First Report of Pestalotiopsis microspora Causing Leaf Spot of Oil Palm (Elaeis guineensis) in China.

Oil palm (Elaeis guineensis) is a widespread, tropical evergreen species that grows in southern China. In November 2012 and July 2013, a new leaf spot was observed on oil palm in Danzhou, Hainan Province, China. A survey of 200 2-year-old oil palm plants revealed that the disease caused serious damage during the typhoon season of July to October in Hainan Province, with 15 to 20% incidence in plants. The spots were initially brown, small, and oval to irregular. Later, they gradually expanded and finally coalesced to form large gray-brown leaf spots surrounded by a dark brown border. Heavily infected leaves became dry and died. Sometimes black acervuli developed on the leaf lesions. Diseased tissues (2 × 2 mm) from lesion margins were surface-disinfested for 10 min with 0.3% NaClO, plated on potato dextrose agar (PDA), and then incubated at 25°C in the dark. Seven Pestalotiopsis isolates (identified by conidial morphological characteristics) were isolated from leaf lesions. These isolates were subcultured by single spore isolation, and a representative isolate was characterized further. The fungus was incubated on PDA at 25°C. After 5 days, the fungus produced circular white colonies. After 10 days, many black conidiomata formed over the mycelia mats. Conidia were fusiform, five-celled with constrictions at the septa, and measured 18.6 to 24.4 × 5.2 to 7.5 μm. The three median cells were light brown to dark brown, and two end cells were colorless. Apical cells had 2 to 4 appendages ranging from 10.4 to 22.6 μm long. Basal cells had 1 appendage ranging from 2.2 to 4.1 μm long. The internal transcribed spacer (ITS) region of the ribosomal DNA was amplified using primers IST4/ITS5, and the 549-bp product of the ITS (GenBank Accession No. KJ019328) showed 100% sequence identity to Pestalotiopsis microspora isolates XSD-42 (EU273522.1) and CBS364.54 (AF377292.1). The pathogenicity of all isolates was tested by inoculation of detached, healthy leaves according to Keith et al. (2). The middle parts of compound leaves with leaflets were cut from 2-year-old oil palm plant. Leaflets were wounded inoculated or unwounded inoculated with mycelial plugs (4 mm in diameter, 30 leaflets per isolate). PDA plugs without mycelia served as controls. All leaves were placed in a growth chamber at 25°C and 90% relative humidity. After 5 days, brown leaf spots appeared on all wounded leaflets, with symptoms similar to those described above. Control leaves and the inoculated leaflets without wound remained symptom free. P. microspora was re-isolated from the infected leaves and confirmed to be the same as the inoculated pathogen through examination of morphology and by conducting an ITS sequence comparison. P. neglecta and P. palmarum were previously reported as the causal agent of Pestalotiopsis leaf spot on oil palm (1). P. microspora was isolated from oil palm in Indonesia (3). To our knowledge, this is the first report of P. microspora on oil palm in China. References: (1) F. O. Aderungboya. Int. J. Pest Manage. 23:305,1977. (2) L. M. Keith et al. Plant Dis. 90:16, 2006. (3) Suwandi et al. Plant Dis. 96:537, 2012.