Lone trees, rows of trees, groves, and hedges (in this study called ''isolated trees and hedges'') dot many Satoyama landscapes in Japan. They have traditionally played an important role in agriculture and horticulture, in the production of firewood, and as windbreaks, boundaries, and screens. Through this investigation of a suburban Satoyama landscape in western Japan, which includes old and new residential areas, public space, farmland, and abandoned land, we aimed to show how isolated trees and hedges are connected with local traditional cultural features, and how this link has impacted their distribution and species composition. By examining land categories, shape patterns, and species compositions of isolated trees and hedges in both traditional and newly developed regions of the study area, we showed how the presence or absence of related traditional cultural features has influenced biodiversity. We further examined the role of traditional management techniques and cultural features in maintaining high numbers of local native species in isolated trees and hedges. Based on our results, we argue that awareness and promotion of the biocultural link in isolated trees and hedges will play an important role in future efforts to preserve the unique local identity and biodiversity of this cultural landscape.