Vardakas, P. | Search bioRxiv

bioRxiv2026

Agroforestry transition increases insect diversity and reorganizes soil function in a Mediterranean orchard

Agricultural intensification has reduced biodiversity and weakened ecosystem functions essential for sustainable crop production. Agroforestry has been proposed as a regenerative strategy to restore these functions through vegetation diversification, yet its ecological effects at the farm scale remain insufficiently documented, particularly in Mediterranean perennial systems. Here, adaptive field monitoring and soil shotgun metagenomics were combined to investigate ecological responses across a 10-year agroforestry transition gradient in a Mediterranean citrus and olive farm. Mature agroforestry plots supported higher insect richness and functional diversity while maintaining stable pollinator communities. Plant-insect interaction analyses suggested that potential pest activity was largely associated with spontaneous and supportive vegetation rather than crops. In contrast, soil communities showed limited changes in overall richness but substantial compositional and functional restructuring, including enrichment of the arbuscular mycorrhizal fungus Rhizophagus and proteins with nutrient-cycling functions in advanced plots. Together, these findings suggest that agroforestry transition can promote functional diversification above and below ground and highlight the value of farmer-led regenerative transitions coupled with integrative ecological monitoring for the development of more resilient agricultural systems.

ecology↗

bioRxiv2021

Bee foraging preferences, microbiota and pathogens revealed by direct shotgun metagenomics of honey

Honeybees (Apis mellifera) continue to succumb to human and environmental pressures despite their crucial role in providing essential ecosystem services. Owing to their foraging and honey production activities, honeybees form complex relationships with species across all domains, such as plants, viruses, bacteria (symbiotic and pathogenic), and other hive pests, making honey a valuable biomonitoring tool for assessing their ecological niche. Thus, the application of honey shotgun metagenomics (SM) has paved the way for a detailed description of the species honeybees interact with, in order to better assess the multiple factors governing their health. Here, we describe the implementation of optimized honey DNA extraction methodology coupled to direct shotgun metagenomics (Direct-SM) analysis, and to a computationally optimised and validated pipeline for taxonomic classification of species detected in honey. By comparing honey collected across 3 harvesting seasons in a stable apiary, we show that Direct-SM can describe the variability of sampled plant species, revealing honeybee behavioural adaptation. In addition, we reveal that Direct-SM can non-invasively capture the diversity of species comprising the core and non-core bacterial communities of the gut microbiome. Finally, we show that this methodology is applicable for the monitoring of pathogens and particularly for the biomonitoring varroa infestation. These results suggest that Direct-SM can accurately and comprehensively describe honeybee ecological niches and can be deployed to assess bee health in the field.

ecology↗