Time-course transcriptome analysis reveals gene co-expression networks and transposable element responses to cold stress in cotton.

Background: Cold stress significantly challenges cotton growth and productivity, yet the genetic and molecular mechanisms underlying cold tolerance remain poorly understood.

Results: We employed RNA-seq and iterative weighted gene co-expression network analysis (WGCNA) to investigate gene and transposable element (TE) expression changes at six cold stress time points (0 h, 2 h, 4 h, 6 h, 12 h, 24 h). Thousands of differentially expressed genes (DEGs) were identified, exhibiting time-specific patterns that highlight a phase-dependent transcriptional response. While the A and D subgenomes contributed comparably to DEG numbers, numerous homeologous gene pairs showed differential expression, indicating regulatory divergence. Iterative WGCNA uncovered 125 gene co-expression modules, with some enriched in specific chromosomes or chromosomal regions, suggesting localized regulatory hotspots for cold stress response. Notably, transcription factors, including MYB73, ERF017, MYB30, and OBP1, emerged as central regulators within these modules. Analysis of 11 plant hormone-related genes revealed dynamic expression, with ethylene (ETH) and cytokinins (CK) playing significant roles in stress-responsive pathways. Furthermore, we documented over 15,000 expressed TEs, with differentially expressed TEs forming five distinct clusters. TE families, such as LTR/Copia, demonstrated significant enrichment in these expression clusters, suggesting their potential role as modulators of gene expression under cold stress.

Conclusions: These findings provide valuable insights into the complex regulatory networks underlying cold stress response in cotton, highlighting key molecular components involved in cold stress regulation. This study provides potential genetic targets for breeding strategies aimed at enhancing cold tolerance in cotton.