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1.
Hammond J 《Phytopathology》1998,88(9):965-971
ABSTRACT Antisera to the cytoplasmic inclusion proteins (CIPs) of bean yellow mosaic (BYMV), clover yellow vein (ClYVV), turnip mosaic (TuMV), sweet potato feathery mottle (SPFMV), and maize dwarf mosaic (MDMV) potyviruses were used to examine the relationships between the CIPs of 18 potyviruses. The antisera to CIPs of BYMV, ClYVV, TuMV, and SPFMV cross-reacted to most or all of the purified CIPs tested in western blot assays. The MDMV CIP antiserum reacted significantly only to the MDMV and sorghum mosaic virus CIPs. Reactivity of antisera to CIPs of dicot-infecting viruses was generally higher with CIPs of other dicot-infecting than with monocot-infecting potyvirus CIPs. Analysis of amino acid sequences of the CI genes of 11 well-characterized potyviruses suggested that epitopes specific for individual potyviruses are primarily in the C-terminal domains of the CIP, whereas epitopes shared among different viruses are clustered in the N-terminal domains. The most highly conserved predicted epitope overlaps the nucleotide binding motif of the N-terminal helicase domain of the CIP. Antibodies to this domain will probably be present in antisera to any potyvirus CIP and contribute to the cross-reactivity observed. Differences in the C-terminal domains may correlate with interactions between the CIP and coat protein necessary for replication and movement.  相似文献   

2.
An isolate of the poty virus turnip mosaic virus (TuMV-Ab) showing severe mosaic symptoms inAbutilon theophrasti from Piedmont (northwestern Italy) in 1993, has been found to be of an unusual pathotype and serotype. The isolate was easily transmitted byAphyis gossiypii and Myzus persicae and was not seed-transmitted inA. Theophrasti. The host range of TuMV-Ab was different from that of another Piedmont isolate of TuMV fromAlliaria officinalis and from a TuMV isolate fromBrassica napus. TuMV-Ab was characterized using the reactions on the fourB. Napus lines S4, R4, 165 and S1 as the rare pathotype 7, found only once previously in Europe. Tests with polyclonal antisera indicated that TuMV-Ab was only distantly related to the two other TuMV isolates. Serological characterization with a panel of 30 monoclonal antibodies showed that TuMV-Ab belonged to one of the less common serotypes (JPN).  相似文献   

3.
ABSTRACT Using a mixture of isolates of Cucumber mosaic virus (CMV) from subgroups I and II as immunogens, 20 mouse hybridoma cell lines secreting monoclonal antibodies were produced. A reliable method for efficient detection and accurate subgrouping of CMV isolates has been developed. Tests with 12 well-characterized strains of CMV and other cucumoviruses demonstrated the presence of epitopes that were virus and subgroup specific. Analyses of 109 accessions of CMV isolates collected from various parts of the world revealed 70% were subgroup I, with 20% identified as subgroup II. Seven isolates (6%) did not react with group-specific antibodies but did react with antibodies that recognized all CMV isolates. Differential reactions among isolates suggested a total of 10 epi-topes were recognized. The antigenic diversity among subgroup II CMVs was greater than for the subgroup I isolates, even though fewer subgroup II isolates were tested.  相似文献   

4.
Host Range and Characterization of Sunflower mosaic virus   总被引:1,自引:0,他引:1  
ABSTRACT Sunflower mosaic is caused by a putative member of the family Potyviridae. Sunflower mosaic virus (SuMV) was characterized in terms of host range, physical and biological characteristics, and partial nucleotide and amino acid sequence. Cells infected with SuMV had cytoplasmic inclusion bodies typical of potyviruses. Of 74 genera tested, only species in Helianthus, Sanvitalia, and Zinnia, all Asteraceae, were systemic hosts. Commercial sunflower hybrids from the United States, Europe, and South Africa were all equally susceptible. The mean length of purified particles is approximately 723 nm. The virus was transmitted by Myzus persicae and Capitphorus elaegni, and also was seedborne in at least one sunflower cultivar. Indirect enzyme-linked immunosorbent assay tests with a broad-spectrum potyvirus monoclonal antibody were strongly positive. SuMV-specific polyclonal antisera recognized SuMV and, to a lesser extent, Tobacco etch virus (TEV). When tested against a panel of 31 potyvirus-differentiating monoclonal antibodies, SuMV was distinct from any potyvirus previously tested. SuMV shared four epitopes with TEV, but had a reaction profile more similar to Tulip breaking virus (TBV). SuMV did not possess epitopes unique only to TBV. The predicted coat protein had a molecular weight of 30.5 kDa. The 3' end of the virus genome was cloned and sequenced. Phylogenetic analysis of the coat protein amino acid sequence revealed that SuMV is a distinct species within the family Potyviridae, most closely related to TEV.  相似文献   

5.
Serological variability of isolates of rice yellow mottle virus (RYMV) collected in Côte d'Ivoire was assessed by immunological tests with polyclonal and monoclonal antibodies (MAbs). Two serotypes (named S1 and S2) were distinguished. The S1 isolates had common epitopes which were absent in S2 isolates, whereas they lacked epitopes shared by S2 isolates. There was no evidence of S1 and S2 mixtures, although S1 and S2 isolates were sometimes found in nearby sites. Serotype S2 was more prevalent in Côte d'Ivoire than S1, and was in a large majority in the centre and the south of the country. By contrast, S1 occurred more widely in the north. S1 isolates were also found in neighbouring countries at the north of Côte d'Ivoire. In tests with monoclonal antibodies, three additional serotypes were found, one in West-Africa and two in East-Africa. Using the primers developed against an S2 isolate from Côte d'Ivoire, all S2 but not the S1 isolates were transcribed and amplified by RT-PCR, and another set of primers was developed to amplify S1 isolates. S1 and S2 have different biological properties, and competition between isolates of the two strains was apparent resulting in S2 dominance over S1. This was assessed using S1 and S2 strain specific MAbs, and it occurred whatever the pattern of inoculation or the rice variety tested. Differences in pathogenicity and virus titre did not account for strain competition, as there was no relation between symptom severity, virus content and serotype of the isolates in Oryza sativa indica cultivars.  相似文献   

6.
豌豆病毒病病原研究   总被引:2,自引:0,他引:2  
 1986年至1990年,从豌豆田中采集了150余份病毒病样本,鉴定出蚕豆萎蔫病毒(BB-WV)、芜菁花叶病毒(TuMV)、马铃薯Y病毒组分离物、黄瓜花叶病毒(CMV)、莴苣花叶病毒(LMV)、大豆花叶病毒(SMV)、豌豆花叶病毒(PMV)、菜豆黄花叶病毒(BYMV)和苜蓿花叶病毒(AMV)等9种病毒。样本中,BBWV所占的比例最高,达59.2%,其次为CMV,占15.5%。BBWV常与CMV复合侵染豌豆,LMV发生也较普遍。田间调查表明,豌豆病毒病发病率因种植地区及品种不同而有差异,平均发病率为12.4%。  相似文献   

7.
ABSTRACT Eight turnip mosaic potyvirus (TuMV) isolates from the Campania region of Italy were characterized. Experiments based on host range and symptomatology indicated that the isolates were biologically different. In addition, the isolates, with the exception of ITA1 and ITA3, were distinguished from each other by using a combination of monoclonal antibodies recognizing the coat protein. Single-strand conformation polymorphism (SSCP) analysis of the coat protein gene revealed that each isolate produced a specific SSCP profile, except for isolates ITA1 and ITA3. This study indicates that (i) even in a small geographical region, there is a great deal of variation in TuMV isolates; (ii) the use of a set of four differential hosts does not always specify the same pathotype in different environments; (iii) the TuMV isolates with the same pathotype on Brassica napus test lines can still differ in host range, symptoms, serology, and SSCP; and (iv) there was perfect correlation between the panel of antibodies and SSCP in differentiating among the isolates; ITA1 and ITA3 were indistinguishable by either assay.  相似文献   

8.
Twelve monoclonal antibodies (MAbs) were obtained by immunizing mice with a French isolate (F1) of wheat yellow mosaic virus (WYMV). Three of these (3D12, 2C1, 6C3) belong to the IgM class and the nine others to the IgG class (3D8, 3H1, 2B8, 1F2, 3C10, 4F12, 3H9, 1G5, 54). In antigen-coated plate (ACP) ELISA and indirect double antibody sandwich (IDAS) ELISA, all MAbs recognize the WYMV (F1) both in the form of purified particles and in wheat leaf extract. The analysis of numerous French isolates of WYMV shows a variable reactivity with MAbs 3D8, 3H1, 2B8, 3C10, 3H9 and 1G5 in IDAS — and ACP-ELISA. The Japanese isolate of WYMV and United States isolates of wheat spindle streak mosaic virus (WSSMV) were detected in IDAS- and ACP-ELISA by ten of the MAbs tested showing that the wheat bymoviruses originating from the three locations share a high epitopic homology. French isolates of barley yellow mosaic virus (BaYMV; pathotypes 1 and 2) were only detected in ACP-ELISA with MAbs 6C3, 3D8, 3H1 and 2B8 whereas the two Japanese strains (I-1, II-1) of MaYMV were recognized with these and also with that of 3C10. In IDAS-ELISA, the two Japanese strains were clearly detected by MAbs, 6C3, 3D8, 3H1, 1F2, 3C10 and 1G5 and the British and Belgian (pathotype 2) isolates only by that of 6C3. Only the Japanese strain of BaYMV, 1-1 could be detected with MAb 3H9 in this ELISA system.  相似文献   

9.
为建立南方菜豆花叶病毒(southern bean mosaic virus,SBMV)的快速、简便、高通量检测技术,加强该病毒的口岸检验检疫,以提纯的SBMV粒子为免疫原免疫BALB/C小鼠,利用杂交瘤技术获得3株杂交瘤细胞株19C3、19H9和20G4,其分泌的SBMV腹水单抗效价均达到10-7,且3个单抗与感染SBMV大豆叶片组织粗提液有强烈的特异性免疫反应,而不与感染南方豇豆花叶病毒(southern cowpea mosaic virus,SCPMV)的豇豆、健康的大豆、毛豆、豌豆、蚕豆和菜豆叶片组织粗提液发生免疫反应。以制备的单抗为核心,建立了检测植物中SBMV的ACP-ELISA和dotELISA两种血清学方法。3个单抗中19H9单抗的检测灵敏度最高,以其建立的ACP-ELISA和dot-ELISA方法检测大豆病叶粗提液的灵敏度分别达到1∶163 840和1∶10 240稀释浓度。利用建立的dot-ELISA方法可从上海口岸截获的大豆种子中检测出SBMV,且该检测结果得到RT-PCR方法验证。表明制备的SBMV单抗及建立的SBMV血清学检测技术可有效用于我国SBMV的口岸检验检疫。  相似文献   

10.
侵染蚕豆的芜菁花叶病毒的鉴定与提纯   总被引:1,自引:0,他引:1  
 在江苏浙江等地的蚕豆苗上出现的红褐色环斑或白色斑驳症状.经鉴定是芜菁花叶病毒侵害的结果.在病组织中的病毒粒体为弯杆状.长度750-800nm。摩擦接种在苋色藜上,接种叶上出现大块黄色枯斑,直径约3毫米,顶叶系统感染,在克氏烟、芜菁、油菜上为系统花叶.在蚕豆苗上的症状,视品种和环境条件而异,在有限生长的品种上多为红褐色环斑.在启东蚕豆上多为白色斑驳,豆荚上也可出现变色斑。对病毒的提纯方法作了改进,用冻融的20%蔗糖溶液可代替常规的10-40%蔗糖梯度液,精提纯的效果相同。  相似文献   

11.
Agglutination of Staphylococcus aureus bacteria particles, conjugated with specific virus antibodies, has been used to identify a wide range of plant viruses in crude sap extracts. The test distinguishes between seven different potyviruses in homologous and heterologous reactions, but does not distinguish different strains of bean yellow mosaic virus. The sensitivity of the virobacterial agglutination (VBA) test compares favourably with virus detection in ISEM particle trapping and local lesion assay and is only slightly less sensitive than direct ELISA tests. The test is more sensitive and simpler to user than latex particle agglutination tests.  相似文献   

12.
Fourteen strains of cucumber mosaic virus (CMV) from Australia have been characterized by their host range and symptomatology. They were classified as subgroup I or II strains by a dot-blot molecular hybridization assay between their total viral RNAs and selected cDNAs. The strains FNY and LNy, both from the USA, were used as the subgroup I- and subgroup II-type strains, respectively. A range of serological tests was used to compare these isolates. Gel immunodiffusion tests, with standard antigens homologous to the antisera prepared against glutaraldehyde-fixed virus of 11 strains, showed that they could be divided into three serogroups on the basis of spur formation in heterologous reactions. Two of the serogroups included either subgroup I or subgroup II isolates, whereas the third serogroup consisted of only one strain (YWA) which was homologous to all the strains tested. Use of heterologous standard antigens in this test failed to show further subgrouping of the antigens. Double-antibody sandwich (DAS) ELISA using polyclonal antibodies to distinct virus strains also placed the 14 strains in the same three serogroups. When eight different monoclonal antibodies (MAbs) were used in indirect ELISA, one of them distinguished subgroup-I strains and another distinguished subgroup-II strains; the YWA strain fell into subgroup II. Other MAbs showed narrower or broader specificity. Thus both molecular hybridization with total RNA and specific MAbs may be useful for separating isolates of CMV into subgroups I and II. Spur formation using heterologous standard antigens to the antisera, as well as being more difficult to interpret, was not a reliable criterion for classification.  相似文献   

13.
Eight provinces of Iran were surveyed during 2003–2008 to find Brassicaceae reservoir weed hosts of Turnip mosaic virus (TuMV). A total of 532 weed samples were collected from plants with virus-like symptoms. The samples were tested for the presence of TuMV by enzyme-linked immunosorbent assay using specific antibodies. Among those tested, 340 samples (64%) were found to be infected with TuMV. Rapistrum rugosum, Sisymberium loeselii, S. irio and Hirschfeldia incana were identified as the Brassicaceae weed hosts of TuMV, and the former two plant species were found to be the most important weed hosts for the virus in Iran. The full-length sequences of the genomic RNAs of IRN TRa6 and IRN SS5 isolates from R. rugosum and S. loeselii were determined. No evidence of recombination was found in both isolates using different recombination-detecting programmes. Phylogenetic analyses of the weed isolates with representative isolates from the world showed that the IRN TRa6 and IRN SS5 isolates fell into an ancestral basal-Brassica group. This study shows for the first time the wide distribution and phylogenetic relationships of TuMV from weeds in the mid-Eurasia of Iran.  相似文献   

14.
从葎草中检出复合侵染的多种病毒   总被引:2,自引:0,他引:2  
采用抗原直接包被酶联免疫吸附测定法(ELISA)对采自重庆近郊的34个葎草样品进行了主要病毒种类的检测。其中马铃薯Y病毒(Potato virus Y, PVY)的侵染最普遍,其阳性检出率达44.12%;马铃薯X病毒(Potato virus X, PVX)的阳性检出率最低,仅为26.47%,其余5种病毒,烟草花叶病毒(Tobacco mosaic virus, TMV)、黄瓜花叶病毒(Cucumber mosaic virus, CMV)、番茄花叶病毒(Tomato mosaic virus, ToMV)、芜菁花叶病毒(Turnip mosaic virus, TuMV)及蚕豆萎蔫病毒2号(Broad bean wilt virus 2, BBWV-2)的阳性检出率均为35.29%。葎草样品受多种病毒的复合侵染现象非常严重,15个阳性样品中病毒复合侵染率为80%,其中75%的样品检测到7种病毒复合侵染。  相似文献   

15.
Pathotype-specific and broad-spectrum resistance to turnip mosaic virus (TuMV) have been identified in the diploid A genome brassica species Brassica rapa. The pathotype-specific resistance is effective against pathotype 1 isolates of TuMV, which are the most common in Europe. It is almost identical in its specificity to that of a mapped resistance gene (TuRB01) present in the A genome of the amphidiploid species Brassica napus. A mutant of a pathotype 1 isolate of TuMV (UK 1M) that is able to overcome TuRB01 also overcame the B. rapa resistance. This, combined with the fact that a single-nucleotide mutation in the cylindrical inclusion gene of TuMV that has been shown to induce a change from avirulence to virulence against TuRB01, had an identical effect on the B. rapa resistance, suggest that the two resistances are conditioned by the same gene. A second source of resistance in B. rapa prevented systemic spread of all TuMV isolates tested. A third source of resistance that appears to provide immunity to, or severely restrict replication of most isolates of TuMV has been characterised. This resistance source also prevented systemic spread of all TuMV isolates tested. Prior to this study, no resistance to pathotype 4 or pathotype 12 isolates of TuMV had ever been identified. For each of these three resistance sources, plant lines that are not segregating for some of the resistance phenotypes and that are presumably homozygous for the genes controlling these phenotypes have been generated. Strategies for further characterising and deploying these resistances in different Brassica species are described.  相似文献   

16.
为调查我国太子参主产区病毒病发生情况,使用RT-PCR法对采自福建省柘荣县,贵州省施秉县、丹寨县和安徽省宣城市的71份具有典型病毒病症状的太子参样品进行检测,并根据CP氨基酸序列对不同地区病毒进行系统进化分析。RT-PCR结果显示,65份样品检出病毒,检出率为91.55%,其中,56份样品检出芜菁花叶病毒Turnip mosaic virus(TuMV),49份样品检出蚕豆萎蔫病毒2号Broad bean wilt virus 2(BBWV2),9份样品检测出黄瓜花叶病毒Cucumber mosaic virus(CMV),检出率分别为78.87%、69.01%和12.68%。未检测到烟草花叶病毒Tobacco mosaic virus(TMV)。序列和系统发育进化分析显示,本研究获得的TuMV、BBWV2和CMV与NCBI数据库中的序列相似度较高,核苷酸相似度分别为98.05%~98.98%、93.61%~94.25%和98.02%~99.17%,氨基酸序列相似度分别为:96.14%~97.47%、97.33%~98.50%和98.00%~100%,分别属于World-B组、Ⅰ-a亚...  相似文献   

17.
Cross reaction often occurs in serological detection for potyviruses that can lead to false positive results. Sequence alignment revealed that the N-terminal sequences of coat proteins (CP) are highly variable among potyviruses. Based on this finding, the preparation of a specific antibody against potyviruses is described here increasing the efficiency of detection and identification. A case study of Turnip mosaic virus (TuMV): a specific fragment “CP-50” (1–50 aa of CP) was obtained by prokaryotic expression and used for producing the polyclonal antibody. Test of specificity indicated that the prepared antibody “Anti-CP-50” strongly reacted with TuMV and is appropriate as a specific anti-TuMV antibody.  相似文献   

18.
Bhat  Varma  Pappu  Rajamannar  Jain  & Praveen 《Plant pathology》1999,48(5):648-654
A potyvirus (eggplant mottle virus, EMoV) causing mosaic mottling in eggplant ( Solanum melongena ) was characterized on the basis of biological, serological and partial nucleotide sequence properties. EMoV infected Chenopodium amaranticolor and members of the Solanaceae. Polyclonal antiserum against EMoV showed antigenic relationship with henbane mosaic potyvirus (HMV) and potato Y potyvirus (PVY). Virus-specific antibodies directed to the N-terminal region of EMoV cross-reacted only with PVY. Determination and comparison of nucleotide sequence of the coat protein (CP) and the 3'-untranslated region (UTR) of EMoV with other potyviruses showed that the level of homology was highest with PVY isolates. Comparative sequence analyses of the CP amino acid and 3'-UTR sequences with distinct PVY isolates placed EMoV within the PVYO subgroup.  相似文献   

19.
Four mouse monoclonal antibodies (MAbs) against potato virus Y (PVY) were produced. MAb 4C1 reacted with four isolates of PVYNTN and only very weakly with one isolate of the necrotic strain of PVY (PVYN). It did not react with other isolates of the ordinary strain of PVY tested. MAb 2C9 reacted with all isolates tested and can be used to produce a specific diagnostic kit for routine PVY detection. Other MAbs had different specificities and reacted with isolates of various strains of PVY. MAbs did not react with seven other members of the Potyvirus group including potato virus A. A MAb-based ELISA, using MAb 4C1, was devised and shown to detect PVYNTN specifically.  相似文献   

20.
Potyviral nuclear inclusion b protein (NIb), the RNA-dependent RNA polymerase, contains three highly conserved regions. Peptides corresponding to these regions were synthesised and used for immunisation. A panel of monoclonal antibodies was obtained. Most of the MAbs reacted with the peptides and a recombinant NIb of PVY in PTA-ELISA. Two of them specifically detected native NIb of potato A, potato V, potato Y, plum pox and turnip mosaic potyviruses in extracts of infected plants in Western blots. Time course experiments revealed that NIb protein can be first detected on the fifth day after infection.  相似文献   

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