An increasing interest in carotenoids as nutritional sources of provitamin A and health-promoting compounds has prompted a significant effort in metabolic engineering of carotenoid content and composition in food crops. The strategy commonly used in plants is to increase the biosynthetic capacity by altering the carotenogenic enzyme activities. The recent isolation of the Or gene from a cauliflower orange mutant has brought a new endeavor for carotenoid enhancement by increasing the sink strength to sequester and store the synthesized carotenoids. Potato as one of the major staple crops usually accumulates low levels of carotenoids. In this chapter, we describe a detailed protocol for metabolic engineering of carotenoids in potato plants with the Or gene and the analysis of the Or transformants.