Abstract <p>Soil salinization severely restricts the sustainable development of the grape industry in arid/semi-arid northwest China. Establishing a standardized salt tolerance evaluation system and screening suitable germplasm are crucial. Fifteen grape germplasm (rootstocks, table/wine grapes, wild <i>Vitis amurensis</i>) were used to construct a salt stress screening system for tissue-cultured leafy single-bud stem segments. Tissue-cultured ‘Cabernet Sauvignon’ (main local cultivar) was treated with 0–100 mM NaCl; the half-inhibitory concentration (IC₅₀ = 31.83 mM) was calculated using probit regression, and 50 mM NaCl (about 1.57 × IC₅₀) was set as the screening concentration. Salt tolerance was evaluated by plant height, root length, rooting rate, survival rate, and salt damage index, with ranking/classification via membership function and principal component analysis. Na⁺/K⁺ contents in aboveground parts and roots of representative germplasm were determined by flame atomic absorption spectrophotometry. Significant differences (<i>P</i> &lt; 0.05) in salt tolerance were observed, with germplasm classified into high/medium/low tolerance levels. Wild <i>V. amurensis</i> ‘Shuangyou’ showed the strongest salt tolerance: its roots accumulated 62.3% more Na⁺ than salt-sensitive ‘Cabernet Sauvignon’, while aboveground parts maintained stable K⁺ levels with a 3.8-fold higher K⁺/Na⁺ ratio. This study clarified grape germplasm salt tolerance potential, identified ‘Shuangyou’ as a high-quality salt-tolerant resource, and established a standardized screening system. The system enables rapid pre-screening of breeding materials, while ‘Shuangyou’ can serve as a donor parent for improving salt tolerance in grape breeding programs. This work provides theoretical and technical support for grape breeding and cultivation in saline-alkali soils.</p>