First Report of Stem Blight of Blueberry Caused by Botryosphaeria dothidea in China.

Abstract

Blueberry (Vaccinium spp.) production in southwest and northeast China has grown to over 100,000 ha in the last 20 years thanks to the fruit's high nutritional and economic value. As blueberry acreage increases, the diversity of diseases and challenges for control are gaining more attention. In August 2010, stem and branch blight occurred on Highbush Blueberries (Vaccinium corymbosum L.) at commercial farms in Lijiang and Zongdian, Yunnan Province (southwestern China), with crop damage ranging from 10 to 15%. Typical symptoms of the disease were blight and dieback on the stems with lesions extending along entire branches. Diseased samples (phloem and xylem sectors in the wood) were washed with running tap water, disinfected with 2% sodium hypochlorite, then 70% alcohol, rinsed in sterile distilled water, plated on potato dextrose agar (PDA), and incubated at 28°C. Fungal isolates developed copious, white aerial mycelium that became dark gray after 4 to 5 days and formed black pycnidia after 18 days. Conidia were hyaline, aseptate, thin walled, fusiform, and measured 21 to 27 × 4 to 6 μm. Identity was confirmed by analysis of the ribosomal DNA internal transcribed spacer region ITSI-5.8S -ITS2 with primers ITS1 and ITS4. BLAST searches showed 99% identity with Botryosphaeria dothidea isolates from GenBank (Accession Nos. AB693904 and JF800139). Representative sequences of B. dothidea from Highbush Blueberries from China were deposited into GenBank (Accession No. JX096631). On the basis of morphological and molecular results, the fungus isolated from diseased Highbush Blueberries stem was confirmed to be B. dothidea. Pathogenicity tests were conducted on 2-year-old blueberry seedlings (Highbush Blueberries). Mycelial plugs (2 to 3 mm in diameter) of B. dothidea from actively growing colonies (PDA) were applied to same-size bark wounds in the center of the stems. Inoculation wounds were wrapped with Parafilm. Control seedlings received sterile PDA plugs. Inoculated and control seedlings (five each) were kept in a greenhouse and watered as needed. After 12 days, all of the inoculated but none of the control blueberry seedlings showed dark vascular stem tissue. B. dothidea was reisolated from symptomatic tissues, thus fulfilling Koch's postulates. No symptoms were visible in the control seedlings. B. dothidea has been reported as a pathogen of sycamore (3), olives (1), and peach (2). However, no research has been conducted on stem blight of blueberry caused by B. dothidea in southwest or mainland China. To our knowledge, this is the first report of B. dothidea on blueberry in southwest China.