Abstract: To establish a rapid determination method for seed viability of Smilax glabra Roxb., an L₁₆(4⁵) orthogonal experimental design was adopted to investigate the effects of soaking temperature, soaking duration, TTC(2,3,5-triphenyltetrazolium chloride) concentration, staining temperature, and staining duration on seed viability testing. Meanwhile, the effects of different storage methods on seed viability were compared. The results showed that all five factors exerted significant effects on the staining result, and the degree of influence followed the order: Soaking temperature > soaking duration > TTC concentration > staining duration > staining temperature. The orthogonal experiment showed that when seeds were soaked at 35 ℃ for 6 h and then stained with 0.10% TTC solution at 45 ℃ for 48 h, the seed embryos were clearly stained, and the viability reached the highest level of 88.33% ± 7.64%. Through range analysis and variance analysis, the optimal single-factor treatment combination was determined as: Seeds soaked at 25 ℃ for 6 h, followed by staining in 0.10% TTC solution at 45 ℃ for 24 h. Further storage trials showed that the seed viability under low-temperature storage was significantly higher than that under low-temperature sand storage. However, room-temperature shade-drying storage was operationally simple, low cost, and more suitable for practical production applications.

Key words: Smilax glabra Roxb., seed viability, TTC (2, 3, 5-triphenyltetrazolium chloride) method, orthogonal experiment