A faster killing effect of plastid-mediated RNA interference on a leaf beetle through induced dysbiosis of the gut bacteria.

The expression of double-stranded RNAs (dsRNAs) from the plastid genome has been proven to be an effective method for controlling herbivorous pests by targeting essential insect genes. However, there are limitations to the efficiency of plastid-mediated RNA interference (PM-RNAi) due to the initial damage caused by insects and their slow response to RNA interference. In this study, we developed transplastomic poplar plants that expressed dsRNAs targeting the ß-Actin (dsACT) and Srp54k (dsSRP54K) genes of Plagiodera versicolora. Feeding experiments showed that transplastomic poplar plants caused significantly higher mortality in P. versicolora larvae compared to nuclear transgenic or wild-type poplar plants. The efficient killing effect of PM-RNAi on P. versicolora larvae was found to be dependent on the presence of gut bacteria. Importantly, we also demonstrated that the foliar application of a gut bacterial strain, Pseudomonas putida, induced dysbiosis in the gut bacteria of P. versicolora larvae, leading to a significant acceleration in the speed-to-kill by PM-RNAi. Overall, our findings suggest that interfering with gut bacteria could be a promising strategy to enhance the effectiveness of PM-RNAi for insect pest control, offering a novel and effective approach for crop protection based on RNAi technology.

Enhanced capacity of a leaf beetle to combat dual stress from entomopathogens and herbicides mediated by associated microbiota.

Herbicides have demonstrated their impact on insect fitness by affecting their associated microbiota or altering the virulence of entomopathogenic fungi toward insects. However, limited research has explored the implications of herbicide stress on the intricate tripartite interaction among insects, associated bacterial communities, and entomopathogens. In this study, we initially demonstrated that associated bacteria confer a leaf beetle, Plagiodera versicolora, with the capability to resist the entomopathogenic fungus Aspergillus nomius infection, a capability sustained even under herbicide glyphosate stress. Further analysis of the associated microbiota revealed a significant alteration in abundance and composition due to glyphosate treatment. The dominant bacterium, post A. nomius infection or following a combination of glyphosate treatments, exhibited strong suppressive effects on fungal growth. Additionally, glyphosate markedly inhibited the pathogenic associated bacterium Pseudomonas though it inhibited P. versicolora's immunity, ultimately enhancing the beetle's tolerance to A. nomius. In summary, our findings suggest that the leaf beetle's associated microbiota bestow an augmented resilience against the dual stressors of both the entomopathogen and glyphosate. These results provide insight into the effects of herbicide residues on interactions among insects, associated bacteria, and entomopathogenic fungi, holding significant implications for pest control and ecosystem assessment.

Gut microbiota facilitate adaptation of invasive moths to new host plants.

Gut microbiota are important in the adaptation of phytophagous insects to their plant hosts. However, the interaction between gut microbiomes and pioneering populations of invasive insects during their adaptation to new hosts, particularly in the initial phases of invasion, has been less studied. We studied the contribution of the gut microbiome to host adaptation in the globally recognized invasive pest, Hyphantria cunea, as it expands its range into southern China. The southern population of H. cunea shows effective adaptation to Metasequoia glyptostroboides and exhibits greater larval survival on Metasequoia than the original population. Genome resequencing revealed no significant differences in functions related to host adaptation between the two populations. The compatibility between southern H. cunea populations and M. glyptostroboides revealed a correlation between the abundance of several gut bacteria genera (Bacteroides, Blautia, and Coprococcus) and H. cunea survival. Transplanting the larval gut microbiome from southern to northern populations enhanced the adaptability of the latter to the previously unsuitable plant M. glyptostroboides. This research provides evidence that the gut microbiome of pioneering populations can enhance the compatibility of invasive pests to new hosts and enable more rapid adaptation to new habitats.

Dynamic Gut Microbiota of Apolygus lucorum Across Different Life Stages Reveals Potential Pathogenic Bacteria for Facilitating the Pest Management.

Insect's gut microbiota has diverse effects on their fitness, and a comprehensive understanding of gut microbiota functions requires analyzing its diversity. Apolygus lucorum is a highly destructive pest that threatens many economically important crops in China. This study investigated the gut microbiota of A. lucorum across its life cycle using both culture-dependent and culture-independent methods. A total of 87 gut bacterial isolates were identified, belonging to 4 phyla, 27 families, and 45 genera, while Miseq sequencing detected 91 amplicon sequence variants (ASVs) assigned to 5 phyla, 28 families, and 39 genera. Proteobacteria and Firmicutes were the predominant phyla, with Staphylococcus and Serratia as the major genera. There were significant differences in the relative abundance of these genera between the nymph and adult stages. Staphylococcus was significantly more abundant in nymphs than it in adults, while Serratia was significantly more abundant in sexually mature adults than in other developmental stages. Notably, Serratia is a common opportunistic pathogen in many insects. Injecting the gut-dominant isolate Serratia marcescens verified its high pathogenicity. Additionally, immune indicators of the bug at different developmental stages supported the hypothesis that Serratia is a pathogen of A. lucorum. This study provides a foundation for understanding the role of gut bacteria in the life history of A. lucorum and developing new pest control strategies based on microbes.

The pivotal roles of gut microbiota in insect plant interactions for sustainable pest management.

The gut microbiota serves as a critical "organ" in the life cycle of animals, particularly in the intricate interplay between herbivorous pests and plants. This review summarizes the pivotal functions of the gut microbiota in mediating the insect-plant interactions, encompassing their influence on host insects, modulation of plant physiology, and regulation of the third trophic level species within the ecological network. Given these significant functions, it is plausible to harness these interactions and their underlying mechanisms to develop novel eco-friendly pest control strategies. In this context, we also outline some emerging pest control methods based on the intestinal microbiota or bacteria-mediated interactions, such as symbiont-mediated RNAi and paratransgenesis, albeit these are still in their nascent stages and confront numerous challenges. Overall, both opportunities and challenges coexist in the exploration of the intestinal microbiota-mediated interactions between insect pests and plants, which will not only enrich the fundamental knowledge of plant-insect interactions but also facilitate the development of sustainable pest control strategies.

Priority control factors for heavy metal groundwater contamination in peninsula regions based on source-oriented health risk assessment.

Peninsula regions in China face serious environmental issues, such as heavy metal (HM) groundwater contamination. However, attempts to investigate the pollution sources and health risks of HM contamination in such regions require considerable resources and costs. Moreover, the priority control factors for groundwater HMs remain unclear. In this study, absolute principal component score/multiple linear regression (APCS/MLR) was used to quantify the groundwater pollution sources of typical peninsular areas in China, and a health risk assessment (HRA) was performed to assess the health risks caused by different sources. The results showed that the concentrations of Mn and Fe were higher than those of other HMs, and HM pollution was high in shallow groundwater. The dominant source of HMs in groundwater was agricultural activities (31.12 %), followed by natural sources (26.33 %), industrial activities (22.47 %), and atmospheric deposition (20.09 %). The non-carcinogenic risks to residents were acceptable, whereas the carcinogenic risks were high. Agricultural sources, atmospheric deposition sources, and Cr and As were identified as the priority control factors for HM groundwater contamination. This study has implications for improving the control of groundwater HM contamination in peninsula regions and ensuring sustainable groundwater development.

Enhanced toxicity of entomopathogenic fungi Beauveria bassiana with bacteria expressing immune suppressive dsRNA in a leaf beetle.

The entomopathogenic fungus is recognized as an ideal alternative to chemical pesticides, nonetheless, its efficacy is often limited by insect's innate immune system. The suppression of the host immunity may overcome the obstacle and promote the toxicity of the fungi. Here, by using an entomopathogenic fungus Beauveria bassiana and immune genes dsRNA-expressing bacteria, we explored the potentially synergistic toxicity of the two agents on a leaf beetle Plagiodera versicolora (Coleoptera: Chrysomelidae). We first determined the susceptibilities of P. versicolora to a B. bassiana 476 strain (hereafter referred to Bb476). And the immune genes were identified based on the transcriptome of Bb476 challenged beetles. Subsequently, five immune genes (PGRP1, Toll1, Domeless，SPN1，and Lysozyme) were targeted by feeding dsRNA-expressing bacteria, which produced a 71.4, 39.0, 72.0, 49.0, and 68.7% gene silencing effect, respectively. Furthermore, we found a significantly increased mortality of P. versicolora when combined the Bb476 and the immune suppressive dsRNAs. Taking together, this study highlights the importance of insect immunity in the defense of entomopathogens and also paves the way toward the development of a more efficient pest management strategy that integrates both entomopathogens and immune suppressive dsRNAs.

How to Effectively Reduce Honey Adulteration in China: An Analysis Based on Evolutionary Game Theory.

Apiculture has been greatly developed in recent years in China. Beekeeping cooperatives and honey manufacturing enterprises have increased rapidly. As a result, a variety of honey products have entered the market, adding vitality to the food economy; however, the adulteration of honey products is on the rise in China. Previous attempts to control the adulteration of honey products mostly relied on technical, product-specific measures, and there was a lack of modeling research to guide the supervision of the honey product industry. In order to help local governments to better control the adulteration of honey products from a management perspective, this paper establishes an evolutionary game model composed of beekeeping cooperatives, honey product enterprises, and local governments. Through stability analysis and model simulation, we found that local government subsidies to cooperatives have little impact on the game system. Local government penalties to cooperatives and price adjustments of unadulterated raw honey by cooperatives are effective management tools to reduce the adulteration behavior of cooperatives. Local government penalties for enterprises are an effective management tool to reduce the adulteration behavior of enterprises. This research provides useful information for government agencies to design appropriate policies/business modes so as to promote sustainability and the healthy development of the honey product industry in China.

Comprehensive treatment strategy for diesel truck exhaust.

At present, diesel vehicles still play an irreplaceable role in the traditional energy field in China. Diesel vehicle exhaust contains hydrocarbons, carbon monoxide, nitrogen oxides, and particulate matter, which can lead to haze weather, photochemical smog, and the greenhouse effect; endanger human health; and damage the ecological environment. In 2020, the number of motor vehicles in China reached 372 million, and the number of automobiles reached 281 million, of which 20.92 million are diesel vehicles, accounting for only 5.6% of the number of motor vehicles and 7.4% of the number of automobiles. Nevertheless, diesel vehicles emitted 88.8% of nitrogen oxides and 99% of particulate matter in total vehicle emissions. Diesel vehicles, especially diesel trucks, have become the top priority of motor vehicle pollution control. However, there are few reviews on the comprehensive treatment of diesel vehicle exhaust. This review provides an overview of exhaust gas composition, hazards, and treatment techniques. Phytoremediation, three-way catalytic conversion, rare earth catalytic degradation, and nanoscale TiO2 catalytic degradation are briefly described.

Efficient Circularly Polarized Electroluminescence from Achiral Luminescent Materials.

Circularly polarized electroluminescence (CP-EL) is generally produced in organic light-emitting diodes (OLEDs) based on special CP luminescent (CPL) materials, while common achiral luminescent materials are rarely considered to be capable of direct producing CP-EL. Herein, near ultraviolet CPL materials with high photoluminescence quantum yields and good CPL dissymmetry factors are developed, which can induce blue to red CPL for various achiral luminescent materials. Strong near ultraviolet CP-EL with the best external quantum efficiencies (ηext s) of 9.0 % and small efficiency roll-offs are achieved by using them as emitters for CP-OLEDs. By adopting them as hosts or sensitizers, commercially available yellow-orange achiral phosphorescence, thermally activated delayed fluorescence (TADF) and multi-resonance (MR) TADF materials can generate intense CP-EL, with high dissymmetry factors and outstanding ηext s (30.8 %), demonstrating a simple and universal avenue towards efficient CP-EL.

Gut bacterial microbiota of Lymantria dispar asiatica and its involvement in Beauveria bassiana infection.

The gut bacterial microbiota of insects has been shown to play essential roles in processes related to physiology, metabolism, and innate immunity. In this study, we firstly performed a broad analysis of the gut bacteria in Lymantria dispar asiatica, one of the most devastating forestry defoliators. We analyzed the bacterial composition among different individuals from lab-reared or wild-collected using 16 s rRNA-sequencing, revealing that the gut bacteria of wild-collected larvae were highly diverse, while lab-reared larvae were only associated with a few genera. We found Lactobacillus sp. present in all the gut samples, which indicates that it is part of the core microbiome in the caterpillar. Further Beauveria bassiana infection-based assays showed that the mortality of non-axenic L. dispar asiatica larvae was significantly higher than that of axenic larvae at 72 h. Moreover, we isolated several bacteria from the hemolymph of the non-axenic larvae infected by B. bassiana, which may be caused by the translocation of gut bacteria from the gut to the hemocoel. Reintroduction of Enterococcus sp., Pseudomonas sp., Enterobacter sp., and Microbacterium sp. into axenic larvae recurred the larval mortality in their non-axenic counterpart. Taken together, our study demonstrates that the gut bacteria of L. dispar asiatica are highly volatile, and different bacteria taxa can promote host infection by entomopathogenic fungus, providing a new strategy for the pest management.

Composition and Diversity of Gut Bacterial Community in Different Life Stages of a Leaf Beetle Gastrolina depressa.

Insect gut bacteria have a significant impact on host biology, which has a favorable or negative impact on insect fitness. The walnut leaf beetle (Gastrolina depressa) is a notorious pest in China, causing severe damage to Juglandaceae trees including Juglans regia and Pterocarya rhoifolia. To date, however, we know surprisingly little about the gut microbiota of G. depressa. This study used a high-throughput sequencing platform to investigate the gut bacterial community of G. depressa throughout its life cycle, including the 1st, 2nd, and 3rd instar larvae, as well as male, female, and pre-pregnant female adults. Our results showed that the diversity of the gut bacterial community in larvae was generally higher than that in adults, and young larvae (1st and 2nd larvae) possessed the most diversified and abundant community. Principal coordinate analysis results showed that the gut microbiota of adults cluster together, which is independent of the 1st and 2nd instar larvae. The main phyla were Proteobacteria and Firmicutes in the microbial community of G. depressa, while the dominant genera were Enterobacter, Rosenbergiella, Erwinia, Pseudomonas, and Lactococcus. The gut bacteria of G. depressa were mostly enriched in metabolic pathways (carbohydrate metabolism and amino acid metabolism) as revealed by functional prediction. This study contributes to a better knowledge of G. depressa's gut microbiota and its potential interactions with the host insect, facilitating the development of a microbial-based pest management strategy.

A life-and-death struggle: interaction of insects with entomopathogenic fungi across various infection stages.

Insects constitute approximately 75% of the world's recognized fauna, with the majority of species considered as pests. Entomopathogenic fungi (EPF) are parasitic microorganisms capable of efficiently infecting insects, rendering them potent biopesticides. In response to infections, insects have evolved diverse defense mechanisms, prompting EPF to develop a variety of strategies to overcome or circumvent host defenses. While the interaction mechanisms between EPF and insects is well established, recent findings underscore that their interplay is more intricate than previously thought, especially evident across different stages of EPF infection. This review primarily focuses on the interplay between EPF and the insect defense strategies, centered around three infection stages: (1) Early infection stage: involving the pre-contact detection and avoidance behavior of EPF in insects, along with the induction of behavioral responses upon contact with the host cuticle; (2) Penetration and intra-hemolymph growth stage: involving the initiation of intricate cellular and humoral immune functions in insects, while symbiotic microbes can further contribute to host resistance; (3) Host insect's death stage: involving the ultimate confrontation between pathogens and insects. Infected insects strive to separate themselves from the healthy population, while pathogens rely on the infected insects to spread to new hosts. Also, we discuss a novel pest management strategy underlying the cooperation between EPF infection and disturbing the insect immune system. By enhancing our understanding of the intricate interplay between EPF and the insect, this review provides novel perspectives for EPF-mediated pest management and developing effective fungal insecticides.

Associated bacteria of a pine sawyer beetle confer resistance to entomopathogenic fungi via fungal growth inhibition.

BACKGROUND: The entomopathogenic Beauveria bassiana is a popular fungus used to control the Japanese pine sawyer, Monochamus alternatus Hope, the key vector of pine wood nematode (Bursaphelenchus xylophilus) that is the causal agent of pine wilt disease, resulting in devastating losses of pines in China and Portugal. However, recent studies have demonstrated that some insect-associated bacteria might decrease fungal toxicity and further undermine its biological control efficacy against M. alternatus. Thus, it is of great significance to uncover whether and how associated bacteria of M. alternatus become involved in the infection process of B. bassiana. RESULTS: Here, we show that axenic M. alternatus larvae died significantly faster than non-axenic larvae infected by four increasing concentrations of B. bassiana spores (Log-rank test, P < 0.001). The infection of B. bassiana significantly changed the richness and structure of the beetle-associated bacterial community both on the cuticle and in the guts of M. alternatus; meanwhile, the abundance of Pseudomonas and Serratia bacteria were significantly enriched as shown by qPCR. Furthermore, these two bacteria genera showed a strong inhibitory activity against B. bassiana (One-way ANOVA, P < 0.001) by reducing the fungal conidial germination and growth rather than regulating host immunity. CONCLUSIONS: This study highlights the role of insect-associated bacteria in the interaction between pest insects and entomopathogenic fungi, which should be taken into consideration when developing microbial-based pest control strategies.

Design, synthesis, and anticancer activity of three novel palbociclib derivatives.

Cancer is one of the most serious diseases threatening human health, so it is particularly important to develop effective tumor-targeting drugs. As the first CDK4/6 inhibitor, palbociclib effectively inhibits tumor proliferation by blocking the cell cycle to the G1 phase. 10-HCPT is a Topo I inhibitor; however, its clinical application has been greatly limited due to its high toxicity. Based on the successful development of double target inhibitors, three novel palbociclib derivatives (HP-1, HP-2, and HP-3) were designed and synthesized from Palbociclib and 10-HCPT, and their biological activities were investigated. At first, the possible binding sites of the three compounds to Topo I and CDK4/6 were predicted by molecular docking. Then, we evaluated the anti-proliferative effects of the three palbociclib derivatives. In general, human lung cancer cells were more sensitive to HP-1, HP-2, and HP-3, especially NCI-H460. In addition, cell cycle arrest and apoptosis induction were investigated by flow cytometry. The three palbociclib derivatives, especially HP-1, had obvious cell cycle arrest phenomenon on NCI-H460 cells and induced apoptosis of NCI-H460 cells significantly. In the end, it was proved that these three drugs had obvious cyclin-dependent kinase inhibitory activities. In short, all the data showed that HP-1, HP-2, and HP-3 could play anti-cancer roles by acting on dual targets and had the characteristics of high efficiencies and low toxicities, which opened up a new idea for the study of palbociclib derivatives.

Multi-omics analysis of the regulatory effects of low-phosphorus stress on phosphorus transport in soybean roots.

The regulatory effects of uneven phosphorus supplies on phosphorus transport in soybean roots are still unclear. To further analyze the regulatory effects of low-phosphorus stress on phosphorus transport in soybean roots and the effects of uneven phosphorus application on the physiological mechanism of phosphorus transport in soybean roots, dual-root soybean plants were prepared via grafting, and a sand culture experiment was performed. From the unfolded cotyledon stage to the initial flowering stage, one side of each dual-root soybean system was irrigated with a low-phosphorus-concentration solution (phosphorus-application [P+] side), and the other side was irrigated with a phosphorus-free nutrient solution (phosphorus-free [P-] side); this setup allowed the study of the effects of different phosphorus supply levels on the expression of genes and proteins and the accumulation of metabolites in soybean roots on the P- side to clarify the method through which phosphorus transport is regulated in soybean roots and to provide a theoretical basis for improving the use rate of phosphorus fertilizer. The results revealed that the unilateral supply of low-concentration phosphorus promoted the uptake of phosphorus by soybean roots and the transport of phosphorus from the P+ side to the P- side. Compared with the normal concentration of phosphorus supply and the phosphorus-free supply, the low concentration phosphorus supply affected the regulation of the metabolic pathways involved in starch and sucrose metabolism, glycolysis, fructose, and mannose metabolism, etc., thereby affecting soybean root phosphorus transport. The low-phosphorus stress inhibited fructose synthesis and sucrose synthase synthesis in the soybean roots and the synthesis of hexokinase (HK) and fructose kinase, which catalyzes the conversion of fructose to fructose-6-phosphate. Low-phosphorus stress promoted the synthesis of sucrose invertase and the conversion of sucrose into maltose by the activity of starch synthase (StS) and stimulated the synthesis of UDPG pyrophosphorylase (UGP) and phosphoglucose isomerase (GP1), which is involved in the conversion of UDP-glucose to glucose-6-phosphate. The phosphorus transport pathway of soybean roots was then affected, which promoted phosphorus allocation to UTP and glucose-6-phosphate. Additionally, low-phosphorus stress hastened glycolysis in the soybean roots and inhibited the synthesis of malic acid, thereby promoting the transport of phosphorus in the roots. In addition, low-phosphorus stress inhibited the synthesis of fructose, mannose, and mannose-1-phosphate and the synthesis of other enzymes involved in phosphorus transport as well as invertase, thereby inhibiting the transport and synthesis of several organic phosphorus-containing compounds.

Comparative analysis of the immune system and expression profiling of Lymantria dispar infected by Beauveria bassiana.

Lymantria dispar is one of the most devastating forest pests worldwide. Fungal biopesticides have great potential as alternatives owing to their high lethality to pests and eco-friendly feature, which is, however, often severely compromised by the pests' innate immunity. A better understanding of the antifungal immune system in L. dispar would significantly facilitate the development of the biopesticide. Here, we investigated phylogenetic characteristics of immunity-related genes as well as the tissue expression patterns in L. dispar after the infection of an entomopathogen Beauveria bassiana using RNA-sequencing data. Results showed most immune genes remain at a low level of response after 24 h post-infection (HPI). Almost all genes in the Toll pathway were significantly up-regulated at 48 HPI, and SPH1, SPN6, Toll6, Toll12, Myd88, pelle, and Drosal were significantly down-regulated at 72 HPI. Immunoblotting analysis revealed that the protein levels of ßGRP3 and PPO1 were significantly upregulated at 24 and 48 HPI, while Myd88 was downregulated at 24 HPI, which was further confirmed by Quantitative real-time PCR experiments. Moreover, the relative content of H2O2, a potent reactive oxygen species (ROS), was significantly increased with the decrease of the total antioxidant capacity, indicating that oxidative stress system positively participates in the clearance of the pathogenic fungus. Together, our study provides detailed genetic characteristics of antifungal immunity as well as profiling of the host defense against entomopathogenic infection, and comprehensive insight into molecular interaction between L. dispar and the entomopathogen.

Analysis of the Humoral Immunal Response Transcriptome of Ectropis obliqua Infected by Beauveria bassiana.

Ectropis obliqua is a destructive masticatory pest in China's tea gardens. Beauveria bassiana as microbial insecticides can effectively control E. obliqua larvae; however, the immune response of this insect infected by B. bassiana are largely unknown. Here, after isolating a highly virulent strain of B. bassiana from E. obliqua, the changes in gene expression among different tissues, including hemocytes and fat bodies, of E. obliqua larvae infected by the entomopathogen were investigated using transcriptome sequencing. A total of 5877 co-expressed differentially expressed genes (DEGs) were identified in hemocytes and fat bodies, of which 5826 were up-regulated in hemocytes and 5784 were up-regulated in fat bodies. We identified 249 immunity-related genes, including pattern recognition receptors, immune effectors, signal modulators, and members of immune pathways. A quantitative real-time PCR analysis confirmed that several pattern recognition receptors were upregulated in hemocytes and fat bodies; however, others were downregulated. The investigated immune effectors (ATT and PPO-1) were suppressed. The results showed that there were tissue differences in the expression of immune genes. This study provides a large number of immunity-related gene sequences from E. obliqua after being infected by B. bassiana, furthering the understanding of the molecular mechanisms of E. obliqua defenses against B. bassiana.

Isolation, Identification, and Analysis of Potential Functions of Culturable Bacteria Associated with an Invasive Gall Wasp, Leptocybe invasa.

Symbioses between invasive insects and bacteria are one of the key drivers of insect invasion success. Gall-inducing insects stimulate host plants to produce galls, which affects the normal growth of plants. Leptocybe invasa Fisher et La Salle, an invasive gall-inducing wasp, mainly damages Eucalyptus plantations in Southern China, but little is known about its associated bacteria. The aim of this study was to assess the diversity of bacterial communities at different developmental stages of L. invasa and to identify possible ecological functions of the associated bacteria. Bacteria associated with L. invasa were isolated using culture-dependent methods and their taxonomic statuses were determined by sequencing the 16S rRNA gene. A total of 88 species belonging to four phyla, 27 families, and 44 genera were identified by phylogenetic analysis. The four phyla were Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes, mainly from the genera Pantoea, Enterobacter, Pseudomonas, Bacillus, Acinetobacter, Curtobacterium, Sphingobium, Klebsiella, and Rhizobium. Among them, 72 species were isolated in the insect gall stage and 46 species were isolated from the adult stage. The most abundant bacterial species were γ-Proteobacteria. We found significant differences in total bacterial counts and community compositions at different developmental stages, and identified possible ecological roles of L. invasa-associated bacteria. This study is the first to systematically investigate the associated bacteria of L. invasa using culture-dependent methods, and provides a reference for other gall-inducing insects and associated bacteria.

Differing Dietary Nutrients and Diet-Associated Bacteria Has Limited Impact on Spider Gut Microbiota Composition.

Spiders are a key predator of insects across ecosystems and possess great potential as pest control agents. Unfortunately, it is difficult to artificially cultivate multiple generations of most spider species. Since gut bacterial flora has been shown to significantly alter nutrient availability, it is plausible that the spiders' microbial community plays a key role in their unsuccessful breeding. However, both the gut microbial composition and its influencing factors in many spiders remain a mystery. In this study, the gut microbiota of Campanicola campanulata, specialists who prey on ants and are widely distributed across China, was characterized. After, the impact of diet and diet-associated bacteria on gut bacterial composition was evaluated. First, two species of prey ants (Lasius niger and Tetramorium caespitum) were collected from different locations and fed to C. campanulata. For each diet, we then profiled the nutritional content of the ants, as well as the bacterial communities of both the ants and spiders. Results showed that the protein and carbohydrate content varied between the two prey ant species. We isolated 682 genera from 356 families in the ants (dominant genera including Pseudomonas, Acinetobacter, Paraburkholderia, Staphylococcus, and Novosphingobium), and 456 genera from 258 families in the spiders (dominated by Pseudomonas). However, no significant differences were found in the gut microbiota of spiders that were fed the differing ants. Together, these results indicate that nutritional variation and diet-associated bacterial differences have a limited impact on the microbial composition of spider guts, highlighting that spiders may have a potentially stable internal environment and lay the foundation for future investigations into gut microbiota.