Insect galls are extended phenotypes built through sustained reprogramming of host development, yet the genomic innovations and effector mechanisms that enable this process remain poorly resolved. Here we generated chromosome-level genomes for the horned gall aphid Schlechtendalia chinensis and its host tree Rhus chinensis, and integrated these resources with morph-resolved salivary proteomics, metabolomics and functional assays. The aphid genome contains an anomalously expanded chromosome 1 spanning 93.61 Mb that carries 33.08% of anchored genes and concentrates 70.47% of genome-wide segmental duplications, together with elevated transposable-element load and multiple tandem gene clusters. We identified a salivary M20-family enzyme, ACY1, that consistent with an auxin-conjugate hydrolase. Loss- and gain-of-function assays show that ACY1 hydrolyzes stored, inactive auxin-amino acid conjugates in host tissue, increasing local active auxin levels. This hormone manipulation reprograms plant cell development to form galls. Our findings demonstrate how concentrated genomic duplication can drive the evolution of an effector that controls host physiology, suggesting a potential principle in the evolution of parasitic traits.

Aging-associated dyslipidemia and chronic low-grade inflammation contribute to atherosclerosis and cardiovascular risk. As a blend of long-chain aliphatic alcohols, policosanol from insect wax (PIW) has been documented to regulate lipid metabolism. However, the effects of PIW on atherosclerosis remain insufficiently characterized. In this study, ApoE-/- mice fed a high-fat diet were concurrently administered PIW (75 and 150 mg/kg) for eight weeks. PIW was associated with weight gain reduction and improvement in lipid profile, particularly a decrease in triglycerides and total cholesterol. PIW also lowered circulating inflammatory biomarkers (IL-6, TNF-α, and C-reactive protein). Histopathological analyses revealed attenuated hepatic injury and reduced aortic lipid deposition and lesion features. In parallel, PIW reduced serum endothelin-1 and oxidized LDL levels and modulated aortic ET-1, MMP-9/TIMP-1 balance, and LOX-1/NF-κB-related protein signals. Notably, as PIW was administered concurrently with high-fat diet induction, these findings should be interpreted within a preventive intervention framework. Collectively, PIW help attenuate HFD-associated atherosclerotic features and hold promise as a functional food ingredient for cardiovascular health and healthy aging.

The microRNAs play an important role in regulating biological posttranscriptional gene expressions to affect different biological processes of hexapod. Ericerus pela, commonly known as the white wax scale insect (hereafter, WWS), is a resource insect with significant economic value with two distinct developmental styles in male and female. In order to obtain the differential expression of miRNAs in the growth and development of WWS, we conducted miRNA sequencing of male and female through high-throughput sequencing. A total of 162 and 63 miRNAs were obtained from the male and female larvae respectively with systematic analysis. Mainly mapping on chromosome 2, the miRNAs were classified into 46 families throughout the whole genome of WWS. Furthermore, 17 differentially expressed miRNAs were obtained and a total of 3092 target genes were predicted and related to postembryonic, gonadal development and wing morphogenesis by enrichment analysis. It was concluded that miRNAs have a pivotal role in the sexual differential development of WWS. Our study is the original report for analyzing the metamorphosis of WWS by miRNAs. It will lay a foundation for further investigation of the miRNAs function in the growth and development of insects.

Eclosion is the critical process of the butterfly transforming from a pupa into an adult. During this process, the morphology of the lepidopteran wings undergoes drastic changes, as the butterfly rapidly transitions from a wingless pupa to an adult with large-scale wings, resulting in a several-fold or even dozen-fold expansion in wing surface area. Although the lepidopteran wings undergo drastic morphological changes within a short period, this process does not cause damage to the butterfly’s body. The reason lies in the pupal developmental stage, where the wing membrane, veins, tracheae, and membranous tissues gradually differentiate into numerous micron-scale foldable units. They are double-layered, foldable structures and capable of providing a several-dozen-fold expansion in surface area—far exceeding the actual requirement for wing expansion—thus supplying sufficient surface area reserves for wing flattening. During eclosion, the foldable units are progressively unfolded under wing expansion forces, with the stored area being utilized as the foldable units are flattened sequentially. At the macroscopic level, this results in rapid expansion of the surface area. The unfolding process of lepidopteran wings is influenced by the structure and arrangement of wing veins, exhibiting a change trend where length increases first, followed by width. Therefore, the morphological changes of lepidopteran wings are the result of cumulative deformations in foldable units. In summary, these foldable units serve not only as the constituent units of the wings but also as functional units, regulating wing deformation and the unfolding process.

As a sessile wax scale insect, Ericerus pela heavily relies on its host plant for nutrition. While E. pela utilizes the nitrogen-poor plant sap as its primary nutrient source, the mechanisms by which this insect overcomes the nitrogen deficiency are poorly understood. In this study, we first confirm the nitrogen fixation capability of E. pela through isotopic tracer experiments and the acetylene reduction assay, which demonstrate that female adults exhibit an efficient nitrogen fixation rate. High-throughput sequencing further revealed 42 nitrogen-fixing bacterial species in the tissues of E. pela, most notably including Rhizobiales and Methylobacterium as the dominant species converting atmospheric nitrogen to ammonia. Several critical genes involved in nitrogen fixation, ammonia transporting, amino acid synthesis, and transportation were determined to be transcriptionally active across different developmental stages of E. pela. In addition, the symbiotic fungus Ophiocordyceps—located in the fat body of E. pela—was found to be capable of synthesizing all amino acids, including the essential amino acids required for the survival of E. pela. Taken together, this study demonstrates that E. pela has evolved a highly effective nitrogen acquisition system driven by symbiotic microorganisms, ensuring a sufficient nitrogen supply and enabling it to thrive on nitrogen-deficient food sources. Our findings reveal a unique evolutionary adaptation in which E. pela leveraged both bacterial nitrogen fixation and fungal amino acid synthesis to bolster its growth and development.

In the original publication [...].

Plant-insect coevolution is exemplified by Schlechtendalia chinensis inducing gallnuts with record-breaking hydrolyzable tannin (HT) concentrations of 74.49% to 32-fold higher than normal leaves. How do plants achieve this extreme defensive chemistry, and how do aphids survive it? We integrated transcriptomics, heterologous gene validation in Arabidopsis, and enzyme assays to investigate both plant HT biosynthesis and aphid detoxification mechanisms throughout gall development. Three key genes: Phosphoglucomutase (PGM), UDP-glucosyltransferase BX9 (BX9), and gallate 1-beta-D-glucosyltransferase (GDG) govern HT biosynthesis, with expression patterns closely tracking tannin accumulation dynamics, and GDG as the rate-limiting enzyme. Arabidopsis transformants showed threefold HT increases. Critically, S. chinensis employs specialized laccases rather than tannase for detoxification, with laccase activity exceeding tannase by 20,000-fold. The aphid genome encodes three laccase genes, with Sc-Lac1 expressed in digestive tissues, achieving 39 to 54% HT degradation. These findings reveal how plants weaponize secondary metabolism while herbivores evolve enzymatic countermeasures. The identified genes enable engineering enhanced defenses or pharmaceutical tannin production, while laccase-based detoxification offers new insights into insect adaptation to chemical defenses. ### Competing Interest Statement The authors have declared no competing interest. Natural Science Foundation of Yunnan Province, 202301AT070178 National Natural Science Foundation of China, 32370551 Expert workstation in Yunnan Province, 202405AF140107

Color and odor are crucial cues for butterflies during foraging and courtship. While most sexual dimorphic butterflies rely more on vision, our understanding of how butterflies with similar coloration use different signals remains limited. This study investigated the visual and olfactory behavioral responses of the similarly colored butterfly Byasa hedistus during foraging and courtship. While visiting artificial flowers of different colors, we found that B. hedistus exhibits an innate color preference, with a sequence of preferences for red, purple, and blue. The frequency of flower visits by B. hedistus significantly increased when honey water was sprayed on the artificial flowers, but it hardly visited apetalous branches with honey water. This proves that locating nectar sources by odor alone is difficult in the absence of floral color guides. During courtship, males are active while females hardly chase; only two models were observed: males chasing males and males chasing females. The courtship process includes four behaviors: slowing approach, straight chasing, hovering, and spinning. B. hedistus cannot distinguish between sexes based on color, as there is no significant difference in color and shape between them. Twenty-three VOCs (>1%) were identified in B. hedistus, with 21 shared by both sexes, while ketones are specific to males. These VOCs are principally represented by cineole, β-pinene, and linalool. When cineole was added to butterfly mimics, many butterflies were attracted to them, but the butterflies did not seem to distinguish between males and females. This suggests that cineole may be the feature VOC for identifying conspecific groups. Adding β-pinene and linalool to mimics induced numerous butterflies to chase, hover, spin around, and attempt to mate with them. This suggests that β-pinene and linalool are crucial cues indicating the presence of females.

The Insect Cell-Baculovirus Expression Vector System (IC-BEVS) is widely used for the generation of a variety of gene products, including proteins, vaccines, and gene therapy vectors; however, it has some limitations, including a constrained host range and low protein yields. In a previous study, we established the RIRI-PA1 cell line, which was derived from Periplaneta americana. This cell line is susceptible to Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infection, which results in a higher yield production of recombinant protein within a short post-infection period of 24-48 h compared to the commonly used engineered cell line Sf21. To elucidate the basis for this phenomenon, we used RNA sequencing and transcriptome analysis of RIRI-PA1 and Sf21 cells infected with AcMNPV-GFP at 24, 72, and 168 h post-infection. Differentially expressed genes (DEGs) were identified in both cell lines. GO, eggNOG, and KEGG annotation analyses were used to identify DEGs and select candidate genes that could regulate recombinant protein expression. The results indicated a significant link between ribosomal pathway regulation and recombinant protein expression. After 24 h of AcMNPV-GFP infection, relatively high levels of protein were produced in RIRI-PA1 cells compared to Sf21 cells, which exhibited lesser enrichment of ribosomal protein-related DEGs (7 : 12). Moreover, a correlation was observed in the gene expression patterns between AcMNPV-GFP infection and recombinant protein synthesis, including genes associated with the ribosome, Toll and Imd signaling, and the cytochrome P450 pathway. Overall, our findings suggested that the ribosomal pathway might be more involved in regulation of protein expression during the early stages of RIRI-PA1 infection. The mechanisms underlying this process could have potential future applications in engineering cell modifications to reduce production time for recombinant proteins and to promote the use of IC-BEVS.

[目的]观察性二型性的白蜡虫Ericerus pela的蜡腺结构特征,探究其泌蜡对雌雄虫生态适应性的意义.[方法]在野外调查各发育阶段白蜡虫在寄主植物上的分布及泌蜡特征;在实验室用电子显微镜观察白蜡虫蜡腺结构及其分泌蜡丝的形态;通过石蜡切片分析雌雄虫表皮厚度;通过转录组数据结合白蜡虫泌蜡及表皮增厚的规律分析雌雄虫泌蜡量和几丁质合成的差异.[结果]在白蜡虫体表共发现6种蜡腺,其中四格腺为1龄雄若虫特有,十格腺为雌成虫特有,2龄雄若虫全身布满管腺这一白蜡虫的主要雄性产蜡器官,表皮五格腺、刺腺和肛门腺是各龄期所共有.雄虫有更多的蜡腺种类和数目,雌若虫的表皮厚度比雄若虫的厚将近35％.蜡酯合成关键酶脂酰辅酶A还原酶(fatty acyl-CoA reductase)基因far在雄成虫中表达量更高,几丁质合成关键酶几丁质合成酶(chitin synthase)基因cs在雌成虫中表达量更高.[结论]白蜡虫雌雄虫泌蜡差异显著,两性蜡腺的种类和数量与其不同的生态适应性对策相关.雄虫具有避光的生态习性,主要通过分泌一层蜡丝覆盖身体,保护雄虫在蜡被下完成变态过程.雌虫喜光,泌蜡少,通过增加表皮厚度的方式来适应环境,雌虫泌蜡是为了保证呼吸通畅,防止排泄的蜜露反流和微生物入侵,保护卵不被捕食,避免卵块粘连和减缓外力对卵的冲击.

The aim of this study was to investigate the structure, chemical composition and gene regulation mechanism of glandular trichome（GT） on Rhus potaninii. The GTs was studied by paraffin section, tissue staining and real-time fluorescence quantification. We found the GTs look like pinecone and consist by a head with12.00 ± 4.14 cells and an acellular stalk. The GTs became thinner after secreting and fall down at later stage. The secretion is composed of lots of polysaccharides, phenolic compounds, and acidic lipids, it could protect the young leaves from drought and pathogen in the surface. Furthermore, we found the selectively expressed genes in the GTs based on transcriptome and identified it by quantitative real-time polymerase chain reaction（RT-qPCR）. The WRKY33 and PTI6 which regulated the plant-pathogen interaction were enhanced in the GTs, meanwhile, SLC36A which regulated the Cyanoamino acid metabolism（Cyanoamino acid has toxic or side effects to insects） was enhanced too.

Telomere and telomerase are crucial factors in cell division and chromosome stability. Telomerase activity in most cells depends on the transcription control by the telomerase reverse transcriptase (TERT). The introduction of an exogenous human TERT (hTERT) in cultured cells could enhance telomerase activity and elongate the lifespan of various cells. Telomere elongation mechanisms vary between insects and are complex and unusual. Whether the use of exogenous hTERT can immortalize primary insect cells remains to be investigated. In this study, we used a recombinant virus expressing hTERT to infect primary cultured cells of Periplaneta americana and evaluated its effects on insect cell immortalization. We found that hTERT was successfully expressed and promoted the growth of P. americana cells, shortening their doubling time. This was due to the ability of hTERT to increase the activity of telomerase in P. americana cells, thus prolonging the telomeres. Our study lays the foundation for understanding the mechanisms of telomere elongation in P. americana, and suggests that the introduction of hTERT into insect cells could be an efficient way to establish certain insect cell lines.

Lac insect is an important resource insect with great commercial value. However, the lack of its genome information restricts the fundamental biological research and applied studies of this species. Here, we first assembled the contig of Kerria lacca using Illumina and Nanopore sequencing by Hi-C and T2T techniques to obtain the genome of K. lacca at the chromosome 0 gap level. The genome of K. lacca was 256.62 Mb and the Scaffold N50 was 28.53 Mb. A total of 56.94 Mb of repeat sequences, constituting 22.19% of the assembled genome, were identified. We annotated 10,696 protein-encoding geneswith 89.74% annotated. By horizontal gene transfer analysis, we obtained a putative gene Isoprenyl diphosphate synthase (IPPS), a key enzyme in the isoprene synthesis pathway in the regulation of lac biosynthesis that transferred horizontally from bacteria to the genome of K. lacca . Meanwhile, we constructed the lac synthesis biosynthetic pathway and screened 25 putative key genes in the synthesis pathway by transcriptome analysis in different developmental stages and tissues. It provides new research ideas to reveal the molecular mechanism of lac biosynthesis and regulation. The high-quality chromosomal-level genome of lac insect will provide a basis for the study of development, genetics, and the evolution of scale insects.

Insect wax is a traditional Chinese medicine, which can be converted into policosanol through a reduction reaction. The efficacy of policosanol from insect wax (PIW) depends on its purity and composition ratio. Plant-derived policosanol has a hypolipidemic effect, however, the effects of PIW on serum lipid levels have not yet been clarified. In this study, we investigated the effects of orally administered PIW on lipid metabolism in rats fed a high-fat diet. We found that PIW administration reduced the serum lipid levels of rats fed a high-fat diet, ameliorated body weight gain, and alleviated liver fat accumulation. This effect could be associated with the upregulation of fatty acid breakdown- and lipid metabolism–related genes, Pparα and Pparβ/δ, downregulation of fatty acid absorption and storage-related genes, Srebp1c, Il6, and Tnfα, and an antioxidant effect. These findings suggest the potential utility of PIW for developing functional foods with lipid-lowering capabilities.

Galls are products of the hyperplasia of host plant structures induced by gall-inducing organisms and have been considered as an extended phenotype of the inducers. There is little evidence regarding the effect of host plants on gall morphology. We hypothesised that the morphology and developmental pattern of galls are different because of the different location of their stimulation, even though two kinds of inducers are close relatives. We observed that horned galls and their leaflets of their host plant, Rhus chinensis required a longer rapid growth stage than fusiform galls and Rhus potaninii leaflets. The distribution of trichomes showed positional dependence. Molecular analysis showed that in the fusiform gall, the target genes that regulate the plastochron of leaflets and serration development were hardly expressed, and CUP-SHAPED COTYLEDON-2 may be a key gene that regulates the formation of the horns. In summary, horned and fusiform galls showed a developmental pattern similar to those of their host plant leaflets. We suggest that the inducing site is important in the morphology and development of galls.

The study investigated the species composition and epiphytic pattern of fungi on the body surface of female Ericerus pela, which would provide a scientific basis for the study of the interaction between E. pela and surface fungi and the exploration of the potential value of surface fungi. TM3000 scanning electron microscope was used to observe the attachment types of fungi and distribution on the body surface of female E. pela. The species of fungi were identified by isolating culture, morphological characteristics and molecular data. The observation found that the fungi on the back of the female E. pela mainly colonized on the body surface of the female E. pela by appressorium in the form of hypha mesh. The colonization of some fungi on the abdomen of the female E. pela was consistent with that on the back of the female E. pela, and some fungi were mixed with the wax filaments secreted by the female E. pela or colonization on the nearby of edge spines of female E. pela. After isolation, culture and purification, 8 strains fungi were obtained from the body surface of the female E. pela, which were identified as Hypoxylon sp., Trichoderma yunnanense, Xylaria brevipes, Papiliotrema flavescens, Rhodosporiobolus odoratus, Cladosporium cladosporioides, Epicoccum nigrum and Colletotrichum acutatum. The colonization of these fungi on the body surface of female E. pela, which was related to the excretion of honeydew by female E. pela. Honeydew provided abundant glycogen, vitamins and other nutrients for these fungi. Among the 8 strains fungi isolated, Trichoderma and yeast may be beneficial to the female E. pela. Trichoderma had antagonistic effect on pathogenic microorganisms, yeast used honeydew through fermentation and produces antibacterial substances, which could reduce the harm of other microorganisms to female E. pela. Other fungi were plant pathogens, which may induce host plant diseases and have potential harm to the insect wax industry. C. cladosporioides and C. acutatum were both insect pathogens and plant pathogens, which may seriously affected the cultivation of E. pela and the development of insect wax industry.

Eclosion is a rapid process of morphological changes in insects, especially for the wings of butterflies. The orange oakleaf butterfly (Kallima inachus) transits from pupae to adults with a 9.3 fold instant increase in the surface area of their wings. To explore the mechanism for the rapid morphological changes in butterfly wings, we analyzed changes in microstructures in the wings of K. inachus. We found that there were lots of micron-sized foldable units in the wings at the pupal stage. The foldable units could provide as much as 31.35 times of increase in wing surface area. During eclosion, foldable units were flattened sequentially and resulted in a rapid increase in wing surface areas. The unfolding process was regulated by the structures and layouts of wing veins. Based on our observation, foldable units play important roles in both deformation and stretching of wings. The foldable units of microstructures may provide mimics for simulating entities of large-deformational bionic structures with practical application.

Kerria lacca (Kerr) is an important lac insect extensively used in industrial products in the form of resin, wax and dye. The scarce knowledge on molecular markers for K. lacca is a barrier in elucidating genetic information. Our study identified a total of 16,921 single-nucleotide polymorphisms (SNPs), and 6231 insertions and deletions (InDels)-of which, intergenic variation accounted for 41.22% and 56.30%, and exonic variation accounted for 39.10% and 17.46%, of SNPs and InDels, respectively. Observation of SNPs suggested that nucleotide substitution frequency and transition to transversion (Ts/Tv) ratio were highest at the late adult stage, 3.97, compared to at the other stages, with a genome-wide Ts/Tv ratio of 2.95. The maximum number of SNPs, 2853 (16.86%), was identified in chromosome 8, while the lowest, 1126 (6.65%), was identified in chromosome 7. The maximum and minimum numbers of InDels were located on chromosome 1 and 7, with 834 (13.38%) and 519 (8.33%), respectively. Annotation showed that highest numbers of exonic and intergenic SNPs were present at the late adult stage, whereas the maximum number of InDels was found at the larval stage. On the basis of gene function, 47 gene variations were screened and 23 candidate genes were identified in associations with lac production. Concluding work will enhance knowledge on molecular markers to facilitate an increase in lac production in K. lacca as well as other lac insects.