Indirect Effects of Temperature on Stink Bug Fitness,via Maintenance of Gut-Associated Symbionts

Impacts of climate change on organisms are already apparent, with effects ranging from the individual to ecosystem scales. For organisms engaged in mutualisms, climate may affect population performance directly or indirectly through mediated effects on their mutualists. We tested this hypothesis for two stink bugs, Acrosternum hilare and Murgantia histrionica, and their gut-associated symbionts. We reared these species at two constant temperatures, 25 and 30°C, and monitored population demographic parameters and the presence of gut-associated symbionts with diagnostic PCR primer sets. Both stink bugs lost their respective gut symbionts within two generations at 30°C. In addition, the insect survivorship and reproductive rates of both A. hilare and M. histrionica at 30°C were lower than at 25°C. Other demographic parameters also indicated a decrease in overall insect fitness at the high temperature. Collectively our data showed that the decrease in host fitness was coupled with, and potentially mediated by, symbiont loss at 30°C. This work illustrates the need to better understand the biology of animal-symbiont associations and the consequences of local climate for the dynamics of these interactions.The effect of climate on organisms, communities, and the environment at large has become a pressing issue for biologists and environmental scientists. Recent studies indicate that previous forecasts were conservative in their predictions for the magnitude of global warming (29). Up-to-date models suggest that the global mean surface temperature will increase by 1.8 to 4°C by the year 2100 (11). The ecological impact of such warming is already apparent (35) in the effects seen on species fitness (24), range shifts (22), species interactions (10), and community structure (32).It is important to note that many macroorganisms live in symbiosis with microbes and that host fitness may be affected indirectly by higher temperatures due to the disruption of mutualistic relationships. Some corals, for example, have symbiotic relationships with photosynthesizing dinoflagellates (zoothanxellae) that provide them with nutrients (31). Higher water temperatures in reef ecosystems, among other factors, induce the expulsion of microbial symbionts by the host, resulting in coral bleaching (15). Therefore, it is plausible that observed effects of climate on species distribution or performance might stem from disruption of symbiotic interactions as much as from direct effects on host biology.Despite the current interest in insect-microbe symbioses, the vast majority of such systems have been poorly studied. A group of insects that has recently received some attention are the true bugs (Hemiptera, Pentatomomorpha). Studies in the early 1900s suggested that mutualistic bacteria colonized a portion of the gut of insects in different pentatomomorphan families (9). More specifically, monocultures of bacteria were present in high densities in the crypt- (or cecum-) bearing organ preceding the hindgut of hosts, with different bacterial taxa associated with different true bug families. Furthermore, studies have shown that these symbionts are mutualistic (1, 8, 13, 14, 26). Among these bug families, the stink bugs (Pentatomidae) have been shown to depend on their gut symbionts (1). Pentatomid symbionts are polyphyletic and closely related to plant-associated bacteria in the genera Erwinia and Pantoea (25). Although the mechanism of symbiont vertical transmission is poorly understood, females seem to smear the surface of eggs with bacteria while ovipositing (3). Aposymbiotic first instars hatch but remain on the surface of eggs and acquire the symbiont by probing on the egg surface, as evidenced by the fact that surface sterilization of egg masses generates aposymbiotic individuals (1, 26, 28).Climate change has already affected stink bug performance and geographic range (19, 20). In Japan, populations of two pentatomid species, Nezara viridula and N. antennata, have shifted northwards and to higher elevations, respectively, over the last 50 years (33). However, we found previously that for one of these species high temperature eliminated gut symbionts, without any clear decrease in host fitness (27). Thus, it remains unclear whether temperature change played a role, either directly or indirectly, in these geographic shifts. To better understand the extent to which temperature mediates stink bug ecology and prevalence of their gut bacteria, we conducted laboratory studies with two pentatomid species, Acrosternum hilare and Murgantia histrionica. We show that high temperature affects the symbiotic relationship, with concomitant reduction in insect fitness.