Although much research on nature-based tourism (NBT) has been conducted in natural areas, such as national parks and other protected areas, studies on NBT in rural areas have been limited. Moreover, few NBT studies, if any, have examined the impact of seasons and/or locations on visitors’ perceptions of NBT. This comes as little surprise, given that naturalness, the fundamental core of NBT, is likely to vary with seasons and locations. To this end, this study examines NBT in a rural area in the Appalachian Region, USA, with a focus on market segmentation, based on data collected from a four-season on-site survey of 2692 visitors. Different from many other tourism market studies where multiple-item measures are used, this study uses a single-item measure on natural observation/study to identify NBT segments. Analysis of variance, Chi-square analysis, and discriminant analysis are used to confirm NBT segments and determine the effectiveness of the use of the single-item approach for segmentation. The results show that the single-item approach is effective to differentiate visitors by their socio-demographics and trip characteristics, with findings being consistent with NBT studies in natural areas. It is worth noting that people’s responses to questions on natural observation/study vary with seasons and locations, suggesting that different perceptions of natural affinity may arise from the differences in seasons and locations with different levels of natural elements. Theoretical, methodological, and managerial implications are discussed.