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俄克拉荷马州大豆田大豆叶脉坏死病毒的首次报告

First Report of Soybean vein necrosis virus in Soybean Fields of Oklahoma.

作者信息

Ali A, Abdalla O A

机构信息

Department of Biological Science, the University of Tulsa, OK 74104.

出版信息

Plant Dis. 2013 Dec;97(12):1664. doi: 10.1094/PDIS-05-13-0515-PDN.

Abstract

Soybean vein necrosis virus (SVNV) causes a new emerging disease of soybean that has been recorded in more than 10 states (1,2,3,4) of the United States, but so far no information is available about its presence in soybean crops of Oklahoma. Surveys of commercial soybean fields were conducted for soybean viruses during summer of 2012. A total of 327 samples were randomly collected from soybean fields in 11 counties. Symptoms typical of SVNV infections including leaf chlorosis and leaf-vein necrosis were observed on some soybean plants in the field (4). All soybean leaf samples were tested against SVNV polyclonal antisera obtained from AC Diagnostics, Inc. (Fayetteville, AR) by dot-immunobinding assay (DIBA) (1). Fifty-three samples reacted positively with SVNV antisera. Total RNA was extracted from three DIBA-positive samples collected from soybean plants in Choctaw County and tested by reverse transcription (RT)-PCR using SVNV-specific primers (forward primer 5'-ATGTTCTCTCTATAATAGCCA and reverse primer 5'-ACCCATAACAATTGATCAAGA-3') that were designed from the available sequence in the GenBank (Accession No. GU722317.1) to amplify a fragment from RNA1. A band of the expected size of 344 bp was observed on a 1% agarose gel in all three samples. The PCR products were purified using QIAquick PCR Purification Kit (QIAGEN, Valencia, CA), cloned (pGEM-T Easy Vector, Promega, Madison, WI) and sequenced in both directions. The consensus sequence of the 344-bp fragment was 99% identical with the corresponding region of RNA 1 of SVNV isolate 'Milan_TN' (Accession No. GU722317.1). These results confirmed the presence of SVNV in soybean fields, which are mostly located in Criage, Choctaw, Hughes, LeFlore, Mayes, Muskogee, McCurtain, Okmulgee, Ottawa, Rogers, and Sequoyah counties of Oklahoma. None of the samples collected from north central or western parts of the state were positive against SVNV. To our knowledge, this is the first report of SVNV in soybean crops in Oklahoma. Soybean is one of the major oil seed crops cultivated on approximately 200,000 hectares annually in Oklahoma and the presence of SVNV could pose a potential threat to the production of soybean in the future. References: (1) J. L. Jacobs and M. I. Chilvers. Plant Dis. 97:1387, 2013. (2) J. Han et al. Plant Dis. 97:693, 2013. (3) D. L. Smith et al. Plant Dis. 97:693, 2013. (4) J. Zhou et al. Virus Genes 43:289, 2011.

摘要

大豆叶脉坏死病毒(SVNV)引发了一种新出现的大豆病害,该病害已在美国10多个州被记录(1,2,3,4),但目前尚无关于其在俄克拉荷马州大豆作物中存在情况的信息。2012年夏季对商业大豆田进行了大豆病毒调查。从11个县的大豆田中随机采集了327个样本。在田间的一些大豆植株上观察到了典型的SVNV感染症状,包括叶片黄化和叶脉坏死(4)。所有大豆叶片样本均通过斑点免疫结合测定法(DIBA)(1),用从AC诊断公司(阿肯色州费耶特维尔)获得的SVNV多克隆抗血清进行检测。53个样本与SVNV抗血清反应呈阳性。从乔克托县大豆植株上采集的3个DIBA阳性样本中提取总RNA,并使用根据GenBank中可用序列(登录号GU722317.1)设计的SVNV特异性引物(正向引物5'-ATGTTCTCTCTATAATAGCCA和反向引物5'-ACCCATAACAATTGATCAAGA-3')通过逆转录(RT)-PCR进行检测,以从RNA1扩增一个片段。在所有3个样本的1%琼脂糖凝胶上均观察到预期大小为344 bp的条带。使用QIAquick PCR纯化试剂盒(QIAGEN,加利福尼亚州瓦伦西亚)对PCR产物进行纯化,克隆(pGEM-T Easy载体,Promega公司,威斯康星州麦迪逊)并进行双向测序。344 bp片段的一致序列与SVNV分离株“米兰_TN”(登录号GU722317.1)的RNA 1相应区域有99%的同一性。这些结果证实了SVNV在大豆田中的存在,这些大豆田大多位于俄克拉荷马州的克里奇、乔克托、休斯、勒弗洛、梅斯、马斯库吉、麦柯廷、奥克穆尔吉、渥太华、罗杰斯和塞阔亚县。从该州中北部或西部采集的样本均未检测出SVNV呈阳性。据我们所知,这是SVNV在俄克拉荷马州大豆作物中的首次报道。大豆是俄克拉荷马州每年种植面积约20万公顷的主要油料作物之一,SVNV的存在可能对未来大豆生产构成潜在威胁。参考文献:(1)J. L. Jacobs和M. I. Chilvers。植物病害97:1387,2013。(2)J. Han等人。植物病害97:693,2013。(3)D. L. Smith等人。植物病害97:693,2013。(4)J. Zhou等人。病毒基因43:289,2011。

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