ABSTRACT Background: Betula platyphylla Sukaczev is one of important tree species due to its ecological and economic value. It is a cold-tolerant tree species which also faces heat stress during summer. In the current study, RNA-Seq profiling of two-month-old B. platyphylla seedlings were conducted utilizing the MGISEQ-2000 platform. Results: In total, 856,347,961 clean reads were obtained from 26 RNA-Seq libraries. Totally, 822,552,820 reads were successfully mapped to B. platyphylla reference genome. Further, a total of 360 and 264 DEGs were discovered under cold and heat exposure, respectively, while a total of 104 DEGs were identified under both cold and heat exposure. It was found that several pathways including response to cold, response to heat, response to temperature stimulus, response to stress, lipid metabolic, jamonic acid and ethylene, even developmental processes were significantly enriched in GO enrichment analysis of cold and heat stress in biological process term. Several transcription factors (TFs), including MYB66, CBF2, bHLH96and bZIP7 take a pivotal role in response to temperature stresses. Furthermore, heat shock proteins were identified under cold and heat stress, respectively, suggesting these genes participate in reducing cold and heat stress detrimental effect by interacting with TFs or other genes related to abiotic stresses, chlorophyll and photosynthesis, osmoprotectants, and phytohormone as well. Conclusion: This study not only underlying B. platyphylla’s molecular mechanism in response to temperature stresses but also provides candidate genes involved in response to temperature stresses. Background value coldtolerant tolerant summer RNASeq RNA Seq twomonthold two month old B MGISEQ2000 MGISEQ 2000 MGISEQ-200 platform Results 856347961 856 347 961 856,347,96 2 libraries Totally 822552820 822 552 820 822,552,82 genome Further 36 exposure respectively 10 stimulus metabolic ethylene term TFs, , (TFs) MYB66 MYB CBF2 CBF bHLHand bHLH bZIP Furthermore photosynthesis osmoprotectants well Conclusion platyphyllas s MGISEQ200 200 MGISEQ-20 85634796 85 34 96 856,347,9 82255282 82 55 822,552,8 3 1 (TFs MYB6 MGISEQ20 20 MGISEQ-2 8563479 8 9 856,347, 8225528 5 822,552, MGISEQ2 MGISEQ- 856347 856,347 822552 822,552 85634 856,34 82255 822,55 8563 856,3 8225 822,5 856, 822,