A Genome-Wide Characterization of the 14-3-3 Protein Family in Ginger Reveals That Zo14-3-3-03 Enhances Salt Tolerance via the ZoSOS2-Mediated Signaling Pathway

Soil salinity limits ginger productivity, but the underlying molecular mechanisms remain largely unclear. The 14-3-3 proteins are conserved regulators in stress signaling. Here, we genome-wide characterized the 14-3-3 family in Zingiber officinale and examined the possible involvement of Zo14-3-3-03 in salt response. A total of 21 Zo14-3-3 genes were identified and classified into four groups with uneven chromosomal distribution. Among them, Zo14-3-3-03 was strongly salt-responsive: transcript levels increased 9.91- to 33.82-fold during 1–7 days of treatment and reached 62.47-fold in leaves at day 14. NaCl treatment elevated GUS expression driven by the Zo14-3-3-03 promoter. Virus-induced gene silencing (VIGS) of Zo14-3-3-03 resulted in silenced plants exhibiting higher malondialdehyde (up to 73.6%), lower antioxidant enzyme activities (SOD, POD, CAT, and APX: 18.9–31.9% reduction), reduced osmolytes (proline, soluble protein, sugars, and ascorbic acid: 23.2–36.2% reduction), excessive reactive oxygen species, and decreased relative water content. Several antioxidant-related genes were significantly downregulated. Protein interaction assays suggested a possible interaction with ZoSOS2, and the expression of SOS2 pathway genes was altered in silenced plants, indicating a potential link to calcium signaling and ion homeostasis. Taken together, these results suggest that Zo14-3-3-03 participates in ginger salt stress response possibly through redox balance, osmotic adjustment, and calcium-mediated pathways which would provide a basis for understanding 14-3-3-mediated stress responses and nominates Zo14-3-3-03 as a candidate requiring deeper validation for salt tolerance improvement in ginger. Nevertheless, due to limited functional validation, its role as a positive regulator and breeding target remains preliminary. Further genetic and mechanistic studies are needed to confirm causality and assess field-level applicability.