Yam (Dioscorea opposita) tuber development is a complex process regulated by various phytohormones, with gibberellin (GA) playing a crucial role. However, the underlying mechanisms and interaction of GA with other phytohormone pathways on yam tuber development remain incompletely understood. This study investigated the regulatory role of GA and its crosstalk with other phytohormones during yam tuber growth through phenotypic, cytological, physiological, and transcriptomic as well as targeted phytohormone metabolomics analyses. The results reveal that exogenous GA promoted tuber enlargement increases vascular bundle and the number and diameter of sieve tubes, and alters the expression of GA anabolism genes and GA signal transduction pathways. Integrated transcriptome and targeted metabolomics analyses revealed coordinated changes in GA and ethylene (ETH) biosynthesis and signaling pathways during tuber development, particularly DELLA-GAI2 acting as a negative regulator of GA signaling. Overexpression of DoDELLA-GAI2 in transgenic tobacco significantly reduced GA level, starch, cytokinin (CTK), and ETH content, as well as aerenchyma tissue growth and parenchyma cell size. Exogenous GA and ethephon treatments increased GA, starch, CTK, and ETH content, and downregulated DoDELLA-GAI2 gene expression. The yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays confirmed a direct interaction between DoDELLA-GAI2 and DoMTCPB, an upstream gene-encoding key enzyme in ETH biosynthesis. DoDELLA-GAI2 acts as a negative regulator of ETH synthesis by interacting with DoMTCPB. GA-induced degradation of DoDELLA-GAI2 relieves this inhibition, promoting ETH production and contributing to tuber growth. Taken together, our findings reveal a novel mechanism based on DoDELLA-GAI2 integrating the GA and ETH signaling processes to regulate tuber development in D. opposita, offering a potential target for improving yam crop productivity.

Yam (Dioscorea cayennensis subsp. rotundata,hereafter referred to as Dioscorea rotundata) is a staple tropical tuber crop with notable nutritional and economic value. Its development and yield depend on efficient sucrose allocation from source tissues. Sucrose transporters (SUTs), a conserved family of membrane proteins, mediate sucrose loading, translocation, and unloading. Although well-studied in model plants and cereals, SUTs in yam remain largely uncharacterized. This study aims to identify and characterize the SUT gene family in yam and explore their roles in sucrose transport and tuber development. We conducted a genome-wide analysis of yam SUT genes, including gene structure, subcellular localization, and phylogeny. Molecular docking was used to predict sucrose-binding residues, and qRT-PCR assessed gene expression across tissues and tuber developmental stages. Eight SUT genes were identified and classified based on sequence similarity and domain structure. Docking analysis revealed key residues involved in sucrose binding and possible conformational shifts influencing transport. Expression profiling showed that most SUT genes, especially in the tuber apex, were progressively upregulated during development, suggesting roles in sucrose unloading and cell expansion. Additionally, functional validation of DrSUT1 in Arabidopsis thaliana confirmed its involvement in sucrose transport, supporting its role in yam sucrose partitioning. Yam SUT genes, especially those highly expressed in sink tissues, are involved in sucrose partitioning and tuber development. These findings provide structural and functional insights into SUT-mediated sugar transport and lay a foundation for improving sucrose utilization and yield in yam and other tuber crops.

This study provides the first comprehensive genome-wide identification and characterization of the SWEET gene family in yam (Dioscorea rotundata), integrating structural bioinformatics, gene expression profiling, and functional validation to explore its roles in sucrose transport and tuber development. A total of 19 SWEET genes were identified and predicted to localize to the plasma membrane, and they showed high phylogenetic conservation with Arabidopsis thaliana, suggesting conserved functions in sugar distribution. Yeast substrate assays revealed that DrSWEET6 and DrSWEET12 are capable of transporting both hexose and sucrose across the plasma membrane, with their expression predominantly observed in the tuber, implicating their involvement in sucrose unloading. Expression profiling indicated high expression levels of the SWEET genes at the tuber apex, which progressively increased during tuber development, underscoring their critical roles in sucrose unloading, cell expansion, and biomass accumulation. These findings provide novel insights into the structural and functional mechanisms of the SWEET-mediated sucrose transport in yam, laying a solid foundation for future crop improvement strategies aiming to optimize sucrose distribution and enhance tuber yield and quality.