First Report of Impatiens necrotic spot virus on Spiderlily in China.
First Report of Impatiens necrotic spot virus on Spiderlily in China.
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DOI：
10.1094/pdis-94-4-0484c
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发表时间：
2010-03
期刊：
影响因子：
4.5
通讯作者：
Y. Liu;Y. Zheng;Y. Li;Z. Y. Li
中科院分区：
文献类型：
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作者：
Y. Liu;Y. Zheng;Y. Li;Z. Y. Li
凤仙花坏死斑病毒（INSV） （Tospovirus属；布尼亚病毒科）是观赏花卉生产中的一种毁灭性疾病(1)。2007 - 2009年，对云南省昆明市10个主要公园和游憩区进行的调查表明，8个公园的蜘蛛百合植物中约有60 ~ 70%存在同心圆斑和坏死斑症状。用免疫条（Agdia Inc.）对有症状植株进行了INSV和番茄斑点枯萎病毒（TSWV）的检测。埃尔克哈特,)。结果表明，只有指定HDL的样品对INSV呈阳性，其他样品对INSV和TSWV均呈阴性。机械接种的雏菊、烟叶、凤仙花和野蔷薇表现出褪绿病变、同心圆和严重坏死，这些都是INSV宿主的典型症状。电镜检查发现舌状病毒样的球形包膜颗粒，直径为90纳米。Primer 5软件（Premier, Canada）设计了14条引物。NC_003625扩增L RNA， NC_003616扩增M RNA， NC_003624扩增S RNA。这些引物以被侵染植物组织中提取的总RNA为模板进行RT -PCR，得到29个250 ~ 900 bp的目标片段。这些片段用载体pMD19 simple-T载体（Takara Bio Inc., Dalian, China）克隆并测序。克隆序列经DNAman （version 2.5; Lynnon Biosoft, Quebec, Canada）软件比对，结果显示rna L、M、S为8776 bp （GenBank Accession No. 5）。GU112505), 4948 bp (GenBank登录号：GU112503)和2875 bp (GenBank Accession No.；分别GU112504)。利用NCBI序列数据库对这些蜘蛛百合INSV序列进行BLAST分析，发现RdRp蛋白（L RNA）与意大利分离株（No. 5）的RdRp蛋白同源性为99.6%。DQ425094)， Nsm蛋白（M RNA）与意大利分离物Nsm蛋白的同源性为99.0%。DQ425095)和一份来自美国(编号：NC_003616)， G1G2多蛋白（M RNA）与意大利分离物（No. 003616）的类似蛋白具有99.9%的同源性。DQ425095)， N蛋白（S RNA）与意大利分离物（No. 425095）的N蛋白具有99.6%的同源性。DQ425096)， NSs蛋白（S RNA）与日本分离物NSs蛋白的同源性为98.7%。AB109100)。据我们所知，这是INSV对蜘蛛百合在中国的第一份报告。参考文献：(1)a.e. Whitfield等。为基础。植物酚类化合物学报，2005,33(3):459 - 459。
Impatiens necrotic spot virus (INSV) (genus Tospovirus; family Bunyaviridae) is a devastating disease in the production of ornamental flowers (1). From 2007 to 2009, a survey of 10 major parks and recreation areas in Kunming, the capital of Yunnan Province, China, indicated that approximately 60 to 70% of Spiderlily (Hymenocallis littoralis Salisb.) plants from eight parks had symptoms of concentric ring spots and necrotic spots. Symptomatic plants were tested for INSV and Tomato spotted wilt virus (TSWV) with an immunostrip (Agdia Inc. Elkhart, IN). Results indicated that only the samples designated HDL were positive for INSV and all other samples were negative for both INSV and TSWV. Mechanically inoculated Emilia sonchifolia, Nicotiana glutinosa, Impatiens balsamina, and N. rustica showed chlorotic lesions, concentric rings, and severe necrosis, symptoms typical for INSV in these hosts. Electron microscope inspection found tospovirus-like spheroidal, enveloped particles that were 90 nm in diameter. Primer 5 software (Premier, Canada) was used to design 14 primers from GenBank Accession No. NC_003625 to amplify the L RNA, nine from NC_003616 to amplify the M RNA, and six from NC_003624 to amplify the S RNA. With total RNA extracted from infected plant tissue as templates in reverse transcription (RT)-PCR, these primers generated 29 target fragments of 250 to 900 bp. These fragments were cloned with the vector pMD19 simple-T vector (Takara Bio Inc., Dalian, China) and sequenced. The sequences of the clones were aligned with the software DNAman (version 2.5; Lynnon Biosoft, Quebec, Canada), showing that RNAs L, M, and S are 8,776 bp (GenBank Accession No. GU112505), 4,948 bp (GenBank Accession No. GU112503), and 2,875 bp (GenBank Accession No. GU112504), respectively. BLAST analysis of these Spiderlily INSV sequences against the NCBI sequence database indicated that the RdRp protein (L RNA) was 99.6% identical with the RdRp protein from an Italian isolate (No. DQ425094), the Nsm protein (M RNA) has 99.0% identity with the Nsm protein from an isolate from Italy (No. DQ425095) and one from the United States (No. NC_003616), the G1G2 polyprotein (M RNA) has 99.9% identity with the analogous protein from an Italian isolate (No. DQ425095), the N protein (S RNA) has 99.6% identity with the N protein from an Italian isolate (No. DQ425096), and the NSs protein (S RNA) has 98.7% identity with the NSs protein from an isolate from Japan (No. AB109100). To our knowledge, this is the first report of INSV on Spiderlily in China. Reference: (1) A. E. Whitfield et al. Annu. Rev. Phytopathol. 43:459, 2005.