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由丁香假单胞菌烟草致病变种引起的大豆野火病,韩国的一种新病害。

Wildfire of Soybean Caused by Pseudomonas syringae pv. tabaci, a New Disease in Korea.

作者信息

Myung I-S, Kim J-W, An S H, Lee J H, Kim S K, Lee Y-K, Kim W G

机构信息

Agricultural Microbiology Division, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA), Suwon 441-707, Korea.

Dongguk University, Seoul 100-715, Korea.

出版信息

Plant Dis. 2009 Nov;93(11):1214. doi: 10.1094/PDIS-93-11-1214A.

Abstract

In 2006 and 2007, a new bacterial disease was observed in field-cultivated soybeans in Boeun District and Munkyung City of Korea. The disease caused severe blighting of soybean (Glycine max) leaves. Soybean leaves in fields showed yellowish spots with brown centers. Brown and dead areas of variable size and shape were surrounded by wide, yellow haloes with distinct margins. Spots might coalesce and affected leaves fell readily. Seven bacterial strains were isolated from chlorotic areas of soybean leaves and all produced white colonies on trypticase soy agar. With the Biolog Microbial Identification System, version 4.2, (Biolog Inc., Hayward, CA) all strains and Pseudomonas syringae pv. tabaci CFBP2106 were identified as P. syringae pv. tabaci with a Biolog similarity index of 0.28 to 0.52 and 0.48 after 24 h. Pathogenicity of the strains (three plants per strain) on soybean leaves at the V5 stage (cv. Hwanggeum) was confirmed by rub inoculation with bacterial suspensions (1 × 10 CFU/ml) in sterile distilled water on the lesions cut 1 cm long on the upper side of the leaves with razor blades and by pinprick on 3-week-old leaves of tobacco (Nicotiana tabacum cv. Samsun) in the greenhouse. Wildfire symptoms on the soybean leaves and faint halos on tobacco leaves were observed 4 days after inoculation. The identification of reisolated bacterial strains was confirmed with the metabolic fingerprintings on Biolog. LOPAT tests (1) and phenotypic characteristics (3) of the strains were similar to those of the CFBP2106. Colonies were levan positive, oxidase negative, potato soft rot negative, arginine dihydrase negative, and tobacco hypersensitivity negative. All strains were gram-negative, aerobic rods with a polar flagellum. Strains were negative for esculin hydrolysis, gelatin liquefaction, urea production, accumulation of poly-β-hydroxy butyrate, starch hydrolysis, ornithine dihydrolase, lysine dihydrolase, growth at 37°C, utilization of geraniol, benzoate, cellobiose, sorbitol, trehalose, l-rhamnose, and adonitol. Positive reactions were catalase and arbutin hydrolysis, utilization of sorbitol, d-arabinose, and dl-serine. The strains were variable in utilization of mannitol, sucrose, and d-arabinose. The 1,472-bp PCR fragments of strains, BC2366 (GenBank Accession No. FJ755788) and BC2367 (No. FJ755789) was sequenced using 16S rDNA universal primers (2). The sequences shared 100% identity with the analogous sequences of P. syringae pv. glycenea (GenBank Accession No. AB001443) available in NCBI databases. Based on the phenotypic, genetic, and pathological characteristics, all strains were identified as P. syringae pv. tabaci. To our knowledge, this is the first report of P. syringae pv. tabaci causing wildfire on soybean in Korea. References: (1) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966. (2) I.-S. Myung et al. Plant Dis. 92:1472, 2008. (3) N. W. Schaad et al., eds. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001.

摘要

2006年和2007年,在韩国槐山郡和闻庆市的田间种植大豆上观察到一种新的细菌性病害。该病害导致大豆(Glycine max)叶片严重枯萎。田间的大豆叶片出现中心褐色的淡黄色斑点。大小和形状各异的褐色坏死区域被宽的、边缘清晰的黄色晕圈包围。斑点可能会融合,受影响的叶片很容易掉落。从大豆叶片的褪绿区域分离出7个细菌菌株,所有菌株在胰蛋白胨大豆琼脂上均产生白色菌落。使用4.2版的Biolog微生物鉴定系统(Biolog公司,美国加利福尼亚州海沃德市),所有菌株和丁香假单胞菌烟草致病变种CFBP2106在24小时后被鉴定为丁香假单胞菌烟草致病变种,Biolog相似性指数为0.28至0.52以及0.48。通过用无菌蒸馏水中的细菌悬液(1×10 CFU/ml)对用剃须刀片在叶片上侧切割的1厘米长的伤口进行涂抹接种,并对温室中3周龄烟草(Nicotiana tabacum cv. Samsun)叶片进行针刺接种,证实了这些菌株(每个菌株接种3株植物)对处于V5阶段(品种“黄金”)的大豆叶片的致病性。接种4天后观察到大豆叶片上出现野火症状以及烟草叶片上出现淡晕圈。通过Biolog上的代谢指纹图谱确认了重新分离的细菌菌株的鉴定。这些菌株的LOPAT试验(1)和表型特征(3)与CFBP2106的相似。菌落为左旋糖阳性、氧化酶阴性、马铃薯软腐阴性、精氨酸双水解酶阴性以及烟草过敏阴性。所有菌株均为革兰氏阴性、需氧杆菌,具极生鞭毛。菌株对七叶苷水解、明胶液化、尿素产生、聚-β-羟基丁酸积累、淀粉水解、鸟氨酸双水解酶、赖氨酸双水解酶、37°C生长、香叶醇利用、苯甲酸盐利用、纤维二糖利用、山梨醇利用、海藻糖利用、L-鼠李糖利用和阿东糖醇利用呈阴性反应。对过氧化氢酶和熊果苷水解、山梨醇利用、D-阿拉伯糖利用和DL-丝氨酸利用呈阳性反应。这些菌株在甘露醇、蔗糖和D-阿拉伯糖利用方面存在差异。使用16S rDNA通用引物(2)对菌株BC2366(GenBank登录号FJ755788)和BC2367(登录号FJ755789)的1472 bp PCR片段进行测序。这些序列与NCBI数据库中丁香假单胞菌大豆致病变种(GenBank登录号AB001443)的类似序列具有100%的同一性。基于表型、遗传和病理特征,所有菌株均被鉴定为丁香假单胞菌烟草致病变种。据我们所知,这是丁香假单胞菌烟草致病变种在韩国引起大豆野火病的首次报道。参考文献:(1)R. A. Lelliott等人,《应用细菌学杂志》29:470,1966年。(2)I.-S. Myung等人,《植物病害》92:1472,2008年。(3)N. W. Schaad等人编,《植物病原细菌鉴定实验室指南》第3版。美国植物病理学会,明尼苏达州圣保罗,2001年。

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