Integrating physiological and transcriptomic analyses explored the regulatory mechanism of cold tolerance at seedling emergence stage in upland cotton (Gossypium hirsutum L.).

Cold stress is one of the major abiotic stressor that profoundly impacts plant growth. Cotton, a widely cultivated variety, is particularly susceptible to cold stress. Unraveling the responses to cold stress is critical for cotton demand. In this investigation, we conducted comparative physiological and transcriptomic analyses of the cold-tolerant variety XLZ16 and cold-sensitive variety XLZ84 at seedling emergence stage under cold stress. Following exposure to cold stress, XLZ16 exhibited a markedly higher growth phenotype and increased activity of antioxidant enzymes, while simultaneously showing reduced cellular oxidative damage and apoptosis. Furthermore, the levels of auxin (IAA), cytokinin (CTK), and salicylic acid (SA) significantly increased during cold stress, whereas the contents of catendorsterol (TY), brassinosterone (CS), and jasmonic acid (JA) significantly decreased. Integrated with stoichiometric analysis, these findings definitively demonstrated significant differences in antioxidant capacity and hormone content between the two varieties during their response to cold stress. A total of 6207 potential cold-responsive differentially expressed genes (DEGs) were identified through transcriptome sequencing analysis. Enrichment analyses of these DEGs revealed that pathways related to "hormones biosynthesis and signaling" as well as "circadian rhythm" were associated with cold response. Notably, the hub gene Gh_D12G2567 (GhJAZ3), encoding jasmonate ZIM-domain (JAZ) proteins, was found to influence the JA signal transduction pathway and regulate cotton growth under cold stress within the MEred module network. Furthermore, suppressing the expression level of GhJAZ3 by virus-induced gene silencing led to the reduction of cold resistance, implying GhJAZ3 as a positive regulator of cold tolerance. This study provides valuable insights into the response mechanisms of cotton under cold stress. It also serves as a reference and foundation for further enhancing cold tolerance of new cotton varieties.