Coordinated Cell-Wall and Starch Maturation Is Associated with Winter-Harvest Quality in Sparganium stoloniferum Tubers.

Sparganium stoloniferum tubers (SL), known medicinally as Sparganii Rhizoma, are commonly considered superior at the winter-harvest stage, when they show the traditional quality traits of heavy weight and firm texture. However, the developmental basis of this quality phenotype remains insufficiently understood. This study aimed to determine how tissue organization, cell-wall architecture, starch deposition, and related transcriptional patterns are associated with winter-harvest quality in SL. By comparing SL at different developmental stages, we found that maturation was accompanied by reduced moisture content, increased tuber density, higher parenchyma cell density, progressive cell-wall thickening, and marked starch accumulation. Laser scanning confocal microscopy (LSCM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) observations further revealed thickened multilamellar cell walls and abundant clustered or compound-like starch bodies in mature SL. Starch isolated from mature SL displayed an A-type crystalline pattern, short-range order, and high gelatinization and pasting temperatures, indicating an ordered and thermally stable starch matrix. Cell-wall Fourier-transform infrared spectroscopy (FTIR) and solid-state nuclear magnetic resonance (NMR) analyses showed a predominantly polysaccharide-rich framework with subtle maturation-associated changes in aromatic- and methoxy-associated wall signals. Transcript-guided pathway analysis, supported by reverse transcription quantitative polymerase chain reaction (RT-qPCR)validation, suggested developmental shifts in carbohydrate metabolism, lipid-related metabolism, and gibberellin-associated transcriptional patterns. Together, these findings indicate that winter-harvest quality in SL is associated with coordinated tissue consolidation, cell-wall maturation, starch deposition, and transcriptional reprogramming, providing a structural and molecular framework for understanding the traditional firm-texture trait of S. stoloniferum.