Polizzi G, Dimartino M, Bella P, Catara V
Dipartimento di Scienze e Tecnologie Fitosanitarie, University of Catania, Via S. Sofia 100, 95123 Catania, Italy.
Plant Dis. 2008 May;92(5):835. doi: 10.1094/PDIS-92-5-0835A.
Bird of Paradise tree (Strelitzia alba (L. f.) Skeels) is an ornamental perennial tropical plant grown in southern Italy. In the summer of 2006 and 2007, a widespread, severe leaf disease was observed on seedlings and 1- to 2-year-old plants in two glasshouses located in eastern Sicily. Disease incidence ranged from 10 to 25%. Symptoms on the leaves consisted of dark brown-to-black stripes of varying length and found between the lateral veins. Lesions sometimes coalesced into a large area of necrotic tissue. Symptomatic tissues were ground in a drop of sterile distilled water (SDW) with a scalpel. Suspensions were streaked on King's medium B (KB), nutrient agar, and yeast extract nutrient agar (2). Isolated strains were gram negative and oxidase positive, non-levan, negative in tobacco hypersensitivity test, white and nonmucoid on yeast dextrose calcium carbonate agar, did not produce fluorescent pigments on KB, and utilized glucose, mannitol, trehalose, arabinose, mannose, and N-acetylglucosamine. Bacterial strains were identified as Ralstonia pickettii by using the Biolog Identification System (MicroLogTM System Release 4.2; Biolog, Inc., Hayward, CA) with a similarity index ranging from 0.52 to 0.67. For an additional confirmation of identity, the small subunit rRNA gene (SSUrDNA) was amplified with primers 530F and Uni 1492R (1). The resulting nucleotide sequence was compared with sequences deposited in GenBank and showed the highest identity (99%) to sequences of R. pickettii strains. Pathogenicity tests were performed on 20 cm tall potted plants. Four S. alba plants were inoculated by infiltrating leaf veins with bacterial suspensions for each of the four isolates (10 CFU ml in SDW) with a 25-gauge needle and syringe. Plants were placed in polyethylene bags 1 day before inoculation and maintained there for 3 days after inoculation. Four control plants were inoculated with SDW. Water-soaked areas in the lateral veins of leaves were observed in all inoculated plants 4 days after inoculation. Within 10 days, dark brown-to-black stripes that coalesced into dark necrotic areas were observed. All isolates induced similar symptoms. Control plants did not show any symptoms. The pathogen was reisolated from symptomatic tissue and identified as R. pickettii by Biolog. A similar disease on S. reginae caused by a Pseudomonas sp. was previously reported from Florida (3). To our knowledge, this is the first record in the world of leaf spot and blight caused by R. pickettii. References: (1) D. J. Lane. 16S/23S rRNA sequencing. Page 115 in: Nucleic Acid Techniques in Bacterial Systematics. E. Stackebrandt and M. Goodfellow, eds. John Wiley and Sons, NY, 1991. (2) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. The American Phytopathological Society, St. Paul, MN, 2001. (3) C. Wehlburg. Plant Dis. Rep. 55:447, 1971.
天堂鸟树(白鹤望兰(L. f.)斯基尔斯)是一种多年生观赏热带植物,生长在意大利南部。2006年和2007年夏天,在西西里岛东部的两个温室中,在幼苗以及1至2年生植株上观察到一种广泛且严重的叶部病害。发病率在10%至25%之间。叶片症状表现为长度各异的深褐色至黑色条纹,位于侧脉之间。病斑有时会融合成一大片坏死组织。用手术刀将有症状的组织在一滴无菌蒸馏水中研磨。将悬液划线接种在King氏培养基B(KB)、营养琼脂和酵母提取物营养琼脂上(2)。分离得到的菌株革兰氏阴性、氧化酶阳性、不产果聚糖、烟草过敏试验呈阴性、在酵母葡萄糖碳酸钙琼脂上呈白色且无黏液、在KB上不产生荧光色素,并利用葡萄糖、甘露醇、海藻糖、阿拉伯糖、甘露糖和N - 乙酰葡糖胺。使用Biolog鉴定系统(MicroLogTM系统版本4.2;Biolog公司,加利福尼亚州海沃德)将细菌菌株鉴定为皮氏伯克霍尔德菌,相似性指数在0.52至0.67之间。为进一步确认身份,用引物530F和Uni 1492R扩增小亚基rRNA基因(SSUrDNA)(1)。将得到的核苷酸序列与GenBank中保存的序列进行比较,结果显示与皮氏伯克霍尔德菌菌株的序列具有最高的同一性(99%)。对20厘米高的盆栽植株进行致病性测试。用25号针头和注射器,将四种分离菌株的细菌悬液(在无菌蒸馏水中浓度为10 CFU/ml)注入四株白鹤望兰植株的叶脉进行接种。接种前1天将植株置于聚乙烯袋中,并在接种后保持3天。四株对照植株接种无菌蒸馏水。接种后第4天,在所有接种植株的叶片侧脉中观察到水渍状区域。10天内,观察到深褐色至黑色条纹融合成深色坏死区域。所有分离菌株引起相似症状。对照植株未表现出任何症状。从有症状的组织中重新分离出病原菌,并通过Biolog鉴定为皮氏伯克霍尔德菌。此前曾有报道称佛罗里达州一种由假单胞菌属引起的类似病害发生在女王鹤望兰上(3)。据我们所知,这是世界上首次关于皮氏伯克霍尔德菌引起叶斑病和叶枯病的记录。参考文献:(1)D. J. 莱恩。16S/23S rRNA测序。见:《细菌系统学中的核酸技术》,第115页。E. 斯塔克布兰特和M. 古德费洛编。约翰威利父子公司,纽约,1991年。(2)N. W. 沙德等人。《植物病原细菌鉴定实验室指南》。美国植物病理学会,明尼苏达州圣保罗,2001年。(3)C. 韦尔堡。《植物病害报告》55:447,1971年。