Rural areas occupy a large part of Serbia’s population; they are home to a significant number of people, and they have a key role in the food production chain. As their comparative advantage, natural resources, biodiversity and unpolluted environment can be defined. Decades of negligence and marginalisation contributed to the deterioration of rural areas in a social, economic and spatial sense. Based on the analyses of the specific characteristics of rural areas within the Region of Southern and Eastern Serbia, this chapter examines opportunities and challenges for their revitalisation in the context of environmental sustainability. Starting from the assumption that the revitalisation of rural areas is a precondition for their development and that rural development can contribute to even regional and overall sustainable development, the impact of the revitalisation of rural areas on ecological transition will also be discussed within this chapter. This chapter aims to investigate how the process of revitalisation of rural areas within the Region of Southern and Eastern Serbia can foster ecological transition and improve environmental sustainability. By revitalising rural areas, it is possible to improve their sustainable development, improve quality of life, and preserve biodiversity, contributing to achieving a better place for life for the rural population.