Lindera benzoin L. or spicebush is an aromatic small native shrub that grows in the moist, understory areas of Appalachia and has potential as a new niche crop for small farmers. This plant was traditionally used as a tea by native cultures and early settlers. The berries can be used for jam and spicing of foods, and may have many health benefits including antioxidant compounds. Native spicebush patches also serve an important role in forest ecosystems around streams and rivers in terms of fruit production for animals, soil erosion control, and enhancing insect biodiversity. Populations of spicebush occur in a variety of light environments from forest interiors to canopy gaps and edge habitats. In many parts of the eastern United States, these edge environments are persistent and may have existed for hundreds of years. Spicebush may serve to hold ecological niches by outcompeting invasive plants compared to those in unchallenged areas. Genetic diversity of spicebush populations in Kentucky has not been examined in various forest regions and locations. The objective of this study is to determine the genetic diversity in six spicebush populations in Kentucky using simple sequence repeat (SSR) DNA marker systems. Leaf samples were collected from spicebush plants in the forests at the Kentucky State University Environmental Education Center (EEC), Cove Spring Park in Frankfort, KY, and at a location near the Kentucky River. Each population contained twenty individuals and two populations were sampled at each site. DNA was extracted using the DNAmite Plant Kit. Primers sb-A7, sb-A115, sb-B105, sb-B122, sb-C4, and sb-C10 labeled with FAM or HEX were used to amplify SSR products, and products were separated with a 3130 Applied Biosystems capillary electrophoresis system. The software program Power Marker was used to examine genetic relationships among the spicebush genotypes. The SSR markers generated showed genetic variation among the spicebush genotypes. A number of selections with unique genotypes will be sampled and propagated for study in the KSU germplasm collection for potential cultivar development.