Li B, Qiu W, Fang Y, Xie G L
Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Ministry of Agriculture, State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China. Supported by the Agricultural Ministry of China (nyhyzx200803010).
Plant Dis. 2009 May;93(5):552. doi: 10.1094/PDIS-93-5-0552B.
In December 2007, stem rot symptoms on orchids (Oncidium Gower Ramsey) were observed at a flower nursery in the Zhejiang Province of China. Initial symptoms were water-soaked lesions starting at the base of the stem. As these lesions expanded and elongated, the stem and leaf tissues became soft and watery. When examined with a microscope, cut edges of symptomatic stem and leaf tissues consistently exhibited bacterial streaming. The bacteria were isolated by streaking on nutrient agar (3). All isolates were gram-negative, facultative, anaerobic rods with peritrichous flagella. Infiltration of tobacco leaves (Nicotiana tabacum cv. Samsun) with the bacterial suspension of 10 CFU/ml resulted in typical hypersensitivity reactions within 24 h. Five representative isolates were further characterized by the Biolog Microbial Identification System, version 4.2 (Biolog Inc., Hayward, CA) and gas chromatography of fatty acid methyl esters (FAME) by the Microbial Identification System (MIDI Inc., Newark, DE) with aerobic bacterial library (TSBA50). The five isolates were identified as Erwinia chrysanthemi (Pectobacterium chrysanthemi) with a Biolog and FAME similarity index of 0.81 to 0.88 and 0.62 to 0.75, respectively. The transfer of P. chrysamthemi to a novel genus, Dickeya gen. nov., was recently proposed (2). The almost complete 16S rDNA sequence from Oncidium isolate SCH-01 (1,604 bp; EMBL Accession No. FM946179) was determined according to the method of Li et al. (1). A subsequent GenBank search showed that this isolate is 98% identical to that of type strain CFBP 1269T of Dickeya dadantii (EMBL Accession No. AF520707) and CFBP 1200T of Dickeya dianthicola (EMBL Accession No. AF520708). Nevertheless, species identification within genus Dickeya is still difficult since only a limited number of strains of each species have been characterized fully. Koch's postulates were completed with the inoculation of Oncidium seedlings with cell suspensions (10 CFU/ml) by a pinprick at the base of the stem. All five representative isolates induced stem rot similar to that observed in natural infections. No symptoms were noted on the control plants inoculated with sterilized distilled water by the same method. The bacterium was reisolated from symptomatic stems of Oncidium plants. To our knowledge, this is the first report of stem rot on Oncidium orchid in Mainland China caused by the bacterium formerly referred to as P. chrysanthemi, now proposed as Dickeya sp. References: (1) B. Li et al. J. Phytopathol. 154:711, 2006. (2) R. Samson et al. Int. J. Syst. Evol. Microbiol. 55:1415, 2005. (3) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society. St. Paul, MN, 2001.
2007年12月,在中国浙江省的一家花卉苗圃中,观察到兰花(文心兰戈氏拉姆齐品种)出现茎腐病症状。最初症状是从茎基部开始的水渍状病斑。随着这些病斑扩大和拉长,茎和叶组织变得柔软且呈水渍状。在显微镜下检查时,有症状的茎和叶组织的切口边缘始终呈现细菌溢菌现象。通过在营养琼脂上划线分离细菌(3)。所有分离菌株均为革兰氏阴性、兼性、厌氧杆菌,具周生鞭毛。用10 CFU/ml的细菌悬浮液浸润烟草叶片(烟草品种Samsun),24小时内会产生典型的过敏反应。使用Biolog微生物鉴定系统4.2版(Biolog公司,加利福尼亚州海沃德)以及微生物鉴定系统(MIDI公司,特拉华州纽瓦克)的脂肪酸甲酯气相色谱法(FAME),并结合需氧细菌文库(TSBA50),对五个代表性分离菌株进行了进一步鉴定。这五个分离菌株通过Biolog和FAME鉴定,分别被鉴定为菊欧文氏菌(菊果胶杆菌),相似性指数分别为0.81至0.88和0.62至0.75。最近有人提议将菊果胶杆菌转移到一个新属,即迪基氏菌属(Dickeya gen. nov.)(2)。根据Li等人(1)的方法,测定了来自文心兰分离菌株SCH - 01的几乎完整的16S rDNA序列(1604 bp;EMBL登录号FM946179)。随后在GenBank中的搜索显示,该分离菌株与达旦迪基氏菌(Dickeya dadantii)模式菌株CFBP 1269T(EMBL登录号AF520707)和石竹迪基氏菌(Dickeya dianthicola)模式菌株CFBP 1200T(EMBL登录号AF520708)的序列相似度为98%。然而,由于每个物种中只有有限数量的菌株得到了充分鉴定,因此在迪基氏菌属内进行物种鉴定仍然困难。通过用细胞悬浮液(10 CFU/ml)在茎基部针刺接种文心兰幼苗,完成了科赫法则验证。所有五个代表性分离菌株均引发了与自然感染中观察到的类似的茎腐病。用相同方法接种灭菌蒸馏水的对照植株未出现症状。从出现症状的文心兰植株茎部再次分离出该细菌。据我们所知,这是中国大陆首次关于由以前称为菊果胶杆菌、现提议为迪基氏菌属某种细菌引起的文心兰茎腐病的报道。参考文献:(1)B. Li等人,《植物病理学杂志》154:711,2006年。(2)R. Samson等人,《国际系统与进化微生物学杂志》55:1415,2005年。(3)N. W. Schaad等人,《植物病原细菌鉴定实验室指南》第三版。美国植物病理学会,明尼苏达州圣保罗,2001年。