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番茄环斑病毒(TmRSV)的病汁液和 PEG 粗提纯液,用温度、甲醛和戍二醛以及超声波进行灭活处理。PEG 粗提纯液经温度(水浴)60℃处理10分钟或使甲醛浓度达0. 3%二种方法后,均丧失侵染性而有抗原性。将其在4℃冰箱中存放,前者抗原性能保持8个月以上,后者只能保持5个月。病汁液不经处理,在4℃冰箱中存放1个月能丧失侵染性,而抗原性也可保持8个月以上。所以阳性对照的灭活处理以病汁液在4℃下存放1个月和 PEG 粗提纯液60℃处理10分钟为最佳方案。 相似文献
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本文报道应用ＲＴ－ＰＣＲ技术检测烟草环斑病毒（ＴＲＳＶ）的。根据ＴＲＳＶＲＮＡ－２序列３末端非编码区设计，合成引物－１和引物－２；用ＴＲＳＶＲＮＡ作模板，引物－２作引物，反转录合成ｃＤＮＡ，并以此作模板进行ＰＣＲ扩增，５－６小时内可得到结果，检测粗提液中ＲＮＡ灵敏度达４００ｐｇ，并且可以直接从受侵染的叶片汁液中检出ＴＲＳＶ，为口岸快速检测疫推出一个更灵敏、准确的分子生物学新方法。值得推广应用。 相似文献
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番茄环斑病毒单克隆抗体细胞株的建立及检测方法的研究 总被引：1，自引：0，他引：1
用纯化的番茄环斑病毒（ＴｍＲＳＶ）免痊Ｂａｌｂ／ｃ小白鼠，取脾细胞与ＳＰ２／０骨髓瘤细胞融合。筛选出６株分泌ＴｍＲＳＶ单克隆抗体（ＭｃＡｂ）杂交瘤细胞株。抗体免疫球旦白分属于１ｇＧ２ａ，ＩｇＧ３和１ｇＭ。用免疫电镜诱捕法检测，６个ＭｃＡｂ与加拿大分离物反应均为阳性，其中ＣＥ３，ＣＧ８，９Ｇ９不与西德分离物反应。属于１ｇＧ３的单抗具有与Ａ旦白结合的特性。 相似文献
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用特异性抗血清制备的抗血清纸片应用于酶联免疫吸咐实验（ＥＬＩＳＡ），取得了良好的结果。目前已将抗血清纸片用于南芥菜花叶病毒（ＡｒＭＶ）、番茄环斑病毒（ＴｏｍＲＳＶ）、烟草环斑病毒（ＴＲＳＶ）、南方菜豆花叶病毒（ＳＢＭＶ）等进境植物危险性病毒的检测，取得与使用冻干保存抗血清相同的效果。应用抗血清纸片方便、经济、便于保存和邮寄。 相似文献
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菊花为常见栽培园艺花卉植物。从世博园采集的来自日本的菊花病株表现轻花叶 ,具有病毒病症状。日本为番茄环斑病毒 (ＴｏＲＳＶ)、南芥菜花叶病毒 (Ａｒ ＭＶ)、烟草环斑病毒 (ＴＲＳＶ)的疫区。这几种病毒的寄主范围广泛 ,易侵染草本花卉植物[1] ,因此用酶联、电镜和生物接种的方法对菊花病标样进行了诊断和鉴定。1 材料和方法1 .1 样品的采集和症状观察 样品采自世博园国际馆 ,并于检疫温室内隔离种植。以健康植物为对照 ,对样品进行症状观察和记录。1 .2 生物侵染实验 采用 0 .0 1ＭＰＢＳ(ｐＨ7.0 ) ,选用相关寄主 ,常规摩擦接种… 相似文献
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香石竹环斑病毒的分子检测 总被引：2，自引：0，他引：2
试验具体研究了两类不同性质的ｃＤＮＡ探针对香石竹环斑病毒的检测效果。毒源由中国农业大学植物病毒室提供，繁殖于苋色藜Ｃｈｅｎｏｐｏｄｉｕｍａｍａｒａｎｔｉｃｏｌｏｒ上，提纯该病毒，然后抽提其核酸ＲＮＡ，将ＲＮＡ多聚腺苷化后，以Ｏｌｉｇｏ（ｄＴ）为引物，反转录合成ｃＤＮＡ，补平后与载体连接，转化大肠杆菌，获得阳性克隆，用３２Ｐ和光敏生物素分别标记ｃＤＮＡ制作探针，与ＣＲＳＶ及其ＲＮＡ进行点杂交，结果表明，两类不同性质的探针在检测ＲＮＡ水平上具有相同的灵敏度，其检测极限均达到１ｎｇ／ｍｌ，并对二类探针进行了比较。 相似文献
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几种危险性病毒抗血清的制备及检测方法的建立 总被引：1，自引：2，他引：1
番茄环斑病毒、烟草环斑病毒和南芥菜花叶病毒用改进的方法提纯后免疫兔子，制备出高质量的抗血清。琼脂扩散效价为１／３２０，用ＳＰＡ－ＥＬＩＳＡ方法检测病汁液可达１０－３～１０－４。３种血清之间无交叉反应，与健康寄主和常见的自然寄主蛋白也无反应。应用３种血清分别对２３种植物１００多个采集的样品和进口种子进行了ＳＰＡ－ＥＬＩＳＡ检测，未发现有上述３种病毒存在 相似文献
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A seed-transmitted virus was isolated from vetch seedlings showing severe stunting, reduced lamina and leaf mottling. Identification
was based on host range, seed transmission, coat protein and nucleic acid analysis, electron microscopy and serology. This
is the first record of vetch as a natural host of artichoke yellow ringspot virus (AYRSV).
http://www.phytoparasitica.org posting Jan. 13, 2002. 相似文献
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Six nepoviruses are transmitted by five Longidorus and three Xiphinema species in Europe. The virus and vector associations are: L. apulus and L. fasciatus with artichoke Italian latent nepovirus, L. attenuatus and L. elongatus with tomato black ring nepovirus, L. elongatus and L. macrosoma with raspberry ringspot nepovirus, X. diversicaudatum with arabis mosaic and strawberry latent ringspot nepoviruses and X. index and X. italiae with grapevine fanleaf nepovirus. The distributions of the viruses and their vectors in Europe and the Mediterranean basin are described. The following points are commented: presence of non-indigenous nematode-transmitted nepoviruses in Europe; occurrence of vector nematodes in European countries in the absence of their associated nepoviruses; the possible vectors of serologically distinct strains of grapevine fanleaf nepovirus. 相似文献
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The purpose of this study was a comprehensive survey on the occurrence and distribution of the viruses affecting almond nursery trees in western Anatolia (Turkey). Three provinces (Aydýn, Balýkesir and Ýzmir) which are well known for their state and commercial nursery plantations of almond (mixed with other stone fruits) were visited and 19 300 and 19 000 almond nursery trees were surveyed in 1992 and 1993, and 56 and 40 leaf samples collected, respectively. The main viral symptoms were chlorotic and necrotic spots, calico, mosaic, narrowing and deformation of the leaves; gummosis, weak growth, dwarfing and short internodes of the trees. DAS-ELISA was used to detect prunus necrotic ringspot ilarvirus (PNRSV), prune dwarf ilarvirus (PDV), raspberry ringspot nepovirus (RRV), peach rosette mosaic nepovirus (PRMV) and tomato ringspot nepovirus (ToRSV) in almond nursery trees in west Anatolia. PNRSV was found to be the most widespread according to ELISA. PDV was the second most important virus, and some samples were infected with both ilarviruses. Fewer samples were found infected with PRMV, RRV and only a few samples were infected with ToRSV. 相似文献
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Masamichi Isogai Koji Ishii Seisaku Umemoto Manabu Watanabe Nobuyuki Yoshikawa 《Journal of General Plant Pathology》2009,75(2):140-143
Virus-like symptoms—red ringspots on stems and leaves, circular blotches or pale spots on fruit—were found on commercial highbush
blueberry (Vaccinium
corymbosum) cultivars Blueray, Weymouth, Duke and Sierra in Japan. In PCR testing, single DNA fragments were amplified from total nucleic
acid samples of the diseased blueberry bushes using primers specific to Blueberry red ringspot virus (BRRV). Sequencing analysis of the amplified products revealed 95.7–97.7% nucleotide sequence identity with the BRRV genome.
This paper is the first report of blueberry red ringspot disease caused by BRRV in Japan.
The nucleotide sequence data reported in this paper are available in the GenBank/EMBL/DDBJ database as accessions AB469884
to AB469893 for BRRV isolates from Japan. 相似文献
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烟草环斑病毒(Tobacco ringspot virus,TRSV)和番茄环斑病毒(Tomato ringspot virus,ToRSV)是我国禁止入境的检疫性有害生物。采用半巢式RT-PCR方法对这两种病毒进行了检测,用Trizol快速提取病毒总RNA,并根据TRSV和ToRSV的外壳蛋白基因设计特异性引物,经过第一轮RT- PCR和第二轮半巢式PCR扩增,TRSV样本分别得到359 bp和206 bp特异性片段,ToRSV样本分别得到340 bp和219 bp特异性片段。半巢式RT-PCR扩增产物的测序表明,TRSV产物序列与GenBank中登录的外壳蛋白基因存在88%～97%的同源性,ToRSV产物序列与GenBank中登录的外壳蛋白基因存在96%～100%的同源性。研究显示,DAS-ELISA与RT-PCR的检测灵敏度相近,而半巢式RT-PCR的检测灵敏度比这两种方法高出10~3倍以上。 相似文献
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E. Choueiri C. Haddad N. Abou Ghanem-Sabanadzovic F. Jreijiri S. Issa A. T. Saad B. Di Terlizzi V. Savino 《EPPO Bulletin》2001,31(4):493-497
Field surveys were carried out in the main peach-growing areas of Lebanon to assess the presence and distribution of viruses and viroids in commercial orchards. Field inspections were made in spring and summer 2000 to observe symptoms of virus and viroid diseases respectively. In total, 950 trees in 95 commercial plantings from three different regions of Lebanon (Bekaa Valley, Mount Lebanon and north Lebanon) were surveyed and sampled. Immunoenzymatic tests (DAS-ELISA) were used to ascertain the presence of the following: Prunus necrotic ring spot ilarvirus (PNRSV), Prune dwarf ilarvirus (PDV), Apple mosaic ilarvirus (ApMV), Apple chlorotic leaf spot trichovirus (ACLSV), Plum pox potyvirus (PPV), Tomato ringspot nepovirus (ToRSV) and Strawberry latent ringspot nepovirus (SLRSV). Peach latent mosaic pelamoviroid (PLMVd) and Hop stunt hostuviroid (HSVd) were identified by molecular hybridization. About 25% of the tested samples were infected by one or more viruses. In particular, the prevailing virus was PNRSV (61.2% of infection), followed by ACLSV (27.1%), PDV (22.4%) and ApMV (2.1%). Mixed infections were about 13%. ToRSV, SLRSV and PPV were not found. HSVd was apparently absent, whereas PLMVd was identified in 34% of the samples examined. This viroid prevailed in certain areas of Mount Lebanon in both native and foreign cultivars. 相似文献