Rodriguez-Salamanca L M, Enzenbacher T B, Derie M L, du Toit L J, Feng C, Correll J C, Hausbeck M K
Department of Plant Pathology, Michigan State University, East Lansing 48824.
Department of Plant Pathology, Washington State University, Mount Vernon 98273.
Plant Dis. 2012 May;96(5):769. doi: 10.1094/PDIS-01-12-0022-PDN.
In July of 2010, dry, oval lesions, each with a salmon-colored center and bleached overall appearance, were observed on the leaves and neck of onions plants growing in production fields of Newaygo, Ottawa, Kent, and Ionia counties, Michigan. Acervuli and setae that are characteristic of Colletotrichum spp. were observed with a dissecting microscope, and elliptical conidia (8 to 23 × 3 to 12 μm) with attenuated ends were observed with a compound microscope. Symptomatic tissues were excised and cultured onto potato dextrose agar amended with 30 and 100 ppm of rifampicin and ampicillin, respectively. The cultures produced pale salmon-colored sporulation after growing for 5 days at 22 ± 2°C and black microsclerotia after 2 weeks. Six isolates were confirmed as C. coccodes based on sequence analysis of the internal transcribed (ITS) region of the ribosomal DNA and a 1-kb intron of the glutamine synthase gene (GS) (2). Sequences were submitted to GenBank (Accession Nos. JQ682644 and JQ682645 for ITS and GS, respectively). Pathogenicity tests were conducted on two- to three-leaved 'Stanley' and 'Cortland' onion seedlings. Prior to inoculation, seedlings were enclosed in clear plastic bags overnight to provide high relative humidity. The bags were removed, and seedlings were sprayed inoculated with a C. coccodes conidial suspension (5 × 10 conidia/ml and 25 ml/plant) in sterile double-distilled water. Control seedlings were sprayed with sterile double-distilled water. Tween (0.01%) was added to the conidial suspension and the water. Plants were enclosed in bags for 72 h postinoculation and incubated in growth chambers at 28°C day/23°C night with a 12-h photoperiod. Sunken, oval lesions were observed on the foliage of the onion seedlings inoculated with C. coccodes 4 days postinoculation. Lesions coalesced and foliage collapsed 7 days postinoculation. Control plants remained asymptomatic. When five leaf samples per replication were detached and incubated in a moist chamber for 3 days at 21 ± 2°C, abundant acervuli and setae were observed on the symptomatic tissue but not on control tissue. C. coccodes was consistently recovered from the onion seedling lesions. Six different Colletotrichum spp. have been reported to cause diseases on onions worldwide (1,4). C. circinans, which causes smudge, is an occasional onion pathogen in Michigan, while C. gloeosporioides has only been reported to be infecting onions in Georgia (3). To our knowledge, this is the first report of C. coccodes infecting and causing disease in onions plants. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , August 6, 2010. (2) J. C. Guerber et al. Mycologia 95:872. 2003. (3) C. Nischwitz et al. Plant Dis. 92:974. 2008. (4) H. F. Schwartz, and K. S. Mohan. Compendium of Onion and Garlic Diseases and Pests, 2nd ed. The American Phytopathological Society, St. Paul, MN. 1995.
2010年7月,在密歇根州纽埃戈、渥太华、肯特和艾奥尼亚县的生产田地里种植的洋葱植株的叶片和颈部,观察到干燥的椭圆形病斑,每个病斑中心呈鲑鱼色,整体外观呈漂白状。用解剖显微镜观察到炭疽菌属特有的分生孢子盘和刚毛,用复式显微镜观察到两端渐细的椭圆形分生孢子(8至23×3至12μm)。将有症状的组织切除并分别接种到添加了30 ppm利福平(Rifampicin)和100 ppm氨苄青霉素(Ampicillin)的马铃薯葡萄糖琼脂上。培养物在22±2°C下生长5天后产生浅鲑鱼色的孢子,2周后产生黑色微菌核。基于核糖体DNA的内部转录间隔区(ITS)和谷氨酰胺合成酶基因(GS)的1 kb内含子的序列分析,6个分离株被确认为块茎炭疽菌(C. coccodes)(2)。序列已提交至GenBank(ITS和GS的登录号分别为JQ682644和JQ682645)。对两叶至三叶期的‘斯坦利’(Stanley)和‘科特兰’(Cortland)洋葱幼苗进行致病性测试。接种前,将幼苗置于透明塑料袋中过夜以提供高相对湿度。移除袋子后,用无菌双蒸水中的块茎炭疽菌分生孢子悬浮液(5×10⁵分生孢子/ml,25 ml/株)对幼苗进行喷雾接种。对照幼苗喷洒无菌双蒸水。在分生孢子悬浮液和水中添加吐温(0.01%)。接种后将植株置于袋中72小时,并在生长室中于28°C日/23°C夜、12小时光周期下培养。接种块茎炭疽菌后4天,在洋葱幼苗的叶片上观察到凹陷的椭圆形病斑。接种后7天,病斑融合,叶片枯萎。对照植株无症状。当每次重复取5片叶样,在21±2°C的湿润培养箱中培养3天时,在有症状的组织上观察到大量的分生孢子盘和刚毛,而对照组织上未观察到。从洋葱幼苗病斑中持续分离到块茎炭疽菌。据报道,全球有6种不同的炭疽菌属真菌可引起洋葱病害(1,4)。引起污斑病的旋孢炭疽菌(C. circinans)是密歇根州偶尔出现的洋葱病原菌,而胶孢炭疽菌(C. gloeosporioides)仅在佐治亚州有报道感染洋葱(3)。据我们所知,这是块茎炭疽菌感染洋葱植株并引起病害的首次报道。参考文献:(1)D. F. Farr和A. Y. Rossman。真菌数据库。美国农业部农业研究局系统真菌学和微生物学实验室。检索自http://nt.ars-grin.gov/fungaldatabases/,2010年8月6日。(2)J. C. Guerber等人。《真菌学》95:872。2003年。(3)C. Nischwitz等人。《植物病害》9:974。2008年。(4)H. F. Schwartz和K. S. Mohan。《洋葱和大蒜病虫害汇编》,第2版。美国植物病理学会,明尼苏达州圣保罗。1995年。