Transcriptomic Insights into the Degree of Polymerization-Dependent Bioactivity of Xylo-Oligosaccharides

Plant cell wall-derived oligosaccharides, such as xylo-oligosaccharides (XOS), serve as key signaling molecules regulating plant growth and immunity. The bioactivity of XOS is closely tied to their degree of polymerization (DP), yet the molecular mechanisms underlying DP-specific effects remain poorly understood. Here, we investigated the transcriptional and phenotypic responses of lettuce (<i>Lactuca sativa</i>) to foliar application of four high-purity XOS variants: xylobiose (XOSY, DP2), xylotriose (XOSB, DP3), xylotetraose (XOSD, DP4), and xylopentose (XOSW, DP5). Phenotypic analyses revealed that high-DP XOS (XOSD and XOSW) significantly enhanced aboveground biomass and root system development, with XOSD showing the most pronounced effects, including a 31.74% increase in leaf area and a 20.71% increase in aboveground biomass. Transcriptomic profiling identified extensive transcriptional reprogramming across treatments, with XOSD eliciting the highest number of differentially expressed genes (DEGs). Functional enrichment analyses indicated that XOSD and XOSW upregulated genes involved in plant hormone signaling, starch and sucrose metabolism, and cell wall biosynthesis, while downregulating photosynthesis-related genes. Notably, MapMan and KEGG pathway analyses revealed that XOSD significantly activated biotic stress-related pathways, including MAPK signaling, β-1,3-glucanase activity, and PR protein pathways. In contrast, XOSY treatment primarily upregulated genes linked to basal immunity, highlighting distinct mechanisms employed by low- and high-DP XOS. These findings demonstrate that XOS with varying DP differentially modulate growth- and immunity-related processes in lettuce. High-DP XOS, particularly XOSD, not only promote plant biomass accumulation but also enhance immune responses, highlighting their potential as biostimulants for sustainable agriculture. This study provides a molecular framework for understanding the DP-specific bioactivity of XOS and their dual role in optimizing plant growth and defense.