Microbiome composition shapes temperature tolerance in a Hawaiian picture-winged <em>Drosophila</em>

Hawaiian picture-winged Drosophila are undergoing rapid biodiversity loss, with twelve species listed as endangered and others in decline. Microbiota are increasingly recognized as contributors to host adaptation and influence stress tolerance, reproduction, and survival. We investigated the role of microbial communities in two Drosophila basisetae populations from Hawaiian rainforests at 900m and 1200m elevation. Microbiome profiling of wild flies by high-throughput amplicon sequencing revealed distinct bacterial and fungal communities between sites. To test microbiome effects on host physiology, we conducted a fully factorial research design with microbiome inoculations in laboratory-reared flies acclimated at 18°C (control) or 24°C (stressful). Flies receiving low-elevation microbiota exhibited higher survival across temperatures, whereas those given high-elevation microbiota produced more eggs, indicating microbiome-mediated differences in survival and reproductive investment. Activ..., Wild-caught flies from both locations were transported to the laboratory at UNLV and immediately placed in separate 1-gallon (∼3.8 l) breeding jars kept in a climate-controlled walk-in chamber and maintained at 18°C and 60–70% RH, the standard rearing temperature for picture-winged Drosophila, with a 12 h:12 h day: night cycle.&nbsp;For microbiome inoculation experiments, virgin female and male flies were first separated by sex within 7 days of eclosion and placed in adult food vials for 3 days at 18°C and then subsequently placed in vials with an antimicrobial diet for 7 days and housed in an incubator at 18°C.After the antimicrobial treatment, male and female D. basisetae from Tom’s Trail and Ola‘a populations were randomly placed (1 or 2 flies per vial) in sterilized glass vials containing a standard diet supplemented with 12.5 μl of a microbiome slurry from either Tom’s Trail or Ola‘a pipetted onto the surface of the food. The vials, along with the flies, were randomly placed in eit..., # Microbiome composition shapes temperature tolerance in a Hawaiian picture-winged *Drosophila* Dataset DOI: [10.5061/dryad.j6q573nt7](10.5061/dryad.j6q573nt7) ## Description of the data and file structure ### Files and variables #### File: Phenotyic_Data_D._basisetae_microbiome_and_temperature_tolerance_revised.csv **Description:** ##### Variables * Block: The data was collected over five time periods that are designated as Block. * FlyID: The identification of the individual fly. * Fly Location: The flies in the experiments were originally collected from two locations in the Hawaiian rainforest - olaa and toms * Sex: F is female flies and M is male flies * Acclimation Temperature: There were two acclimation temperatures 18 Celsius and 24 Celsius. * Microbe Treatment: There were two inoculation treatments of microbes (bacteria and fungi) from olaa and toms. * Survived\_ Acclimation: The flies that survived the acclimation period are designated Y and the flies that did no...,