Wolbachia are common heritable endosymbionts that influence many aspects of ecology and evolution in various insects, yet Wolbachia ‐mediated intracellular metabolic responses to temperature stress have been largely overlooked. Here, we introduced the Wolbachia strain wLhui from the invasive Liriomyza huidobrensis (Blanchard) into a Drosophila Schneider 2 cell line (S2) and investigated the metabolite profile of wLhui‐infected (S2_wLhui) and uninfected cell lines (S2_wu) under short‐term exposure to either high (37°C), moderate (27°C), or low (7 and 17°C) temperatures. We find that Wolbachia infection, temperature stress, and their interactions significantly affect cellular metabolic profiles. Most significantly, when comparing the changes in metabolites between S2_wLhui and S2_wu, glycerophospholipids, amino acids, and fatty acids associated with metabolic pathways, microbial metabolism in diverse environments, and other pathways were significantly accumulated at either low or high temperatures. Our findings suggest Wolbachia ‐induced cellular physiological responses to short‐term temperature stress, which may in turn affect the fitness and adaptive ability of its host as an invasive species.

Microbe–microbe interactions within a host drive shifts in the host’s microbiota composition, profoundly influencing host physiology, ecology, and evolution. Among these microbes, the maternally inherited endosymbiont Wolbachia is widespread in the invasive pest Liriomyza huidorbrensis (Diptera: Agromyzidae). However, its influence on the host microbiota remains largely unexplored. In the study presented herein, we investigated the bacterial communities of Wolbachia wLhui-infected (wLhui+) and -uninfected lines (wLhui−) of L. huidorbrensis using 16S rRNA gene high-throughput sequencing. For both leaf-miner lines, Bacteroidota was the dominant phylum (relative abundance: 59.18%), followed by Pseudomonadota (36.63%), Actinomycetota (2.42%), and Bacillota (0.93%). We found no significant differences in alpha-diversity indices between the wLhui+ and wLhui− lines (p > 0.05). However, principal coordinates analysis revealed significant differences in microbiota composition between the wLhui+ and wLhui− lines (PERMANOVA: p < 0.001), explaining 76.70% of the variance in microbiota composition. Correlation network analysis identified robust negative and positive associations between Wolbachia and several genera, suggesting that Wolbachia shapes microbial composition through competitive or cooperative interactions with specific taxa. Overall, our study suggests that Wolbachia plays a key role in shaping the leaf-miner microbiome, potentially affecting host fitness.

Arthropods harbor complex microbiota that play a pivotal role in host fitness. While multiple factors, like host species and diet, shape microbiota in arthropods, their impact on community assembly in wild insects remains largely unknown. In this study, we surveyed bacterial and fungal community assembly in nine sympatric wild insect species that share a common citrus fruit diet. Source tracking analysis suggested that these insects acquire some bacteria and fungi from the citrus fruit with varying degrees. Although sharing a common diet led to microbiota convergence, the diversity, composition, and network of both bacterial and fungal communities varied significantly among surveyed insect groups. Null model analysis indicated that stochastic processes, particularly dispersal limitation and drift, are primary drivers of structuring insect bacterial and fungal communities. Importantly, the influence of each community assembly process varied strongly depending on the host species. Thus, we proposed a speculative view that the host specificity of the microbiome and mycobiome assembly is widespread in wild insects despite sharing the same regional species pool. Overall, this research solidifies the importance of host species in shaping microbiomes and mycobiomes, providing novel insights into their assembly mechanisms in wild insects. IMPORTANCE: Since the microbiome has been shown to impact insect fitness, a mechanistic understanding of community assembly has potentially significant applications but remains largely unexplored. In this paper, we investigate bacterial and fungal community assembly in nine sympatric wild insect species that share a common diet. The main findings indicate that stochastic processes drive the divergence of microbiomes and mycobiomes in nine sympatric wild insect species. These findings offer novel insights into the assembly mechanisms of microbiomes and mycobiomes in wild insects.

The bacterial communities of arthropods play important roles in the growth and development of their hosts, such as by providing nutrition or protecting against parasites and pathogens. However, for Tetranychus truncatus, a major crop pest, very little is known about its bacterial community. To understand the effects of the bacterial community on T. truncatus, mites were fed bean leaves that had been soaked in different concentrations of tetracycline hydrochloride (5 mg to 5 g per L). Antibiotic treatment had a negative impact on the fitness of T. truncatus, decreasing their fecundity and longevity. However, we could not exclude the possibility that the decrease in fecundity was due to a toxic effect of the antibiotic. 16S rRNA analyses showed that the antibiotic treatment did not significantly affect the diversity of bacterial species, but it did significantly change their relative abundances. Furthermore, predicted functions of microbiota were also changed with antibiotic treatment. Our research revealed that the antibiotic induced change of bacterial communities mostly decreased the fecundity of T. truncatus, although we couldn’t totally exclude the direct toxin of antibiotics on spider mites.

Heritable endosymbiont Wolbachia is prevalent among arthropods, serving multiple functions for their hosts. However, the role of Wolbachia in mediating thermal preference selection remains largely unexplored. In this study, we utilized a custom-built thermal gradient to evaluate the thermal preference (Tp) of 1367 individuals of the invasive leaf-miner Liriomyza huidobrensis with or without Wolbachia wLhui from Yunnan and Xinjiang populations. Under meticulously controlled conditions and with a vast sample size, we found no significant difference in the mean Tp between wLhui-infected and uninfected leaf miners from either population when host age and sex were not considered. Furthermore, generalized linear model (GLM) analysis revealed no significant correlation between average Tp and age, sex, or Wolbachia infection, nor interactions among these factors, except in the Xinjiang population, where Tp was strongly associated with host age. Finally, we discuss the ecological implications of these findings and propose future research directions on Wolbachia-mediated host Tp in the leaf miner. Overall, our findings do not provide evidence that Wolbachia significantly affects the thermal preference of L. huidobrensis. Further studies across different systems are needed to investigate the complex interactions between Wolbachia and insect thermal behavior.