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中国阴沟肠杆菌引起的桑细菌性枯萎病

Bacterial Wilt of Mulberry (Morus alba) Caused by Enterobacter cloacae in China.

作者信息

Wang G F, Praphat K, Xie G L, Zhu B, Li B, Liu B, Zhou Q

机构信息

State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China.

Biotechnology Research Institute, Fujian Academy of Agricultural Science, Fuzhou 350003, China.

出版信息

Plant Dis. 2008 Mar;92(3):483. doi: 10.1094/PDIS-92-3-0483B.

DOI:10.1094/PDIS-92-3-0483B
PMID:30769704
Abstract

In August of 2006, a new bacterial disease was noted in Hangzhou mulberry orchards of Zhejiang Province, China where bacterial wilt of mulberry caused by Ralstonia solanacearum was previously reported (3). In the summer, the disease caused severe wilt, especially on 1- or 2-year-old mulberry plants, that resulted in premature plant death. Leaf wilt symptoms generally started on older leaves at the bottom of the plant and spread to the younger leaves. The leaves of infected plants became withered and dry, turned dark brown, and eventually the plants became defoliated. The root xylem of infected plants was moist and discolored with brown stripes. The phloem was asymptomatic, however, in severe infections, the phloem was decayed. The observation of wilting proceeding from the bottom of the plant to the top distinguishes this disease from bacterial wilt caused by R. solanacearum. Five bacterial strains isolated from infected mulberry plants showed characteristics similar to those of the standard reference strain of Enterobacter cloacae subsp. cloacae IBJ0611from China, but differed from R. solanacearum IBJ35, E. cancerogenus LMG2693, and E. cloacae subsp. dissolvens LMG2683 from the University of Gent, Belgium in phenotypic tests, including the Biolog Identification System version 4.2 (Biolog Inc., Hayward CA), pathogenicity tests, transmission electron microscopy (TEM,KYKY-1000B, Japan) observation, and gas chromatographic analysis of fatty acid methyl esters (FAMEs) using the Microbial Identification System (MIDI Company, Newark, DE) with the aerobic bacterial library (TABA50). Isolates were gram negative, facultative anaerobic, rod shaped, 0.3 to 1.0 × 1.0 to 3.0 μm with peritrichous flagella. Colonies on nutrient agar were light yellow, smooth, circular, entire, and convex with no green fluorescent diffusible pigment on King's medium B (3). Weak hypersensitive reaction was observed on tobacco 3 days after inoculation. All five strains were identified as E. cloacae with Biolog similarity of 0.662 to 0.863 and FAMEs similarity of 0.632 to 0.701. Inoculation of 10 6-month-old intact mulberry plants of cv Husang with cell suspensions containing 10 CFU/ml by pinprick at the base of the stem reproduced symptoms observed in natural infections. No symptoms were noted on the two control plants inoculated by the same method but with sterilized distilled water. The bacterium was reisolated from the symptomatic mulberry plants. E. cloacae has been reported from the United States as the cause of internal yellowing of papaya fruits (1) and rhizome rot of edible ginger (2). To our knowledge, this is the first report of mulberry wilt caused by E. cloacae in China. References: (1) K. Nishijima et al. Plant Dis. 71:1029, 1987. (2) K. Nishijima et al. Plant Dis. 88:1318, 2004. (3) L. Xu et al. Acta Phytophylacica. Sin. 34:141, 2007.

摘要

2006年8月,在中国浙江省杭州的桑园中发现了一种新的细菌性病害,此前该地区曾报道过由青枯雷尔氏菌引起的桑树青枯病(3)。夏季,该病导致严重萎蔫,尤其是1年生或2年生的桑树,导致植株过早死亡。叶片萎蔫症状通常从植株底部的老叶开始,蔓延至幼叶。受感染植株的叶片枯萎干燥,变成深褐色,最终植株落叶。受感染植株的根木质部潮湿,有褐色条纹变色。韧皮部无症状,然而,在严重感染时,韧皮部会腐烂。这种从植株底部向上发展的萎蔫现象可将此病与青枯雷尔氏菌引起的青枯病区分开来。从受感染的桑树中分离出的5个细菌菌株,其特征与中国阴沟肠杆菌阴沟亚种标准参考菌株IBJ0611相似,但在表型测试中与比利时根特大学的青枯雷尔氏菌IBJ35、致癌肠杆菌LMG2693和阴沟肠杆菌溶解亚种LMG2683不同,这些测试包括Biolog鉴定系统4.2版(Biolog公司,加利福尼亚州海沃德)、致病性测试、透射电子显微镜(TEM,KYKY - 1000B,日本)观察以及使用微生物鉴定系统(MIDI公司,特拉华州纽瓦克)和需氧细菌文库(TABA50)对脂肪酸甲酯(FAMEs)进行气相色谱分析。分离菌株为革兰氏阴性、兼性厌氧、杆状,大小为0.3至1.0×1.0至3.0μm,有周生鞭毛。营养琼脂上的菌落为浅黄色、光滑、圆形、边缘整齐且凸起,在King氏培养基B上无绿色荧光扩散性色素(3)。接种后3天在烟草上观察到微弱的过敏反应。所有5个菌株经Biolog鉴定相似度为0.662至0.863,FAMEs相似度为0.632至0.701,均被鉴定为阴沟肠杆菌。用含有10 CFU/ml的细胞悬浮液通过在6个月大的完整湖桑品种植株茎基部针刺接种,再现了自然感染中观察到的症状。用相同方法但接种无菌蒸馏水的2株对照植株未出现症状。从有症状的桑树植株中重新分离出该细菌。在美国,阴沟肠杆菌已被报道为番木瓜果实内部变黄(1)和食用生姜根茎腐烂(2)的病因。据我们所知,这是中国首次报道由阴沟肠杆菌引起的桑树萎蔫病。参考文献:(1)K. Nishijima等人,《植物病害》71:1029,1987。(2)K. Nishijima等人,《植物病害》88:1318,2004。(3)L. Xu等人,《植物保护学报》34:141,2007。

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