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1.
In the last 10 years, more than a thousand almond trees have been analysed by the DAS-ELISA method in the Valencia region. The most frequent virus infecting unselected almond trees was prune dwarf ilarvirus (PDV) (62%), followed by prunus necrotic ringspot ilarvirus (PNRSV) (36%), apple mosaic ilarvirus (ApMV) (14%) and apple chlorotic leafspot trichovirus (ACLSV) (2%). Infection levels of selected trees were 26% for PDV, 15% for PRSV, 0% for ApMV and 5% for ACLSV.  相似文献   

2.
Twenty-four isolates of Chilli veinal mottle virus (ChiVMV) from China, India, Indonesia, Taiwan and Thailand were analysed to determine their genetic relatedness. Pathogenicity of virus isolates was confirmed by induction of systemic mosaic and/or necrotic ringspot symptoms on Capsicum annuum after mechanical inoculation. The 3' terminal sequences of the viral genomic RNA were determined. The coat protein (CP) coding regions ranged from 858 to 864 nucleotides and the 3' untranslated regions (3'UTR) from 275 to 289 nucleotides in length. All isolates had the inverted repeat sequence GUGGNNNCCAC in the 3'UTR. The DAG motif, conserved in aphid-transmitted potyviruses, was observed in all isolates. All 24 isolates were considered as belonging to ChiVMV because of their high CP amino acid and nucleotide identity (more than 94·8 and 89·5%, respectively) with the reported ChiVMV isolates including the pepper vein banding virus (PVBV), the chilli vein-banding mottle virus (CVbMV) and the CVbMV Chiengmai isolate (CVbMV-CM1). Based on phylogenetic analysis, ChiVMV isolates including all 24 isolates tested, PVBV, CVbMV and CVbMV-CM1 can be classified into three groups. In addition, a conserved region of 204 amino acids with more than 90·2% identity was identified in the C terminal of the CP gene of ChiVMV and Pepper veinal mottle virus (PVMV), and may explain the serological cross reaction between these two viruses. The conserved region may also provide useful information for developing transgenic resistance to both ChiVMV and PVMV.  相似文献   

3.
Two tospovirus isolates collected from tomato and bell pepper in Thailand were studied. The isolates induced severe necrotic mottling and/or necrotic spots and rings on the leaves and fruits of the respective plants as confirmed by back-inoculation. A polyclonal antiserum raised against its nucleocapsid (N) protein reacted only with an extract from plants infected with the homologous virus. Analysis of the nucleocapsid (N) gene sequence and its deduced amino acid sequence (Mw ∼31 kDa) showed 99% amino acid sequence homology with that of Tomato necrotic ring virus (TNRV). The nucleotide sequence of the 5΄ untranslated region and intergenic region flanking the N gene revealed typical features of the S RNA segment of tospoviruses. Mechanical inoculation of the virus on some plant species showed that most of the tested solanaceous species were susceptible to this virus. The biological, serological and molecular data presented here indicate that both isolates are identical to TNRV, a recently described tospovirus species in Thailand.  相似文献   

4.
Investigations were carried out in the main stone-fruit growing areas of Lebanon to assess the phytosanitary condition of commercial orchards. The presence of virus and virus-like diseases and their identification was ascertained through: (i) field surveys, (ii) sap transmission to herbaceous hosts, (iii) graft transmission to woody indicators; and (iv) ELISA and IEM tests. The mean infection level was 25%. ranging from 5% in apricot to 45% in cherry. The following viruses were identified: apple chlorotic leaf spot trichovirus (ACLSV), prunus necrotic ringspot (PNRSV) and prune dwarf (PDV) ilarviruses. Plum pox potyvirus (PPV), apple mosaic ilarvirus (ApMV), and six nepoviruses tested for (SLRV, TBRV, RRV, CLRV, ArMV and ToRSV) were not encountered.  相似文献   

5.
Field surveys were carried out in the main stone-fruit-growing areas of East Anatolia (Turkey) to assess the sanitary status of varietal collections, mother blocks and commercial orchards. The presence of virus and virus-like diseases was ascertained by enzyme-linked immunosorbent assay (ELISA), sap transmission to herbaceous hosts, graft transmission to peach cv. GF305 and molecular hybridization tests. A total of 1019 samples was tested by ELISA (859 apricot, 120 cherry, 21 almond and 19 peach). The sanitary status of apricot was extremely satisfactory, as the infection level was less than 0.3%. Cherry and almond, however, showed 21% and 33% infection respectively. The viruses identified were apple chlorotic leaf spot trichovirus (ACLSV), prune dwarf ilarvirus (PDV) and prunus necrotic ringspot ilarvirus (PNRSV). The commonest virus was PDV. Plum pox potyvirus (PPV), apple mosaic ilarvirus (ApMV) and the nepoviruses tomato black ring (TBRV), raspberry ringspot (RpRSV), strawberry latent ringspot (SLRV), cherry leaf roll (CLRV), arabis mosaic (ArMV) and tomato ringspot (ToRSV) were not encountered. Peach latent mosaic viroid (PLMVd) and hop stunt viroid (HSVd) were not detected either.  相似文献   

6.
The major and most widespread virus-induced disorder of almond is a complex disease called mosaic. The disease is characterized by a variety of symptoms ranging from bright chrome-yellow (calico) to chlorotic discolorations, localized necrosis of the leaf blade, leaf curling, bud failure, fasciations, rosetting, stunting and bushy growth. Three ilarviruses, apple mosaic (ApMV), prune dwarf (PDV) and prunus necrotic ringspot (PNRSV), are associated with almond mosaic throughout the Mediterranean, and are involved to different extents in its aetiology. Apple chlorotic leaf spot trichovirus (ACLSV) may also be present in mosaic-affected plants, and is sometimes associated with a chlorotic leafroll condition. Two epoviruses, tomato ringspot (ToRSV) and tomato black ring (TBRV), have been reported from the USA and Europe as the agents of diseases called yellow bud mosaic and enations, respectively. Plum pox potyvirus (PPV) and another filamentous virus, prunus latent virus (PLV), can also infect almond. Brown line and decline, a putative phytoplasma disease, occurs in California, together with the leaf scorch induced by the xylem-limited bacterium Xylella fastidiosa. A stem pitting lot associated with nepovirus infection and a graft-union necrosis apparently not caused by phytoplasmas have been reported from Italy. Infections by hop stunt viroid (HSVd) were recently reported from Spain.  相似文献   

7.
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   

8.
Surveys were carried out in the traditional areas of almond and apricot cultivation in Tunisia to assess the phytosanitary status of these species in varietal collections, mother block stands and commercial orchards. The presence of virus and virus-like diseases was ascertained through field surveys, sap transmission to herbaceous hosts, and ELISA and IEM tests. The mean infection level in almond was 34%. compared with only 4.7% in apricot. The following viruses were identified: apple chlorotic leaf spot trichovirus (ACLSV), apple mosaic (ApMV), prune dwarf (PDV) and prunus necrotic ringspot (PNRSV) ilarviruses. Plum pox potyvirus (PPV) was not detected. Likewise, none of the six nepoviruses tested for (SLRV, TBRV, RRV. CLRV. ArMV and ToRSV) was encountered.  相似文献   

9.
Coat protein (CP) sequences of 17 Ilarvirus isolates were obtained from hops at three farms in Tasmania, Australia. Phylogenetic analysis of these sequences and additional database sequences indicated several Apple mosaic virus (ApMV) isolate clusters distinct from Prunus necrotic ringspot virus (PNRSV): one containing isolates from apple; one containing a single isolate from almond; a third containing Australian hop isolates of the 'apple' serotype and a German isolate of unknown origin; and a fourth containing Australian hop isolates of the 'intermediate' serotype. Isolates from hop, pear and prune from the Czech Republic either formed a fifth grouping, or were divergent members of the 'intermediate' serotype group. Deduced amino acid (aa) residue differences between the coat proteins of the two hop isolate serotype groups were highlighted as possible regions of serological differentiation. No evidence for coinfection of plants with both serotypes was found. Tests of ApMV-infected hop buds using the Shirofugen flowering cherry assay revealed a possible differentiation of the two strains based on hypersensitivity. Because of serological similarities to PNRSV, these viruses have commonly been reported as strains of PNRSV. However, this study shows ilarviruses from Australian hops are strains of ApMV, but distinct from those infecting Malus spp.  相似文献   

10.
Surveys were carried out in the main stone-fruit growing areas of Albania to assess the phytosanitary status of Prunus in conimercial orchards and varietal collections. The presence of virus and virus-like diseases and their identification was ascertained through field observations, sap transmission to herbaceous hosts, graft transmission to woody indicators, ELISA and IEM tests. The mean infection level was 42%. In particular, infections in apricot and almond were 12 and 16%, respectively, i.e. lower than in plum and cherry (47 and 56%, respectively). The following viruses were identified: plum pox potyvirus (PPV). apple chlorotic leaf spot trichovirus (ACLSV). prunus necrotic ringspot (PNRSV) and prune dwarf (PDV) ilarviruses. PPV infection was very severe in plum, and limited in apricot and peach. Apple mosaic ilarvirus (ApMV), and six nepoviruses tested for (SLRV, TBRV, RRV, CLRV, ArMV and ToRSV) were not encountered in Primus.  相似文献   

11.
 应用DAS-ELISA和RT-PCR方法从褪绿和银色斑驳的西瓜叶片中检测到病毒分离物（WSMoV-YN）,感病样品能与WSMoV/GBNV复合抗血清（Agdia）呈阳性反应。获得WSMoV N蛋白的多克隆抗体,抗体能与WSMoV血清组成员CaCV和TZSV反应,但不能与INSV、TSWV、HCRV和GYSV反应。为明确引起该病害的病毒种类,采用Tospovirus通用引物对样品的总RNA进行RT-PCR扩增,获得长度为3 554 nt的S RNA全序列,经Blastn比对分析与WSMoV中国台湾分离物同源性最高,为95.8%,其N和NSs蛋白氨基酸序列同源性分别为99%和97.6%。构建系统进化树发现,西瓜银灰斑驳病毒云南分离物（WSMoV-YN）与其他WSMoV聚为一支。确定引起云南西瓜病害的病毒为WSMoV。  相似文献   

12.
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13.
A putative virus-induced disease showing chlorotic ringspots on leaves of Phalaenopsis orchids has been observed in Taiwan for several years. A virus culture, 91-orchid-1, isolated from a Phalaenopsis orchid bearing chlorotic ringspot symptoms was established in Chenopodium quinoa and Nicotiana benthamiana, and characterized serologically and biologically. The virus reacted slightly with the antiserum of Watermelon silver mottle virus (WSMoV) but not with those of Tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV) and Groundnut ringspot virus (GRSV). Isometric particles measuring about 70–100 nm were observed. Inoculation with isolated virus was conducted to confirm that 91-orchid-1 is the causal agent of chlorotic ringspot disease of Phalaenopsis orchids. To determine the taxonomic relationships of the virus, the conserved region of L RNA and the complete nucleocapsid gene (N gene) were cloned and sequenced. The sequence of conserved region of L RNA shares 83.8, 82.5, 64.4 and 64.9% nucleotide identities and 96.5, 97.7, 67.3 and 67.6% amino acid identities with those of Peanut bud necrosis virus (PBNV), WSMoV, TSWV and INSV, respectively, indicating that 91-orchid-1 is a tospovirus related to WSMoV. The complete nucleotide sequence of the N gene determined from a cDNA clone was found to be 828 nucleotides long encoding 275 amino acids. Sequence analyses of the N gene showed that 91-orchid-1 is an isolate of Capsicum chlorosis virus (CaCV) which has been reported to infect tomato and capsicum plants in Australia and Thailand. 91-orchid-1 is therefore designated as CaCV-Ph. To our knowledge, this is the first formal report of a tospovirus infecting Phalaenopsis orchids.  相似文献   

14.
河南甜樱桃病毒病害调查及病原检测   总被引：1，自引：0，他引：1  
在河南省郑州市、巩义市、荥阳市、新郑市选择具有代表性的甜樱桃生产园对病毒病发生情况进行调查,采集表现为疑似病毒病症状的样本65份,利用7种病毒引物进行RT-PCR检测。5种病毒检测结果呈阳性,分别是李属坏死环斑病毒(Prunus necrotic ringspot virus,PNRSV)、李矮缩病毒(Prune dwarf virus,PDV)、樱桃绿环斑驳病毒(Cherry green ring mottle virus,CGRMV)、樱桃坏死锈斑病毒(Cherry necrotic rusty mottle virus,CNRMV)及樱桃病毒A(Cherry virus A,CVA);序列分析结果表明,5种病毒扩增片段与GenBank中注册的相应病毒核苷酸序列均具有较高的一致性;样本病毒检出率为100%,其中13份样本为单独侵染,其余52份样本均为多病毒复合侵染,占比高达80%,复合侵染比例随着侵染病毒种类的增多逐渐降低;病毒侵染组合与叶片表型症状无明显对应关系。  相似文献   

15.
Although Prune dwarf virus (PDV) is one of the most common viruses infecting stone fruits (Prunus spp.), most of the molecular diversity studies have so far focused on the capsid protein (CP) gene. An alternative diagnostic primer pair targeting the movement protein (MP) gene was used in the present work, increasing then polyvalence of PDV detection in cherry samples from Slovakia as compared to the previously published primers amplifying 5′MP-intergenic region-CP-3′UTR fragment. The nearly complete sequence of the RNA3 from 24 Slovak PDV isolates was determined, together with 19 partial sequences of the MP gene for other isolates. Even though the isolates originated from a geographically limited region, analyses showed a high degree of nucleotide sequence diversity between isolates. No clear-cut phylogenetic grouping based on host or geographical origin could be observed. The topology of the MP and CP trees were incongruent for some isolates. Highly supported recombination signals were detected in five PDV isolates, with breakpoints located at different positions in the central part of RNA3, within or close to the intergenic region. Taken together these results extend our understanding of the variability of PDV and for the first time highlight the contribution of recombination to the evolutionary history of this virus.  相似文献   

16.
For the identification and analysis of new RNA plant viruses infecting fruit trees, an initial step often involves the laborious procedure of isolation and cDNA synthesis and cloning from purified viral dsRNA. For subsequent RT-PCR detection of these and other viruses from tissue with high phenolic and polysaccharide concentrations, a simple and efficient extraction protocol for viral nucleic acid is also important. A method for rapid cDNA cloning from small amounts of purified dsRNA using a modification of degenerate oligo primed polymerase chain reaction mbox(DOP-PCR), and a modification of a protocol for effective extraction of viral RNA for use in RT-PCR are presented. Both methods were used to analyze a number of mottling diseases described in cherry. The causal agents for two of these diseases have been previously described, Cherry green ring mottle virus, a tentative member of the foveaviruses, and Cherry mottle leaf virus, a member of the trichoviruses. For the diseases cherry rusty mottle and cherry necrotic rusty mottle, data are presented identifying viruses associated with each disease. Viruses associated with cherry rusty mottle, cherry necrotic rusty mottle and European isolates of cherry mottle leaf diseases, are closely related to Cherry green ring mottle virus and can be tentatively included in the foveavirus genus. An additional virus, related to cherry green ring mottle virus, was discovered by RT-PCR cloning and appears to be a common latent virus of cherry. Finally, isolates of cherry necrotic mottle disease could be assayed positive by RT-PCR for a virus  相似文献   

17.
Symptoms induced in rose by single isolates of the cherry serotype of prunus necrotic ringspot virus (PNRSV) and an apple serotype (apple mosaic virus; ApMV) were characteristically different, and appeared at different times throughout the growing season according to the ambient temperature. These features remained discrete, even in roses infected by both viruses and were shown by immunospecific electron microscopy to be a reliable indication of infection by either virus.
However, cross-protection between the two isolates was not reciprocal; mixed infections were established only when roses were simultaneously graft-inoculated with ApMV and PNRSV, or when PNRSV-infected roses were supei-infected with ApMV. The significance of these results in relation to the possible natural occurrence of mixed infections in rose or of isolates of intermediate serotype is discussed.  相似文献   

18.
The genomic regions encoding the putative movement protein (MP), coat protein (CP) and intergenic region (IGR) of seven Spanish isolates of the Parietaria mottle virus which infects tomato plants (PMoV-T) were sequenced. Values for the genetic diversity of the PMoV-T isolates were 0.056, 0.047 and 0.013 for the CP, MP genes and IGR, respectively. Nucleotide and amino acid sequence comparison of the seven PMoV-T isolates with those of PMoV revealed significant differences. All of them had a cytosine deletion at position 1366, also confirmed in an Italian tomato isolate, which involves a start codon for the CP gene different from that for the PMoV sequence, resulting in a CP 16 amino acids shorter than the PMoV CP. The certainty of a cytosine deletion only associated to the tomato isolates or the possibility of a mistake in the PMoV published sequence are the two hypotheses that could explain this difference. Structural motifs highly conserved in Ilarviruses were identified in PMoV-T MP and CP. A stable hairpin structure is proposed for IGR, by the initiation site for subgenomic RNA 4 synthesis. Phylogenetic analysis of CP and MP amino acid sequences showed that Spanish PMoV-T isolates form a separate group from PMoV and other members of the Ilarvirus genus. Comparative analysis with different PMoV isolates including tomato isolates from other regions and isolates from different hosts are necessary to confirm this differentiation.  相似文献   

19.
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20.
A complete sequence for the RNA 3 of Prunus necrotic ringspot virus (PNRSV) is described (Genbank Accession U57046). Primers from this sequence were used to amplify both the movement protein and coat protein genes of 3 other isolates of PNRSV originating from different host species and geographic locations. Comparisons of these sequences with those of other published sequences for PNRSV and the closely related apple mosaic virus (ApMV) showed that both the movement proteins and coat proteins of isolates of PNRSV are extensively conserved irrespective of either the original host or the geographic origin. The movement protein and coat protein of ApMV and PNRSV are sufficiently conserved to suggest that these two viruses may have evolved from a common ancestor. The amino acid sequence of the two coat proteins shows areas of similarity and difference that would explain the serological continuum reported to occur among isolates of these two viruses. Nevertheless, the movement protein and coat protein of the two viruses are sufficiently different so that ApMV and PNRSV should be considered to be distinct viruses.  相似文献   

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