Internal ice formation leads to wholesale changes in ionic, osmotic and pH homeostasis, energy metabolism, and mechanical damage, across a small range of temperatures, and is thus an abiotic stressor that acts at a distinct, physiologically-relevant, threshold. Insects that experience repeated freeze-thaw cycles over winter will cross this stressor threshold many times over their lifespan. Here we examine the effect of repeatedly crossing the freezing threshold on short-term physiological parameters (metabolic reserves and cryoprotectant concentration) as well as long-term fitness-related performance (survival and egg production) in the freeze-tolerant goldenrod gall fly Eurosta solidaginis. We exposed overwintering prepupae to a series of low temperatures (-10, -15, or -20 °C) with increasing numbers of freezing events (3, 6, or 10) with differing recovery periods between events (1, 5, or 10 days). Repeated freezing increased sorbitol concentration by about 50% relative to a single freezing episode, and prompted prepupae to modify long chain triacylglycerols to acetylated triacylglycerols. Long-term, repeated freezing did not significantly reduce survival, but did reduce egg production by 9.8% relative to a single freezing event. Exposure temperature did not affect any of these measures, suggesting that threshold crossing events may be more important to fitness than the intensity of stress in E. solidaginis overwintering.
Katie E. Marshall and Brent J. Sinclair