Adult Transitions in Tuta absoluta

by Kangkang Xu, Yue Wang, Shuyan Yan, Fanghao Wan, Guy Smagghe and Wenjia Yang

Insects 2026, 17(1), 114; https://doi.org/10.3390/insects17010114 - 20 Jan 2026

Cited by 2 | Viewed by 910

Abstract

Chitinases are crucial in the molting process of insects and represent potential targets for the development of RNA interference (RNAi)-based insecticidal strategies. In this study, we identified and characterized 11 chitinase genes (TaChts) in T. absoluta, each harboring at least [...] Read more.

Chitinases are crucial in the molting process of insects and represent potential targets for the development of RNA interference (RNAi)-based insecticidal strategies. In this study, we identified and characterized 11 chitinase genes (TaChts) in T. absoluta, each harboring at least one GH18 catalytic domain. Phylogenetic analysis placed these proteins into 11 established groups (I-X and h), revealing conserved lineage-specific patterns. Spatiotemporal expression profiling showed that most TaChts peak during key developmental transitions. TaCht1, TaCht2, TaCht3, TaCht5, TaCht6, TaCht7, TaCht10, TaCht-h, and TaIDGF were predominantly expressed in the integument, whereas TaCht8 and TaCht11 showed gut-enriched expression, indicating functional specialization. Stage-tailored RNAi assays demonstrated that silencing TaCht5, TaCht7, TaCht10, and TaIDGF caused pronounced mortality in both larvae and pupae. In larvae, delivery of dsRNA using carbon quantum dots (CQDs) disrupted molting: knockdown of TaCht7, TaCht10, and TaIDGF prevented shedding of the old cuticle, whereas TaCht5 silencing produced severe cuticular shrinkage and blackening. Hematoxylin and eosin staining revealed detachment of epidermal cells from the cuticle, providing cytological evidence of disrupted cuticle–epidermis remodeling. In pupae, microinjection of dsRNA induced cuticular collapse, melanization, and eclosion failure due to persistent adhesion of the pupal case, accompanied by marked reductions in chitinase activity. Together, our findings highlight the critical roles of four key chitinases in endocuticular turnover and metamorphic progression in T. absoluta, offering mechanistic insight into chitin-mediated developmental processes. The identified genes represent strong candidates for RNAi-based, species-specific pest management strategies against this globally significant insect. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

31 pages, 10340 KB

Open AccessArticle

Silencing the cyp314a1 and cyp315a1 Genes in the Aedes albopictus 20E Synthetic Pathway for Mosquito Control and Assessing Algal Blooms Induced by Recombinant RNAi Microalgae

by Xiaodong Deng, Changhao He, Chunmei Xue, Dianlong Xu, Juncai Li and Xiaowen Fei

Insects 2025, 16(10), 1033; https://doi.org/10.3390/insects16101033 - 7 Oct 2025

Viewed by 1412

Abstract

As one of the key vectors for the transmission of Dengue fever, Aedes albopictus is highly ecologically adaptable. The development of environmentally compatible biological defence and control technologies has therefore become an urgent need for vector biological control worldwide. This study constructed and [...] Read more.

As one of the key vectors for the transmission of Dengue fever, Aedes albopictus is highly ecologically adaptable. The development of environmentally compatible biological defence and control technologies has therefore become an urgent need for vector biological control worldwide. This study constructed and used double-stranded RNA (dsRNA) expression vectors targeting the cyp314a1 and cyp315a1 genes of Ae. albopictus to transform Chlamydomonas reinhardtii and Chlorella vulgaris, achieving RNA interference (RNAi)-mediated gene silencing. The efficacy of the RNAi recombinant algal strain biocide against Ae. albopictus was evaluated by administering it to Ae. albopictus larvae. The results showed that the oral administration of the cyp314a1 and cyp315a1 RNAi recombinant C. reinhardtii/C. vulgaris strains was lethal to Ae. albopictus larvae and severely affected their pupation and emergence. The recombinant algal strains triggered a burst of ROS (Reactive Oxygen Species) in the mosquitoes’ bodies, resulting in significant increases in the activities of the superoxide dismutase (SOD), peroxiredoxin (POD) and catalase (CAT), as well as significant upregulation of the mRNA levels of the CME pathway genes in larvae. In the simulated field experiment, the number of Ae. albopictus was reduced from 1000 to 0 in 16 weeks by the RNAi recombinant Chlorella, which effectively controlled the population of mosquitoes. Meanwhile, the levels of nitrogen (N), phosphorus (P), nitrate, nitrite, ammonia and COD (Chemical Oxygen Demand) in the test water decreased significantly. High-throughput sequencing analyses of 18S rDNA and 16S rDNA showed that, with the release of RNAi recombinant Chlorella into the test water, the biotic community restructuring dominated by resource competition caused by algal bloom, as well as the proliferation of anaerobic bacteria and the decline of aerobic bacteria triggered by anaerobic conditions, are the main trends in the changes in the test water. This study is an important addition to the use of RNAi recombinant microalgae as a biocide. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

15 pages, 2320 KB

Open AccessArticle

Elimination of Ultraviolet Light-Mediated Attraction Behavior in Culex Mosquitoes via dsRNA-Mediated Knockdown of Opsins

by Xinyi Liu, Guoqiang Zhao, Hui Liu, Yuxuan Mao, Meng Xu, Jing Wu, Lijiao Li, Zongzhao Zhai and Pa Wu

Insects 2025, 16(10), 997; https://doi.org/10.3390/insects16100997 - 25 Sep 2025

Cited by 1 | Viewed by 1911

Abstract

It is widely recognized that mosquitoes are attracted to ultraviolet (UV) light traps in field and semi-field trials. However, the specific characteristics of mosquito behavioral responses to UV light remain poorly defined. Moreover, the molecular mechanisms underlying their phototactic behavior remain unexplored. Here, [...] Read more.

It is widely recognized that mosquitoes are attracted to ultraviolet (UV) light traps in field and semi-field trials. However, the specific characteristics of mosquito behavioral responses to UV light remain poorly defined. Moreover, the molecular mechanisms underlying their phototactic behavior remain unexplored. Here, we characterized mosquito photobehavior under UV light in a laboratory setting using three experimental apparatuses. Our findings indicate that mosquitoes exhibit strong attraction to low-intensity UV light, yet show no preference between high-intensity UV light and darkness. Video recordings and automatic analyses of photobehavior under low-intensity UV light revealed that mosquitoes preferred the window illuminated by UV light over an unilluminated window and were more active when exposed to UV light. Through RNA interference (RNAi)-mediated knockdown of opsins highly expressed in the adult stage of Culex quinquefasciatus, we identified CqOpsin3, CqOpsin5, and CqOpsin6 as crucial mediators of UV phototaxis. This study provides methods for characterizing mosquito photobehavior under UV light in the laboratory, and represents the first mechanistic investigation into UV light-mediated attractive behavior in mosquitoes. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Graphical abstract

17 pages, 5194 KB

Open AccessArticle

Transcriptome and Functional Analyses Revealed the Carboxylesterase Genes Involved in Pyrethroid Resistance in Anopheles sinensis (Diptera: Culicidae)

by Yiyun Wei, Xinyao Gu, Fengling Si, Xiaojie Chen, Liang Qiao, Hongxing Yan and Bin Chen

Insects 2025, 16(9), 938; https://doi.org/10.3390/insects16090938 - 5 Sep 2025

Cited by 1 | Viewed by 1546

Abstract

Carboxylesterases (CCEs) have been demonstrated to be involved in pyrethroid resistance in insect species. This study aims to investigate CCE-mediated resistance mechanisms in Anopheles sinensis, a major malaria vector. Through comparative transcriptomics of a deltamethrin-resistant strain (CQ-LR) versus susceptible strain (WX-LS) of [...] Read more.

Carboxylesterases (CCEs) have been demonstrated to be involved in pyrethroid resistance in insect species. This study aims to investigate CCE-mediated resistance mechanisms in Anopheles sinensis, a major malaria vector. Through comparative transcriptomics of a deltamethrin-resistant strain (CQ-LR) versus susceptible strain (WX-LS) of An. sinensis, we identified differentially expressed CCE genes across five developmental stages, five tissues, and three time points post-blood-meal. Four candidate genes (AsAe9, AsAe10, AsAce2, AsUn5) showed significantly upregulated expression. Subsequent qRT-PCR validation across four field-derived resistant strains (WX-LR, AH-LR, YH-LR, CQ-LR) and the susceptible strain confirmed significant upregulation of AsAe9, AsAe10, AsAce1, AsAce2, and AsBe4 in more than two resistant populations. RNAi-based functional validation showed that silencing AsAe10 or AsBe4 in the WX-LR strain significantly decreased knockdown time and raised 24 h mortality upon diagnostic deltamethrin exposure, with AsAe10 silencing having the strongest effect. This study identifies CCE genes involved in deltamethrin resistance in An. sinensis, providing valuable insights into the resistance mechanisms of pyrethroid and a theoretical basis for mosquito resistance management. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Graphical abstract

12 pages, 1515 KB

Open AccessArticle

Expression of Heat Shock Protein 90 Genes Induced by High Temperature Mediated Sensitivity of Aphis glycines Matsumura (Hemiptera: Aphididae) to Insecticides

by Xue Han, Yulong Jia, Changchun Dai, Xiaoyun Wang, Jian Liu and Zhenqi Tian

Insects 2025, 16(8), 772; https://doi.org/10.3390/insects16080772 - 28 Jul 2025

Cited by 4 | Viewed by 1386

Abstract

Soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is a major pest of soybean fields. While high-temperature stress induced by global warming can initially suppress aphid populations, these pests may eventually adapt, leading to more severe infestations and crop damage. Heat shock proteins (HSPs), [...] Read more.

Soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is a major pest of soybean fields. While high-temperature stress induced by global warming can initially suppress aphid populations, these pests may eventually adapt, leading to more severe infestations and crop damage. Heat shock proteins (HSPs), which are upregulated in response to heat stress to protect aphid development, also confer tolerance to other abiotic stressors, including insecticides. To investigate the role of HSPs in insecticide resistance in A. glycines, we analyzed the expression profiles of three AgHsp90 genes (AgHsp75, AgHsp83, and AgGrp94) following exposure to high temperatures and insecticides. Functional validation was performed using RNA interference (RNAi) to silence AgHsp90 genes. Our results demonstrated that AgHsp90 genes were significantly upregulated under both heat and insecticide stress conditions. Furthermore, after feeding on dsRNA of AgHsp90 genes, mortality rates of A. glycines significantly increased when exposed to imidacloprid and lambda-cyhalothrin. This study provides evidence that AgHsp90 genes play a crucial role in mediating thermal tolerance and insecticide resistance in A. glycines. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

14 pages, 1508 KB

Open AccessArticle

Effects of Trehalase on the Gene Expression of the Reproductive Regulation Pathway Network and Triglyceride Metabolism in Nilaparvata lugens (Stål)

by Bin Tang, Yuxin Ge, Yongkang Liu, Liwen Guan, Ye Han, Yang Zhu, Gao Hu and Yan Wu

Insects 2025, 16(7), 725; https://doi.org/10.3390/insects16070725 - 16 Jul 2025

Cited by 2 | Viewed by 1335

Abstract

The high reproductive performance of Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) is one of the key factors contributing to its serious damage. Studies have demonstrated that trehalose hydrolysis, catalyzed by trehalase (TRE), plays an important role in the reproductive regulation of N. lugens. [...] Read more.

The high reproductive performance of Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) is one of the key factors contributing to its serious damage. Studies have demonstrated that trehalose hydrolysis, catalyzed by trehalase (TRE), plays an important role in the reproductive regulation of N. lugens. However, it remains unclear how TRE regulates the reproduction of N. lugens. To address this question, the current study was designed to investigate the effects of TRE on the reproductive regulatory network of N. lugens. Specifically, RNA interference (RNAi) was conducted by injecting double-stranded RNA (dsRNA) targeting the TRE genes, and a trehalase inhibitor, validamycin, was used to suppress TRE activity. Subsequently, the relative expression levels of key genes involved in the reproductive regulatory pathway, as well as the triglyceride content in the fat body and ovary, were determined. The results indicated that both dsTREs and validamycin treatment decreased the relative expression of Vitellogenin (Vg) without affecting the expression of its receptor, VgR. However, their impact on the reproductive regulatory network differed: dsTREs injection did not affect the relative expression of JHAMT and Met in the juvenile hormone signaling pathway, but resulted in the upregulation of USP and TOR, and inhibition of InR2 and S6K. In contrast, validamycin injection inhibited the expression of Met, USP, EcR, InR1, TOR, and S6K. Both dsTREs and validamycin inhibited the expression of Fas, but only validamycin decreased the ovarian triglyceride content. These findings suggest that the inhibition of TRE leads to alterations in reproductive regulatory signals, thereby reducing the reproductive capacity of N. lugens. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

14 pages, 5582 KB

Open AccessArticle

Silencing Miniature Gene Disrupts Elytral and Hindwing Structures in Leptinotarsa decemlineata

by Man-Hong Cheng, Kai-Yun Fu, Wei Zhou, Ji-Feng Shi and Wen-Chao Guo

Insects 2025, 16(7), 700; https://doi.org/10.3390/insects16070700 - 8 Jul 2025

Viewed by 1337

Abstract

The Colorado potato beetle (Leptinotarsa decemlineata, CPB) is a major pest in potato crops, notorious for its rapid dispersal and insecticide resistance, which are enabled by its robust elytra and flight-capable hindwings. The Miniature (Mi) gene, encoding a protein [...] Read more.

The Colorado potato beetle (Leptinotarsa decemlineata, CPB) is a major pest in potato crops, notorious for its rapid dispersal and insecticide resistance, which are enabled by its robust elytra and flight-capable hindwings. The Miniature (Mi) gene, encoding a protein with a zona pellucida (ZP) domain, is involved in wing development and cuticle integrity, yet its functional role in beetles remains underexplored. In this study, we cloned and characterized the LdMi gene in the CPB and investigated its function using RNA interference (RNAi), morphological analyses, and spectroscopy. LdMi encodes a 146.35 kDa transmembrane protein with a conserved ZP domain, clusters with coleopteran homologs, and exhibits relative conservation across insect species. Expression profiling showed high LdMi transcript levels in the hindwings, the elytra, and the pupal stages. RNAi knockdown in fourth-instar larvae resulted in severe eclosion defects, including malformed wings and reduced adult weight. Scanning electron microscopy (SEM) revealed disrupted elytral patterns and deformed hindwing veins in knockdown individuals. Spectroscopic analyses using Fourier-transform infrared (FTIR) and Raman spectroscopy indicated a reduction in protein–chitin crosslinking and diminished hydrogen bonding, suggesting compromised cuticular integrity. These results highlight the essential role of LdMi in cuticle formation and the surface morphology of the elytra and hindwings, offering new insights into ZP domain proteins in insects. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

16 pages, 2174 KB

Open AccessArticle

Development of a Stage- and Species-Specific RNAi System for Molecular Insights in Trichogramma Wasps

by Zelong Yang, Yan Lu, Zhuo Jiang, Xilin Jiao, Han Lin, Wanning Jiang, Wenmei Du, Xue Zhang, Zhao Peng, Junjie Zhang, Xiao Wang and Ying Hu

Insects 2025, 16(7), 673; https://doi.org/10.3390/insects16070673 - 27 Jun 2025

Cited by 1 | Viewed by 1463

Abstract

Trichogramma wasps, egg parasitoids widely used to control lepidopteran pests, have long eluded in-depth molecular mechanistic studies due to their minute size and genetic tool scarcity. While previous RNAi efforts were restricted to T. dendrolimi, we developed the first cross-species RNAi system [...] Read more.

Trichogramma wasps, egg parasitoids widely used to control lepidopteran pests, have long eluded in-depth molecular mechanistic studies due to their minute size and genetic tool scarcity. While previous RNAi efforts were restricted to T. dendrolimi, we developed the first cross-species RNAi system for both T. dendrolimi and the previously intractable T. ostriniae. Temporal expression profiling identified white and laccase 2 as stage-specific RNAi targets, peaking during prepupal/pupal stages, which were tested across species and developmental stages using microinjection and soaking dsRNA delivery methods. Survival analysis prioritized soaking for T. dendrolimi prepupae/pupae, while microinjection was essential for T. ostriniae to bypass prepupal mortality during soaking. Concentration-dependent RNAi targeting the white gene achieved 85.61% transcript reduction in T. dendrolimi via soaking and 89.36% in T. ostriniae via microinjection at 2000 ng/μL, correlating with 64.06% and 32.09% white-eyed pupae, causing a significant reduction in eye pigments. For the laccase 2 gene, soaking at 2000 ng/μL induced 88.35% transcript reduction in T. dendrolimi and 73.31% in T. ostriniae, leading to incomplete cuticle tanning and sclerotization. This study resolves the long-standing challenge of genetic manipulation in Trichogramma wasps, providing a universally applicable framework to decipher parasitoid–host interactions at the molecular scale, which is useful for sustainable pest management strategies. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

16 pages, 1963 KB

Open AccessArticle

Characterization and Functional Analysis of Small Heat Shock Protein Genes (Hsp22.2 and Hsp26.7) in Sitodiplosis mosellana Diapause

by Qitong Huang, Qian Ma, Xiaobin Liu, Keyan Zhu-Salzman and Weining Cheng

Insects 2025, 16(7), 649; https://doi.org/10.3390/insects16070649 - 20 Jun 2025

Cited by 1 | Viewed by 1408

Abstract

Small heat shock proteins (sHsps) play crucial roles in organismal adaptation to stress tolerance. Sitodiplosis mosellana, a devastating insect wheat pest, undergoes long obligatory larval diapause to survive temperature extremes during summer and winter. To elucidate the function of sHsps in this [...] Read more.

Small heat shock proteins (sHsps) play crucial roles in organismal adaptation to stress tolerance. Sitodiplosis mosellana, a devastating insect wheat pest, undergoes long obligatory larval diapause to survive temperature extremes during summer and winter. To elucidate the function of sHsps in this process, two sHsp-encoding genes (SmHsp22.2 and SmHsp26.7) were characterized from S. mosellana, and their responsiveness to diapause and thermal stress, as well as their roles in cold stress, was analyzed. Both SmHsp22.2 and SmHsp26.7 possessed the canonical α-crystallin domain and lacked introns. Quantitative PCR indicated significant upregulation of SmHsp22.2 and SmHsp26.7 during diapause, especially in summer and winter. Notably, SmHsp22.2 exhibited higher expression in summer relative to winter, whereas SmHsp26.7 showed the opposite profile. Moreover, short-term heat shock (≥35 °C) in over-summering larvae or cold shock (≤−10 °C) in over-wintering larvae was found to trigger transcriptional upregulation of both genes, while prolonged temperature extremes (i.e., 45–50 °C or −15 °C) did not elicit a comparable response. RNA interference-mediated knockdown of both genes significantly increased the mortality of S. mosellana larvae under cold stress. These findings indicate the importance of both SmHsps in diapause and environmental adaptation in S. mosellana. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Graphical abstract

12 pages, 1087 KB

Open AccessArticle

Activation of BmToll9-1 in Silkworm (Bombyx mori) Larval Midgut by Escherichia coli and Regulation of Growth

by Jisheng Liu, Weijian Chen, Minchun Lai, Jiahua Chen and Luc Swevers

Insects 2025, 16(6), 621; https://doi.org/10.3390/insects16060621 - 11 Jun 2025

Cited by 3 | Viewed by 2774

Abstract

Insects rely on their innate immune system to defend against pathogens, and the Toll signaling pathway plays an important role in immune regulation. Our previous studies have shown that BmToll9-1 functions as a positive regulator in the Toll pathway. This study seeks to [...] Read more.

Insects rely on their innate immune system to defend against pathogens, and the Toll signaling pathway plays an important role in immune regulation. Our previous studies have shown that BmToll9-1 functions as a positive regulator in the Toll pathway. This study seeks to elucidate the role of BmToll9-1, as a sensor to bacterial challenge, in modulating larval development and downstream Toll signaling pathways. Silkworm larvae were subjected to infection with either Gram-negative Escherichia coli or Gram-positive Staphylococcus aureus bacteria following silencing of BmToll9-1 by RNA interference (RNAi). This bacterial challenge triggered a compensatory re-induction of BmToll9-1 expression, which resulted in the recovery of larval weight and size to levels observed in untreated controls. Furthermore, upon bacterial infection of BmToll9-1-silenced larvae, there was an up-regulation in the expression of both signaling genes in the Toll pathway and downstream effector genes, with a marked preference for Gram-negative bacteria. These results highlight the involvement of BmToll9-1 in the Toll signaling pathway as a positive regulator, influencing silkworm development. Additionally, BmToll9-1 and BmToll9-2 were cross-validated to be genetically distinct genes, even though they were confirmed to be functionally analogous in the silkworm. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

14 pages, 2253 KB

Open AccessArticle

Risk Assessment of RNAi-Based Potential Pesticide dsNlAtg3 and Its Homologues for Nilaparvata lugens and Non-Target Organisms

by Kai Li, Tongtong Chen, Yuliang Li, Kai Sun, Kun Pang, Xiaoping Yu and Peiying Hao

Insects 2025, 16(2), 225; https://doi.org/10.3390/insects16020225 - 19 Feb 2025

Cited by 5 | Viewed by 2143

Abstract

The brown planthopper (Nilaparvata lugens) is an insect pest of rice, which mainly feeds on the phloem sap of the leaf sheath. RNA interference (RNAi) has application prospects in pest control, but it is necessary to select target genes and design [...] Read more.

The brown planthopper (Nilaparvata lugens) is an insect pest of rice, which mainly feeds on the phloem sap of the leaf sheath. RNA interference (RNAi) has application prospects in pest control, but it is necessary to select target genes and design suitable dsRNA fragments for RNAi so that it can achieve effective pest control and avoid risks to non-target organisms. NlAtg3 is a key protein in the autophagy pathway of N. lugens. Three kinds of dsRNA fragments of the NlAtg3 gene (dsNlAtg3-474×1, dsNlAtg3-138×3 and dsNlAtg3-47×10) were designed to compare the RNAi efficiency and specificity against the target insect N. lugens and non-target organisms through microinjection. The results showed that the fragment dsNlAtg3-474×1 showed strong inhibitory effects on the survival of N. lugens, which resulted in the survival rate decreasing to zero on the fifth day, while the survival rate of a closely related species, Sogatella furcifera, dropped to 2.22%. In contrast, dsNlAtg3-47×10 specifically designed against N. lugens only showed slight or no inhibitory effects on S. furcifera and other non-target organisms such as Drosophila melanogaster, but still showed good lethal effects against N. lugens, with the survival rate dropping to 18.89% on the ninth day. In addition, after being fed N. lugens injected with dsNlAtg3-47×10 fragments, the survival rate of the natural enemies Dolomedes sulfureus and Tytthus chinensis did not show significant change, compared with those treated with the dsGFP control. Our results suggest that the NlAtg3 gene can serve as a potential target for controlling N. lugens. Moreover, by designing suitable RNAi fragments, it is possible to avoid harm to non-target organisms while effectively inhibiting the target insect N. lugens. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

Review

Jump to: Editorial, Research

18 pages, 1810 KB

Open AccessReview

Odorant Binding Proteins in Tribolium castaneum: Functional Diversity and Emerging Applications

by Lei Wang, Yujie Lu and Zongpei Zhao

Insects 2025, 16(12), 1250; https://doi.org/10.3390/insects16121250 - 10 Dec 2025

Viewed by 1162

Abstract

Tribolium castaneum, the red flour beetle, is both a major pest of stored products and a valuable genetic model. Odorant Binding Proteins (OBPs), traditionally associated with olfaction, are now recognized as multifunctional, contributing to detoxification, immunity, and reproduction. This review synthesizes recent [...] Read more.

Tribolium castaneum, the red flour beetle, is both a major pest of stored products and a valuable genetic model. Odorant Binding Proteins (OBPs), traditionally associated with olfaction, are now recognized as multifunctional, contributing to detoxification, immunity, and reproduction. This review synthesizes recent advances in the molecular structure, gene expression, and functional characterization of T. castaneum OBPs—particularly TcOBPC11, TcOBPC12, TcOBPC17, and TcOBP7G. Experimental evidence, including RNA interference, ligand-binding assays, and expression profiling, supports their role in defense against xenobiotics. Comparative genomic analyses reveal lineage-specific expansions and adaptive evolution, especially in Minus-C OBPs. Translational applications include RNA interference (RNAi)-based pest control, OBP-targeting repellents, and biosensors. Outstanding challenges remain, including structural resolution and functional redundancy. Future research integrating CRISPR, single-cell transcriptomics, and structural biology will be critical to decode OBP regulatory networks and leverage their potential in pest management and biotechnology. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

25 pages, 2052 KB

Open AccessReview

Perspectives of RNAi, CUADb and CRISPR/Cas as Innovative Antisense Technologies for Insect Pest Control: From Discovery to Practice

by Hemant Kumar, Nikita Gal’chinsky, Verma Sweta, Nikita Negi, Roman Filatov, Anamika Chandel, Jamin Ali, Vol Oberemok and Kate Laikova

Insects 2025, 16(7), 746; https://doi.org/10.3390/insects16070746 - 21 Jul 2025

Cited by 8 | Viewed by 3458

Abstract

Pest management is undergoing a transformative shift with the development of the cutting-edge antisense technologies: RNA interference (RNAi), contact unmodified antisense DNA biotechnology (CUADb), and the CRISPR-associated proteins (CRISPR/Cas). These approaches function by facilitating sequence-specific pairing of nucleic acids followed by nuclease-mediated cleavage, [...] Read more.

Pest management is undergoing a transformative shift with the development of the cutting-edge antisense technologies: RNA interference (RNAi), contact unmodified antisense DNA biotechnology (CUADb), and the CRISPR-associated proteins (CRISPR/Cas). These approaches function by facilitating sequence-specific pairing of nucleic acids followed by nuclease-mediated cleavage, offering exceptional precision for targeted pest control. While RNA-guided mechanisms such as RNAi and CRISPR/Cas were initially characterized in non-insect systems, primarily as innate defenses against viral infections, the DNA-guided CUADb pathway was first identified in insect pests as a functional pest control strategy. Its broader role in ribosomal RNA (rRNA) biogenesis was recognized later. Together, these discoveries have revealed an entirely new dimension of gene regulation, with profound implications for sustainable pest management. Despite sharing a common principle of sequence-specific targeting RNAi, CUADb, and CRISPR/Cas differ in several key aspects, including their mechanisms of action, target specificity, and applicability. Rather than serving as universal solutions, each technology is likely to be optimally effective against specific pest groups. Moreover, these technologies allow for rapid adaptation of control strategies to overcome target-site resistance, ensuring long-term efficacy. This review summarizes the core functional characteristics, potential applications, and current limitations of each antisense technology, emphasizing their complementary roles in advancing environmentally sustainable pest control. By integrating foundational biological discoveries with applied innovations, this work provides a new perspectives on incorporating antisense-based strategies into next-generation integrated pest management systems. Full article

(This article belongs to the Special Issue RNAi in Insect Physiology)

► Show Figures

Figure 1

Journal Menu

Journal Browser

RNAi in Insect Physiology

Share This Special Issue

Special Issue Editors

Special Issue Information

Keywords

Benefits of Publishing in a Special Issue

Related Special Issue

Published Papers (15 papers)

Editorial

Research

Review

Further Information

Guidelines

MDPI Initiatives

Follow MDPI