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Some cold-water fishes are particularly sensitive to the water temperature increasing caused by current global warming. However, the alterations in the physiology and behavior of infraspecific populations living in heterogeneous landscapes in response to water temperature increasing were significantly different. Consequently, understanding the impact of temperature increasing on different populations may be crucial for the conservation of cold-water fishes in the context of global warming. The burbot is the only freshwater specie in Gadiformes. To better understand the differences of different populations of burbot under similar thermal stress, Lota lota was selected as the research objects. Firstly, RNA-seq was applied to identify the transcriptomic responses of Heilongjiang population exposed to three temperature gradients (0 °C, 18 °C and 28 °C). Compared with 0 °C, 4216 and 12,657 genes were significantly differentially expressed at 18 °C and 28 °C, respectively. Meanwhile, 49 genes were significantly differentially expressed in three temperature pairs and these genes were presumed to involve in stress response process, immunologic process, reproductive process, development process, material metabolism process, signal transduction process, spermatogenesis process and cell apoptosis process. The response differences of two L. lota populations to similar thermal stress were compared and the results showed that they have different gene expression responses (the number of differentially expression genes and biological processes). The lower annual temperature of the Heilongjiang River might make it more sensitive to temperature increasing. Based on the comparative transcriptome analyses, 12 orthologous genes were considered as the potential regulators of L. lota preference for cold-water environment and these genes are potentially related to the immunologic process, reproductive process, development process, signal transduction process, and cell apoptosis process. Those results can provide basic information for the rational development of conservation strategies of different L. lota populations under the background of global warming. [Display omitted] • Some biological process-related genes were found to be involved in the thermal response of burbot. • The thermal response ability of different Lota lota populations was different. • The lower annual temperature of the Heilongjiang River might make Heilongjiang population more sensitive to temperature increasing. [ABSTRACT FROM AUTHOR]