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韩国木槿上由多主棒孢引起的叶斑病的首次报道

First Report of Leaf Spot Caused by Corynespora cassiicola on Rose of Sharon in Korea.

作者信息

Seo S T, Park J H, Cho S E, Shin H D

机构信息

Division of Forest Diseases and Insect Pests, Korea Forest Research Institute, Seoul 130-712, Korea.

Division of Environmental Science and Ecological Engineering, Korea University, Seoul 136-701, Korea.

出版信息

Plant Dis. 2013 Jun;97(6):847. doi: 10.1094/PDIS-11-12-1099-PDN.

Abstract

Rose of Sharon, Hibiscus syriacus L., is a flowering shrub in the family Malvaceae planted as the national flower of South Korea. In September 2012, previously unknown leaf spots with premature defoliation were observed on dozens of Rose of Sharon plants growing in the shaded area in a park of Dongducheon, Korea. The same symptoms were found on Rose of Sharon in several localities of Korea in 2012. The symptoms usually started as small, dark brown to grayish leaf spots, eventually causing leaf yellowing with significant premature defoliation. The diseased leaves retained for a while green color at the margin of the spots. Representative samples (n = 5) were deposited in the Korea University Herbarium (KUS). Conidiophores of the fungus observed microscopically on the leaf spots were erect, brown to dark brown, single or in clusters, amphigenous but mostly hypophyllous, and measured 80 to 400 × 5 to 10 μm. Conidia were borne singly or in short chains, ranging from cylindrical to broadest at the base and tapering apically, straight to slightly curved, pale olivaceous brown, 2 to 16 pseudoseptate, 50 to 260 × 9 to 20 μm, each with a conspicuous thickened hilum. On potato dextrose agar, single-spore cultures of two isolates were identified as Corynespora cassiicola (Berk. & M.A. Curtis) C.T. Wei on the basis of morphological and cultural characteristics (1,2). Two monoconidial isolates were preserved at the Korean Agricultural Culture Collection (KACC46956 and KACC46957). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequences of 520 bp were deposited in GenBank (Accession Nos. KC193256, KC193257). A BLAST search in GenBank revealed that the sequences showed 100% identity with those of numerous C. cassiicola isolates from diverse substrates. To conduct a pathogenicity test, a conidial suspension (ca. 2 × 10 conidia/ml) was prepared in sterile water by harvesting conidia from 2-week-old cultures of KACC46956, and the suspension was sprayed onto the leaves of three healthy 2-year-old plants. Inoculated plants were kept in humid chambers for the first 48 h and thereafter placed in the glasshouse. After 10 days, typical leaf spot symptoms developed on the leaves of all three inoculated plants. C. cassiicola was reisolated from the lesions, confirming Koch's postulates. Control plants treated with sterile water remained symptomless. C. cassiicola is cosmopolitan with a very wide host range (1,2). Though Corynespora hibisci Goto was recorded to be associated with brown spot disease of H. syriacus in Japan (4), there is no previous record of C. cassiicola on H. syriacus (3). To our knowledge, this is the first report of Corynespora leaf spot on Rose of Sharon in Korea. According to our field observations in Korea, this disease was found in August and September, following a prolonged period of moist weather. Severe infection resulted in leaf yellowing and premature defoliation, reducing tree vigor and detracting the beauty of green leaves. References: (1) L. J. Dixon et al. Phytopathology 99:1015, 2009. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonw. Mycol. Inst., Kew, UK, 1971. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved November 22, 2012. (4) K. Goto. Ann. Phytopathol. Soc. Japan 12:14, 1942.

摘要

木槿(Hibiscus syriacus L.)是锦葵科的一种开花灌木,被定为韩国的国花。2012年9月,在韩国东豆川市一个公园的阴凉处生长的几十株木槿上,发现了以前未知的叶斑病并伴有早期落叶现象。2012年在韩国的几个地方,木槿上也发现了同样的症状。症状通常始于小的、深褐色至灰色的叶斑,最终导致叶片变黄并伴有严重的早期落叶。患病叶片在病斑边缘会有一段时间保持绿色。代表性样本(n = 5)保存在韩国大学植物标本馆(KUS)。在叶斑上显微镜观察到的真菌分生孢子梗直立,褐色至深褐色,单生或丛生,两面生但大多生于叶下面,大小为80至400×5至10μm。分生孢子单生或成短链,从圆柱形到基部最宽且顶端渐细,直或稍弯曲,浅橄榄褐色,有2至16个假隔膜,50至260×9至20μm,每个都有一个明显加厚的脐点。在马铃薯葡萄糖琼脂上,根据形态和培养特征,两个分离株的单孢培养物被鉴定为多主棒孢(Corynespora cassiicola (Berk. & M.A. Curtis) C.T. Wei)(1,2)。两个单孢分离株保存在韩国农业文化收藏中心(KACC46956和KACC46957)。使用DNeasy植物小量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。用引物ITS1/ITS4扩增rDNA的完整内部转录间隔区(ITS)并进行测序。得到的520 bp序列保存在GenBank(登录号KC193256、KC193257)。在GenBank中进行的BLAST搜索显示,这些序列与来自不同底物的众多多主棒孢分离株的序列具有100%的同一性。为了进行致病性测试,通过从KACC46956的2周龄培养物中收集分生孢子,在无菌水中制备分生孢子悬浮液(约2×10个分生孢子/ml),并将该悬浮液喷洒到三株健康的2年生植物的叶片上。接种的植物在最初48小时置于湿度箱中,之后放在温室中。10天后,所有三株接种植物的叶片上都出现了典型的叶斑症状。从病斑中重新分离出多主棒孢,证实了柯赫氏法则。用无菌水处理的对照植物没有症状。多主棒孢分布广泛,寄主范围非常广(1,2)。虽然在日本记录到日本棒孢(Corynespora hibisci Goto)与木槿的褐斑病有关(4),但以前没有多主棒孢在木槿上的记录(3)。据我们所知,这是韩国首次报道木槿上的多主棒孢叶斑病。根据我们在韩国的实地观察,这种病害在潮湿天气持续较长时间后的8月和9月出现。严重感染导致叶片变黄和早期落叶,降低了树木活力并损害了绿叶的美观。参考文献:(1) L. J. Dixon等人,《植物病理学》99:1015,2009年。(2) M. B. Ellis,《半知菌亚门》,英国皇家植物园邱园真菌研究所,1971年。(3) D. F. Farr和A. Y. Rossman,《真菌数据库》,系统真菌学和微生物学实验室,在线出版物,美国农业部农业研究局,2012年11月22日检索。(4) K. Goto,《日本植物病理学会年报》12:14,1942年。

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