Microbes in bee-plant networks: Composition characterization and their ecological implications

Abstract

Honey bees are vital pollinators in ecosystems around the world, and microbes play key roles in connecting bees and plants. Collectively, microbes, bees and plants form intricate tripartite interactions networks. Through a co-evolution, many bee- and plant-associated microbes have developed functions that benefit their hosts, including promoting growth, enhancing pathogen resistance, and aiding digestion. Microbiome associated with different hosts tend to be specific, and within bee-microbe-plant networks, both beneficial and pathogenic microbes are dynamically transmitted among hosts. In the current study, honey bee corbicular samples were collected over a two-year period from beehives at Justus-Liebig-University Giessen. Corbicular pollen is able to reflect both microbes and plants encountered by honey bees during foraging activity. Plant and microbial communities in honey bee corbicular pollen were profiled using 16S rRNA gene and plant ITS2 metabarcoding. The results indicated that the corbicular pollen microbiome exhibited clear seasonal variations, and was affected by multiple environmental factors as well as choices of forage plants. Co-occurrence network analysis further revealed specific plant-microbe associations and identified several hub plant taxa that may serve as hotspots for microbial transmissions. Following this study, we characterized bacterial and fungal microbiome of flowers from a highly insect-visited hub plant, bramble (genus Rubus), using 16S rRNA gene and fungal ITS2 metabarcoding. The data showed that insect visitation increased microbial loads on flowers and enriched specific microbial groups including fermentative and pathogenic microbes, highlighting the role of bramble flowers as hotspots for microbial transmission. In addition, insect visitation altered floral microbiome structure, potentially through the introduction of several hub microbial taxa and by increasing the centralization of the microbial interaction networks. Honey bees were collected from beehives, and common bacterial, fungal and viral honey bee pathogens were screened. The expression levels of several immunity-related genes (defensin-1, lysozyme-like, vitellogenin, glucose oxidase) and the composition of the bee microbiome were examined to assess honey bee health status. Black queen cell virus (BQCV) was detected in almost all individuals, while Vairimorpha pathogens were only partially detected. Paenibacillus larvae, Melissococcus plutonius, Kashmir bee virus (KBV), Acute Bee Paralysis Virus (ABPV), Chronic bee paralysis virus (CBPV), Deformed Wing Virus (DWV), and Sacbrood bee virus (SBV) were not detected in the samples. The data indicated that BQCV and Vairimorpha infections had no significant impact on the expression of immunity-related gene. Since the microbiome composition was assessed at the hive level and BQCV was present in every hive, its potential influence on the microbiome remains to be further clarified. In addition, a bacterial isolate from birch pollen was phenotypically, genotypically, and chemotaxonomically characterized using a polyphasic approach. Based on 16S rRNA gene phylogeny and comparative genomic analysis, the isolate was identified as a novel species of the genus Robbsia. The bacterium was rod-shaped, non-motile, facultative anaerobic and grew optimally at 28 °C and pH 6–7. Unlike its closest relative Robbsia andropogonis, which is a known phytopathogen, the isolate exhibited no phytopathogenic traits such as flagellum formation, rhizobitoxine production, or induction of plant hypersensitive response. The proposed and accepted name of the isolate is Robbsia betulipollinis Bb-Pol-6T.