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由密执安棒形杆菌密执安亚种引起的番茄细菌性溃疡病在墨西哥下加利福尼亚半岛的发生情况

Bacterial Canker Caused by Clavibacter michiganensis subsp. michiganensis on Tomato in the Baja California Peninsula of Mexico.

作者信息

Holguín-Peña R J, Vázquez-Juárez R C, Rueda-Puente E O

机构信息

Laboratorio de Biología Molecular de Plantas, Centro de Investigaciones Biológicas del Noroeste, La Paz, B.C.S. 23000, Mexico.

Universidad de Sonora, Santa Ana, Sonora 84600, Mexico.

出版信息

Plant Dis. 2006 Dec;90(12):1550. doi: 10.1094/PD-90-1550A.

Abstract

During the 2005 tomato-growing (Lycopersicon esculentum Mill.) season, an apparent bacterial disease with cankers on the stems and bird's eye lesions on the fruit appeared in commercial fields and greenhouses in the San Quintin and San Simon areas (a 60-mile long coastal plain) near the central region of the Baja California Peninsula of Mexico. The disease was found in midseason, especially when plants were flowering, and the mature and ripe stage. Incidence ranged from 4 to 46%, which represented an important loss in field and greenhouse production. Symptomatic plants showed reddish brown cavities in the stem, discoloration, and water soaking of vascular tissue. Diseased tissues were washed with phosphate buffer and placed on semiselective Clavibacter medium (SCM) (1), and a gram-positive, nonmotile, nonspore-forming, aerobic, curved rod bacterium was consistently isolated and morphologically characterized. Twenty-eight isolates were identified as Clavibacter michiganensis subsp. michiganensis by polymerase chain reaction (PCR) technology with primers CMM5/CMM6 to amplify a fragment of approximately 6.2 kb (2). The isolates were also identified by REP-PCR (repetitive extragenic palindromic PCR) genomic fingerprinting techniques (3) with REP and BOX primer sets (4). Pathogenicity tests consisting of three replicates of 4-week-old tomato seedlings (cv. Tequila) were performed by spraying (twice, 2 days apart) inocula at 10 CFU/ml. Control seedlings were sprayed with sterile water. Inoculated plants previously covered in polyethylene bags were incubated in a growth chamber at 25°C for 48 h. Within 3 weeks, symptoms of reddish-brown cavities, water-soaked lesions, and asymmetrical wilting appeared on inoculated plants and were similar to those symptoms observed in the field. No symptoms were observed on control plants. Confirmation of the causal agent was done by culturing the bacteria on SCM and PCR analysis. Occurrence of the disease in San Quintin Valley is relevant because the disease is one of the five most serious tomato diseases in the peninsula. Moreover, the potential spread of the pathogen by tomato seedlings represents a permanent risk to other pathogen-free areas in the peninsula. Although bacterial canker has been observed in Baja California (Punta Colonet, Vicente Guerrero, San Quintin, and San Simon), to our knowledge, this is the first confirmation of C. michiganensis subsp. michiganensis in Baja, Mexico. References: (1). C. Alarcon et al. Phytopathology 88:306, 1998. (2) J. Dreier et al. Phytopathology 85:462, 1995. (3) F. J. Louws et al. Phytopathology 88:862, 1998. (4) J. Versalovic et al. Methods Mol. Cell. Biol. 5:25, 1994.

摘要

在2005年番茄(Lycopersicon esculentum Mill.)种植季节,墨西哥下加利福尼亚半岛中部地区附近的圣昆廷和圣西蒙地区(一片60英里长的沿海平原)的商业田地和温室中出现了一种明显的细菌性病害,其茎上有溃疡,果实上有鸟眼状病斑。该病在生长季中期被发现,尤其是在植株开花以及成熟和熟透阶段。发病率在4%至46%之间,这在田间和温室生产中造成了重大损失。有症状的植株茎部出现红棕色空洞、变色以及维管组织水浸状。将患病组织用磷酸盐缓冲液冲洗后置于半选择性棒状杆菌培养基(SCM)上(1),持续分离出一种革兰氏阳性、不运动、不形成芽孢、需氧的弯曲杆菌,并对其进行形态学特征鉴定。通过聚合酶链反应(PCR)技术,使用引物CMM5/CMM6扩增出一段约6.2 kb的片段(2),将28个分离株鉴定为密执安棒状杆菌密执安亚种。还通过重复外显子回文PCR(REP-PCR)基因组指纹技术(3),使用REP和BOX引物对(4)对分离株进行了鉴定。致病性测试以4周龄番茄幼苗(品种Tequila)进行三次重复,通过喷雾(两次,间隔2天)接种菌液,浓度为10 CFU/ml。对照幼苗喷洒无菌水。先前用聚乙烯袋覆盖的接种植株在生长室中于25°C下培养48小时。在3周内,接种植株上出现了红棕色空洞、水浸状病斑和不对称萎蔫症状,与在田间观察到的症状相似。对照植株未观察到症状。通过在SCM上培养细菌并进行PCR分析来确认病原菌。圣昆廷谷出现这种病害具有重要意义,因为该病是该半岛最严重的五种番茄病害之一。此外,病原菌通过番茄幼苗传播的可能性对半岛其他无病地区构成了长期风险。尽管在下加利福尼亚(蓬塔科洛内特、比森特·格雷罗、圣昆廷和圣西蒙)已观察到细菌性溃疡病,但据我们所知,这是墨西哥下加利福尼亚首次确认密执安棒状杆菌密执安亚种。参考文献:(1)C. 阿拉尔孔等人,《植物病理学》88:306,1998年。(2)J. 德赖尔等人,《植物病理学》85:462,1995年。(3)F. J. 洛斯等人,《植物病理学》88:862,1998年。(4)J. 韦尔萨洛维奇等人,《分子细胞生物学方法》5:25,1994年。

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