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苜蓿花叶病毒侵染加利福尼亚罗勒(罗勒属)的首次报道

First Report of Alfalfa mosaic virus Infecting Basil (Ocimum basilicum) in California.

作者信息

Wintermantel W M, Natwick E T

机构信息

USDA-ARS, Salinas, CA.

University of California Cooperative Extension, Holtville, CA.

出版信息

Plant Dis. 2012 Feb;96(2):295. doi: 10.1094/PDIS-06-11-0516.

DOI:10.1094/PDIS-06-11-0516
PMID:30731822
Abstract

Basil (Ocimum basilicum L.) plants collected from three fields in Imperial County, CA in May, 2011 were found to be exhibiting yellowing, chlorotic sectors and spots on leaves, resulting in unmarketable plants. Dodder (Cuscuta spp.) was present in one of the fields, but was not visibly associated with symptomatic plants. Total nucleic acid was extracted from four symptomatic and three asymptomatic basil plants, as well as from the dodder plant with the RNeasy Plant Mini Kit (Qiagen, Valencia, CA). Nucleic acid extracts were tested by reverse transcription (RT)-PCR for the presence of Alfalfa mosaic virus (AMV) using primers designed to amplify a 350-nt region of the AMV coat protein gene (3). RT-PCR produced bands of the expected size in extracts from all symptomatic plants and the dodder sample. No amplification was obtained from symptomless plants. A 350-nt band amplified from one plant was gel-extracted, sequenced (TACGen, Richmond, CA), and confirmed to be AMV by comparison to sequences available in GenBank (Accession No. K02703). Although serological tests on an initial basil sample were negative for AMV by ELISA using antiserum produced against AMV by R. Larsen, USDA-ARS, Prosser, WA (unpublished), AMV was confirmed by ELISA and RT-PCR in symptomatic Nicotiana benthamiana, N. clevelandii, and Malva parviflora plants following mechanical transmission from basil source plants. The fields with AMV infections were located at opposite ends of the production region from one another, indicating widespread dispersal of AMV in the region. All AMV positive plants were adjacent to alfalfa. Two additional basil plantings in shade houses open to the outside environment did not have AMV symptomatic plants and were also confirmed negative by RT-PCR, but these plantings were at the extreme north end of Imperial Valley agriculture and well away from any alfalfa fields. At the time the basil plantations were sampled for AMV, no aphids were found in any plantations, but during the several weeks prior to finding the AMV-positive plants, cowpea aphid, Aphis craccivora Koch; pea aphid, Acyrthosiphon pisum Harris; blue alfalfa aphid, Acyrthosiphon kondoi Shinji; and spotted alfalfa aphid, Therioaphis maculata Buckton were colonizing Imperial Valley alfalfa fields, producing winged adults. AMV is transmitted by at least 14 aphid species (1), and most aphid populations increase during the late spring in this important desert agricultural region. The acquisition of AMV by dodder suggests the parasitic plant may serve as a vector of AMV within basil fields, although further study will be necessary for clarification. Significant acreage of basil is grown in the Imperial Valley. This acreage is surrounded by extensive and increasing alfalfa production totaling 55,442 ha (137,000 acres) in Imperial County and representing a 21% increase in acreage over 2009 for the same region (2). To our knowledge, this is the first report of basil infected by AMV in California. The proximity of basil production to such a large alfalfa production region warrants the need for enhanced efforts at aphid management in basil production to reduce vector populations and reduce transmission to basil crops. References: (1) E. M. Jaspars and L. Bos. Alfalfa mosaic virus. No. 229 in: Descriptions of Plant Viruses. Commonw. Mycol. Inst./Assoc. Appl. Biol., Kew, England, 1980. (2) C. Valenzuela. Imperial County California Crop and Livestock Report, 2010. (3) H. Xu and J. Nie. Phytopathology 96:1237, 2006.

摘要

2011年5月从加利福尼亚州帝国县三块田地采集的罗勒(Ocimum basilicum L.)植株出现叶片发黄、褪绿斑块和斑点,导致植株无法上市。菟丝子(Cuscuta spp.)出现在其中一块田地中,但未发现其与有症状植株有明显关联。使用RNeasy Plant Mini试剂盒(Qiagen,加利福尼亚州瓦伦西亚)从4株有症状和3株无症状的罗勒植株以及菟丝子植株中提取总核酸。核酸提取物通过逆转录(RT)-PCR检测苜蓿花叶病毒(AMV)的存在,使用设计用于扩增AMV外壳蛋白基因350 nt区域的引物(3)。RT-PCR在所有有症状植株和菟丝子样本的提取物中产生了预期大小的条带。无症状植株未获得扩增条带。从一株植物中扩增出的350 nt条带进行凝胶提取、测序(TACGen,加利福尼亚州里士满),并通过与GenBank中可用序列(登录号K02703)比较确认为AMV。尽管使用美国农业部农业研究局普罗瑟(华盛顿州)的R. Larsen生产的抗血清对初始罗勒样本进行的ELISA血清学检测对AMV呈阴性(未发表),但在从罗勒源植物机械接种后,通过ELISA和RT-PCR在有症状的本氏烟草、克利夫兰烟草和小花锦葵植株中证实了AMV。感染AMV的田地位于产区的两端,表明AMV在该地区广泛传播。所有AMV阳性植株都与苜蓿相邻。另外两个向外部环境开放的荫棚中的罗勒种植没有AMV有症状植株,RT-PCR也证实为阴性,但这些种植位于帝国谷农业的最北端,远离任何苜蓿田。在对罗勒种植园进行AMV采样时,在任何种植园中都未发现蚜虫,但在发现AMV阳性植株的前几周,豆蚜、豌豆蚜、蓝苜蓿蚜和斑点苜蓿蚜正在帝国谷苜蓿田中定殖并产生有翅成虫。AMV由至少14种蚜虫传播(1),在这个重要的沙漠农业地区,大多数蚜虫种群在晚春时数量增加。菟丝子获取AMV表明这种寄生植物可能在罗勒田中充当AMV的传播媒介,尽管需要进一步研究来阐明。帝国谷种植了大量罗勒。这片种植面积被广泛且不断增加的苜蓿生产所包围,帝国县苜蓿生产总面积达55442公顷(137000英亩),与2009年相比,该地区种植面积增加了21%(2)。据我们所知,这是加利福尼亚州罗勒感染AMV的首次报道。罗勒生产与如此大面积的苜蓿生产区域相邻,因此有必要加强罗勒生产中的蚜虫管理,以减少传毒昆虫数量并减少向罗勒作物的传播。参考文献:(1)E. M. Jaspars和L. Bos。苜蓿花叶病毒。载于《植物病毒描述》第229号。英联邦真菌研究所/应用生物协会,英国基尤,1980年。(2)C. Valenzuela。加利福尼亚州帝国县作物和牲畜报告,2010年。(3)H. Xu和J. Nie。植物病理学96:1237,2006年。

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