Final version of the chapter at: http://dx.doi.org/10.1007/978-1-4020-6352-7_28.Since the 1970s, natural resources the world over have been progressively re-evaluated in accordance with the principles of sustainable agriculture. The European chestnut, Castanea sativa Mill., a hardwood species belonging to the family Fagaceae, has not been unaffected by this process. It is an important resource in many parts of the world because of its economic and environmental role in many agroforestry systems, and in Europe it has been gaining in value as a source of timber and nut production and due to the contribution of chestnut groves to the landscape (Bounous, 2005).Like the American chestnut C. dentata, C. sativa has been plagued for more than a century by ink disease and chestnut blight, caused by the fungi Phytophthora cinnamomi and Cryphonectria parasitica, respectively. A great deal of research on chestnut focuses on the development of vegetative propagation systems capable of satisfying the demand for elite genotypes that provide both high-quality timber and/or nuts and resistance to these diseases. Since chestnut is a difficult-to-root species, grafting is the most frequent conventional propagation technique, although methods for layering and cutting have recently been improved and are widely used in nurseries to propagate ink-disease-resistant Euro-Japanese hybrids. However, as an alternative to conventional vegetative propagation methods, efforts are being made to establish reliable in vitro regeneration systems that allow clonal propagation. The two major systems are based on embryogenesis or on micropropagation of axillary shoots.Thanks are given to Spanish Ministry of Education and Science (project AGL2005-00709) and Xunta de Galicia, Spain (project PGIDIT03RFO40001PR) for financial support.Peer reviewed