Integrative taxonomy, phylogenetics and historical biogeography of subgenus Aeschyntelus Stål, 1872 (Hemiptera: Heteroptera: Rhopalidae).

The subgenus Aeschyntelus includes six species that show variations in body color and shape, thus making it difficult to identify them based on morphological identification alone. To date, no genetic study has evaluated species within this genus. Herein, we collected 171 individuals from 90 localities of Rhopalus and employed an integrative taxonomic approach that incorporated morphological data, mitochondrial genomic data (COI, whole mitochondrial data) and nuclear genomic data (18S + 28S rRNAs, nuclear genome-wide SNPs) to delineate species boundaries. Our analyses confirmed the status of nine described species of Rhopalus and proposed the recognition of one new species known as Rhopalus qinlinganus sp. nov., which is classified within the subgenus Aeschyntelus. Discrepancies arising from nuclear and mitochondrial data suggest the presence of mito-nuclear discordance. Specifically, mitochondrial data indicated admixture within Clade A, comprising R. kerzhneri and R. latus, whereas genome-wide SNPs unambiguously identified two separate species, aligning with morphological classification. Conversely, mitochondrial data clearly distinguished Clade B- consisting of R. sapporensis into two lineages, whereas genome-wide SNPs unequivocally identified a single species. Our study also provides insights into the evolutionary history of Aeschyntelus, thus indicating that it likely originated in East Asia during the middle Miocene. The development of Aeschyntelus biodiversity in the southwestern mountains of China occurred via an uplift-driven diversification process. Our findings highlight the necessity of integrating both morphological and multiple molecular datasets for precise species identification, particularly when delineating closely related species. Additionally, it reveals the important role of mountain orogenesis on speciation within the southwestern mountains of China.

The ncRNA-mediated regulatory networks of defensins and lysozymes in Riptortus pedestris: involvement in response to gut bacterial disturbances.

Insects depend on humoral immunity against intruders through the secretion of antimicrobial peptides (AMPs) and immune effectors via NF-κB transcription factors, and their fitness is improved by gut bacterial microbiota. Although there are growing numbers of reports on noncoding RNAs (ncRNAs) involving in immune responses against pathogens, comprehensive studies of ncRNA-AMP regulatory networks in Riptortus pedestris, which is one of the widely distributed pests in East Asia, are still not well understood under feeding environmental changes. The objective of this study employed the whole-transcriptome sequencing (WTS) to systematically identify the lncRNAs (long noncoding RNA) and circRNAs (circular RNA) and to obtain their differential expression from the R. pedestris gut under different feeding conditions. Functional annotation indicated that they were mainly enriched in various biological processes with the GO and KEGG databases, especially in immune signaling pathways. Five defensin (four novel members) and eleven lysozyme (nine novel members) family genes were identified and characterized from WTS data, and meanwhile, phylogenetic analysis confirmed their classification. Subsequently, the miRNA-mRNA interaction network of above two AMPs and lncRNA-involved ceRNA (competing endogenous RNA) regulatory network of one lysozyme were predicted and built based on bioinformatic prediction and calculation, and the expression patterns of differentially expressed (DE) defensins, and DE lysozymes and related DE ncRNAs were estimated and selected among all the comparison groups. Finally, to integrate the analyses of WTS and previous 16S rRNA amplicon sequencing, we conducted the Pearson correlation analysis to reveal the significantly positive or negative correlation between above DE AMPs and ncRNAs, as well as most changes in the gut bacterial microbiota at the genus level of R. pedestris. Taken together, the present observations provide great insights into the ncRNA regulatory networks of AMPs in response to rearing environmental changes in insects and uncover new potential strategies for pest control in the future.

Fabrication and application of carrier-free and carrier-based nanopesticides in pest management.

Pesticides are widely used for pest control to promote sustained and stable growth of agricultural production. However, indiscriminate pesticide usage poses a great threat to environmental and human health. In recent years, nanotechnology has shown the ability to increase the performance of conventional pesticides and has great potential for improving adhesion to crop foliage, solubility, stability, targeted delivery, and so forth. This review discusses two types of nanopesticides, namely, carrier-free nanopesticides and carrier-based nanopesticides, that can precisely release necessary and sufficient amounts of active ingredients. At first, the basic characterization and preparation methods of these two distinct types of nanopesticides are briefly summarized. Subsequently, current applications and future perspectives on scientific examples and strategies for promoting the usage efficacy and reducing the environmental risks of these nanopesticides were also described. Overall, nanopesticides can promote higher crop yields and lay the foundation for sustainable agriculture and global food security.

Identification and expression patterns of somatic piRNAs and PIWI genes in Riptortus pedestris (Hemiptera: Alydidae).

RNA interference (RNAi)-based gene silencing is a feasible and sustainable technology for the management of hemipteran pests by double-stranded RNA involvement, including small-interfering RNA, microRNA, and Piwi-interacting RNA (piRNA) pathways, that may help to decrease the usage of chemical insecticides. However, only a few data are available on the somatic piRNAs and their biogenesis genes in Riptortus pedestris, which serves as a significant pest of soybean (Glycine max). In this study, two family members of the PIWI gene were identified and characterized in R. pedestris, containing Argonaute3 (RpAgo3) and Aubergine (RpAub) genes with conserved protein domains, and their clusters were validated by phylogenetic analysis. In addition, they were widely expressed in all developmental stages of the whole body of R. pedestris and had lower expression levels in R. pedestris guts under different rearing conditions based on previous transcriptome sequencing. Furthermore, abundant clean reads were filtered to a total number of 45,998 piRNAs with uridine bias at the first nucleotide (nt) position and 26-32 nt in length by mapping onto the reference genome of R. pedestris according to our previous whole-transcriptome sequencing. Finally, our data revealed that gut bacterial changes were significantly positively or negatively associated with differentially expressed piRNAs among the five comparison groups with Pearson correlation analysis. In conclusion, these findings paved new avenues for the application of RNAi-based biopesticides for broad-spectrum hemipteran pest control.

Identifying a potentially invasive population in the native range of a species: The enlightenment from the phylogeography of the yellow spotted stink bug, Erthesina fullo (Hemiptera: Pentatomidae).

The yellow spotted stink bug (YSSB), Erthesina fullo (Thunberg, 1783) is an important Asian pest that has recently successfully invaded Europe and an excellent material for research on the initial stage of biological invasion. Here, we reported the native evolutionary history, recent invasion history, and potential invasion threats of YSSB for the first time based on population genetic methods [using double digest restriction-site associated DNA (ddRAD) data and mitochondrial COI and CYTB] and ecological niche modelling. The results showed that four lineages (east, west, southwest, and Hainan Island) were established in the native range with a strong east-west differentiation phylogeographical structure, and the violent climate fluctuation might cause population divergence during the Middle and Upper Pleistocene. In addition, land bridges and monsoon promote dispersal and directional genetic exchanging between island populations and neighboring continental populations. The east lineage (EA) was identified as the source of invasion in Albania. EA had the widest geographical distribution among all other lineages, with a star-like haplotype network with the main haplotype as the core. It also had a rapid population expansion history, indicating that the source lineage might have stronger diffusion ability and adaptability. Our findings provided a significant biological basis for fine tracking of invasive source at the lineage or population level and promote early invasion warning of potential invasive species on a much subtler lineage level.

Integrated analysis of miRNA profiles and gut bacterial changes in Altica viridicyanea following antibiotic treatment.

The gut bacteria involves in insect homeostasis by playing essential roles in host physiology, metabolism, innate immunity, and so forth. microRNAs (miRNAs) are endogenous small noncoding RNAs that posttranscriptionally regulate gene expression to affect immune or metabolic processes in insects. For several non-model insects, the available knowledge on the relationship between changes in the gut bacteria and miRNA profiles is limited. In this study, we investigated the gut bacterial diversity, composition, and function from Altica viridicyanea feeding on normal- and antibiotic-treated host plants using 16S rRNA amplicon sequencing; antibiotics have been shown to affect the body weight and development time in A. viridicyanea, suggesting that the gut bacteria of the normal sample were more diverse and abundant than those of the antibiotic-fed group, and most of them were involved in various physical functions by enrichment analysis. Furthermore, we executed small RNA transcriptome sequencing using the two experimental groups to obtain numerous sRNAs, such as piRNAs, siRNAs, and known and novel miRNAs, by data mapping and quality control, and furthermore, a total of 224 miRNAs were identified as significantly differentially expressed miRNAs, of which some DEMs and their target genes participated in immune- and metabolism-related pathways based on GO and KEGG annotation. Besides, regarding the regulatory roles of miRNA and target genes, a interaction network of DEM-target gene pairs from eight immune- or metabolism-related signaling pathways were constructed. Finally, we discovered that DEMs from above pathways were significantly positively or negatively correlated with gut bacterial alterations following antibiotic treatment. Collectively, the observations of this study expand our understanding of how the disturbance of gut bacteria affects miRNA profiles in A. viridicyanea and provide new valuable resources from extreme ranges for future studies on the adaptive evolution in insects.

Changes in the gut bacterial community affect miRNA profiles in Riptortus pedestris under different rearing conditions.

Insects possess complex and dynamic gut microbial system, which contributes to host nutrient absorption, reproduction, energy metabolism, and protection against stress. However, there are limited data on interactions of host-gut bacterial microbiota through miRNA (microRNA) regulation in a significant pest, Riptortus pedestris. Here, we performed the 16S rRNA amplicon sequencing and small RNA sequencing from the R. pedestris gut under three environmental conditions and antibiotic treatment, suggesting that we obtained a large amount of reads by assembly, filtration and quality control. The 16S rRNA amplicon sequencing results showed that the abundance and diversity of gut bacterial microbiota were significantly changed between antibiotic treatment and other groups, and they are involved in metabolism and biosynthesis-related function based on functional prediction. Furthermore, we identified different numbers of differentially expressed unigenes (DEGs) and differentially expressed miRNAs (DEMs) based on high-quality mappable reads, which were enriched in various immune-related pathways, including Toll-like receptor, RIG-I-like receptor, NOD-like receptor, JAK/STAT, PI3K/Akt, NF-κB, MAPK signaling pathways, and so forth, using GO and KEGG enrichment analysis. Later on, the identified miRNAs and their target genes in the R. pedestris gut were predicted and randomly selected to construct an interaction network. Finally, our study indicated that alterations in the gut bacterial microbiota are significantly positively or negatively associated with DEMs of the Toll/Imd signaling pathway with Pearson correlation analysis. Taken together, the results of our study lay the foundation for further deeply understanding the interactions between the gut microbiota and immune responses in R. pedestris through miRNA regulation, and provide the new basis for pest management in hemipteran pests.

The lncRNA-mediated ceRNA network of Altica viridicyanea is involved in the regulation of the Toll/Imd signaling pathway under antibiotic treatment.

Long noncoding RNAs (lncRNAs) play significant roles in the regulation of mRNA expression or in shaping the competing endogenous RNA (ceRNA) network by targeting miRNA. The insect gut is one of the most important tissues due to direct contact with external pathogens and functions in the immune defense against pathogen infection through the innate immune system and symbionts, but there are limited observations on the role of the lncRNA-involved ceRNA network of the Toll/Imd pathway and correlation analysis between this network and bacterial microbiota in the Altica viridicyanea gut. In this research, we constructed and sequenced six RNA sequencing libraries using normal and antibiotic-reared samples, generating a total of 17,193 lncRNAs and 26,361 mRNAs from massive clean data by quality control and bioinformatic analysis. Furthermore, a set of 8,539 differentially expressed lncRNAs (DELs) and 13,263 differentially expressed mRNAs (DEMs), of which related to various immune signaling pathways, such as the Toll/Imd, JAK/STAT, NF-κB, and PI3K-Akt signaling pathways, were obtained between the two experimental groups in A. viridicyanea. In addition, numerous GO and KEGG enrichment analyses were used to annotate the DELs and their target genes. Moreover, six Toll family members and nineteen signal genes from the Toll/Imd signaling pathway were identified and characterized using online tools, and phylogenetic analyses of the above genes proved their classification. Next, a lncRNA-miRNA-mRNA network of the Toll/Imd pathway was built, and it contained different numbers of DEMs in this pathway and related DELs based on prediction and annotation. In addition, qRT-PCR validation and sequencing data were conducted to show the expression patterns of the above DELs and DEMs related to the Toll/Imd signaling pathway. Finally, the correlated investigations between DELs or DEMs of the Toll/Imd signaling pathway and most changes in the gut bacterial microbiota revealed significantly positive or negative relationships between them. The present findings provide essential evidence for innate immune ceRNAs in the beetle gut and uncover new potential relationships between innate immune pathways and the gut bacterial microbiota in insects.

Out of East Asia: Early Warning of the Possible Invasion of the Important Bean Pest Stalk-Eyed Seed Bug Chauliops fallax (Heteroptera: Malcidae: Chauliopinae).

The short stay at the beginning of the invasion process is a critical time for invasive species identification and preventing invasive species from developing a wider distribution and significant economic impact. The stalk-eyed seed bug Chauliops fallax is an important agricultural pest of soybean and was first reported to occur outside East Asia. Here, we reported the native evolutionary history, recent invasion history, and potential invasion threats of C. fallax for the first time based on population genetic methods and ecological niche modelling. The results showed that four native East Asian genetic groups (EA, WE, TL, and XZ) were well supported, showing an east-west differentiation pattern consistent with the geographical characteristics of three-step landforms in China. Two main haplotypes existed: Hap1 might have experienced a rapid northwards expansion process after the LGM period, and Hap5 reflected local adaptation to the environment in southeastern China. The Kashmir sample was found to come from the recent invasion of populations in the coastal areas of southern China. Ecological niche modelling results suggested that North America has a high risk of invasion, which might pose a serious threat to local soybean production. In addition, with future global warming, the suitable habitat in Asia will move towards the higher latitude region and gradually deviate from the soybean planting area, which indicates the threat of C. fallax to soybean production in Asia will decrease in the future. The results could provide new insights into the monitoring and management of this agricultural pest in the early invasion stage.

Mitochondrial introgression and mito-nuclear discordance obscured the closely related species boundaries in Cletus Stål from China (Heteroptera: Coreidae).

Accurate taxonomy and delimitation are of great importance for pest control strategies and management programs. Here, we focus on Cletus (Insecta: Hemiptera: Coreidae), which includes many crop pests. The species boundaries still conflict and only cytochrome c oxidase subunit I (COI) barcoding has been previously used for molecular studies. We generated new mitochondrial genome and nuclear genome-wide SNPs to explore the species boundaries of 46 Cletus samples from China using multiple species delimitation approaches. All results recovered a monophyly with high support, except for two closely related species in clade I - C. punctiger and C. graminis. Mitochondrial data demonstrated admixture in clade I, while genome-wide SNPs unambiguously identified two separate species, which were confirmed by morphological classification. Inconsistent nuclear and mitochondrial data indicated mito-nuclear discordance. Mitochondrial introgression is the most likely explanation, and more extensive sampling and more comprehensive data are needed to ascertain a pattern. Accurate species delimitation will shed light on species status; thus, an accurate taxonomy is of particular concern, as there is a pressing need to implement precise control of agricultural pests and to perform further research on diversification.

The fecal shield is a double-edged sword for larvae of a leaf beetle.

Larvae of some leaf beetles carry masses of feces covering parts or all of the body, which is called a "fecal shield". In general, the shield is thought to be a defense structure against natural enemies. However, some studies have suggested that defense effectiveness varies depending on the natural enemy. In this study, we used a fecal retention leaf beetle Ophrida xanthospilota (Coleoptera: Chrysomelidae), and 2 local generalist predators (an ant, Camponotus japonicus (Hymenoptera: Formicidae) and a stinkbug, Arma custos [Hemiptera: Pentatomidae]) as a system to test the hypothesis that the fecal shield of O. xanthospilota plays different roles in predation behavior of different predators and can provide multiple chemical communication signals in predator-prey interactions. Prey bioassays showed that the fecal shield of O. xanthospilota larvae repelled the ant C. japonicus while attracting the stinkbug A. custos. The results also strongly demonstrated that hexane extracts of the fecal shield significantly repelled C. japonicus, while dichloromethane (DCM) extracts did not inhibit ant predation. Interestingly, DCM extracts attracted A. custos, but hexane extracts did not. Therefore, we suggest that the fecal shield is a double-edged sword for the larvae of O. xanthospilota. Our results also indicated that the risk-benefit tradeoff of an insect should be estimated at a community level involving multiple enemies (predators and parasites) and herbivores, rather than in a single prey-predator pair.

The first discovery of Tc1 transposons in yeast.

Background: Identification of transposons without close homologs is still a difficult task. IS630/Tc1/mariner transposons, classified into a superfamily, are probably the most widespread DNA transposons in nature. Tc1/mariner transposons have been discovered in animals, plants, and filamentous fungi, however, not in yeast. Results: In the present study, we report the discovery of two intact Tc1 transposons in yeast and filamentous fungi, respectively. The first one, named Tc1-OP1 (DD40E), represents Tc1 transposons in Ogataea parapolymorpha. The second one, named Tc1-MP1 (DD34E), represents Tc1 transposons in the Rhizopodaceae and Mucoraceae families. As a homolog of Tc1-OP1 and Tc1-MP1, IS630-AB1 (DD34E) was discovered as an IS630 transposon in Acinetobacter spp. Conclusion: Tc1-OP1 is not only the first reported Tc1 transposon in yeast, but also the first reported nonclassical Tc1 transposon. Tc1-OP1 is the largest of IS630/Tc1/mariner transposons reported to date and significantly different from others. Notably, Tc1-OP1 encodes a serine-rich domain and a transposase, extending the current knowledge of Tc1 transposons. The phylogenetic relationships of Tc1-OP1, Tc1-MP1 and IS630-AB1 indicated that these transposons had evolved from a common ancestor. Tc1-OP1, Tc1-MP1 and IS630-AB1 can be used as reference sequences to facilitate the identification of IS630/Tc1/mariner transposons. More Tc1/mariner transposons will be identified in yeast, following our discovery.

Chromosome-level genome of the bean bug Megacopta cribraria in native range, provides insights into adaptation and pest management.

Megacopta cribraria, a bean pest causing tremendous economic losses in Asia, was discovered in North America in 2009. Although M. cribraria has become the focus of research on biological invasion and pest management, the lack of genomic resources limits in-depth studies. Here, we report the first chromosome-level genome of M. cribraria using Illumina, PacBio, and Hi-C data. The assembled genome size was 699.65 Mb, with a contig N50 of 1.43 Mb and a scaffold N50 of 109.27 Mb. >97.51 % of bases were successfully anchored to six chromosomes. Through genome annotation, a total of 13,308 coding genes were predicted, 96.3 % of which were successfully accessed function. Expanded gene families were involved in proteolysis, protein metabolism and nitrogen metabolism reflected the underlying genome basis for host adaptation during evolution. Transcriptome analysis revealed different gene expression patterns in antenna, mouthpart, head, leg, wing, and carcass body of the adult M. cribraria, respectively. Moreover, the expression profiles of the odorant receptor genes indicated the potential target genes for pest control. The high-quality chromosome-level genome will benefit further research on the adaptation, evolution, and population genetics of the M. cribraria that will assist in the pest management and tracking the biological invasion routes.

The anthropogenic effect of land use on population genetics of Malcus inconspicuus.

Since the beginning of the Holocene era, human activities have seriously impacted animal habitats and vegetative environments. Species that are dependent on natural habitats or with narrow niches might be more severely affected by habitat changes. Malcus inconspicuus is distributed in subtropical China and highly dependent on the mountain environment. Our study investigated the role of the mountainous landscape in the historical evolution of M. inconspicuus and the impact of Holocene human activities on it. A phylogeographical approach was implemented with integrative datasets including double-digest restriction site-associated DNA (ddRAD), mitochondrial data, and distribution data. Three obvious clades and an east-west phylogeographical pattern were found in subtropical China. Mountainous landscape has "multifaceted" effects on the evolutionary history of M. inconspicuus, it has contributed to population differentiation, provided glacial refuges, and provided population expansion corridors during the postglacial period. The effective population size (Ne) of M. inconspicuus showed a sharp decline during the Holocene era, which revealed a significantly negative correlation with the development of cropland in a hilly area at the same time and space. It supported that the species which are highly dependent on natural habitats might undergo greater impact when the habitat was damaged by agricultural activities and we should pay more attention to them, especially in the land development of their distribution areas.

Species boundary and phylogeographical pattern provide new insights into the management efforts of Megacopta cribraria (Hemiptera: Plataspidae), a bean bug invading North America.

BACKGROUND: Correct identification together with information on distribution range, geographical origin and evolutionary history are the necessary basis for the management and control of invasive species. The bean bug Megacopta cribraria is a crucial agricultural pest of soybean. Recently, M. cribraria has invaded the United States and spread rapidly, causing severe reductions in soybean yields. However, the species boundary and phylogeographical pattern of this invasive bean bug are still unclear. RESULTS: The results of different species delimitation methods (Automatic Barcode Gap Discovery, Assemble Species by Automatic Partitioning, Bayesian Poisson Tree Processes and Bayesian Phylogenetics and Phylogeography) strongly demonstrated that M. cribraria and Megacopta punctatissima represent the same species. M. punctatissima should not be considered a distinct species but rather a variety of M. cribraria. Phylogenetic analyses revealed three well-supported clades (Southeast Asia [SEA], East Asia continent [EAC] and Japan [JA]) with distinct geographical structures in the M. cribraria-M. punctatissima complex. The SEA clade was at the base of the phylogenetic tree, and the sister relationship between the EAC clade and JA clade was strongly supported. The split between the EAC clade and JA clade occurred at approximately 0.71 Ma, corresponding to the submergence period of the East China Sea land bridge. CONCLUSION: This study clarified the species boundary between M. cribraria and its closely related species and revealed the phylogeographical pattern and evolutionary history of M. cribraria. The species delimitation and phylogeography results achieved in this study could provide new insights into the monitoring and management of this agricultural pest. © 2022 Society of Chemical Industry.

Mitogenomics of Chinch Bugs from China and Implications for Its Coevolutionary Relationship with Grasses.

Blissidae (the Chinch bug) is a group with high species richness in Lygaeoidea, but there are only a few descriptions of mitochondrial genomes available. We obtained mitogenomes from 10 species of eight genera from Blissidae through second-generation sequencing technology. The length of the mitochondrial genome (excluding the control region) is between 14643 and 14385 bp; the content of AT is between 74.1% and 77.9%. The sequence of the evolution rate of protein coding genes was as follows: ND5 > ATP8 > ND6 > ND2 > ND4 > ND4L > ND1 > ATP6 > ND3 > COIII > COII > CYTB > COI. The mitogenomic structure of Blissidae is highly conservative. Gene rearrangement is only found in Pirkimeru japonicus (PiGXBS1), which is formed as the duplication of tRNA-H. The intergenic spacer between ND4 and tRNA-H, which form an obvious stem-and-loop structure, was found in all samples in this study. The phylogenetic trees generated by BI and ML indicated that Blissidae can be divided into three major clades: Clade A (only included Macropes); Clade B ((Pirkimerus + Bochrus) + Iphicrates); and Clade C ((Ischnodemus + Capodemus) + (Cavelerius + Dimorphopterus)). The divergence within the Blissidae began at about 56 Ma. At the genus level, the divergence was concentrated at 30−51 Ma, slightly later than the diversification of Poaceae. The consistency of divergence time between Blissidae and Poaceae might hint at the coevolutionary relationship between them, but further molecular and biological evidence is still needed to prove it.

Parallel metatranscriptome analysis reveals degradation of plant secondary metabolites by beetles and their gut symbionts.

Switching to a new host plant is a driving force for divergence and speciation in herbivorous insects. This process of incorporating a novel host plant into the diet may require a number of adaptations in the insect herbivores that allow them to consume host plant tissue that may contain toxic secondary chemicals. As a result, herbivorous insects are predicted to have evolved efficient ways to detoxify major plant defences and increase fitness by either relying on their own genomes or by recruiting other organisms such as microbial gut symbionts. In the present study we used parallel metatranscriptomic analyses of Altica flea beetles and their gut symbionts to explore the contributions of beetle detoxification mechanisms versus detoxification by their gut consortium. We compared the gut meta-transcriptomes of two sympatric Altica species that feed exclusively on different host plant species as well as their F1 hybrids that were fed one of the two host plant species. These comparisons revealed that gene expression patterns of Altica are dependent on both beetle species identity and diet. The community structure of gut symbionts was also dependent on the identity of the beetle species, and the gene expression patterns of the gut symbionts were significantly correlated with beetle species and plant diet. Some of the enriched genes identified in the beetles and gut symbionts are involved in the degradation of secondary metabolites produced by plants, suggesting that Altica flea beetles may use their gut microbiota to help them feed on and adapt to their host plants.

Soil-derived bacteria endow Camellia weevil with more ability to resist plant chemical defense.

BACKGROUND: Herbivorous insects acquire their gut microbiota from diverse sources, and these microorganisms play significant roles in insect hosts' tolerance to plant secondary defensive compounds. Camellia weevil (Curculio chinensis) (CW) is an obligate seed parasite of Camellia oleifera plants. Our previous study linked the CW's gut microbiome to the tolerance of the tea saponin (TS) in C. oleifera seeds. However, the source of these gut microbiomes, the key bacteria involved in TS tolerance, and the degradation functions of these bacteria remain unresolved. RESULTS: Our study indicated that CW gut microbiome was more affected by the microbiome from soil than that from fruits. The soil-derived Acinetobacter served as the core bacterial genus, and Acinetobacter sp. was putatively regarded responsible for the saponin-degradation in CW guts. Subsequent experiments using fluorescently labeled cultures verified that the isolate Acinetobacter sp. AS23 can migrate into CW larval guts, and ultimately endow its host with the ability to degrade saponin, thereby allowing CW to subsist as a pest within plant fruits resisting to higher concentration of defensive chemical. CONCLUSIONS: The systematic studies of the sources of gut microorganisms, the screening of taxa involved in plant secondary metabolite degradation, and the investigation of bacteria responsible for CW toxicity mitigation provide clarified evidence that the intestinal microorganisms can mediate the tolerance of herbivorous insects against plant toxins. Video Abstract.