Rungnapha K, Yu S H, Xie G L
State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China. Supported by the National Science Foundation of China (30671397) and the Agricultural Ministry of China (nyhyzx07-056).
Plant Dis. 2008 Jul;92(7):1135. doi: 10.1094/PDIS-92-7-1135B.
In December 2006, a rot symptom of unknown etiology was observed on stems of plants (Euphorbia pulcherrima cv. Fu-xing) at a flower nursery in the Zhejiang Province of China where we had previously reported leaf spot of poinsettia caused by Xanthomonas campestris (2). Chlorotic spots anywhere along the stem and purplish black petioles were the first noticeable symptoms. The spots rapidly coalesced, forming large irregular chlorotic areas. Petioles turned black and shriveled and affected leaves wilted. Infected tissues were soft and water soaked. Ten bacterial strains were isolated from the diseased samples and five were selected for identification. They were similar to those of the standard reference strains of Pectobacterium chrysanthemi (Dickeya sp.), LMG 2804 from Belgium and ZUPB20056 from China, in phenotypic tests based on the Biolog Microbial Identification System, version 4.2 (Biolog Inc., Hayward, CA), pathogenicity tests, gas chromatography of fatty acid methyl esters (FAME) using the Microbial Identification System (MIDI Inc, Newark, DE) with aerobic bacterial library (TABA50), and transmission electron microscopy (TEM,KYKY-1000B, Japan). All strains tested were gram-negative facultative anaerobic rods measuring 1.5 to 3.6 × 0.6 to 1.1 μm, with peritrichous flagella. Colonies were gray-white and slightly raised with smooth margins on nutrient agar. They were negative for trehalose and positive for phosphatase production and reducing substances from sucrose. A hypersensitive reaction was observed on tobacco cv. Benshi, 24 h after inoculation. All five isolates, LMG 2804, and ZUPB20056 were identified as P. chrysanthemi (Dickeya sp.) with a Biolog similarity index of 0.58 to 0.83, 0.68, and 0.72 and a FAME similarity index of 0.52 to 0.80, 0.59, and 0.70, respectively. Identification as P. chrysanthemi (Dickeya sp.) was confirmed by PCR with specific primers used by Nassar et al (3). Koch's postulates were completed with the inoculation of 12 4-month-old intact poinsettia plants of cv. Fu-xing with cell suspensions containing 10 CFU/ml by a pinprick at the base of the stem. All five strains induced stem infection similar to those observed in natural infections. No symptoms were noted on the two control plants inoculated with sterilized distilled water by the same method. The bacterium was reisolated from symptomatic stems of poinsettia plants. P. chrysanthemi (Dickeya sp.) was first reported in United States as the cause of bacterial stem rot of poinsettia in 1972 (1). To our knowledge, this is the first report of poinsettia stem rot caused by P. chrysanthemi (Dickeya sp.) in China. The disease cycle and the control strategies of the bacterial stem rot of poinsettia in the regions are being further studied. References: (1) H. A. J. Hoitink et al. Plant Dis. Rep. 56:480, 1972. (2) B. Li et al. Plant Pathol. 55:293, 2006. (3) A. A. Nassar et al. Appl. Environ. Microbiol. 62:2228, 1996.
2006年12月,在中国浙江省的一家花卉苗圃中,我们在一品红(Euphorbia pulcherrima cv. Fu-xing)植株的茎上观察到一种病因不明的腐烂症状,此前我们曾报道过由野油菜黄单胞菌引起的一品红叶斑病(2)。茎上任何部位出现的褪绿斑点和紫黑色叶柄是最初明显的症状。这些斑点迅速融合,形成大的不规则褪绿区域。叶柄变黑并枯萎,受影响的叶片萎蔫。受感染的组织柔软且呈水渍状。从患病样本中分离出10株细菌菌株,选取其中5株进行鉴定。基于Biolog微生物鉴定系统4.2版(Biolog公司,美国加利福尼亚州海沃德市)的表型测试、致病性测试、使用微生物鉴定系统(MIDI公司,美国特拉华州纽瓦克市)和需氧细菌文库(TABA50)进行的脂肪酸甲酯气相色谱分析(FAME)以及透射电子显微镜观察(TEM,KYKY - 1000B,日本)显示,它们与菊果胶杆菌(Dickeya sp.)的标准参考菌株、来自比利时的LMG 2804和来自中国的ZUPB20056相似。所有测试菌株均为革兰氏阴性兼性厌氧杆菌,大小为1.5至3.6×0.6至1.1μm,具周生鞭毛。在营养琼脂上,菌落呈灰白色,边缘光滑且略凸起。它们对海藻糖呈阴性反应,对磷酸酶产生和蔗糖还原物质呈阳性反应。接种后24小时,在烟草品种Benshi上观察到过敏反应。所有5株分离物、LMG 2804和ZUPB20056经鉴定均为菊果胶杆菌(Dickeya sp.),其Biolog相似性指数分别为0.58至0.83、0.68和0.72,FAME相似性指数分别为0.52至0.80、0.59和0.70。使用Nassar等人(3)所用的特异性引物通过PCR确认了菊果胶杆菌(Dickeya sp.)的鉴定结果。通过用含有10 CFU/ml的细胞悬浮液在12株4月龄的一品红品种Fu-xing完整植株的茎基部进行针刺接种,完成了柯赫氏法则验证。所有5株菌株均诱导出与自然感染中观察到的类似的茎部感染症状。通过相同方法接种灭菌蒸馏水的两株对照植株未出现症状。从出现症状的一品红植株茎部重新分离出该细菌。菊果胶杆菌(Dickeya sp.)于1972年在美国首次被报道为一品红细菌性茎腐病的病因(1)。据我们所知,这是中国首次报道由菊果胶杆菌(Dickeya sp.)引起的一品红茎腐病。该地区一品红细菌性茎腐病的病害循环和防治策略正在进一步研究中。参考文献:(1)H. A. J. Hoitink等人,《植物病害报告》56:480,1972年。(2)B. Li等人,《植物病理学》55:293,2006年。(3)A. A. Nassar等人,《应用与环境微生物学》62:2228,1996年。