NF-YB, a subfamily of Nuclear Factor Y (NF-Y) transcription factor, play crucial role in many biological processes of plant growth and development and abiotic stress responses, and they can therefore be good candidate factors for breeding stress-resistant plants. However, the NF-YB proteins have not yet been explored inLarix kaempferi, a tree species with high economic and ecological values in northeast China and other regions, limiting the breeding of anti-stressL. kaempferi.In order to explore the roles of NF-YB transcription factors inL. kaempferi, we identified 20LkNF-YBfamily genes fromL. kaempferifull-length transcriptome data and carried out preliminary characterization of them through series of analyses on their phylogenetic relationships, conserved motif structure, subcellular localization prediction, GO annotation, promotercis-acting elements as well as expression profiles under treatment of phytohormones (ABA, SA, MeJA) and abiotic stresses (salt and drought). TheLkNF-YBgenes were classified into three clades through phylogenetic analysis and belong to non-LEC1 type NF-YB transcription factors. They have 10 conserved motifs; all genes contain a common motif, and their promoters have various phytohormones and abiotic stress relatedcis-acting elements. Quantitative real time reverse transcription PCR (RT-qPCR) analysis showed that the sensitivity of theLkNF-YBgenes to drought and salt stresses was higher in leaves than roots. The sensitivity ofLKNF-YBgenes to ABA, MeJA, SA stresses was much lower than that to abiotic stress. Among theLkNF-YBs,LkNF-YB3showed the strongest responses to drought and ABA treatments. Further protein interaction prediction analysis forLkNF-YB3revealed thatLkNF-YB3interacts with various factors associated with stress responses and epigenetic regulation as well as NF-YA/NF-YC factors. Taken together, these results unveiled novelL. kaempferiNF-YB family genes and their characteristics, providing the basic knowledge for further in-depth studies on their roles in abiotic stress responses ofL. kaempferi.