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Although Quercus brantii Lindl. has shown suitable genetic variation in Iran, which is required for a sustainable forest ecosystem, but during the last decade, it came across some difficulties which are considered as oak decline. This research investigated seedling characteristics of four plant populations of the Quercus brantii species to assess genetic potentials of the populations to base a cornerstone for complimentary studies. Seeds were collected on ten single trees for each of plant population located in Kurdistan, Lorestan, Ilam, and Fars provinces in Iran. Seeds were sown in pots at greenhouse conditions to produce at least 30 single progeny seedlings from each population with three replications were studied. Vegetative characteristics such as plant height, leaf length, leaf width, and plant vigor were studied on the single plants. Data were analyzed based on a double nested statistical model, and then the data were analyzed on each population separately. Results indicated significant differences between the studied populations based on seedling height and plant vigor. Plant within population, showing possible differences between the single trees within the populations, was also significant for seedling height, leaf length and leaf width. In population based analysis, the populations showed different behaviors, so as there was not significant effects between single plants in Kurdistan population for plant height and leaf traits; whereas, in Lorestan and Ilam populations significant effects for the mentioned traits were observed. Although the populations showed different characteristics at seedling stage, but it is sought that there are still enough genetic variability and the high heritability that might be used in future breeding projects.