Liriodendron chinense is a tree species of the Magnoliaceae family, an ancient relict plant mainly used for landscaping and timber production due to its excellent material properties and ornamental value. The cytokinin oxidase/dehydrogenase (CKX) enzyme regulates cytokinin levels and plays an important role in plant growth, development, and resistance. However, too-high or too-low temperatures or soil drought can limit the growth of L. chinense, representing a key issue for research. Here, we identified the CKX gene family in the L. chinense genome and examined its transcriptional responses to cold, drought, and heat stresses. A total of five LcCKX genes, distributed on four chromosomes and divided into three phylogenetic groups, were identified across the whole L. chinense genome. Further analysis showed that multiple hormone- and stress-responsive cis-acting elements are located in the promoter regions of LcCKXs, indicating a potential role of these LcCKXs in plant growth, development, and response to environmental stresses. Based on existing transcriptome data, LcCKXs, especially LcCKX5, were found to transcriptionally respond to cold, heat, and drought stresses. Furthermore, quantitative reverse-transcription PCR (qRT-PCR) showed that LcCKX5 responds to drought stress in an ABA-dependent manner in stems and leaves and in an ABA-independent manner in roots. These results lay a foundation for functional research on LcCKX genes in the resistance breeding of the rare and endangered tree species of L. chinense.

Somatic embryogenesis (SE) is a crucial strategy for in vitro regeneration in woody plants, yet its efficiency is affected by multiple factors, and the underlying regulatory mechanisms remain insufficiently understood. In this study, callus tissues from two Liriodendron sino-americanum genotypes involving different hybrid combinations, ON-LoS and TN-LoS, were treated with varying concentrations (0, 0.005, 0.01, 0.05 mg/L) of exogenous zeatin (ZT) to evaluate its regulatory effect on SE. Treatment with 0.01 mg/L ZT significantly promoted SE in ON-LoS but suppressed it in TN-LoS, indicating that ZT elicited divergent regulatory effects on SE between the two genotypes. To explore the molecular basis of this divergence, transcriptome analysis was conducted at the early stage of SE. Differentially expressed genes (DEGs) were significantly enriched in hormone signaling, particularly in the cytokinin (CK) and brassinosteroid (BR) signaling pathways, as well as biosynthetic and redox-related pathways. In particular, given the established role of cell cycle-related gene CYCD3 (Lchi20922) in promoting cell division, CYCD3 was markedly upregulated by ZT in ON-LoS but downregulated in TN-LoS. These results indicate that ZT regulates SE efficiency through differential modulation of CYCD3 expression in distinct genotypes. This study enhances our understanding of the molecular basis of SE regulation in Liriodendron sino-americanum and offers a theoretical framework for improving regeneration efficiency in woody plants.