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阿根廷玉米斯图尔特枯萎病由斯氏泛菌引起的首次报道

First Report of Stewart's Wilt of Maize in Argentina Caused by Pantoea stewartii.

作者信息

Orio A G Albarracín, Brücher E, Plazas M C, Sayago P, Guerra F, De Rossi R, Ducasse D A, Guerra G D

机构信息

Laboratorio de Fitopatología, Microbiología y Biología Molecular, Facultad de Ciencias Agropecuarias, Universidad Católica de Córdoba, Av. Armada Argentina 3555, Córdoba (5017), Argentina.

出版信息

Plant Dis. 2012 Dec;96(12):1819. doi: 10.1094/PDIS-07-12-0668-PDN.

DOI:10.1094/PDIS-07-12-0668-PDN
PMID:30727295
Abstract

Stewart's wilt is a serious disease of corn (Zea mays L.) caused by the bacterium Pantoea stewartii subsp. stewartii (Pss). Typical symptoms of infected fields and dent corn are longitudinal streaks with irregular or wavy margins, which are parallel to the veins and may extend the length of the leaf. These pale to green yellow lesions become dry and brown as the disease progresses producing a leaf blight (4). During the growing seasons 2010 to 2011 and 2011 to 2012, symptoms of bacterial leaf blight of corn were observed in central Argentina maize fields, with an incidence of 54% in Córdoba province. To identify the pathogen, leaves from 10 symptomatic maize plants per field were collected from 15 fields covering a representative geographical area. High populations of morphologically uniform bacteria were isolated from leaf tissues by conventional methods using King's medium B agar (2). Ten representative facultatively anaerobic gram-negative, non-fluorescing, non-motile, catalase positive and oxidase negative rod-shaped and yellow-pigmented bacterial isolates were evaluated further. The biochemical profile obtained was: fermentative metabolism, negative indol, acetoin and hydrogen sulfide production, negative gelatin hydrolysis (22°C), positive acid production from D-glucose and lactose, negative gas production from D-glucose, and negative nitrate reduction (1). All the isolates produced a 300-bp band with PCR using the species specific primer pair PST3581/PST3909c (3). The Pss ATCC 8199 and Pseudomonas fluorescens ATCC 13525 strains were used as positive and negative controls for the PCR assays, respectively. The pathogenicity test was performed by stem inoculation of five to ten P2069 YR maize plants (one to two leaf growth stage) grown in growth chamber. Plants were inoculated by syringe with a 10 to 10 cell/ml bacterial suspension and kept in a humid chamber at 25 to 27°C. Plants inoculated with Pss ATCC 8199 or with sterile water were used as positive and negative control treatments, respectively. The development of symptoms similar to those originally found in the field was observed on all the plants inoculated with the different isolates at 7 to 10 days post inoculation. In addition, symptoms on inoculated plants were similar to those observed for the positive control treatment. No symptoms were found on negative controls. Koch's postulates were fulfilled since bacteria isolated from symptomatic tissue had identical characteristics to isolates used to inoculate plants and to the reference Pss strain for biochemical tests and PCR reaction mentioned above. To our knowledge, this is the first report of P. stewartii subsp. stewartii isolated from diseased maize in Argentina. References: (1) J. G. Holt et al. Page 179 in: Bergey's manual of determinative bacteriology. Williams and Wilkins, Baltimore, MD, 1994. (2) OEPP/EPPO. Bulletin OEPP/EPPO Bulletin, 36: 111, 2006. Pantoea stewartii subsp. stewartii diagnostic. (3) A. Wensing et al. Appl. Environ. Microbiol. 76:6248, 2010. (4) D. G. White Page 4 in: Compendium of corn disease. The American Phytopathology Society, 1999.

摘要

斯图尔特枯萎病是由斯图尔特泛菌斯图尔特亚种(Pss)引起的玉米(Zea mays L.)的一种严重病害。受感染田地和马齿型玉米的典型症状是纵向条纹,边缘不规则或呈波浪状,与叶脉平行,可能延伸至叶片全长。随着病情发展,这些从浅到绿黄色的病斑会变干并变成褐色,导致叶枯(4)。在2010年至2011年以及2011年至2012年的生长季节,在阿根廷中部的玉米田中观察到玉米细菌性叶枯病症状,科尔多瓦省的发病率为54%。为了鉴定病原菌,从覆盖代表性地理区域的15块田地中,每块田地采集10株有症状玉米植株的叶片。通过使用King氏培养基B琼脂的常规方法(2)从叶片组织中分离出大量形态一致的细菌。对10株具有代表性的兼性厌氧革兰氏阴性、无荧光、不运动、过氧化氢酶阳性和氧化酶阴性的杆状且产黄色色素的细菌分离株进行了进一步评估。获得的生化特征如下:发酵代谢,吲哚、3-羟基丁酮和硫化氢产生阴性,明胶水解(22°C)阴性,D-葡萄糖和乳糖产酸阳性,D-葡萄糖产气阴性,硝酸盐还原阴性(1)。使用物种特异性引物对PST3581/PST3909c进行PCR时,所有分离株均产生一条300 bp的条带(3)。Pss ATCC 8199菌株和荧光假单胞菌ATCC 13525菌株分别用作PCR检测的阳性和阴性对照。通过对生长在生长室中的5至10株P2069 YR玉米植株(一至二叶期)进行茎接种来进行致病性测试。用注射器向植株接种10⁶至10⁷个细胞/毫升的细菌悬液,并将其置于25至27°C的潮湿培养箱中。分别以接种Pss ATCC 8199或无菌水的植株作为阳性和阴性对照处理。接种后7至10天,在所有接种不同分离株的植株上均观察到与最初在田间发现的症状相似的症状发展。此外,接种植株上的症状与阳性对照处理中观察到的症状相似。阴性对照未发现症状。科赫法则得到满足,因为从有症状组织中分离出的细菌与用于接种植株的分离株以及上述生化测试和PCR反应的参考Pss菌株具有相同的特征。据我们所知,这是首次从阿根廷患病玉米中分离出斯图尔特泛菌斯图尔特亚种的报告。参考文献:(1)J.G.霍尔特等人。载于:《伯杰氏鉴定细菌学手册》第179页。威廉姆斯和威尔金斯出版社,马里兰州巴尔的摩,1994年。(2)OEPP/EPPO。《OEPP/EPPO公报》,36:111,2006年。斯图尔特泛菌斯图尔特亚种诊断。(3)A.温辛等人。《应用与环境微生物学》,76:6248,2010年。(4)D.G.怀特。载于:《玉米病害简编》第4页。美国植物病理学会,1999年。