Populus alba ‘Berolinensis’ is a fast-growing, high-yielding species with strong biotic and abiotic stress resistance, and widely planted for timber, shelter belts and aesthetic purposes. In this study, molecular development is explored and the important genes regulating xylem formation in P. alba ‘Berolinensis’ under artificial bending treatments was identified. Anatomical investigation indicated that tension wood (TW) was characterized by eccentric growth of xylem and was enriched in cellulose; the degree of lignification was lower than for normal wood (NW) and opposite wood (OW). RNA-Seq-based transcriptome analysis was performed using developing xylem from three wood types (TW, OW and NW). A large number of differentially expressed genes (DEGs) were screened and 4889 counted. In GO and KEGG enrichment results, genes involved in plant hormone signal transduction, phenylpropanoid biosynthesis, and cell wall and secondary cell wall biogenesis play major roles in xylem development under artificial bending. Eight expansin (PalEXP) genes were identified from the RNA-seq data; four were differentially expressed during tension wood formation. Phylogenetic analysis indicated that PalEXLB1 belongs to the EXPB subfamily and that the other PalEXPs are members of the EXPA subfamily. A transcriptional regulatory network construction showed 10 transcription factors located in the first and second layers upstream of EXP, including WRKY, ERF and bHLH. RT‒qPCR analysis in leaves, stems and roots combined with transcriptome analysis suggests that PalEXPA2, PalEXPA4 and PalEXPA15 play significant regulatory roles in cell wall formation during tension wood development. The candidate genes involved in xylem cell wall development during tension wood formation marks an important step toward identifying the molecular regulatory mechanism of xylem development and wood property improvement in P. alba ‘Berolinensis’.