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得克萨斯州玉米上戈斯细菌性萎蔫病和叶枯病(密执安棒形杆菌内布拉斯加亚种)的首次报道

First Report of Goss's Bacterial Wilt and Leaf Blight (Clavibacter michiganensis subsp. nebraskensis) of Corn in Texas.

作者信息

Korus K A, Timmerman A D, French-Monar R D, Jackson T A

机构信息

Department of Plant Pathology, University of Nebraska-Lincoln 68583-0722.

Department of Plant Pathology and Microbiology, Texas AgriLife Extension Service-Texas A&M System, Amarillo 79106.

出版信息

Plant Dis. 2011 Jan;95(1):73. doi: 10.1094/PDIS-07-10-0541.

Abstract

In September 2009, the University of Nebraska-Lincoln Plant and Pest Diagnostic Clinic received leaf samples of hybrid corn (Zea mays L.) displaying long, necrotic lesions with wavy margins. The lesions had discontinuous water-soaked spots that are indicative of Goss's bacterial wilt and leaf blight. The symptomatic leaves were submitted from Dallam County, located in the Texas Panhandle (northwest Texas). According to the USDA Farm Service Agency and the National Agricultural Statistics Service, in 2009 Dallam County had 54,025 ha planted to corn. This is approximately 19% of the total corn planted in the 26 counties in the Texas Panhandle and 6% of the total corn planted in the state of Texas. Extracts from the infected leaf tissue tested positive for Clavibacter michiganensis subsp. nebraskensis with a commercially available ELISA test (Neogen Inc., Scotland, UK). Isolation from the infected tissue onto CNS selective media (1) resulted in round, dark orange, mucoid colonies that tested gram positive with the Gram-stain test. BLAST nucleotide sequence alignments of the amplified 500-bp 16S rRNA region of the suspect culture's genome (2) revealed a 96% similarity for C. michiganensis subsp. nebraskensis (NCBI BLAST Accession No. U09381.1). To fulfill Koch's postulates, three sweet corn plants (Golden Cross Bantam) at growth stage V3 to V4 were inoculated in the greenhouse with a suspension of approximately 1 × 10 CFU/ml from suspect cultures grown on CNS for 5 days. Wounds approximately 6.5 cm long were created with sterile scissors on the fifth leaf from the bottom running parallel to the veins on either side of the midrib at the leaf apex. The leaf apex was dipped into 150 ml of the inoculum suspension for 5 s. Approximately 6 days after inoculation, discontinuous, water-soaked spots consistent with the symptoms on the original symptomatic leaves appeared on all the inoculated leaves near the site of infection. Colonies consistent with C. michiganensis subsp. nebraskensis (dark orange, mucoid) were reisolated onto CNS, completing Koch's postulates. To our knowledge, this is the first report of Goss's bacterial wilt and leaf blight on corn in Texas and because it is a residue-borne pathogen, the probability of it becoming a resident disease is relatively high. References: (1) D. C. Gross and A. K. Vidaver. Phytopathology 69:82, 1979. (2) X. Li and S. H. De Boer. 1995. Phytopathology 85:837, 1995.

摘要

2009年9月,内布拉斯加大学林肯分校植物与害虫诊断诊所收到了杂交玉米(玉米属)的叶片样本,这些样本上出现了边缘呈波浪状的长坏死病斑。病斑上有不连续的水渍状斑点,这是戈斯细菌性萎蔫病和叶枯病的特征。有症状的叶片是从位于得克萨斯州狭长地带(得克萨斯州西北部)的达拉姆县送来的。根据美国农业部农场服务局和国家农业统计局的数据,2009年达拉姆县种植玉米54025公顷。这约占得克萨斯州狭长地带26个县玉米种植总面积的19%,占得克萨斯州玉米种植总面积的6%。用市售的酶联免疫吸附测定试剂盒(Neogen公司,英国苏格兰)对感染叶组织提取物进行检测,结果显示密歇根棒形杆菌内布拉斯加亚种呈阳性。将感染组织接种到CNS选择性培养基上(1),长出圆形、深橙色、黏液状菌落,革兰氏染色检测为革兰氏阳性。对可疑培养物基因组中扩增出的500bp 16S rRNA区域进行BLAST核苷酸序列比对(2),结果显示与密歇根棒形杆菌内布拉斯加亚种的相似度为96%(美国国立生物技术信息中心BLAST登录号:U09381.1)。为了验证科赫法则,在温室中,用在CNS上培养5天的可疑培养物制成的约1×10CFU/ml的悬浮液,接种了3株处于V3至V4生长阶段的甜玉米植株(金十字矮脚鸡品种)。用无菌剪刀在从底部数第五片叶子上,于叶尖处沿着叶脉两侧平行于中脉剪出约6.5厘米长的伤口。将叶尖浸入150毫升接种悬浮液中5秒。接种后约6天,在所有接种叶片的感染部位附近出现了与原始有症状叶片症状一致的不连续水渍状斑点。从接种叶片上重新分离出了与密歇根棒形杆菌内布拉斯加亚种一致的菌落(深橙色、黏液状),接种到CNS上,从而完成了科赫法则验证。据我们所知,这是得克萨斯州玉米上首次报道戈斯细菌性萎蔫病和叶枯病,而且由于它是一种残体传播的病原菌,它成为当地常驻病害的可能性相对较高。参考文献:(1)D.C.格罗斯和A.K.维达弗。《植物病理学》69:82,1979年。(2)李X和德布尔S.H.。1995年。《植物病理学》85:837,1995年。

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