First Report of Leaf Blight on Duying Caused by Phyllosticta anacardiacearum in China

Duying (Elaeocarpus glabripetalus Merr.; Elaeocarpaceae) is widely cultivated as an ornamental tree of commercial importance in southern China. From 2003 to 2008, severe outbreaks of Duying leaf blight occurred in the Hangzhou area, Zhejiang Province. Disease incidence was greater than 20% and mainly infected young leaves and shoots in the spring and autumn. Severely infected leaves and shoots died and eventually led to branch dieback. The overall growth decline of affected trees occurs over 4 to 6 years before tree death. Infection symptoms are characterized by grayish, round, semicircular- or irregular-shaped spots (5 mm to 5 cm long) with dark brown borders and the appearance of black, granular pycnidia within the dead leaf tissues. The primary infection zones are commonly observed on the leaf margins and apices, are brown, up to 2 mm in diameter, and often surrounded by a yellow zone. Pycnidia were globose and 122 to 127 μm (average 123.5 μm) in diameter. A fungus was consistently isolated from symptomatic tissues on potato dextrose agar (PDA). Ash-black pycnidia appeared on PDA after 10 days. Ascospores developed on modified PDA (1 liter of PDA + 20 g of Duying leaves) after 18 days. Conidiogenous cells were cylindrical to obpyriform. The hyaline conidia were obovoid and guttulate, 10 to 13 × 6 to 8 μm (average 11.5 × 7.5 μm), and usually surrounded by a mucilaginous sheath with a hyaline apical appendage that was 5 to 8 μm long. Pseudothecia were solitary and subglobose with long necks. Asci were 45 to 70 × 7.5 to 12 μm (average 62.5 × 10.8 μm). Ascospores were 12 to 13 × 4 to 5 μm with rounded apices and hyaline, mucilaginous, apical caps. The fungus was morphologically identified as Phyllosticta anacardiacearum van der Aa (teleomorph Guignardia mangiferae A. J. Roy). This identification was also confirmed by the China General Microbiological Culture Collection Center (CGMCC). Six representative fungal isolates were identified by sequencing the internal transcribed spacer (ITS) region of the rDNA and comparing the sequences with those in GenBank using BLAST searches. The ITS sequences of six cultures (GenBank Accession Nos. EU821356-EU821361) showed 100% identity with the ITS sequences of an isolate of a Phyllosticta sp. (GenBank Accession No. AF532314) (2) and G. mangiferae (GenBank Accession No. AY277717) (1). To fulfill Koch's postulates, a conidial suspension (10⁶ conidia per ml) collected from PDA cultures (isolate phy01) was used to spray inoculate leaves of potted 3-year-old Duying trees. Inoculated trees were kept for 48 h under a polyethylene sheet cover and grown at 10 to 15°C in a greenhouse. A total of 30 leaves of five healthy trees were inoculated with the pathogen. In addition, five 3-year-old trees were sprayed with sterile water to serve as uninoculated controls. After 10 to 14 days, inoculated leaves showed infection symptoms resembling those observed on Duying trees naturally infected with P. anacardiacearum. The pathogen was reisolated from the margins of necrotic tissues, but not from controls. To our knowledge, this is the first report of leaf blight on E. glabripetalus caused by P. anacardiacearum in China. Reference: (1) F. R. Katia et al. Mycol. Res. 108:45, 2004. (2) A. K. Pandey et al. Mycol. Res. 107:439, 2003.