Poplar is an important afforestation and urban greening species. Poplar leaf development occurs in stages, from young to mature and then from mature to senescent; these are accompanied by various phenotypic and physiological changes. However, the associated transcriptional regulatory network is relatively unexplored. We first used principal component analysis to classify poplar leaves at different leaf positions into two stages: developmental maturity (the stage of maximum photosynthetic capacity); and the stage when photosynthetic capacity started to decline and gradually changed to senescence. The two stages were then further subdivided into five intervals by gene expression clustering analysis: young leaves, the period of cell genesis and functional differentiation (L1); young leaves, the period of development and initial formation of photosynthetic capacity (L3–L7); the period of maximum photosynthetic capacity of functional leaves (L9–L13); the period of decreasing photosynthetic capacity of functional leaves (L15–L27); and the period of senescent leaves (L29). Using a weighted co-expression gene network analysis of regulatory genes, high-resolution spatiotemporal transcriptional regulatory networks were constructed to reveal the core regulators that regulate leaf development. Spatiotemporal transcriptome data of poplar leaves revealed dynamic changes in genes and miRNAs during leaf development and identified several core regulators of leaf development, such as GRF5 and MYB5. This in-depth analysis of transcriptional regulation during leaf development provides a theoretical basis for exploring the biological basis of the transcriptional regulation of leaf development and the molecular design of breeding for delaying leaf senescence.

Long non-coding RNA (lncRNA) plays an important regulatory role in plant growth and development, but its systematic identification and analysis in Eucalyptus has not yet been reported. Cold stress has a huge impact on the survival and yield of Eucalyptus seedlings, but the regulatory mechanism of lncRNA in Eucalyptus in response to cold stress is still unclear. In this study, the transcriptomes of young leaves of Eucalyptus urophylla S.T.Blake under low-temperature treatment and restoration were analyzed by RNA-seq. A total of 11,394 lncRNAs and 46,276 mRNAs were identified, of which 300 were differentially expressed lncRNAs (DE_lncRNAs) and 5606 were differentially expressed target genes of lncRNAs under cold stress, with the total number of target genes of DE_lncRNAs being 1681. A total of 677 differentially expressed transcription factors (TFs) were also identified, mainly including ERF, MYB and the NAC transcription factor family. Gene ontology (GO) analysis of the differentially expressed genes (DEGs) and target genes of DE_lncRNAs was mostly related to the response to cold stress and external stimuli. Furthermore, lncRNA–miRNA–mRNA regulatory networks were constructed, and 22 DE_lncRNAs were predicted to be targets or targeting mimics of 20 miRNAs. A qRT-PCR was used to verify the relative expression of genes in the regulatory EuGBF3-EUC_00002677-MSTRG.7690 network, and it matched the transcriptome data, indicating that it may play an important role in the response to cold stress in E. urophylla. This study provides a new insight into lncRNA and its regulatory network under abiotic stress, especially cold stress in E. urophylla.