In the summer of 2009 in Zhejiang, Jiangsu, Guangxi, Shanxi, Beijing, Tianjin and Hebei Provinces, a decline of grapevines (Vitis vinifera) was identified with symptoms of dieback on one or more shoots, accompanied by elongated oval-shaped cankers in the wood, shrivelling and rot of fruit clusters. A total of 121 diseased samples were collected from affected areas and from these samples more than 1000 infected tissue pieces were used for pathogen isolation. Tissue from the lesion margins was surface-sterilized by placing in 75% ethanol for one min, then rinsing with sterilized water (three times) before placing on potato dextrose agar plates incubated at 28°C for 5 days (Niekerk et al., 2004). Hyphae from the colony margins were transferred to fresh agar plates to obtain pure isolates.
Botryosphaeria dothidea was isolated from 44 of the diseased samples. Colonies of B. dothidea were white, becoming grey or dark brown with age. Pycnidia started to develop after 10 days. Conidia measured 17–32 × 4–9 μm. Some isolates were used for ITS sequence analysis to confirm the morphological identification. The primers ITS1 and ITS4 were used to amplify part of the nuclear rDNA operon by PCR (White et al., 1990). Ten ITS sequences were obtained (GenBank Accession Nos. GU226844 to GU226850, GU226852, GU226854 and GU226855). All sequences showed 99–100% homology with B. dothidea (Urbez-Torres et al., 2006). This is the first report of grapevine canker associated with B. dothidea in China.