The assessment of seed quality and physiological potential is essential in seed production and crop breeding. In the process of rapid detection of seed viability using tetrazolium (TZ) staining, it is necessary to spend a lot of labour and material resources to explore the pretreatment and staining methods of hard and solid seeds with physical barriers. This study explores the TZ staining methods of six hard seeds (Tilia miqueliana, Tilia henryana, Sassafras tzumu, Prunus subhirtella, Prunus sibirica, and Juglans mandshurica) and summarizes the TZ staining conditions required for hard seeds by combining the difference in fat content between seeds and the kinship between species, thus providing a rapid viability test method for the protection of germplasm resources of endangered plants and the optimization of seed bank construction. The TZ staining of six species of hard seeds requires a staining temperature above 35 °C and a TZ solution concentration higher than 1%. Endospermic seeds require shorter staining times than exalbuminous seeds. The higher the fat content of the seeds, the lower the required incubation temperature and TZ concentration for staining, and the longer the staining time. And the closer the relationship between the two species, the more similar their staining conditions become. The TZ staining method of similar species can be predicted according to the genetic distance between the phylogenetic trees, and the viability of new species can be detected quickly.

种子质量和生理潜力的评估在种子生产和作物育种中至关重要。在使用四唑 （TZ） 染色快速检测种子活力的过程中，需要花费大量的人力和物力来探索具有物理屏障的坚硬和固体种子的预处理和染色方法。本研究探讨了 6 种硬质种子（椴树、椴树、黄樟、毛李、西伯利亚李和胡桃楸）的 TZ 染色方法，并通过结合种子间脂肪含量的差异和物种之间的亲缘关系，总结了硬种子所需的 TZ 染色条件，从而为濒危植物种质资源保护和种子库建设的优化。六种硬种子的 TZ 染色需要 35 °C 以上的染色温度和高于 1% 的 TZ 溶液浓度。胚乳种子需要比蛋白种子更短的染色时间。种子的脂肪含量越高，染色所需的孵育温度和 TZ 浓度越低，染色时间越长。这两个物种之间的关系越密切，它们的染色条件就越相似。根据系统发育树之间的遗传距离，可以预测相似物种的 TZ 染色方法，并且可以快速检测新物种的生存能力。