Park J H, Han K S, Hong S H, Shin H D
Division of Environmental Science and Ecological Engineering, Korea University, Seoul 136-701, Korea.
Horticultural & Herbal Crop Environment Division, National Institute of Horticultural & Herbal Science, Suwon 441-440, Korea.
Plant Dis. 2012 Dec;96(12):1827. doi: 10.1094/PDIS-08-12-0755-PDN.
Erigeron strigosus Muhl. ex Willd., known as daisy fleabane, is native to North America and was accidently introduced to Korea in the 1990s (2). It is increasingly invasive in natural and managed ecosystems throughout Korea. In June 2011, a leaf spot was first observed on daisy fleabanes growing wild in Hongcheon County of Korea. A voucher specimen was deposited in the Korea University Herbarium (KUS-F25759). Symptoms developed on lower leaves as small, distinct, reddish brown lesions, which enlarged progressively and turned into pale, dull brown spots surrounded by dark purplish-brown margins. Black pycnidia became visible in the lesions. Pycnidia were epigenous, occasionally hypogenous, scattered, dark brown to rusty brown, globose, embedded in host tissue or partly erumpent, 60 to 160 μm in diameter, with ostioles measuring 10 to 30 μm in diameter. Conidia were straight to mildly curved or even flexuous, guttulate, hyaline, 30 to 75 × 1.5 to 2 μm, and one- to seven-septate. Based on the morphological characteristics, the fungus was consistent with Septoria erigerontis Peck (3,4). Conidia were harvested from cirrhi of pycnidia on leaf lesions with a drop of sterile water and then directly streaked onto water agar media using a bacterial loop. Isolates were incubated at 24°C for 48 h. Germinating conidia were individually transferred to potato dextrose agar (PDA) plates. An isolate was deposited in the Korean Agricultural Culture Collection (Accession No. KACC46120). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1/ITS4 primers and sequenced. The resulting sequence of 505 bp was deposited in GenBank (Accession No. JX480493). A GenBank BLAST search was conducted with the 505-bp sequence showing 100% identity with the sequences of S. erigerontis ex Erigeron annuus (EF535638, GU269862). Pathogenicity was tested by spraying leaves of three potted plants with a conidial suspension (2 × 10 conidia/ml) harvested from a 4-week-old PDA culture. Control leaves were sprayed with sterile distilled water. The plants were placed in a dew chamber at 26°C in darkness and continuous dew for the first 24 h and then moved to a greenhouse bench. After 7 days, leaf spot symptoms identical to those observed in the field developed on the leaves inoculated with the fungus. No symptoms were observed on control plants. S. erigerontis was reisolated from the lesions of inoculated plants, fulfilling Koch's postulates. A leaf spot disease of E. strigosus associated with S. erigerontis has been reported in the United States and Canada (1). To our knowledge, this is the first report of leaf spot on E. strigosus caused by S. erigerontis outside of North America as well as in Korea. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication. ARS, USDA, Retrieved June 2, 2012. (2) S. H. Park. Colored Illustrations of Naturalized Plants of Korea. Ilchokak Publishers, Seoul, Korea, 1995. (3) M. J. Priest. Fungi of Australia: Septoria. ABRS/CSIRO Publishing, Melbourne, Australia, 1997. (4) E. Radulescu et al. Septoriozele din Romania. Ed. Acad. Rep. Soc. Romania, Bucuresti, Romania, 1973.
糙叶飞蓬(Erigeron strigosus Muhl. ex Willd.),俗称雏菊飞蓬,原产于北美,于20世纪90年代意外传入韩国(2)。它在韩国各地的自然和人工管理生态系统中入侵性越来越强。2011年6月,在韩国洪川郡野生的雏菊飞蓬上首次观察到叶斑病。一份凭证标本保存在韩国大学植物标本馆(KUS-F25759)。症状出现在下部叶片上,表现为小的、明显的红棕色病斑,病斑逐渐扩大,变成淡褐色、暗淡的斑点,周围有深紫褐色边缘。在病斑中可见黑色分生孢子器。分生孢子器生于表面,偶尔生于叶下,散生,深褐色至锈褐色,球形,埋生于寄主组织中或部分突破表皮,直径60至160μm,孔口直径10至30μm。分生孢子直或微弯甚至弯曲,具油滴,透明,30至75×1.5至2μm,有1至7个隔膜。根据形态特征,该真菌与飞蓬壳针孢(Septoria erigerontis Peck)一致(3,4)。用一滴无菌水从叶斑上的分生孢子器的分生孢子梗上采集分生孢子,然后用接种环直接划线接种到水琼脂培养基上。分离物在24°C下培养48小时。将萌发的分生孢子分别转移到马铃薯葡萄糖琼脂(PDA)平板上。一个分离物保存在韩国农业菌种保藏中心(保藏号KACC46120)。使用DNeasy植物微量DNA提取试剂盒(Qiagen公司,加利福尼亚州瓦伦西亚)提取基因组DNA。使用ITS1/ITS4引物扩增rDNA的内部转录间隔区(ITS)并进行测序。得到的505bp序列保存在GenBank(保藏号JX480493)。对505bp序列进行GenBank BLAST搜索,结果显示与来自一年蓬(Erigeron annuus)的飞蓬壳针孢序列(EF535638,GU269862)具有百分之百的同一性。通过用从4周龄的PDA培养物中收获的分生孢子悬浮液(2×10分生孢子/ml)喷洒三盆植物的叶片来测试致病性。对照叶片喷洒无菌蒸馏水。将植物置于26°C的保湿箱中,黑暗处理24小时,保持连续结露,然后转移到温室试验台上。7天后,接种真菌的叶片上出现了与田间观察到的相同的叶斑症状。对照植物上未观察到症状。从接种植物的病斑中重新分离出飞蓬壳针孢,符合柯赫氏法则。在美国和加拿大已报道了与飞蓬壳针孢相关的糙叶飞蓬叶斑病(1)。据我们所知,这是北美以外以及韩国首次报道由飞蓬壳针孢引起的糙叶飞蓬叶斑病。参考文献:(1)D.F.法尔和A.Y.罗斯曼。真菌数据库。系统真菌学与微生物学实验室,在线出版物。美国农业部农业研究局,2012年6月2日检索。(2)朴成焕。韩国归化植物彩色图鉴。Ilchokak出版社,韩国首尔,1995年。(3)M.J.普里斯特。澳大利亚的真菌:壳针孢属。ABRS/CSIRO出版社,澳大利亚墨尔本,1997年。(4)E.拉杜列斯库等人。罗马尼亚的壳针孢属病害。罗马尼亚科学院出版社,罗马尼亚布加勒斯特,1973年。