“…Daily thermal range does not alter defensive mutualism, which is only affected by changes in mean temperature 3 The potentially interactive impacts of daily thermal range and mean temperature might affect the adaptive value of heatprotective mutualism Full factorial designs (controlling for mean temperature, daily thermal range and host infection status) combining fitness assays with molecular measurements (symbiont abundance, holobiont physiological profile) to assess the efficiency of symbiont-mediated responses to daytime or night-time thermal stress (aphids, whiteflies) What could be the impacts of extreme thermal events (heat waves, cold spells)? Facultative symbionts can improve the host ability to withstand heat shock 9,17,18 , while few data are available for cold shock tolerance 10,19 The holobiont resilience might be challenged by more complex patterns of extreme thermal events, while the stress tolerance of some biological models (holobionts from tropical origins) is understudied Experimental designs controlling for timing of stress application, frequency, number and temporal sequence of stress and recovery on various models involving heat-protective facultative symbionts or symbionts with free-living capacities (aphids, whiteflies, heteropterans) Field samplings to track natural variations of symbiotic communities following an extreme thermal event What could be the impacts of seasonal fluctuations? The composition of insect microbiome is highly dynamic across the seasons 20,21,22 Seasonal temperature fluctuations might directly or indirectly (through diapause induction and termination) impact microbiome and holobiont physiology, the latter being poorly understood Experimental approaches involving thermal gradient reflecting seasonality, linking microbiome dynamics to variations in host/ holobiont traits (behaviour, physiology, performance) in species with well-marked seasonal strategies (e.g.…”