Acquisition of an obligate environmental symbiont may be limited in the arboreal environment.

Many eukaryotic organisms have environmentally acquired microbial symbionts. In animals, microbes commonly occupy the gut and may supply critical nutrients. The leaf-footed bug, Leptoglossus zonatus (Coreidae), is a true bug that is dependent upon ingestion of the free-living, soilborne bacterium Caballeronia early in development for growth and reproduction. In 2019 and 2020, we tested the ability of second instar L. zonatus to acquire Caballeronia in the canopy of pomegranate trees where L. zonatus are often found. We compared the acquisition rate of Caballeronia in nymphs left to forage for the symbiont to bugs fed Caballeronia in advance. Additionally, we aimed to determine whether the microhabitat of potential symbiont sources influenced acquisition success. We hypothesized that the acquisition rate would be heterogeneous among treatments. In 2019, ∼30% of experimental bugs acquired Caballeronia, compared to 75% of those fed the symbiont. In 2020, only about 4% of experimental bugs acquired any symbiont. The symbiont composition of caged bugs differed, and strain diversity was reduced relative to wild bugs. We concluded that Caballeronia is present in the canopy environment, but nymphs may fail to acquire it in the fragments of habitat represented by caged branches, suggesting a cost to host dependency on environmentally acquired symbionts.