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1.
以采自河南的真菌传小麦花叶病毒(FWMV-C)为材料,抽提病毒RNA,合成互补DNA(cDNA)。对杂交筛选所得cDNA克隆进行亚克隆及序列分析,结果表明,亚克隆pGSI含有一个长度为891个核苷酸的不完整开放阅读框架(ORF)和长度为258个核苷酸的3'末端非编码区(NTR),并带有Poly(A)尾序。此段序列与大麦黄花叶病毒(BaYMV)及法国报道的一种小麦梭条花叶病毒(WSSMV-F)的RNA-13'末端分别具有67.6%和69.9%的同源性。由所测序列编码区(1-891nt)可推导产生296个氢基酸,并与WSSMV-F及BaYMV外壳蛋白氨基酸序列分别具有75.9%和71%的同源性。此结果表明,FWMV-C为另外一种不同于WSSMV-F的大麦黄花叶病毒组(Baymovirus)病毒,所测基因组片段应为RNA-13'末端序列,其中可能包括了病毒全长外壳蛋白编码区域。 相似文献
2.
为明确何杰金病(Hodgkin’s disease,HD)瘤细胞中是否存在人巨细胞病毒(human cytomegalovirus,HCMV)感染,采用显微切割技术结合PCR方法检测HD组织及其瘤细胞Hodgkin/Reed-Sternberg(H/RS)中的HCMV核酸;采用免疫组织化学催化信号扩增(catalysed signal amplification,CSA)法检测HD中HCMV的立即 相似文献
3.
本文对延安市150例男性高血粘滞综合证患者的卡森粘度(CassonViscosity,CV、ηc)、卡森屈服值(Cassonyieldstress,CY,Tc)与另外十四项血流变指标的相关性进行了研究,36例男健康人作对照,以初步观察CV、CY的内在规律与临床意义,结果表明病例组CV与ηa(230S-1)、HRV、TK、IR呈正相关(P<0.01或<0.005);CY与ηa(230S、46S、5.75n)、MRV、LRV、AI呈正相关(P<0.01)与ESR负相关;CV与ηa(5.75S)、MRV、LRV、AI、Fib,CY呈负相关(P<0.01);对照组例数少,相关性差,待后观察。 相似文献
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抗芜菁花叶病毒转基因甘蓝型油菜的研究 总被引:31,自引:0,他引:31
以子叶柄为材料,建立了甘蓝型油菜(BrassicanapusL.)双低品种的再生体系。通过子叶柄与农杆菌(AgrobacteriumtumefaciensLBA4404)共培养,将表达载体pBTu中芜菁花叶病毒外壳蛋白(TuMV-CP)基因以整合方式导入甘蓝型油菜,然后用卡那霉素进行筛选,获得了油菜再生植株。经PCR特异性扩增、点杂交和Southern印迹分析,证明再生植株基因组DNA中整合了TuMV-CP基因。攻毒实验表明,有TuMV-CP基因插入的工程油菜对TuMV均有不同程度的抗性。 相似文献
6.
竹节花黄斑驳病毒启动了在棉花维管组织中是一个强启动子 总被引:1,自引:0,他引:1
用竹节花黄斑驳病毒(Commelina Yellow Mottle Virus,CoYMV)启动子-gus嵌合基因和CaMV35S-gus嵌合基因通过根癌土壤杆菌(Agrobacterium tumefaciens(Smith et Townsend)Conn)LBA4404介导分别转入棉花(Gossypium hirsutum L.cv.Jingmian7,J7G),诱导再生了棉花转基因植株。G 相似文献
7.
我国真菌传线状小麦花叶病毒病病原初步鉴定为小麦黄花叶病毒(WYMV) 总被引:2,自引:0,他引:2
由禾谷多粘菌(Polymyxagraminis)传播的线状小麦花叶病毒有两种,一种是加拿大首先报道的小麦梭条斑花叶病毒(WSSMV),另一种是日本报道的小麦黄花叶病毒(WYMV)。这两种病毒粒子形态以及血清学性质非常相似,但核酸序列存在一定差异。经反转录聚合酶链反应(RTPCR)和单链构象多态性分析(SSCP),明确我国广泛发生的禾谷多粘菌传线状小麦花叶病毒都是小麦黄花叶病毒,但供试的8个分离物RNA1和RNA2序列存在差异,无一彼此完全相同 相似文献
8.
侵染半夏的两种病毒的分离纯化和初步鉴定 总被引:10,自引:0,他引:10
用自然感染的半夏(Pinelliaternata)为材料,经粗提纯后检查到一种线状病毒和一种球状病毒,用两种方法对担提纯样品中的病毒粒子进行了分离纯化。10%-70%连续甘油梯度80000g离心150分钟获得两条病毒带,经紫外吸收测定均为强的核蛋白吸收峰,病毒粒子检查分别为球状和线状病毒粒子,线状病毒经浓缩收集为均一成份.与芋花叶病毒(Dasheemmosaicvirus,DMV)抗体有强的阳性反应。粗提纯样品经0.8%琼脂糖凝胶电脉分离为一条蛋白带,该条带回收后经紫外吸收测定为核蛋白吸收峰,电镜下检查为均一的球状病毒,以TMV为对照、醋酸铀(UAC)负染后测得该球状病毒(pinelliasphericalvirus1.PsV-1)的大小为31.7nm;戊二醛固定后磷钨酸(PTA)负染测得PsV—1的大小为34.0nm。各组分经SDS-聚丙烯酸胺凝焦电泳分析测得线状病毒和球状病毒的外壳蛋白分子量分别为20KD和28KD。初步确定线状病毒为DMV.球状病毒PsV—1为侵梁天南星科半夏的一种新病毒。 相似文献
9.
汉滩病毒A9株M基因片段在重组痘苗病毒中的表达及鉴定 总被引:7,自引:0,他引:7
为发展国产化肾综合征出血热(HFRS)病毒基因工程疫苗,选择了汉滩病毒A9株M基因片段为目的基因,构建转染质粒pJSBA9M。以携带Lac基因的重组病毒为亲本,使表达载体pJSB-A9M上的M片段与痘苗病毒内的Lac基因重组,将Lac置换成A9M片段。用蓝白斑法筛选重组痘苗病毒,经PCR扩增证实A9M片段重组入痘苗病毒基因组内。重组痘苗病毒感染的Vero E6细胞,用抗糖蛋白单克隆抗体(HCO2、 相似文献
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番茄曲叶病及其血清学和PCR测定 总被引:11,自引:0,他引:11
我国曾报道的番茄病毒病有多种,其中最常见的是黄瓜花叶病毒(CMV)和烟草花叶病毒(TMV)引起的花叶病。柯冲等(1964)在大陆首次报道烟粉虱(Bemisia tabaci)传播的番茄病毒病——番茄黄顶病,此病在50~60年代曾在广州市郊流行,造成大面积减产。Green等(1984)报道台湾发生番茄黄曲叶病,此病与日本的番茄黄矮病(Tomato yellow dwarf)相似,并且与烟草曲叶病毒(TLCV)有血清学关系。印度、委内瑞拉等国也曾报道发生由烟粉虱传播的番茄曲叶病和番茄黄曲叶病。1991和1992年秋,在广西南宁市郊发现一种症状表现为植株矮缩,叶片向上向内卷曲,叶背面产生耳状或杯状增生物,对光看有时可见叶脉呈墨绿色,不结果或少结果的番茄病害。1992年秋广西农业科学院的番茄试验地发病率高达6.8%,对当地秋番茄生产构成了威胁。作者对病害症状、传播、血清学反应及PCR分析等方面与烟草曲叶病毒进行了比较研究,证实了该病的病原与烟草曲叶病毒有很高的同源性。现将研究结果简报如下。 相似文献
12.
M. M. SWANSON J. K. BROWN B. T. POULOS B. D. HARRISON 《The Annals of applied biology》1992,121(2):285-296
The relationships among fifteen isolates of whitefly-transmitted geminiviruses (WTGs) from North, Central and South America and six from other continents were assessed (a) in nucleic acid hybridisation tests with sulphonated DNA probes for eight of the viruses, and/or (b) in triple-antibody-sandwich ELISA with panels of monoclonal antibodies (MAbs) to particles of African cassava mosaic virus (ACMV) and Indian cassava mosaic virus (ICMV). Probes specific for DNA-A of four American viruses, abutilon mosaic (AbMV), bean golden mosaic (BGMV), squash leaf curl (SLCV) and tomato golden mosaic (TGMV), detected virtually all the American viruses but reacted weakly if at all with ICMV, ACMV or tomato yellow leaf curl virus from Thailand (TYLCV-T). Conversely, the probe for ACMV DNA-A did not detect any of the American viruses, and that for TYLCV-T DNA-A reacted weakly with SLCV and TGMV0020but did not detect the others. In contrast, probes specific for DNA-B of the four American viruses or ACMV detected only the homologous virus, except for slight reactions between the AbMV DNA-B probe and both chino del tomate virus (CdTV)-DNA and SLCV-DNA. However, a probe for DNA-B of bean calico mosaic virus (BCMoV) reacted weakly with BGMV-PR DNA, and a probe for DNA-B of CdTV from Mexico detected several American viruses. Six out of 17 MAbs specific for ACMV and six out of 10 MAbs specific for ICMV reacted with one or other of the 14 American virus isolates tested. Two and-ACMV MAbs reacted with all, and one anti-ACMV MAb and two anti-ICMV MAbs reacted with nearly all the American viruses, one anti-ACMV MAb reacted with about half the American viruses and six other MAbs reacted with only one or two of them. Of the American viruses, CdTV and AbMV were the least closely related to the others. The epitope profiles of BCMoV, BGMV, cotton leaf crumple virus, serrano golden mosaic virus and SLCV were virtually indistinguishable. TGMV, potato yellow mosaic virus (PYMV) and an euphorbia virus had profiles intermediate between those of the BGMV cluster and AbMV-CdTV. In general, the epitope profiles and the results of hybridisation tests with DNA-A probes show that the similarities among the American viruses are greater than those between the American viruses and the viruses from other continents; the hybridisation tests with DNA-B probes show that substantial differences exist between individual American viruses. In America, geminivirus evolution seems to have proceeded convergently from different progenitor viruses, or divergently from one ancestral form, with DNA-B diverging to a greater extent than DNA-A and its particle-protein gene. 相似文献
13.
Occurrence of whitefly-transmitted geminiviruses in crops in Burkina Faso, and their serological detection and differentiation 总被引:1,自引:0,他引:1
G KONATÉ N BARRO D FARGETTE M M SWANSON B D HARRISON 《The Annals of applied biology》1995,126(1):121-129
Whitefly-transmitted geminiviruses were found to be associated with four diseases of crop plants in Burkina Faso: cassava mosaic, okra leaf curl, tobacco leaf curl and tomato yellow leaf curl. Tomato yellow leaf curl is an economically serious disease, reaching a high incidence in March, following a peak population of the vector whitefly, Bemisia tabaci, in December. Okra leaf curl is also a problem in the small area of okra grown in the dry season but is not important in the main period of okra production in the rainy season. The geminiviruses causing these four diseases, African cassava mosaic (ACMV), okra leaf curl (OLCV), tobacco leaf curl (TobLCV) and tomato yellow leaf curl (TYLCV) viruses, were each detected in field-collected samples by triple antibody sand-wich-ELISA with cross-reacting monoclonal antibodies (MAbs) to ACMV. Epitope profiles obtained by testing each virus isolate with panels of MAbs to ACMV, OLCV and Indian cassava mosaic virus enabled four viruses to be distinguished. ACMV and OLCV had similar but distinguishable profiles. The epitope profile of TobLCV was the same as that of one form of TYLCV (which may be the same virus) and was close to the profile of TYLCV from Sardinia. The other form of TYLCV reacted with several additional MAbs and had an epitope profile close to that of TYLCV from Senegal. Only minor variations within each of these four types of epitope profile were found among geminivirus isolates from Burkina Faso. Sida acuta is a wild host of OLCV. 相似文献
14.
Serological detection and antigenic variation of two whitefly-transmitted geminiviruses: tobacco leaf curl and croton yellow vein mosaic viruses 总被引:1,自引:0,他引:1
A panel of 25 monoclonal antibodies (MAbs) raised against particles of two heterologous whitefly-transmitted geminiviruses (begomoviruses) was used in triple antibody-sandwich ELISA (TAS-ELISA) to determine the detectability and epitope profiles of 26 Indian isolates of tobacco leaf curl virus (TLCV) and 13 of croton yellow vein mosaic virus (CYVMV). Stock cultures of the two viruses had indistinguishable epitope profiles although they differ in symptomatology and particle stability. Their epitope profiles also strongly resembled those of Indian isolates of bhendi (okra) yellow vein mosaic and Indian cassava mosaic (ICMV) viruses. TLCV isolates from Andhra Pradesh, Gujarat and Karnataka States differed slightly in epitope profile: they reacted with at least eight out of 10 MAbs raised to ICMV but only one to four out of 15 MAbs raised to African cassava mosaic virus (ACMV). Virus isolates serologically indistinguishable from TLCV were detected in symptom-bearing weeds (Acanthospermum hispidum, Ageratum conyzoides, Euphorbia geniculata, Parthenium hysterophorus) found in leaf curl-affected tobacco fields and shown previously to be experimental hosts of TLCV. Indian TLCV isolates had small, consistent differences in epitope profile from Pakistani isolates but large differences from isolates from Burkina Faso, Malawi or Uganda. Isolates from the three African countries reacted with four or five of the ACMV MAbs but only one or two of the ICMV MAbs, and there were small but consistent inter-country differences. CYVMV isolates from three Indian States showed less epitope variation than did Indian isolates of TLCV. TAS-ELISA with MAb SCR 18 was a more sensitive test for detecting Indian TLCV isolates than was double antibody-sandwich ELISA with polyclonal antibodies. 相似文献
15.
B. D. HARRISON V. MUNIYAPPA M. M. SWANSON I. M. ROBERTS D. J. ROBINSON 《The Annals of applied biology》1991,118(2):299-308
Particles resembling those of geminiviruses were found by immunosorbent electron microscopy in extracts of plants infected in India with bhendi yellow vein mosaic, croton yellow vein mosaic, dolichos yellow mosaic, horsegram yellow mosaic, Indian cassava mosaic and tomato leaf curl viruses. All these viruses were transmitted by Bemisia tabaci whiteflies, all reacted with at least one out of ten monoclonal antibodies to African cassava mosaic virus (ACMV), and all reacted with a probe for ACMV DNA-1, but scarcely or not at all with a full-length probe for ACMV DNA-2. Most of the viruses were distinguished by their host ranges when transmitted by whiteflies, and the rest could be distinguished by their pattern of reactions with the panel of monoclonal antibodies. Horsegram yellow mosaic virus was distinguished from Thailand mung bean yellow mosaic virus by its lack of sap transmissibility, ability to infect Arachis hypogaea, failure to react strongly with the probe for ACMV DNA-2 and its pattern of reactions with the monoclonal antibodies. Structures resembling a ‘string of pearls’, but not geminate particles, were found in leaf extracts containing malvastrum yellow vein mosaic virus. Such extracts reacted with two of the monoclonal antibodies, suggesting that this whitefly-transmitted virus too is a geminivirus. All seven viruses from India can therefore be considered whitefly-transmitted geminiviruses. 相似文献
16.
M. M. SWANSON A. VARMA V. MUNIYAPPA B. D. HARRISON 《The Annals of applied biology》1992,120(3):425-433
Geminiviruses associated with yellow or golden mosaic diseases of legume crops in two regions of India were compared by testing their reactivity with 27 monoclonal antibodies (MAbs) prepared to the particles of African cassava mosaic (ACMV) or Indian cassava mosaic (ICMV) viruses. The viruses fell into two main groups. Group 1 comprised isolates of dolichos yellow mosaic virus; these reacted with three or four ACMV MAbs and four ICMV MAbs. Group 2 comprised isolates of horsegram yellow mosaic virus, together with isolates from blackgram, cowpea, French bean, pigeonpea, soybean, Indigofera hirsuta and probably also isolates from mungbean. These reacted with three or four ACMV MAbs but with few or no ICMV MAbs. Isolates within each group differed slightly in epitope profile, depending on the source species (Group 2) or geographical origin (Groups 1 and 2). Isolates from lima bean resembled those in Group 2 but had some antigenic differences, and their status is uncertain. The poor detectability of geminivirus isolates in mungbean may reflect a low virus concentration in this species. 相似文献
17.
The Indian cassava mosaic virus (ICMV) was transmitted by the whitefly Bemisia tabaci and sap inoculation. ICMV was purified from cassava and from systemically infected Nicotiana benthamiana leaves. Geminate particles of 16–18 × 30 nm in size were observed by electron microscopy. The particles contained a single major protein of an estimated molecular weight of 34,000. Specific antiserum trapped geminate particles from the extracts of infected cassava and N. benthamiana plants in ISEM test. The virus was detected in crude extracts of infected cassava, ceara rubber, TV. benthamiana and N. tabacum cv. Jayasri plants by ELISA. ICMV appeared serologically related to the gemini viruses of Acalypha yellow mosaic, bhendi yellow vein mosaic, Croton yellow vein mosaic, Dolichos yellow mosaic, horsegram yellow mosaic, Malvastrum yellow vein mosaic and tobacco leaf curl. 相似文献
18.
Imran Amin Basavaprabhu L Patil Rob W Briddon Shahid Mansoor Claude M Fauquet 《Virology journal》2011,8(1):1-24
Background
Whitefly-transmitted geminiviruses (begomoviruses) are a major limiting factor for the production of numerous dicotyledonous crops throughout the world. Begomoviruses differ in the number of components that make up their genomes and association with satellites, and yet they cause strikingly similar phenotypes, such as leaf curling, chlorosis and stunted plant growth. MicroRNAs (miRNAs) are small endogenous RNAs that regulate plant growth and development. The study described here was aimed at investigating the effects of each virus encoded gene on the levels of developmental miRNAs to identify common trends between distinct begomoviruses.Results
All genes encoded by four distinct begomoviruses (African cassava mosaic virus [ACMV], Cabbage leaf curl virus [CbLCuV], Tomato yellow leaf curl virus [TYLCV] and Cotton leaf curl virus/Cotton leaf curl betasatellite [CLCuV/CLCuMB]) were expressed from a Potato virus X (PVX) vector in Nicotiana benthamiana. Changes in the levels of ten miRNAs in response to the virus genes were determined by northern blotting using specific miRNA probes. For the monopartite begomoviruses (TYLCV and CLCuMV) the V2 gene product was identified as the major symptom determinant while for bipartite begomoviruses (ACMV and CbLCuV) more than one gene appears to contribute to symptoms and this is reflected in changes in miRNA levels. The phenotype induced by expression of the βC1 gene of the betasatellite CLCuMB was the most distinct and consisted of leaf curling, vein swelling, thick green veins and enations and the pattern of changes in miRNA levels was the most distinct.Conclusions
Our results have identified symptom determinants encoded by begomoviruses and show that developmental abnormalities caused by transient expression of begomovirus genes correlates with altered levels of developmental miRNAs. Additionally, all begomovirus genes were shown to modulate miRNA levels, the first time this has been shown to be the case. 相似文献19.
Identification and Molecular Characterization of a New Geminivirus Betasatellite Infecting Malvastrum coromandelianum in China
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Yafei Tang Zifu He Zhenguo Du Xiaoman She Guobing Lan 《Journal of Phytopathology》2016,164(11-12):1105-1110
The complete nucleotide sequence of a satellite molecule associated with Malvastrum leaf curl Guangdong virus (MLCuGdV) infecting M. coromandelianum plants exhibiting leaf curl symptoms in a suburb of Guangzhou, Guangdong Province of China, is described and analysed. The molecule has typical features of betasatellites, containing a single ORF in the complementary‐sense strand, an A‐rich region, the satellite‐conserved region and a stem–loop structure. Compared with the geminivirus betasatellites in GenBank database, this molecule shows the highest nucleotide sequence identity of 71.9% with Tomato leaf curl Philippine betasatellite isolate Laguna1 (ToLCPB, AB307732). Phylogenetic analysis indicates that it is more related to isolate Laguna 1 and Laguna 2 of ToLCPB. According to the proposed species demarcation threshold of betasatellites (78% nucleotide identity), it is a novel betasatellite species, for which we propose the name Malvastrum leaf curl Guangdong betasatellite (MLCuGdB). 相似文献
20.
Abutilon mosaic virus (AbMV), a bipartite geminivirus of the genus Begomovirus, has been vegetatively propagated for many years in Abutilon sellovianum in which it is strictly phloem-restricted. Using in situ hybridization and immunological analyses, the tissue tropism of AbMV in the laboratory host Nicotiana benthamiana was compared with that of two other bipartite begomoviruses, African cassava mosaic virus (ACMV) and tomato golden mosaic virus (TGMV). Analysis of the first systemically infected leaves and longitudinal sections of axillary and flower buds revealed that all three viruses are initially confined to the vascular traces, although both ACMV and TGMV are later detectable in nearly all tissue types. In contrast, AbMV remained strictly phloem-limited in this host throughout the course of infection. The ability of ACMV and TGMV to move out of N. benthamiana phloem tissues is correlated with the development of severe symptoms in comparison with the mild symptoms associated with AbMV infection. It was also demonstrated that Sida micrantha mosaic virus, a virus that is closely related to AbMV, is phloem-limited in Malva parviflora even though it induces severe leaf curl, stunting and necrosis in this host. The present data demonstrate that bipartite begomoviruses can exhibit strikingly different patterns of tissue tropism. 相似文献