Using thermal death time models to analyze cold stress resistance across Drosophila species

O_LIChill-susceptible insects such as Drosophila are vulnerable to progressive disruption of ion and water homeostasis during cold stress, and low temperature exposure is a key factor affecting their physiology and distribution. Comparative studies of cold tolerance traditionally use simple or single-condition assays for interspecific comparisons, but the emergence of thermal death time (TDT) models offers a comprehensive framework to assess cold tolerance across different stress intensities and durations. C_LIO_LIHere we construct TDT curves for six Drosophila species, spanning boreal to tropical habitats, using Lt50 estimates across a range of stressful low temperatures (Lt50 ranging from [~] 20 min to 2 days). For all species, the TDT curves provided good fits to the log(Lt50) vs. temperature data (R2 = 0.87 - 0.99). C_LIO_LITDT curves from all species had steep slopes demonstrating that cold injury rate has a high thermal sensitivity such that small changes in temperature have profound effects on survival duration. The interspecific similarity of TDT slopes indicates that a conserved physiological dysfunction underlies cold injury across species. Further, additive accumulation of cold-induced injury in split-dose experiments suggests that acute and moderate cold damage represent the same underlying physiological dysfunction occurring at different rates. C_LIO_LIThe TDT curve intercepts (species-specific tolerance thresholds) differed markedly between boreal, temperate, and tropical species and correlated strongly with their habitat temperature. Data from the present study and meta-analysis of published data find that the inherent species cold tolerance decreases by [~] 0.45 {degrees}C for each {degrees}C colder the winter environment of the species is. When also considering the cold acclimation cues in cold climates we argue the experienced level of cold stress intensity is similar across environments inhabited by the Drosophila genus. This suggests that cold tolerance is important in shaping the fundamental niche of both boreal and tropical species. C_LIO_LIOverall, the TDT analysis of Drosophila at low temperature provides a powerful and predictive tool for quantifying insect cold tolerance. This approach enables detailed cross-species comparisons that allows for both ecological and physiological inference. Thus, TDT curves offer relevant approximations of insect cold resistance that could help predict insect responses to climatic change. C_LI