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.

Heat shock factors (Hsfs) play a crucial role in plant defense processes. However, the distribution and functional characteristics of Hsf genes in the relict plant Liriodendron chinense are still unclear. In this study, a total of 19 LcHsfs were identified and divided into three separate subgroups, comprising 10 LcHsfA, 7 LcHsfB, and 2 LcHsfC genes, respectively, based on their phylogenetic tree and the presence/absence of conserved protein domains. Whole-genome duplication and segmental duplication led to an expansion of the LhHsf gene family. The promoters of LcHsf genes are enriched for different types of cis-acting elements, including hormone responsive and abiotic-stress-responsive elements. The expression of LcHsfA3, LcHsfA4b, LcHsfA5, LcHsfB1b, and LcHsfB2b increased significantly as a result of both cold and drought treatments. LcHsfA2a, LcHsfA2b, and LcHsfA7 act as important genes whose expression levels correlate strongly with the expression of the LcHsp70, LcHsp110, and LcAPX genes under heat stress. In addition, we found that transiently transformed 35S:LcHsfA2a seedlings showed significantly lower levels of hydrogen peroxide (H₂O₂) after heat stress and showed a stronger thermotolerance. This study sheds light on the possible functions of LcHsf genes under abiotic stress and identifies potentially useful genes to target for molecular breeding, in order to develop more stress-resistant varieties.

Pinus elliottii, a key economic conifer in southern China, requires efficient propagation methods to meet demand for elite germplasm in resin and timber production. While somatic embryogenesis-based plant regeneration has been successfully achieved in Pinus elliottii, large-scale production remains challenging. Our results demonstrate that the genotype of Pinus elliottii significantly influences the induction rate of embryogenic callus. During somatic embryo maturation, the liquid–solid induction method increased the number of mature embryos by 25.85 times. Maturation efficiency was further enhanced by a 3-week pretreatment followed by the application of 9 mg/L ABA, 0.5 mg/L PSK, and 6 mg/L COS. Additionally, the incorporation of activated carbon significantly promoted both the maturation and germination of somatic embryos. In the somatic embryo maturation stage, 1 g/L activated carbon induced 288.67 mature embryos per gram of embryogenic callus, resulting in a total of 1452 embryos. During germination, the application of 4 g/L activated carbon achieved a germination rate of 63%, and the survival rate of somatic embryo-derived seedlings reached 85%. This study not only identifies the optimal conditions for somatic embryogenesis in Pinus elliottii but also establishes an efficient protocol for somatic embryo maturation induction, providing a crucial scientific foundation for the rapid propagation and seedling production of Pinus elliottii.

The F-box gene family is one of the largest gene families in plants, and it plays a crucial role in regulating plant development, reproduction, cellular protein degradation, and response to biotic and abiotic stresses. Despite their significance, a comprehensive analysis of the F-box gene family in Liriodendron chinense and other magnoliaceae species has not been reported. In this study, we report for the first time the identification of 144 full-length F-box genes in L. chinense. Based on specific domains and phylogenetic analyses, these genes were divided into 10 distinct subfamilies. We further analyzed their gene structure, conserved domain and chromosome distribution, genome-wide replication events, and collinearity. Additionally, based on GO analysis, we found that F-box genes exhibit functional specificity, with a significant proportion of them being involved in protein binding (GO:0005515), suggesting that F-box genes may play an important role in gene regulation in L. chinense. Transcriptome data and q-PCR results also showed that F-box genes are involved in the development of multiple tissues in L. chinense, regulate the somatic embryogenesis of Liriodendron hybrids, and play a pivotal role in abiotic stress. Altogether, these findings provide a foundation for understanding the biological function of F-box genes in L. chinense and other plant species.

Somatic embryogenesis represents a critical biotechnology platform for large-scale propagation and germplasm conservation of Pinus elliottii. However, low

Somatic embryogenesis (SE) is a core technology for the clonal propagation of forest tree species. However, the low frequency of normal embryos and high rate of abnormal embryos during SE maturation induction in slash pine (Pinus elliottii) severely constrain its commercial application. In this study, comparative transcriptome analysis between normal embryos (NE) and abnormal embryos (AE) revealed that glutathione metabolism and riboflavin metabolism pathways may be involved in the regulation of somatic embryo maturation. Based on these findings, different concentration gradients of reduced glutathione (GSH) and riboflavin were supplemented into the somatic embryo maturation medium to verify their effects. Single-factor experiments showed that the 0.5 mM GSH treatment yielded 158.33 normal embryos per gram of embryogenic callus with a normal embryo ratio of 42.20%, while the 7.5 µM riboflavin treatment yielded 100 normal embryos per gram with a normal embryo ratio of 18.27%. Further 2 × 2 orthogonal combination experiments demonstrated that the synergistic treatment of 0.5 mM GSH and 7.5 µM riboflavin achieved the optimal effect, producing 333.33 normal embryos per gram of embryogenic callus with a normal embryo ratio of 72.88%. Both maturation rate and induction efficiency were significantly superior to single treatments and other combinations. These results demonstrate that GSH and riboflavin can synergistically improve the normal embryo ratio and accelerate somatic embryo maturation in slash pine. The underlying mechanisms involving metabolic and hormonal pathways warrant further physiological and biochemical investigation. This study provides a technical reference for optimizing the somatic embryogenesis system of slash pine.