The western flower thrips, Frankliniella occidentalis, is the principal thrips vector of Orthotospovirus tomatomaculae (order Bunyavirales, family Tospoviridae), a devastating plant‐pathogenic virus commonly referred to as tomato spotted wilt virus (TSWV). The larval gut is the gateway for virus transmission by F. occidentalis adults to plants. In a previous report, gut expression at the transcriptome level was subtle but significant in response to TSWV in L1s. Since it has been well documented that the relationship between the expression of mRNA and associated protein products in eukaryotic cells is often discordant, we performed identical, replicated experiments to identify and quantify virus‐responsive larval gut proteins to expand our understanding of insect host response to TSWV. While we documented statistically significant, positive correlations between the abundance of proteins (4189 identified) and their cognate mRNAs expressed in first and second instar guts, there was virtually no alignment of individual genes identified to be differentially modulated by virus infection at the transcriptome and proteome levels. Predicted protein–protein interaction networks associated with clusters of co‐expressed proteins revealed wide variation in correlation strength between protein and cognate transcript abundance, which appeared to be associated with the type of cellular processes, cellular compartments and network connectivity represented by the proteins. In total, our findings indicate distinct and dynamic regulatory mechanisms of transcript and protein abundance (expression, modifications and/or turnover) in virus‐infected gut tissues. This study provides molecular candidates for future functional analysis of thrips vector competence and underscores the necessity of examining complex virus‐vector interactions at a systems level.

The western flower thrips, Frankliniella occidentalis , poses a significant challenge in global agriculture as a notorious pest and a vector of economically significant orthotospoviruses. However, the limited availability of genetic tools for F. occidentalis hampers the advancement of functional genomics and the development of innovative pest control strategies. In this study, we present a robust methodology for generating heritable mutations in F. occidentalis using the CRISPR/Cas9 genome editing system. Two eye‐colour genes, white ( Fo‐w ) and cinnabar ( Fo‐cn ), frequently used to assess Cas9 function in insects were identified in the F. occidentalis genome and targeted for knockout through embryonic microinjection of Cas9 complexed with Fo‐w or Fo‐cn specific guide RNAs. Homozygous Fo‐w and Fo‐cn knockout lines were established by crossing mutant females and males. The Fo‐w knockout line revealed an age‐dependent modification of eye‐colour phenotype. Specifically, while young larvae exhibit orange‐coloured eyes, the colour transitions to bright red as they age. Unexpectedly, loss of Fo‐w function also altered body colour, with Fo‐w mutants having a lighter coloured body than wild type, suggesting a dual role for Fo‐w in thrips. In contrast, individuals from the Fo‐cn knockout line consistently displayed bright red eyes throughout all life stages. Molecular analyses validated precise editing of both target genes. This study offers a powerful tool to investigate thrips gene function and paves the way for the development of genetic technologies for population suppression and/or population replacement as a means of mitigating virus transmission by this vector.

The western flower thrips ( Frankliniella occidentalis ) is a significant agricultural pest, causing severe global yield losses due to extensive feeding damage and the transmission of plant pathogenic viruses. Despite recent advancements in RNA interference (RNAi) in thrips species, its application has been mostly limited to the adult stage. Given the crucial role of first instar larval thrips in acquiring and transmitting orthotospoviruses, achieving gene silencing in these larvae is critical for studying virus entry and acquisition. While thoracic and abdominal injections have proven effective in adult thrips, the low post‐injection survival rate hinders their use in larval thrips. This study addresses this challenge by presenting a microinjection methodology to deliver dsRNA into the hemolymph of first instar larval thrips through the coxa, the first proximal segment of the foreleg. This method significantly improved larval survival rate by preventing detrimental damage to the internal tissues. Significant knockdown of V‐ATPase‐B , cytochrome P450 ( CYP3653A2 ), and apolipophorin‐II/I ( ApoLp‐II/I ) transcripts was confirmed after 48 and/or 72 h post injection (hpi), corresponding to the first and second instar larval stages, respectively. Silencing CYP3653A2 or ApoLp‐II/I significantly increased larval mortality. These findings demonstrate proof‐of‐principle of gene silencing and associated silencing phenotype (mortality) for first instar larval thrips and highlight the essential role of CYP3653A2 and ApoLp‐II/I in larval vitality. Our RNAi‐based tool offers an opportunity to investigate the molecular mechanisms of thrips‐orthotospovirus interactions, as the virus must be acquired by young larval thrips for successful transmission to plants, thus presenting potential targets for thrips pest management.

We examined water balance characteristics and the influence of desiccating conditions on the physiology and behavior of adult western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). Western flower thrips are globally invasive and likely to contend with shifts in water availability across their expansive geographic range. Basic water balance characteristics, including water mass and dry mass, were established for adult males and females, revealing a distinct sexual dimorphism wherein females are larger, and males retain a larger percentage of their body mass as water. Males lose relative water mass more quickly and their survival times are shorter compared to females. RNA‐seq analysis identified significant enrichment of a multitude of factors including carbohydrate transport and metabolism in dehydrated males and females. This was validated by altered glycogen levels, suggesting a rapid depletion in glycogen during dehydration. The probability of thrips feeding significantly increases when desiccation occurs, potentially to replenish water content and nutrient reserves. Our results establish the fundamental water balance characteristics of adult thrips and indicate that dehydration significantly influences the survivorship and feeding behavior of thrips; all of which being crucial factors that contribute to their capacity as vectors for plant pathogens.

Curly top disease, caused by beet curly top virus (BCTV), is among the most serious viral diseases affecting sugar beets in western USA. The virus is exclusively transmitted by the beet leafhopper (BLH, Circulifer tenellus) in a circulative and non-propagative manner. Despite the growing knowledge on virus-vector interactions, our understanding of the molecular interactions between BCTV and BLH is hampered by limited information regarding the virus impact on the vector and the lack of genomic and transcriptomic resources for BLH. This study unveils the significant impact of BCTV on both the performance and transcriptome response of BLHs. Viruliferous BLHs had higher fecundity than non-viruliferous counterparts, which was evident by upregulation of differentially expressed transcripts (DETs) associated with development, viability and fertility of germline and embryos in viruliferous insects. Conversely, most DETs associated with muscle movement and locomotor activities were downregulated in viruliferous insects, implying potential behavioural modifications by BCTV. Additionally, a great proportion of DETs related to innate immunity and detoxification were upregulated in viruliferous insects. Viral infection also induced notable alterations in primary metabolisms, including energy metabolism, namely glucosidases, lipid digestion and transport, and protein degradation, along with other cellular functions, particularly in chromatin remodelling and DNA repair. This study represents the first comprehensive transcriptome analysis for BLH. The presented findings provide new insights into the multifaceted effects of viral infection on various biological processes in BLH, offering a foundation for future investigations into the complex virus-vector relationship and potential management strategies for curly top disease.

Beet curly top virus (BCTV) has emerged as a major threat to hemp production in the western United States. Despite this concern, little is known about BCTV biology in hemp. This study investigated the incidence and genetic variability of BCTV strains, as well as the impact of BCTV infection on cannabinoid profiles, potential for seed transmission, and vector survival on hemp. Field surveys across five states revealed four distinct BCTV strains, with BCTV-CO and BCTV-Wor being the most prevalent. In addition, BCTV-PeCT was detected for the first time in hemp samples from Oregon, Colorado, and New Mexico, whereas BCTV-PeYD was detected exclusively in New Mexico. Genetic analysis showed high nucleotide diversity and widespread recombination among hemp-associated BCTV strains, consistent with active genetic exchange in BCTV. Furthermore, BCTV was detected in surface-disinfected seeds (77% in the Elite genotype, 18% in the 791 genotype) and in a small number of seedlings in grow-out experiments (0.98% in Elite, 0.87% in 791). Additional seed dissection experiments in Elite revealed BCTV presence in 41% of embryos, 72% of endosperm, and 5% of seed coats, demonstrating potential for seed transmission in hemp rather than surface contamination. BCTV infection reduced cannabinoid levels in one hemp genotype but not in another, indicating genotype-specific effects of BCTV on cannabinoid production. Finally, beet leafhoppers, the exclusive vector of BCTV, were unable to survive on hemp beyond 7 days, indicating that the insect cannot complete its life cycle on hemp. These findings provide a foundational understanding of BCTV evolutionary dynamics and host interactions in hemp, with implications for disease management. [ ] Copyright © 2026 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .