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Acute lymphoblastic leukemia (ALL) is a malignant disease of the hematologic system. The current treatment is based on chemotherapeutic drugs, which are becoming less effective due to drug resistance. TNF-related apoptosis-inducing ligand (TRAIL) is an apoptotic protein used to treat cancer that does not affect healthy cells. In recent years, however, ALL cells (e.g., U937) have become more resistant to TRAIL. A novel andrographolide derivative (AND7) with high efficiency and low toxicity was synthesized and combined with TRAIL after the optimal combination ratio was screened using U937 cells. We used peripheral blood mononuclear cells (PBMCs) from patients before the initial treatment of ALL as a model and PBMCs from healthy subjects as a control to determine the mechanism underlying ALL treatment. AND7/TRAIL combination treatment was shown to prevent the original TRAIL-resistant cells from activating the caspase-8/caspase-3 pathway through DR4/DR5 and promote apoptosis via expression of ROS and the apoptotic gene, P53, to achieve an anti-cancer effect. Notably, this study demonstrated that the AND7/TRAIL combination enhanced the anti-cancer effect of AND7 and improved TRAIL resistance. Therefore, the AND7/TRAIL combination is promising for treating ALL and lays the foundation for clinical research.

Abstract Continuous cropping has a negative effect on soybean yield. In this study, a positioning experiment was conducted starting in 2015, with three treatments: maize–soybean rotation (SMR), 2-year maize, 2-year soybean rotation cropping (SC2), and 8-year soybean continuous cropping (SC8). We determined soybean yields (2015–2022) and analyzed soil microbial communities, functions, and metabolites composition in the 0–20 cm tillage layer using metagenomics technology and GC–MS technology during soybean flowering in 2022. Results indicated that continuous cropping (SC8) significantly reduced soybean yield compared to crop rotation (SMR) during the experimental period, while SC8 showed higher yield than SC2 in 2022. Compared to SMR, SC8 significantly increased soil N content and significantly decreased pH and TP, AP, and AK content. However, the pH and AK contents of SC8 were significantly higher than those of SC2. LeFSe analysis showed that Friedmanniella, Microlunatus, Nitrososphaera, Rubrobacter, Geodermatophilus, Nitriliruptor were enriched in SC8. Gaiella, Sphaerobacter, Methyloceanibacter were enriched in SC2. Sphingomonas, Cryobacterium, Marmoricola, Haliangium, Arthrobacter, Ramlibacter, Rhizobacter, Pseudolabrys, Methylibium, Variovorax were enriched in SMR. And the relative abundance of Cryobacterium, Marmoricola, Haliangium, Arthrobacter, Ramlibacter, Rhizobacter, Methylibium, Variovorax was significantly positively correlated with yield, while the relative abundance of Gaiella and Sphaerobacter was significantly negatively correlated with yield. SC8 significantly increased the abundance of genes in nitrogen metabolism and significantly decreased the abundance of genes related to phosphorus and potassium metabolism compared with SMR. However, the abundance of genes in potassium metabolism was significantly higher in SC8 than in SC2. Metabolomic analysis showed that compared to SMR, SC8 decreased the abundance of carbohydrates, ketones, and lipid. However, the abundance of carbohydrates, ketones, and lipid was significantly higher in SC8 than in SC2. Mantel test showed that soil pH and AK significantly affected soil microbial community, function, and metabolite composition. Correlation analysis showed significant correlation between soil metabolites and microorganisms, metabolic functions. Graphical Abstract

The long-standing arms race between plants and herbivores has led to the evolution of highly diverse mutual adaptation strategies, among which microbes play an indispensable role [...]

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.

Abstract 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.

Abstract 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

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.

The harlequin ladybird, Harmonia axyridis, is a valuable asset in integrated pest management (IPM); however, issues related to low-temperature storage and transportation have resulted in low hatching rate, while the use of pesticides may lead to non-target effects against this natural enemy during field application. Fluctuating thermal regimes (FTR) have been shown to be beneficial during the low-temperature storage, and the type and concentration of insecticides used are crucial for field application of H. axyridis. Despite this, little research has been conducted on the effects of FTR on the hatching rate of ladybird eggs, and the impact of pesticides on their egg viability remains unclear. To address these gaps, we investigated the effects of different thermal temperatures, recovery frequencies (the number of changes in temperature conditions per unit time), and recovery durations (the duration of the treated temperature condition) on egg hatching under constant low-temperature conditions. We also examined the toxicity and safety of seven commonly used insecticides on egg hatching. Our results indicate that the temperature during FTR application did not significantly affect egg hatching, but the interaction between temperature and recovery frequency can significantly affect egg hatching. Moreover, the recovery frequency and recovery duration had a significant impact on hatching. Under specific conditions, the hatching rate of eggs subjected to FTR was similar to that of eggs stored at 25 °C. Furthermore, we found that matrine (a kind of alkaloid pesticide isolated from Sophora flavescens) had low toxicity to ladybird eggs and is a safe pesticide for use in conjunction with this natural enemy. The study provides valuable information on effectively managing H. axyridis by taking into account both storage temperature and pesticide exposure.

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.

Abstract Background RNA interference (RNAi) has emerged as an efficient tool to control insect pests. When insects ingest double-stranded RNAs (dsRNAs) targeted against essential genes, strong gene silencing and mortality can be induced. To exert their function, dsRNA molecules must pass through the insect’s gut and enter epithelial cells and/or the hemolymph. Gut bacteria are known to play multifarious roles in food digestion and nutrition, and confer protection against pathogens and parasites. Whether there is a cross talk between gut bacteria and ingested dsRNAs and whether the microbiome affects RNAi efficiency are unknown. Results Here, using a leaf beetle gut microbiota system, we investigated whether gut bacteria interact with dsRNA molecules and how the gut microbiota affects RNAi responses in insects. We first showed that the leaf beetle Plagiodera versicolora (Coleoptera) is highly susceptible to RNAi. We then demonstrated that ingestion of dsRNAs by non-axenic P. versicolora larvae results in (i) significantly accelerated mortality compared with axenic larvae, and (ii) overgrowth and dysbiosis of the gut microbiota. The latter may be caused by bacterial utilization of dsRNA degradation products. Furthermore, we found that Pseudomonas putida, a gut bacterium of P. versicolora, acts as major accelerator of the death of P. versicolora larvae by transitioning from commensal to pathogenic lifestyle. Conclusions The present study illuminates the complex interplay between lethal dsRNA, the insect host, and its gut microbiota. The ingestion of dsRNA by the leaf beetle caused a dysbiosis of gut bacterial community, and the dsRNA degradation products by host insect preferentially promoted the growth of an entomopathogenic bacterium, which accelerated dsRNA lethality to the insect. Our findings reveal a synergistic role of the gut microbiota in dsRNA-induced mortality of pest insects, and provide new insights in the mechanisms of RNAi-based pest control. Video abstract

Chilo suppressalis is a major global host-specific pest of rice and water oat crops, having caused severe yield damage and great economic loss. The pest occasionally feeds on other plants, preventing them from completing a full life cycle. Dietary differences may shape the gut microbiota of the pest and may further influence its performance. In order to test this hypothesis, an assembly of the gut bacterial community of C. suppressalis larvae reared on rice, water oat, corn, and an artificial diet were investigated using an amplicon-based next-generation sequencing approach. The larval body size and feeding rate on each diet type were analyzed in parallel. We found that the size and feeding rate of C. suppressalis varied among the different dietary regimes, as a low feeding rate was found for rice and corn diets, whereas rice-fed larvae had the biggest body size and corn-fed larvae had the lowest body size. Further high-throughput sequencing results showed that the artificial-diet-fed larvae had the lowest bacterial diversity among all the samples, and the corn-fed larvae presented the most diversified microbial community. Further analysis revealed that the bacterial genera Enterococcus, Sphingobacterium, Klebsiella, Gluconobacter, Serratia, and Lactococcus possessed high abundance in C. suppressalis larvae, and the varied abundances contributed to the differences in community structure. The microbial function classification suggested that metabolic function categories significantly increased while the larvae were feeding on their preferred diet (rice and water oat) and decreased when on an artificial diet. This study expands our understanding of the microbe–insect interaction of C. suppressalis larvae in response to changes in diet, and is an essential step towards the future development of potential microbial-based pest management strategies.

Given the multiple roles of associated microbiota in improving animal host fitness in a microbial environment, increasing numbers of researchers have focused on how the associated microbiota keeps stable under complex environmental factors, especially some biological ones. Recent studies show that associated microbiota interacts with pathogenic microbes. However, whether and how the interaction would influence microbiota stability is limitedly investigated. Based on the interaction among Delia antiqua, its associated microbiota, and one pathogen Beauveria bassiana, the associated microbiota's response to the pathogen was determined in this study. Besides, the underlying mechanism for the response was also preliminarily investigated. Results showed that B. bassiana neither infect D. antiqua larvae nor did it colonize inside the associated microbiota, and both the bacterial and fungal microbiota kept stable during the interaction. Further experiments showed that bacterial microbiota almost completely inhibited conidial germination and mycelial growth of B. bassiana during its invasion, while fungal microbiota did not inhibit conidial germination and mycelial growth of B. bassiana. According to the above results, individual dominant bacterial species were isolated, and their inhibition on conidial germination and mycelial growth of B. bassiana was reconfirmed. Thus, these results indicated that bacterial instead of fungal microbiota blocked B. bassiana conidia and stabilized the associated microbiota of D. antiqua larvae during B. bassiana invasion. The findings deepened the understanding of the role of associated microbiota–pathogen microbe interaction in maintaining microbiota stability. They may also contribute to the development of novel biological control agents and pest management strategies.

Plagiodera versicolora (Coleoptera: Chrysomelidae) is one of the most destructive pests of the Salicaceae worldwide, which has established complex interactions with surrounding organisms. Uncovering the molecular mechanisms of some antagonistic interactions would facilitate the development of environmentally friendly pest insect management strategies. Suitable reference genes are essential for reliable qPCR and gene expression analysis in molecular studies; however, a comprehensive assessment of reference genes in P. versicolora is still lacking. In this study, the stability of seven housekeeping genes (including Actin, EF1A, α-tubulin, RPL13a, RPS18, RPL8 and UBC) was investigated under both biotic (developmental stages, tissues, sex and pathogen treatment) and abiotic (RNA interference treatment, temperature treatment) conditions. The geNorm, NormFinder, BestKeeper, and ΔCt programs were used to analyze gene expression data. The RefFinder synthesis analysis was applied to suggest a handful of appropriate reference genes for each experimental condition. RPS18 and EF1A were the most reliable reference genes in different development stages; RPS18 and RPL8 were most stable in female and male adults, different tissues, different temperatures, and pathogen treatment; α-tubulin and RPL13a were most stable after dietary RNAi treatment. The research provides a strong basis for future research into the molecular biology of P. versicolora.

The wide-spread culture of transgenic Bt cotton resisting the infamous cotton bollworms has reduced the adoption of broad-spectrum insecticides to a large extent. Consequently, the non-targeted insect Adelphocoris suturalis Jakovlev has become a major cotton pest in China. Entomopathogenic microbes show promising results for controlling this pest in the future, but A. suturalis innate immune responses to these pathogens are poorly understood. Here, we used the entomopathogenic fungus Beauveria bassiana and the Gram-negative pathogenic bacteria Enterobactor cloacae to infect A. suturalis nymphs, followed by high throughput RNA-seq to analyze the immune transcriptomes of A. suturalis in response to the two pathogens. A total of 150 immunity-related genes were identified, including pattern recognition receptors, extracellular signal modulators, signal pathways (Toll, IMD, JNK, and JAK/STAT), and response effectors. Further quantitative real-time PCR analysis demonstrated that B. bassiana and E. cloacae were recognized by different receptors (GNBP and PGRP, respectively); activated Toll pathway and IMD pathway respectively; and both induced expression of the effector gene Defensin. However, melanization is suppressed in B. bassiana-infected nymphs. Collectively, this study provides a transcriptomic snapshot of the A. suturalis immune system, and at the genetic level, gains multifaceted insights of the immune response to fungal and Gram-negative bacterial pathogens. Ultimately this work pioneers the study of molecular mechanisms underlying immune interactions between A. suturalis and its pathogens and assists in the development of novel mitigation strategies to control this pest.

ABSTRACT Colonization resistance, i.e., the protective effects of associated microbiota for the animal host against pathogen infection, has been studied widely over the last 100 years. However, few molecules mediating colonization resistance have been identified. In the symbiosis formed by Delia antiqua and its associated microbes, six bacteria protect larvae from infection with the entomopathogen Beauveria bassiana, providing an ideal model to investigate the chemical mechanism for colonization resistance. Subsequently using this symbiotic system, we first compared effects of the six bacterial species, and one control bacterium (Klebsiella oxytoca) that showed no antifungal effects, on B. bassiana and its infection of D. antiqua. Second, metabolomic profiles of the six bacteria and K. oxytoca were compared to identify candidate metabolites that may prevent infection. Third, the concentrations of candidate metabolites in situ from axenic and nonaxenic larvae were determined. Finally, effects of artificial metabolite cocktails on B. bassiana and its infection of D. antiqua larvae were determined. Results showed that compared to K. oxytoca, the six bacteria produced a metabolite cocktail showing inhibitory effects on conidial germination, mycelial growth of B. bassiana, and fungal infection. Our work revealed novel molecules that mediate colonization resistance, which could help in developing chemical mechanisms of colonization resistance. Moreover, this work may aid in discovery and expansion of new bioactive antibiotics, promoting development of prophylactic and therapeutic approaches for treating infectious diseases. IMPORTANCE The protection of associated microbiota for their animal hosts against pathogen infection has been studied widely over the last 100 years. However, how those microbes protect the animal host remains unclear. In former studies, body surface microbes of one insect, Delia antiqua, protected the insect larvae from infection with the entomopathogen Beauveria bassiana. By comparing the metabolites produced by microbes that protect the insect and microbes that cannot protect the insect, the question of how the microbes protect the insect is answered. It turns out that body surface bacteria produce a metabolite cocktail that inhibits colonization of B. bassiana and consequently protects the insect. This work reveals novel molecules with antifungal activity, which may aid in discovery and expansion of new prophylactic and therapeutic natural chemicals for treating infectious diseases.

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.

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.

RNA interference (RNAi) has emerged as a novel and feasible strategy for pest management. Methods for cost-effective production and stable delivery of double-stranded RNA (dsRNA) to the target insects are crucial for the wide application of RNAi for pest control. In this study, we tested the expression of dsRNA in RNaseIII-deficient Escherichia coli HT115 which was then fed to Plagiodera versicolora larvae, an insect pest of Salicaceae plants worldwide. By targeting six potential genes, including actin (ACT), signal recognition particle protein 54k (SRP54), heat shock protein 70 (HSC70), shibire (SHI), cactus (CACT), and soluble N-ethylmaleimide-sensitive fusion attachment proteins (SNAP), we found that feeding bacteria-expressed dsRNA successfully triggered the silencing of the five target genes tested and the suppression of ACT and SRP54 genes caused significant mortality. Our results suggest that the oral delivery of bacteria-expressed dsRNA is a potential alternative for the control of P. versicolora, and that ACT and SRP54 genes are the potent targets.

Insects harbor a community of gut bacteria, ranging from pathogenic to obligate mutualistic organisms. Both biotic and abiotic factors can influence species composition and structure of the insect gut bacterial communities. Dendroctonus valens is a destructive forest pest in China. To overcome host pine defenses, beetles mass-attack the pine to a threshold density that can exhaust pine defenses. The intensity of pine chemical defenses and carbohydrate concentrations of pines can be influenced by beetle attack, both of which are known factors that modify beetle’s gut microbiota. However, little is known to what extent variation exists in the beetle’s gut communities, and host monoterpenes and carbohydrates at different attack densities. In this study, the gut bacterial microbiota of D. valens at low and high attack densities were analyzed, and monoterpenes and carbohydrates in host pine phloem were assayed in parallel. The results showed that no significant changes of gut bacterial communities of the beetles and concentrations of D-glucose, D-pinitol, and D-fructose in pine phloem were found between low and high attack densities. The concentrations of α-pinene, β-pinene, limonene at high attack densities were significantly higher than those at low attack densities. Our results suggested that different attack densities of D. valens influence monoterpenes concentration of host pines’ phloem but have no significant impact on gut bacterial community structures of D. valens and carbohydrate concentration of host trees’ phloem in early attack phase. Similar gut bacterial community structures of D. valens between low and high attack densities might be due to the quick adaptation of gut microbiota to high monoterpenes concentrations.

Dendroctonus valens, an invasive bark beetle, has caused severe damage to Chinese forests. Previous studies have highlighted the importance of the gut microbiota and its fundamental role in host fitness. Culture-dependent and culture-independent methods have been applied in analyzing beetles’ gut microbiota. The former method cannot present a whole picture of the community, and the latter mostly generates short read lengths that cannot be assigned to species. Here, the PacBio sequencing system was utilized to capture full-length 16S rRNA sequences in D. valens gut throughout its ontogeny. A total of eight phyla, 55 families, 102 genera, and 253 species were identified. Bacterial communities in colonized beetles have the greatest richness but the lowest evenness in all life stages, which is different from those in young larvae. Pseudomonas sp., Serratia liquefaciens possess high abundance throughout its ontogeny and may serve as members of the core bacteriome. A phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) analysis predicted that gut microbiota in larvae are rich in genes involved in carbohydrate, energy metabolism. Gut microbiota in both larvae and colonized beetles are rich in xenobiotics and terpenoids biodegradation, which are decreased in dispersal beetles. Considering that the results are based mainly on the analysis of 16S rRNA sequencing and PICRUSt prediction, further confirmation is needed to improve the knowledge of the gut microbiota in D. valens and help to resolve taxonomic uncertainty at the species level.

The exposure to plant defense chemicals has negative effects on insect feeding activity and modifies insect gut microbial community composition. Dendroctonus valens is a very destructive forest pest in China, and harbors a large diversity and abundance of gut microorganisms. Host pine defensive chemicals can protect the pines from attack by the holobiont. In this study, boring length of D. valens feeding on 0 mg/g α-pinene and 9 mg/g α-pinene concentration in phloem media for 6 and 48 h were recorded, and their gut bacterial communities were analyzed in parallel. Nine milligram per gram α-pinene concentration significantly inhibited boring length of D. valens and altered its gut microbial community structure after 6 h. The inhibition of boring length from 9 mg/g α-pinene in diets ceased after 48 h. No significant differences of the bacterial communities were observed between the beetles in 0 and 9 mg/g α-pinene concentration in phloem media after 48 h. Our results showed that the inhibition of the feeding behavior of D. valens and the disturbance to its gut bacterial communities in 9 mg/g α-pinene concentration in phloem media after 6 h were eliminated after 48 h. The resilience of gut bacterial community of D. valens may help the beetle catabolize pine defense chemical.