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1.
根据鉴别寄主反应，粒体形态及血清学等试验从美国进口悬钩子上的病毒分离物鉴定为烟草花叶病毒。摩擦接种普通烟表现为花叶，在心叶烟，昆诺阿黎，苋色黎接种叶上表现为局部枯斑，电镜粒体为杆状，约３００ｎｍ，免疫电镜中用ＴＭＶ抗血清诱捕修饰法制片铜网能捕获大量病毒粒体，并在粒体表面有一层抗体外套，ＰＡＳ－ＥＬＩＳＡ中与ＴＭＶ抗血清也呈阳性反应。悬钩子中病毒浓度很低，只有经浓缩后才能对病毒进行鉴定。文中还讨论了 相似文献
2.
百合黄瓜花叶病毒及其检测 总被引：3，自引：0，他引：3
应用酶联免疫吸附法(ELISA)检测了浙江丽水和杭州的东方百合、亚洲百合等栽培品种上黄瓜花叶病毒(CMV),64个田间样品中有26个带毒,检出率为40.6%;dsRNA检测结果与已知CMV-FQS株系的电泳条带相同,但百合组织中CMVdsRNA含量较低;电镜观察,受CMV侵染的样品中大多含有线状病毒,复合侵染率为35.9%;寄主反应测定显示从百合植株上获得的CMV株系均不能通过汁液摩擦接种侵染昆诺藜、苋色藜、普通烟、心叶烟等6科10种指示植物。 相似文献
3.
为了解湖南省马铃薯种薯质量和主要病毒病发生情况,2019年-2020年马铃薯秋作和冬作期间,对长沙、益阳、湘潭、澧临等马铃薯生产区的155个马铃薯样品,运用反转录-聚合酶链式反应(RT-PCR)和双抗体夹心酶联免疫吸附检测(DAS-ELISA)技术,筛查6种主要马铃薯病毒,包括马铃薯X病毒Potato virus X(PVX)、马铃薯Y病毒Potato virus Y(PVY)、马铃薯M病毒Potato virus M(PVM)、马铃薯S病毒Potato virus S(PVS)、马铃薯A病毒Potato virus A(PVA)、马铃薯卷叶病毒Potato leaf roll virus(PLRV)。检测结果表明:6种马铃薯病毒病在湖南均有不同程度的发生,单一和两种病毒复合感染植株占比最高,其次是3种病毒复合感染,存在极少数植株复合感染4～5种病毒病情况。在秋作马铃薯中,PVY检出率达到29.41%;PVS和PVA检出率均为27.94%;PVM、PVX、PLRV的检出率分别为20.59%、19.12%、17.65%。在冬作马铃薯中,PVX检出率最高,达到31.03%;其次是PLRV,... 相似文献
4.
应用A蛋白免疫电镜技术对几种蔬菜作物的长叶车前草花叶病毒(RMV)、烟草花叶病毒(TMV)的检测比常规的电镜负染技术至少要灵敏近10,000倍,比普通的免疫电镜技术要灵敏10~100倍,抗血清浓度稀释至400,000倍,病毒粗提液稀释到1,000,000倍或病毒提纯液浓度为2.5×10-7 mg/ml时均能观察到病毒粒子。同时由于抗体分子能特异性地结合到病毒粒子表面,能够很容易地区分不同的病毒种类。
应用此项技术作者检测了杭州地区几种常见的蔬菜病毒病,对大白菜(Brassica campestris L.),蕃茄(Lycopersicum esculentum Mill),榨菜(Brassica juncea var.tsatsai Mao),雪里(Brassicajuncea var.multiceps Tsen et Lee)等作物发病期的叶或果的浸提液用A蛋白免疫电镜进行快速检测,结果在大白菜,番茄等病株上检测到较多的病毒粒子,且大多能被RMV抗血清修饰,同时在番茄病株检测到的少量病毒粒子亦能被TMV抗血清修饰,而榨菜及雪里等病株上检测到的病毒粒子很少能被RMV及TMV抗血清所修饰,另外,病毒粒子的含量以番茄病果中为最高,要比其它几种蔬菜作物叶子中高约10~30倍。 相似文献
应用此项技术作者检测了杭州地区几种常见的蔬菜病毒病,对大白菜(Brassica campestris L.),蕃茄(Lycopersicum esculentum Mill),榨菜(Brassica juncea var.tsatsai Mao),雪里(Brassicajuncea var.multiceps Tsen et Lee)等作物发病期的叶或果的浸提液用A蛋白免疫电镜进行快速检测,结果在大白菜,番茄等病株上检测到较多的病毒粒子,且大多能被RMV抗血清修饰,同时在番茄病株检测到的少量病毒粒子亦能被TMV抗血清修饰,而榨菜及雪里等病株上检测到的病毒粒子很少能被RMV及TMV抗血清所修饰,另外,病毒粒子的含量以番茄病果中为最高,要比其它几种蔬菜作物叶子中高约10~30倍。 相似文献
5.
在国内、外首次选用金黄色葡萄球菌(Staphylococc aureus)的No.180菌株用于病毒细菌协同凝集试验(Virobacterial agglutination test简称VBA)检测了来自马铃薯的4种不同形态粒子的病毒:马铃薯X病毒(PVX)、马铃薯丫病毒(PVY)、烟草花叶病毒(TMV)和马铃薯卷叶病毒(PLRV),其灵敏度达2.7~6.1ng/ml,检出病汁液的最大稀释度达104~105(PLRV1:500),较国内、外采用的Cown I菌株的灵敏度提高5~10倍。与血清学方法相比,VBA在2~3分种内就可获得结果,无假阳性反应,其灵敏度显著高于间接酶联法和免疫电镜,而接近于A蛋白酶联法。经抗血清致敏的菌体在4℃下保存4个月,对VBA检测的灵敏度没有影响。用VBA对采自田间的93个马铃薯病样进行检测,它们大多受2~3种病毒(PVX、PVY及PLRV)的复合侵染;和用间接酶联法及鉴别寄主检测的结果趋向一致。室内和田间试验均表明VBA灵敏度高、特异性强、快速简便和经济,尤其适合在基层单位中推广应用。 相似文献
6.
在国内、外首次选用金黄色葡萄球菌(Staphylococc aureus)的No.180菌株用于病毒细菌协同凝集试验(Virobacterial agglutination test简称VBA)检测了来自马铃薯的4种不同形态粒子的病毒:马铃薯X病毒(PVX)、马铃薯丫病毒(PVY)、烟草花叶病毒(TMV)和马铃薯卷叶病毒(PLRV),其灵敏度达2.7～6.1ng/ml,检出病汁液的最大稀释度达10~4～10~5(PLRV1:500),较国内、外采用的Cown I菌株的灵敏度提高5～10倍。与血清学方法相比,VBA在2～3分种内就可获得结果,无假阳性反应,其灵敏度显著高于间接酶联法和免疫电镜,而接近于A蛋白酶联法。经抗血清致敏的菌体在4℃下保存4个月,对VBA检测的灵敏度没有影响。用VBA对采自田间的93个马铃薯病样进行检测,它们大多受2～3种病毒(PVX、PVY及PLRV)的复合侵染;和用间接酶联法及鉴别寄主检测的结果趋向一致。室内和田间试验均表明VBA灵敏度高、特异性强、快速简便和经济,尤其适合在基层单位中推广应用。 相似文献
7.
为探究利用CRISPR/Cas13a系统获得抗马铃薯X病毒（potato virus X,PVX）马铃薯的可行性,通过设计靶向PVX中TGBp1基因的小向导RNA（small guide RNA,sgRNA）,构建CRISPR/Cas13a基因编辑载体,以马铃薯栽培种Désirée为受体材料进行稳定遗传转化,并通过机械摩擦接种法鉴定转基因植株对PVX的抗性。结果显示,成功构建了靶向PVX的PVX-Cas13a载体,并获得了表达Cas13a的转基因马铃薯植株,对其中3个高表达Cas13a的株系进行PVX抗性鉴定,发现在接种PVX 10~20 d后,转基因植株系统叶上无明显发病症状,且PVX积累量均显著低于未接种PVX对照。表明靶向PVX的CRISPR/Cas13a系统能够有效抑制该病毒的积累,这为马铃薯抗PVX育种提供了一条有效策略。 相似文献
8.
我国马铃薯病毒主要有马铃薯Y病毒(PVY)、马铃薯X病毒(PVX)、马铃薯S病毒(PVS)、马铃薯卷叶病毒(PLRV),常发生复合侵染。根据GenBank中4种马铃薯病毒的外壳蛋白(coat protein,CP)基因全长设计引物,通过RT-PCR扩增得到4种病毒CP基因全长片段,测序结果显示序列同源性96%以上;针对4种病毒CP基因的保守序列分别设计引物,在一个PCR体系中同步对4种病毒进行扩增,得到421、202、516、330bp的特异性条带,优化建立了能同步检测PVY、PVX、PVS和PLRV的多重RT-PCR检测体系。检测结果证明优化后的多重RT-PCR体系能在田间样品中快速、高效地检测出4种病毒。 相似文献
9.
应用Ａ蛋白免疫电镜技术对几种蔬菜作物的长叶车前草花叶病毒（ＲＭＶ）、烟草花叶病毒（ＴＭＶ）的检测比常规的电镜负染技术至少要灵敏近１０，０００倍，比普通的免疫电镜技术要灵敏１０～１００倍，抗血清浓度稀释至４００，０００倍，病毒粗提液稀释到１，０００，０００倍或病毒提纯液浓度为２．５×１０－７ｍｇ／ｍｌ时均能观察到病毒粒子。同时由于抗体分子能特异性地结合到病毒粒子表面，能够很容易地区分不同的病毒种类。应用此项技术作者检测了杭州地区几种常见的蔬菜病毒病，对大白菜（ＢｒａｓｉｃａｃａｍｐｅｓｔｒｉｓＬ．），蕃茄（ＬｙｃｏｐｅｒｓｉｃｕｍｅｓｃｕｌｅｎｔｕｍＭｉｌ），榨菜（Ｂｒａｓｉｃａｊｕｎｃｅａｖａｒ．ｔｓａｔｓａｉＭａｏ），雪里（Ｂｒａｓｉｃａｊｕｎｃｅａｖａｒ．ｍｕｌｔｉｃｅｐｓＴｓｅｎｅｔＬｅｅ）等作物发病期的叶或果的浸提液用Ａ蛋白免疫电镜进行快速检测，结果在大白菜，番茄等病株上检测到较多的病毒粒子，且大多能被ＲＭＶ抗血清修饰，同时在番茄病株检测到的少量病毒粒子亦能被ＴＭＶ抗血清修饰，而榨菜及雪里等病株上检测到的病毒粒子很少能被ＲＭＶ及ＴＭＶ抗血清所修饰，另外，病毒粒子的含量以番茄病果中为最高，要比 相似文献
10.
11.
Keiji Takami Hiroyuki Okubo Shuichi Yamasaki Minoru Takeshita Yoichi Takanami 《Journal of General Plant Pathology》2006,72(6):391-392
Momordica charantia L. plants systemically infected with Cucumber mosaic virus (CMV) were found in Oita Prefecture. The virus isolated from the host plant was characterized by biological, serological,
and molecular biological methods. The purified virus was used to mechanically inoculate the host and produced green mottle,
green mosaic, and/or chlorotic spots in the noninoculated upper leaves of the host. The virus was identified as an isolate
of CMV containing genomic RNA3 derived from subgroup IA by several lines of evidence based on electron microscopy, serological
detection, host range, symptoms, and the entire nucleotide sequence of RNA3. 相似文献
12.
H. LECOQ M. RAVELONANDRO C. WIPF-SCHEIBEL M. MONSION B. RACCAH J. DUNEZ 《EPPO Bulletin》1994,24(3):555-559
Transgenic Nicotiana benthamiana plants expressing the coat protein of an aphid-transmissible strain of plum pox potyvirus (PPV-D) were infected with an aphid non-transmissible strain of another potyvirus, zucchini yellow mosaic potyvirus (ZYMV-NAT). Non-viruliferous Myzus persicae could acquire and transmit ZYMV-NAT from these plants but not from infected N. benthamiana control plants (not transformed, or transformed by the vector alone). Immunosorbent electron microscopy experiments using the decoration technique revealed that ZYMV-NAT virus particles in the infected transgenic plants expressing the PPV coat protein could be coated not only with ZYMV antibodies but also, on segments of the particles, with PPV antibodies. This suggests that aphid transmission of ZYMV-NAT occurred through heterologous encapsidation, and reveals a potential risk of releasing genetically engineered plants expressing viral coat proteins into the environment. 相似文献
13.
用聚丙烯酰胺凝胶电泳病毒衣壳蛋白快速诊断瓜类作物上的三种病毒Ⅰ 总被引：1，自引：0，他引：1
本试验以发生在我国葫芦科作物上的三种主要病毒WMV-2,CMV和SqMV为研究对象,尝试用病毒衣壳蛋白亚基的SDS——聚丙烯酰胺凝胶电泳诊断瓜类作物病毒。试验结果说明,对病株叶片或其他组织进行初步的澄清和病毒浓缩处理,可以检测出病毒衣壳蛋白亚基谱带。根据病毒衣壳蛋白亚基的分子量差异,可以区别这三种病毒。这种方法有其他常规诊断方法(包括生物学测定,电镜观察和血清学方法等)所不具有的优点。病毒浓度的检测限度在1-50μg/ml水平。 相似文献
14.
A. Alfaro‐Fernández V. Medina M. C. Córdoba‐Sellés M. I. Font J. Jornet M. C. Cebrián C. Jordá 《Plant pathology》2010,59(2):231-239
Optical and electron microscopy studies were carried out to investigate the cytopathology induced in tomato leaves infected by Tomato torrado virus (ToTV), a new picorna‐like virus associated with the ‘Torrado’ disease. Infected leaves, showing typical Torrado disease symptoms were surveyed in commercial greenhouses in the main tomato production areas of Spain. The effect of the co‐infection of ToTV with other viruses which commonly infect tomato crops was also studied. Ultra‐thin sections of ToTV‐infected tomato leaves did not show a strong cellular alteration. However, crystalline arrays of isometric virus‐like particles (VLPs) of 20–30 nm in the inclusion bodies were observed in phloem parenchyma cells of the infected tissues. Tissues co‐infected by ToTV and either Tomato chlorosis virus (ToCV) or Pepino mosaic virus (PepMV) presented more severe cellular alterations. The most deleterious consequences for tomato cells were found in triple infections of ToTV, PepMV and Tomato spotted wilt virus (TSWV), where characteristic cell wall overgrowth was distinguishable, together with a large amount of necrotic cells. 相似文献
15.
新疆哈密瓜病毒病毒源的分离和鉴定 总被引：2，自引：0，他引：2
新疆哈密瓜是驰名中外的特产,是我国重要的外销产品之一。五十年代末,六十年代初,由于病毒病的感染,使哈密瓜产量降低,含糖量下降,严重地影响了哈密瓜的产量和质量。 1980～1981年,在新疆石河子,吐鲁番和昌吉等地进行调查时,在哈密瓜和其它瓜类上采集病毒病标样19个,通过鉴别寄主,血清学和电子显微镜的鉴定,把19个标样分为两大类。一类寄主范围广,除侵染葫芦科外,还侵染豆科、茄科、藜科、菊科等多种植物,与黄瓜花叶病毒(CMV)血清发生明显沉淀反应,电镜下观察为球形粒体,确定为CMV的一个株系,另一类寄主范围狭窄,除侵染葫芦科外,还侵染豆科、藜科。电镜下形态为长线条状。与WM-V-2血清发生反应,应为西瓜花叶病毒2号(WMV-2)。WMV-2是哈密瓜病毒中发生最严重的毒源。占田间病株90％以上。 相似文献
16.
A potyvirus was isolated from Datura stramonium, Lycopersicon esculentum (tomato) and Solanum nigrum in the Yemen. It was transmitted mechanically and by Myzus persicae in a non-persistent manner. Its flexuous rod-shaped particles had a mean length of 719 nm and some of its pinwheel inclusion bodies in infected Nicotiana clevelandii leaves were unusual in that they were dichotomously branched. The virus infected various solanaceous species, but the symptoms it induced were distinct from those of pepper veinal mottle (PVMV) and potato Y viruses. Its particles were purified from N. glutinosa and their coat protein had an atypically high molecular mass a potyvirus of 41·5 kDa. They showed a distant serological relationship to those of PVMV and potato virus V in ISEM decoration tests, but did not react with antisera to particles of any other potyvirus tested. The virus has been tentatively named tomato mild mottle virus. 相似文献
17.
Nata&#x;a Mehle Maja Kova
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Maru&#x;a Pompe Novak &#x;pela Baebler Hana Kre
i
Stres Kristina Gruden Maja Ravnikar 《Physiological and Molecular Plant Pathology》2004,64(6):19
The aim of this work was to correlate the appearance of the symptoms, multiplication and spread of virus after mechanical inoculation of potato (Solanum tuberosum L.) cultivars showing different levels of susceptibility and sensitivity to Potato virus YNTN (PVYNTN). The potato cultivars used were the resistant cultivar Sante and susceptible cultivars Igor, Pentland squire and Désirée. The spread of the virus PVYNTN in infected plants was monitored using different methods: DAS-ELISA, tissue printing, immuno-serological electron microscopy and real-time PCR. In all three susceptible cultivars, the virus was detected in the inoculated leaves 4–5 days after inoculation. From there virus spread rapidly, first into the stem, then more or less simultaneously to the upper leaves and roots. Real-time PCR was shown to be very sensitive and enabled viral RNA to be detected in non-inoculated leaves of susceptible cultivar Igor earlier than other methods. Therefore, for exact studies of plant–virus interaction, a combination of methods which detect viruses on the basis of their different properties (coat protein, morphology or RNA) should be used to monitor the spread of viruses. 相似文献
18.
侵染花椰菜的芜菁花叶病毒及RT-PCR扩增外壳蛋白基因研究 总被引：4，自引：0，他引：4
从杭州市郊区及本校附近菜园地表现严重花叶症状的花椰菜上分离到一种线状病毒,接种6科31种鉴别寄主,能侵染5科22种植物。病组织超薄切片可见风轮状内含体和片层聚集状内含体。感病的芥菜叶片,用0.5mol/L磷酸钾缓冲液(pH7.5)匀浆后汁液加4% PEG-8000沉淀3h,5%~40%甘油梯度离心纯化病毒,可获得较高的产量。提纯的病毒均为一弯曲的线状颗粒,长度740nm左右。病毒提纯液于264nm处有一吸收峰,为典型的核蛋白紫外吸收。TuMV-H分离株抗血清包被的铜网能大量吸附病毒颗粒。用特异的PCR引物扩增其外壳蛋白基因,可得-0.9Kb的片段。 相似文献
19.
Tetsuo MAOKA Shinya TSUDA Tomio USUGI Chiyoichi NODA Mabito IWASAKI 《Journal of General Plant Pathology》2002,68(1):89-93
Losses in serological reactivity of Papaya leaf distortion mosaic virus (PLDMV) were demonstrated. An antibody, IgG-papaya,
raised against PLDMV purified from papaya (Carica papaya L.) did not react with virus particles in Cucumis metuliferus leaf extracts in ELISA or SSEM-PAG (serologically specific electron microscopy using protein A-gold). In addition, IgG-papaya
and IgG- Cucumis raised against PLDMV purified from C. metuliferus did not react with virus particles in papaya leaf extracts after western blotting. From results of electrophoresis, the coat
protein (CP) of PLDMV purified from papaya had degraded and migrated in two bands. Similar degradation was also observed when
virus purified from C. metuliferus was treated with papain. These results indicated that the CP of PLDMV purified from papaya was degraded during the purification
process by papain in the host plant. IgG-papaya was reactive to papain-degraded CP, while IgG-Cucumiswas reactive to both intact and degraded CP. Modified serological methods using antipain (a protease inhibitor) or papain
were established to detect PLDMV.
Received 16 February 2001/ Accepted in revised form 26 September 2001 相似文献
20.
B. Sripathi Rao 《国际虫害防治杂志》2013,59(4):461-463
Abstract An intensive survey of cultivated plants throughout the island of Montserrat resulted in the identification of a number of viruses. Squash mosaic virus was found in cantaloupe, pumpkin and squash. There were cases of multiple infection of pumpkin and squash by cucumber and squash mosaic viruses. Potato virus Y was recovered from pepper and tomato. Bean yellow mosaic and cowpea mosaic viruses were found in red kidney bean and string bean while cowpea mosaic virus was found in cowpea and broad bean (Jamaica bean). Other viruses that were previously identified on the basis of field symptomatology were confirmed by host range and serology. Viruses were not recovered from sea island cotton or from sweet potato, although field symptoms on the sweet potato are similar to those caused by sweet potato mosaic virus. The leaf tissues of crop plants were inoculated to a wide range of indicator plants. Some of those that reacted systemically were examined by electron microscopy for virus particles and the sap extracted from them was used in serodiagnostic assays for virus identification. This is the first attempt to identify positively the viruses of crop plants on the island. 相似文献