Adaramodu, O. M.; Rosario, J. G.; Nordgren, E. C.; Paul, M.; Bhat, S. S.; Nguyen, H.; Ashraf, A.; Kim, J.; Helliker, B.; Gregory, B. D.

The transition from juvenile to adult phase (JA) is a key developmental process in plants, driven by conserved pathways that shape growth and stress responses. In monocots such as sorghum, this transition influences traits linked to environmental resilience, yet the regulation of micro-anatomical features such as trichomes and bulliform cells remains poorly understood. Here, we use single-nucleus RNA sequencing (snRNA-seq) to generate high-resolution gene expression maps of juvenile and adult sorghum leaves. Contrary to the traditional developmental model, we find that trichomes are present in both juvenile and adult leaves, with stage-dependent differences in gene expression. We also observe that bulliform cells, typically considered adult-specific, are present in juvenile leaves. At the transcriptomic level, cell populations cluster more strongly by developmental stage than by cell type, with juvenile cells grouping with other juvenile cells and adult cells with adult counterparts, indicating a dominant effect of developmental state on gene expression. In addition, we identify enrichment of dhurrin biosynthetic genes in trichome-associated cells as well as elevated expression of aquaporin genes in these cells, pointing to a potential role for trichomes in coordinating defense and water-related processes. Together, these findings refine current models of sorghum development and suggest that key epidermal cells and their associated functions may be established earlier than previously known. Collectively, this study provides a single nucleus resolved framework for understanding how developmental stage shapes gene expression in sorghum leaves.