Botryosphaeria spp. recently have been identified as important grapevine pathogens worldwide. To date, Botryosphaeria rhodina has been the only species associated with cankers on Vitis vinifera in California. A field survey of 166 vineyards in 21 counties was conducted in order to determine the occurrence of other Botryosphaeria spp. in California. In all, 1,735 samples of cankered trunks, cordons, and spurs were collected. Botryosphaeria spp. were the most common fungi isolated from grapevine cankers in California. Morphological identification along with phylogenetic analysis of the internal transcribed spacer region (ITS1-5.8S-ITS2) of the nuclear ribosomal DNA (rDNA) and a partial sequence of the beta-tubulin gene showed that at least seven Botryosphaeria spp. occur on grapevines in California: B. australis, B. dothidea, B. lutea, B. obtusa, B. parva, B. rhodina, and B. stevensii. Botryosphaeria spp. were found in grapevine cankers in all grape-growing regions surveyed in California, whereas incidence and distribution varied with location. Grapevine cankers in California have been associated mainly with Eutypa dieback. However, the frequent recovery of Botryosphaeria spp. from cankers in this study indicates that the role of these fungi in grapevine health needs to be more carefully considered.