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Morphological characters (six traits) and isozymes (four systems, fi ve loci) were used to discriminate between Spanish chestnut cultivars (Castanea sativa Mill.) from the Iberian Peninsula. A total of 701 accessions (representing 168 local cultivars) were analyzed from collections made between 1989 and 2003 in the main chestnut growing areas: 31 were from Andalucia (12 cultivars), 293 from Asturias (65 cultivars), 25 from Castilla-Leon (nine cultivars), four from Extremadura (two cultivars) and 348 from Galicia (80 cultivars). Data were synthesized using multivariate analy- sis, principal component analysis, and cluster analysis. A total of 152 Spanish cultivars were verifi ed: 58 cultivars of major importance and 94 of minor importance, of which 18 had high intracultivar variation. Thirty-seven cultivars were clustered into 14 synonymous groups. Six of these were from Galicia, one from Castilla-Leon (El Bierzo), four from Asturias, one from Asturias and Castilla-Leon (El Bierzo), and two from Asturias, Castilla-Leon (El Bierzo), and Galicia. The chestnut cultivars from Galicia and Asturias were undifferentiated in genetic terms, indicating that they are not genetically isolated. Overall, chestnut cultivars from southern Spain showed the least variation. Many (58%) of Spanish cultivars produced more than 100 nuts/kg; removing this low market-value character will be a high priority. The data obtained will be of use in chestnut breeding programs in Spain and elsewhere. Growers have selected chestnut cultivars since Roman times. Traditionally, chestnut stands are derived by grafting selected cultivars onto planted seedlings. In the Middle Ages huge plan- tations were established for forestry with the dual purpose of timber and nut production (Pereira-Lorenzo and Ramos-Cabrer, 2004). Some of these plantations can still be found in Tuscany (Italy), Galicia (Spain), Tras-Os-Montes (Portugal), and Aveyron and Cevennes (France). Monasteries established most of these extensive chestnut orchards. There are 187 cultivars documented in Spain (Pereira-Lorenzo et al., 2001a, 2001b). There is considerable need to defi ne precisely chestnut cultivar variation for improved planting regimes and chestnut breeding. Breviglieri (1955) made the fi rst comprehensive study of chestnut variation using morphological characteristics in the "Scheda Castanografi ca." Later, the International Union for the Protection of New Varieties of Plants (UPOV) published a chestnut guide for distinguishing chestnut cultivars and suggested some

Use of Highly Discriminating Morphological Characters and Isozymes in the Study of Spanish Chestnut Cultivars