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Insects
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RNAi in Insect Physiology
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RNAi in Insect Physiology

A special issue of Insects (ISSN 2075-4450). This special issue belongs to the section "Insect Molecular Biology and Genomics".

Deadline for manuscript submissions: 15 December 2025 | Viewed by 3433

Special Issue Editors

Dr. Jisheng Liu

E-Mail Website
Guest Editor
School of Life Sciences, Guangzhou University, Guangzhou 510006, China
Interests: insect physiology; molecular biology; RNA interference; innate immunity; Bombyx mori; Tribolium castaneum; pest management
Dr. Luc Swevers

E-Mail Website
Guest Editor
Insect Molecular Genetics and Biotechnology Group, National Centre for Scientific Research “Demokritos”, Institute of Biosciences and Applications, 15341 Athens, Greece
Interests: insect antiviral immunity; insect virology and immunology; RNA interference
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Since the discovery of the role of dsRNA to trigger gene silencing in nematodes in 1998, RNA interference (RNAi) has evolved into a potent tool for gene functional studies in fundamental research. In this process, long double-stranded RNA (dsRNA) molecules are processed into small interfering RNAs (siRNAs) that carry out specific gene silencing via hybridization to complementary target viral and cellular RNAs. In addition to siRNAs, microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs) act as participants in distinct RNAi pathways. All three small RNA pathways collectively contribute to gene silencing effects in eukaryotic cells. Importantly, due to the specific gene silencing mechanism, RNAi has recently been adopted as a novel pest control approach.

This Special Issue invites contributions on various facets of RNAi research in insects, with a particular focus on gene functional studies and pest control applications. Both original research articles and literature reviews are welcome to showcase advancements in RNAi research in the field of entomology.

Dr. Jisheng Liu
Dr. Luc Swevers
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Insects is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • insects
  • RNAi
  • dsRNA
  • small RNAs
  • siRNAs
  • miRNAs
  • piRNAs
  • gene functional studies
  • biopesticide
  • pest control

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Published Papers (5 papers)

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Research

14 pages, 5582 KiB  
Article
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
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)
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16 pages, 2174 KiB  
Article
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
Viewed by 153
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)
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16 pages, 1963 KiB  
Article
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
Viewed by 420
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)
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12 pages, 1087 KiB  
Article
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
Viewed by 1181
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)
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14 pages, 2253 KiB  
Article
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
Viewed by 811
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)
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