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由菊欧文氏菌引起的细菌性茎腐病在佛罗里达州田间种植番茄上的发生情况。

Occurrence of Bacterial Stem Rot, Caused by Erwinia chrysanthemi, on Field-Grown Tomato in Florida.

作者信息

Chellemi D O, Dankers H A, Hill K, Cullen R E, Simone G W, Gooch M D, Allingham J E

机构信息

USDA-ARS, Horticultural Research Laboratory, Ft. Pierce, FL 34945.

University of Florida-IFAS, NFREC, Quincy 32351.

出版信息

Plant Dis. 1998 Jul;82(7):831. doi: 10.1094/PDIS.1998.82.7.831C.

Abstract

In September 1997, wilted 4-week-old tomato (Lycopersicon esculentum Mill.) plants were observed in a commercial production field in St. Lucie County, FL. Closer inspection of affected plants revealed hollow stems and petioles with dark, water-soaked lesions. Diseased tissue was macerated and streaked onto nutrient agar (NA) and crystal violet pectate (CVP) agar. After incubation for 2 days at 30°C, isolates produced pits on the CVP agar. Isolates were transferred onto NA and the incubation and transfer procedure was performed two additional times to obtain pure cultures. Suspensions of bacterial cells were injected into tomato and tobacco leaves to test for a hypersensitive or pathogenic reaction. Isolates produced collapsed necrotic tissue on tomato while no reaction was observed on tobacco. Tests for differentiating species and subspecies in the 'carotovora' group of Erwinia were conducted following the protocol of Dickey and Kelman (1). With known cultures of E. carotovora subsp. carotovora and E. chrysanthemi as controls, the isolate from tomato was determined to function as a facultative anaerobe, utilize asparagine as a sole source of carbon and nitrogen, and give positive reactions for pectate degradation, phosphatase, and growth at 37°C. Known cultures of E. carotovora subsp. carotovora, E. chrysanthemi, and the tomato isolate were grown on trypticase soy broth agar for 24 h at 28°C and their cellular fatty acids derivatized to fatty acid methyl esters (FAMEs). Statistical analyses of FAME profile data (MIDI Microbial Identification System, Newark, DE, version 3.60) identified the tomato isolate as Erwinia chrysanthemi. Pathogenicity was determined by inoculating 50-day-old tomato plants (cv. SunPride) with a suspension of E. chrysanthemi obtained from nutrient broth plates incubated at 24°C for 60 h. Three plants each were inoculated with the E. chrysanthemi identified from tomato, sterile water, and known cultures of E. chrysanthemi and E. carotovora subsp. carotovora by placing a drop at the junction of the petiole and stem and passing a sterile needle through the drop into the stem. Plants were maintained in a greenhouse. Dark, water-soaked cankers were observed on the stems of plants inoculated with E. chrysanthemi, including the tomato isolate and E. carotovora subsp. carotovora, after 7 days. No symptoms were observed on plants inoculated with sterile water. Reisolation of the pathogen and identification was performed with tissue from one of the symptomatic inoculated plants. Analyses of FAMEs confirmed E. chrysanthemi as the causal agent. This is the first report of E. chrysanthemi causing a vascular disease of field-grown tomato in Florida. Reference: (1) R. S. Dickey and A. Kelman. 1988. Pages 44-59 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. N. W. Schaad, ed. American Phytopathological Society, St. Paul, MN.

摘要

1997年9月,在佛罗里达州圣卢西县的一个商业生产田地里,发现了4周龄的番茄(Lycopersicon esculentum Mill.)植株枯萎。对受影响植株的进一步检查发现茎和叶柄中空,有深色的水渍状病斑。将患病组织研磨并划线接种到营养琼脂(NA)和结晶紫果胶酸盐(CVP)琼脂上。在30°C下培养2天后,分离菌在CVP琼脂上产生凹陷。将分离菌转移到NA上,并再重复进行两次培养和转移程序以获得纯培养物。将细菌细胞悬液注射到番茄和烟草叶片中,以测试过敏反应或致病反应。分离菌在番茄上产生了坏死组织塌陷,而在烟草上未观察到反应。按照迪基和凯尔曼(1)的方案,对欧文氏菌属“胡萝卜软腐菌”组中的种和亚种进行鉴别试验。以胡萝卜软腐欧文氏菌亚种胡萝卜软腐菌和菊欧文氏菌的已知培养物作为对照,确定从番茄中分离出的菌株为兼性厌氧菌,以天冬酰胺作为唯一碳源和氮源,对果胶酸盐降解、磷酸酶和在37°C下生长呈阳性反应。将胡萝卜软腐欧文氏菌亚种胡萝卜软腐菌、菊欧文氏菌的已知培养物以及从番茄中分离出的菌株在胰蛋白胨大豆肉汤琼脂上于28°C培养24小时,并将其细胞脂肪酸衍生为脂肪酸甲酯(FAMEs)。对FAME谱数据(MIDI微生物鉴定系统,特拉华州纽瓦克,版本3.60)进行统计分析,确定从番茄中分离出的菌株为菊欧文氏菌。通过用从在24°C下培养60小时的营养肉汤平板上获得的菊欧文氏菌悬液接种50日龄的番茄植株(品种为SunPride)来测定致病性。分别用从番茄中鉴定出的菊欧文氏菌、无菌水以及菊欧文氏菌和胡萝卜软腐欧文氏菌亚种胡萝卜软腐菌的已知培养物接种三株植物,方法是在叶柄和茎的交界处滴一滴菌液,并用无菌针穿过菌液刺入茎中。植株置于温室中。7天后,在接种了菊欧文氏菌(包括从番茄中分离出的菌株和胡萝卜软腐欧文氏菌亚种胡萝卜软腐菌)的植株茎上观察到深色的水渍状溃疡。接种无菌水的植株未出现症状。从一株有症状的接种植株上取组织进行病原菌的再分离和鉴定。对FAMEs的分析证实菊欧文氏菌为病原菌。这是菊欧文氏菌引起佛罗里达州田间种植番茄维管束病害的首次报道。参考文献:(1)R. S. 迪基和A. 凯尔曼。1988年。载于:《植物病原细菌鉴定实验室指南》。N. W. 沙德编。美国植物病理学会,明尼苏达州圣保罗。

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