First Report of Verticillium Wilt of Grapevine (Vitis vinifera) Caused by Verticillium dahliae in China.

Abstract

Verticillium dahliae Kleb. causes Verticillium wilt in large numbers of crops all over the world. Common symptoms caused by the pathogen include yellowing, wilting of leaves, and discoloration in vascular tissue of the stem. In June 2007, symptoms of Verticillium wilt were observed in a grapevine (Vitis vinifera) field in the Shihezi Region of Xinjiang. To isolate the pathogen, stem segments (0.5 cm long) were surface sterilized with 1% HgCl2 for 1 min and then dipped in 70% ethanol for 10 s. The sterilized tissues were rinsed with sterile water and incubated in the dark for 7 days at 25°C on potato dextrose agar (PDA) medium. The fungus growing from the diseased tissue showed dark colonies that produced verticillate conidiophores with two to three layers with colorless, ovoid, unicellular conidia and small, black microsclerotia, which are characteristics of V. dahliae (3). To confirm its identity, ribosomal DNA fragments (regions ITS1, 5.8S rDNA, and ITS4) were PCR amplified with primer pair ITS1/ITS4 (4) and sequenced (GenBank Accession No. FJ475122). Sequential analysis revealed that the rDNA region of the fungus isolated from grapevine was identical to that of a Greek strain of V. dahliae (GenBank Accession No. AF104926). Furthermore, the specific fragment (1,500 bp) of nondefoliating pathotype of V. dahliae (1) was PCR amplified from 24 grapevine isolates of V. dahliae collected in Xinjiang, indicating that the V. dahliae pathogen from Xinjiang is a nondefoliating pathotype. To verify the causal role of the isolated fungus, pathogenicity assays were conducted on 1-year-old seedlings of the Centennial seedless cultivar. Trimmed roots were submerged in a conidial suspension (1 × 106 conidia/ml) for 30 min and sterile tap water was used as a control. The seedlings were transplanted into a pot containing 2:1 sterile mixture of peat/perlite (vol/vol). Plants were grown in a greenhouse at 25°C. Six Verticillium isolates were found to cause the same symptoms as in fields 50 days after inoculation. V. dahliae was successfully reisolated from the stems of inoculated plants. Control seedlings inoculated in sterile tap water remained healthy. Because grapevine (Vitis vinifera) is an economically important crop for fruit and winemaking material in Xinjiang, Verticillium wilt poses a threat. The disease has been previously reported in the United States (2), but to our knowledge, this is the first report from China.