Telomeres and telomerase have important biological functions and can protect chromosome ends. In this study, sex- and season-dependent changes in telomere length and telomerase activity in ash and willow were analyzed. A statistical analysis showed that the telomere lengths of male and female trees differed significantly (P < 0.05). In ash, the telomere lengths of female trees were shorter than those of male trees. In willow, the telomere lengths of female trees were longer than those of male trees. During the annual developmental cycle, the telomere lengths of male and female ash and willow increased from April to May (P < 0.05), remained stable from May to August (P > 0.05), and decreased significantly in September and October (P < 0.05). Additionally, telomerase activities could be detected in both male and female ash and willow trees from April to October. Our results show that the telomere lengths changed according to season and sex in ash and willow. Telomere length did not have a direct positive correlation with telomerase activity.