First Asian Report of Leaf Spot of Ambrosia trifida Caused by Septoria epambrosiae.

Abstract

Ambrosia trifida L., known as giant ragweed, is native to North America and was introduced in Korea in the 1970s (4). It is now widely naturalized, and since 1999, has been designated as one of 11 most 'harmful nonindigenous plants' by the Korean Ministry of Environment because of its adverse effects on native plants. Various strategies to eradicate this noxious weed have been unsuccessful (4). In June 2008, leaf spot symptoms on this weed were found in Inje, Korea. Hundreds of giant ragweed growing along stream banks contained leaf spots with leaf yellowing and premature defoliation. Leaf lesions were 1 to 5 mm in diameter, angular to irregular, dark brown without a distinct margin, later becoming pallid with a brown margin. Between 2008 and 2011, the authors observed the same symptoms in Dongducheon, Yangku, Namyangju, and Pocheon, Korea. Voucher specimens have been housed in the herbarium of Korea University. Numerous black pycnidia were formed on the lesion. Pycnidia were amphigenous, globose, dark brown, ostiolate, and measured 80 to 130 μm in diameter. Conidia were filiform, straight to mildly curved, eguttulate, hyaline, 18 to 36 × 1.5 to 2.5 μm, one to three septate, subtruncate at the base, and tapering to a rounded apex. Single-conidial isolations onto potato dextrose agar formed dark grayish colonies. Pycnidia matured after 5 weeks when plates were incubated under fluorescent illumination for 12-h photoperiods at 25°C. On the basis of morphological and cultural characteristics, the fungus was identified as Septoria epambrosiae D.F. Farr (2). Three isolates were deposited in the Korean Agricultural Culture Collection (KACC). Preliminary morphological identification of the fungal isolates was confirmed by molecular data. The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1/ITS4 primers and sequenced. The resulting sequences of 449 bp obtained from the three isolates were identical to each other. They showed 100% similarity when compared with a sequence of S. epambrosiae (GenBank No. AF279582). The nucleotide sequence of a representative isolate (KACC43850) was deposited in GenBank (No. JN695498). Pathogenicity was confirmed by inoculating the leaves of three seedlings with a conidial suspension (~2 × 105 conidia/ml). Three noninoculated seedlings served as controls. Plants were maintained in a glasshouse at 100% relative humidity for 48 h. After 6 days, typical leaf spots, identical to the one observed in the field, started to develop on the leaves of the inoculated plants. No symptoms were observed on the control plants. The fungus was successfully reisolated from the symptomatic plants, fulfilling Koch's postulates. A leaf spot disease associated with S. epambrosiae has previously been recorded on A. artemisiifolia in Hungary (1-3) and A. trifida in North America (2,3). To our knowledge, this is the first report of S. epambrosiae on giant ragweed in Asia. Because of its potential as a biocontrol agent, further studies are needed.