Metabolomic Investigations Reveal Properties of Natural Low-Temperature Adaptation Strategies in Five Evergreen Trees

In northern China’s arid and semi-arid regions, evergreen trees demonstrate significant cold tolerance to natural low-temperature stress during winter. However, the metabolic strategies and their associated properties underlying their overwintering adaptation remain incompletely elucidated. This study aims to reveal the metabolic properties of natural low-temperature adaptation strategies in five evergreen trees through metabolomic analysis and to identify key metabolites and their dynamic variation patterns. The GC-TOF-MS platform was used to investigate seasonal differential metabolites in five evergreen trees across January, April, July, and October and further explore core differentially expressed metabolites responsive to low-temperature stress. The results demonstrated that the seasonal changes in the chlorophyll content of five evergreens exhibited distinct patterns, that significant differences were observed between Juniperus sabina L. and Picea meyeri R., Ammopiptanthus mongolicus M., Buxus sinica var. parvifolia M.Cheng, and Pinus tabuliformis C., and that no significant differences were found among the other tree species. A total of 427 metabolites were detected in the metabolome; when assessing seasonal dynamics, it was found that the types of differentially expressed metabolites in the five evergreens underwent significant changes. In spring, the differentially expressed metabolites included some carbohydrates, alcohols, organic acids, and lipids. During summer and autumn, the largest number of differentially expressed metabolites accumulated, mainly including carbohydrates, organic acids, and amino acid compounds. In winter, while Picea meyeri primarily accumulated carbohydrates, the remaining four species mainly accumulated organic acids, along with a small number of alcohols, phenylpropanoids, and polyketides. Three shared carbohydrate metabolites, L-threose, galactinol, and gluconic lactone, were commonly downregulated across all species. Additionally, coniferous trees collectively accumulated 3,6-anhydro-D-galactose, showing downregulation. The KEGG enrichment analysis of winter-accumulated metabolites revealed significant associations with the pentose phosphate pathway, amino acid metabolism, phenylpropanoid biosynthesis, the tricarboxylic acid cycle, and ascorbate–aldarate metabolism pathways. Through comparative analysis with the summer growth season, we ultimately identified the core differentially expressed metabolites of the five evergreens, providing potential metabolic markers for the breeding of cold-tolerant species. In summary, these findings provide critical metabolomic insights into how plants adapt to low temperatures, significantly enhancing our understanding of the metabolic foundations of cold tolerance in evergreen species.