First Report of Brown Spot Needle Blight on Pinus thunbergii Caused by Lecanosticta acicola in Korea.

Pinus thunbergii Parl., known as black pine, is a pine native to coastal areas of Japan and Korea. Because of its resistance to pollution and salt, it is planted as windbreakers along the coast. In March 2010, needle blight symptoms were found on several trees of black pine in Naju, southern Korea. Further surveys in 2010 and 2011 showed that these symptoms are rather common but disease incidence is less than 1%. Small, circular grayish green spots first appeared on the needles. The spots developed into brown bands reaching 1 to 2 mm long, sometimes with yellow margins. Dark olivaceous to dark grayish stromata were erumpent and conspicuous on the brown lesions in the later stage of disease development. Conidiophores were simple or occasionally branched, 1- to 2-septate, pale brown to olivaceous brown, and smooth walled. Conidia (n = 30) were olivaceous brown to grayish brown, verrucose, thick-walled, mildly curved, allantoid to fusiform, one- to five-septate (mostly three-septate), and 20 to 45 × 3.5 to 5 μm. Morphological characteristics of the fungus were consistent with those of Lecanosticta acicola (Thüm.) Syd. (anamorph of Mycosphaerella dearnessii M.E. Barr), previously known as the causal agent of brown spot needle blight of pines (2,4). The teleomorph was not observed. On potato dextrose agar, single-spore cultures of three isolates were obtained from conidia sporulating on needles. An isolate was preserved at the Korean Agricultural Culture Collection (Accession No. KACC44982). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA) and the complete internal transcribed spacer (ITS) region of rDNA was amplified and sequenced with the primers ITS1/ITS4. The resulting ITS sequence of 543 bp was deposited in GenBank (Accession No. JQ245448). A GenBank BLAST search produced an exact match for the sequences of M. dearnessii (= L. acicola) on P. mugo Tura from Lithuania (HM367708) and P. radiata D. Don from France (GU214663), with 100% sequence similarity. To conduct a pathogenicity test, a conidial suspension (approx. 2 × 10⁵ conidia/ml) was prepared by harvesting conidia from 5-week-old cultures of KACC44982 and sprayed onto the needles of five 3-year-old healthy seedlings. Five noninoculated seedlings of the same age served as controls. Inoculated and noninoculated plants were kept in humid chambers for 48 h in a glasshouse. After 28 days, typical leaf spot symptoms started to develop on the needles of inoculated plants. The fungus, L. acicola, was reisolated from those lesions, confirming Koch's postulates. No symptoms were observed on control plants. The disease has been previously reported on several species of Pinus in the Americas (1) and recently in China (3), Japan (4), and Europe (2). To our knowledge, this is the first report of the Lecanosticta-Pinus association in Korea. Occurrence of the disease in Korea is a new threat to the health of black pine, especially in nursery plots. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.arsgrin.gov/fungaldatabases/ December 2011. (2) L. Jankovsky et al. Plant Protect. Sci. 45:16, 2009. (3) C. Li et al. J. Nanjing Inst. For. 1986:11, 1986. (4) Y. Suto and D. Ougi. Mycoscience 39:319, 1998.