A potyvirus isolated from solanaceous hosts

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.     

Introgression of a Tombusvirus Resistance Locus from Nicotiana edwardsonii var. Columbia to N. clevelandii

ABSTRACT A new variety of Nicotiana, N. edwardsonii var. Columbia, was evaluated for its capacity to serve as a new source for virus resistance genes. Columbia was developed from a hybridization between N. glutinosa and N. clevelandii, the same parents used for the formation of the original N. edwardsonii. However, in contrast to the original N. edwardsonii, crosses between Columbia and either of its parents are fertile. Thus, the inheritance of virus resistance genes present in N. glutinosa could be characterized by using Columbia as a bridge plant in crosses with the susceptible parent, N. clevelandii. To determine how virus resistance genes would segregate in interspecific crosses between Columbia and N. clevelandii, we followed the fate of the N gene, a single dominant gene that specifies resistance to Tobacco mosaic virus (TMV). Our genetic evidence indicated that the entire chromosome containing the N gene was introgressed into N. clevelandii to create an addition line, designated N. clevelandii line 19. Although line 19 was homozygous for resistance to TMV, it remained susceptible to Tomato bushy stunt virus (TBSV) and Cauliflower mosaic virus (CaMV) strain W260, indicating that resistance to these viruses must reside on other N. glutinosa chromosomes. We also developed a second addition line, N. clevelandii line 36, which was homozygous for resistance to TBSV. Line 36 was susceptible to TMV and CaMV strain W260, but was resistant to other tombusviruses, including Cucumber necrosis virus, Cymbidium ringspot virus, Lettuce necrotic stunt virus, and Carnation Italian ringspot virus.     

Significance of the heterologous encapsidation of zucchini yellow mosaic potyvirus in transgenic plants expressing plum pox potyvirus capsid protein1

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.     

Characterization of an isolate of beet mosaic virus from South Kazakhstan

A filamentous virus isolated from a sugar-beet plant showing systemic mosaic collected in South Kazakhstan was identified as an isolate of beet mosaic virus (BMV-K). BMV-K was transmitted by the green peach aphid Myzus persicae in a non-persistent manner, and by sap inoculation to 11 out of 19 species from seven families tested. The virus could not be transmitted to Nicotiana tabacum, N. debneyi, N. glutinosa and N. clevelandii, cither mechanically or with M. persicae. The thermal inactivation point of BMV-K in sugar-beet sap was 55-60 C, dilution end point 1:1000 and longevity in vitro 2 days at 20 C. A purification procedure produced 1-5-3 mg of purified virus from 100 g of infected Stellaria media plants. Purified virus contained a single protein species of molecular weight 34 700 Da. In ELISA tests, BMV-K reacted positively with BMV-specifc antisera obtained from Japan. Germany and Portugal. By competitive DAS- ELISA, the virus isolate was shown to be closely serologically related to all the three isolates of BMV, and very distantly related to bean yellow mosaic and soy bean mosaic viruses.     

Occurrence of potyvirus and potexvirus infections in black bryony (Tamus communis) in Devon, UK

Leaf samples of black bryony (Tamus communis L.) from Devon, UK, showing various virus-like symptoms contained potyvirus-like particles (normal length c. 790 nm) and cytoplasmic cylindrical (pinwheel) inclusions. In immunoelectron microscopy, particles of most samples reacted with antiserum to dioscorea greenbanding mosaic virus, a potyvirus isolate from Dioscorea rotundata in Togo which is related to yam mosaic virus from the Ivory Coast. Potyvirus particles were not transmitted by sap or aphids (Myzuspersicae) from infected black bryony to black bryony seedlings or Nicotiana benthamiana. One sample from a symptomless plant of black bryony contained a potexvirus which formed massed virion aggregates in the cytoplasm of cells of black bryony, Nicotiana benthamiana and N. megalosiphon. Virions of the potexvirus (normal length 553 nm) contained a coat protein with an apparent molecular weight of 27.7 kd. The potexvirus differed from an Italian potexvirus isolate from black bryony by a serological differentiation index of 4 and gave only weak or no reactions with 23 other antisera to potexviruses, including dioscorea latent virus. The potexvirus caused systemic symptoms in only a few host plants and could be transmitted back to black bryony in which it caused no symptoms. It is provisionally named tamus latent virus.     

侵染番红花的芜菁花叶病毒

 从表现花叶症状的德国进口番红花上获得一线状病毒分离物SA,线状病毒粒子长度为700~800nm。其在人工接种的11科39种植物中能侵染6科14种植物,在克里芙兰烟上产生系统坏死,在小白菜、花椰菜等十字花科植物上产生系统花叶或为隐症,在昆诺藜等藜科植物上为局部侵染。在番红花病叶、球茎及克里芙兰烟病叶组织中观察到卷轴状和片层状聚集的风轮状内含体。免疫吸附和免疫修饰电镜观察结果显示,SA与芜菁花叶病毒((TuMV)具有紧密的血清学关系。根据这些特征将SA鉴定为TuMV的一个分离物。这是TuMV侵染番红花的首次报道。     

Differentiation between two potyviruses inAlstroemeria

Alstroemeria mosaic virus (AlMV) is one of the viruses known to occur inAlstroemeria spp. Its detection in DAS-ELISA needed improvement. The often simultaneous presence of a second potyvirus has been mentioned by various authors. The recently detected virus inAlstroemeria, tentatively namedAlstroemeria streak virus [AlSV; Wong, 1992] was multiplied in indicator plants and had a host range similar to that of AlMV, although the symptoms in these hosts were less severe. Both viruses reacted with antisera prepared in the Netherlands and in Great Britain to AlMV-isolates purified from infectedAlstroemeria plants, and fromNicotiana clevelandii, respectively. Where AlSV occurs separately, distinction from AlMV is possible by its negative reaction with potyvirus group-specific monoclonal antibodies.     

Isolation and characterization of a potyvirus associated with bushy dwarf symptom in chickpea, Cicer arietinum, in India

A potyvirus that induced stunting and a characteristic bushy appearance at the apical region, due to proliferation of terminal branches with narrowed, reduced and deformed leaflets, was isolated from chickpea in India. The virus was sap-transmissible to 14 species of Chenopodiaceae, Leguminosae, Solanaceae and Malvaceae; Chenopodium amaranticolor was a good local lesion host. Virus particles, trapped by immunosorbent electron microscopy and stained with uranyl acetate, were 710 ×10 nm long. Purified virus preparations contained a single polypeptide species of 32,500 Da and one nucleic acid species of 3.1 · 10⁶ Da. The virus was serologically related to soybean mosaic, azuki bean mosaic and peanut mottle viruses but not to clover yellow vein, pea seed-borne mosaic and bean yellow mosaic viruses.
On the basis of these properties, the virus was identified as a previously undescribed potyvirus in chickpea, for which the name chickpea bushy dwarf virus is proposed.     

Biological and Molecular Characterization of a New Carlavirus Isolated from an Aconitum sp

ABSTRACT A new virus was isolated from symptomless Aconitum napellus plants. The virus, for which the name Aconitum latent virus (AcLV) is proposed, has flexuous particles 640 nm in length. The experimental host range was limited to Nicotiana clevelandii. Electron microscopy studies of ultrathin sections of infected A. napellus tissues revealed the presence of elongated virus particles. No inclusion bodies characteristic of potyvirus infection were observed. AcLV was purified from naturally infected A. napellus by cesium chloride step gradient centrifugation. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis of dissociated purified virus preparations, a major protein component with a molecular mass of 35 kDa was observed. Diagnostic antibodies that could specifically bind to virus particles were produced. The 5' terminus (620 nucleotides) of the viral RNA was cloned and sequenced. It comprised 71 nucleotides from the untranslated 5' terminus and 549 nucleotides of an open reading frame encoding 183 amino acids. Comparison of the predicted amino acid sequence with those of other plant viruses revealed 40 to 60% identity with several carlaviruses. Based on particle morphology, absence of inclusion bodies in ultrathin sections, the relative molecular weight of the coat protein, the nucleotide sequence, and predicted amino acid homology, it is suggested that this virus belongs to the carlavirus group.     

大豆花叶病毒病的鉴定

 1978年到1979年在江苏南京和扬州进行了大豆上发生最普遍的一种花叶病毒病的鉴定工作。系统地观察了病害的症状。发病初期表现明脉,并发展为斑驳花叶,叶缘自下呈波纹状卷曲,以后出现皱缩或沿叶脉两侧呈泡状突起等症状。病株根系发育差,产生的根瘤少而小,并常常形成无绒毛的豆荚和褐色斑纹的种子,病株有恋青情况。
病株种子的传毒率可高达58.1%。测定的6种蚜虫中有桃蚜(Myzus per-sicae)、大豆蚜(Aphis glycine),首蓿蚜(Aphis craccivera)、棉蚜(Aphis gossgpii)、菜蚜(Rhopalosiphuns pseudobrassicae)能传病,而麦长管蚜(Sitobion avenae)不能传病。
这种病毒的寄主范围很窄,测定的14种植物中只能侵染大豆,不能侵染其它豆科植物,也不能侵染烟草、心叶烟、千日红、百日菊、曼陀罗和小藜等。
抗性测定的致死温度为65~68℃,稀释限点是1:2000~1:10,000倍,体外保毒期6天。
用浸出法和磷钨酸负染的电子显微镜观察为大小±750×13nm的线条状病毒粒体。
根据以上性状,这种大豆病毒病的病原鉴定为大豆花叶病毒(SMV)属马铃薯Y病毒群(Poly virus group)。     

Characterization of a new potyvirus isolated from peanut (Arachis hypogaea)

During a survey of viruses of peanuts in South Africa a mechanically transmissible virus was isolated from a plant exhibiting chlorotic ringspots and blotches on the leaves. Typical potyvirus-like flexuous particles were detected by electron microscope examination. Pinwheel-shaped and laminated inclusions in ultrathin sections, reaction with a monoclonal antibody directed to a potyvirus common epitope, a single 33 kDa coat protein and aphid transmission using Myzus persicae all confirmed that the virus was a subdivision II member of the Potyviridae. Host range studies suggested that the virus was none of the previously reported potyviruses of peanuts or of subdivision II potyviruses. The serological relationships of the virus were studied using a range of 17 antisera to potyviruses in ELISA and immunosorbent electron microscopy (ISEM). The isolate reacted weakly with antisera to plum pox virus and bean yellow mosaic virus in ISEM only. Nucleotide sequence of a 624 bp DNA product was obtained following immuno-capture with a potyvirus common epitope antiserum, cDNA synthesis and PCR amplification with potyvirus specific primers which amplify the 3' untranslated region and a part of the coat protein gene. The sequence was only distantly related to a number of potyviruses, whether amino acid or nucleotide sequences were used for comparisons. It is proposed that the virus be named peanut chlorotic blotch virus and be accepted as a new member of the genus Potyvirus in the family Potyviridae.     

Identification and characterization of a potyvirus causing chlorotic spots on <Emphasis Type="Italic">Phalaenopsis</Emphasis> orchids

A putative virus-induced disease showing chlorotic spots on leaves of Phalaenopsis orchids was observed in central Taiwan. A virus culture, phalaenopsis isolate 7-2, was isolated from a diseased Phalaenopsis orchid and established in Chenopodium quinoa and Nicotiana benthamiana. The virus reacted with the monoclonal antibody (POTY) against the potyvirus group. Potyvirus-like long flexuous filament particles around 12–15 × 750–800 nm were observed in the crude sap and purified virus preparations, and pinwheel inclusion bodies were observed in the infected cells. The conserved region of the viral RNA was amplified using the degenerate primers for the potyviruses and sequence analysis of the virus isolate 7-2 showed 56.6–63.1% nucleotide and 44.8–65.1% amino acid identities with those of Bean yellow mosaic virus (BYMV), Beet mosaic virus (BtMV), Turnip mosaic virus (TuMV) and Bean common mosaic virus (BCMV). The coat protein (CP) gene of isolate 7-2 was amplified, sequenced and found to have 280 amino acids. A homology search in GenBank indicated that the virus is a potyvirus but no highly homologous sequence was found. The virus was designated as Phalaenopsis chlorotic spot virus (PhCSV) in early 2006. Subsequently, a potyvirus, named Basella rugose mosaic virus isolated from malabar spinach was reported in December 2006. It was found to share 96.8% amino acid identity with the CP of PhCSV. Back-inoculation with the isolated virus was conducted to confirm that PhCSV is the causal agent of chlorotic spot disease of Phalaenopsis orchids in Taiwan. This is the first report of a potyvirus causing a disease on Phalaenopsis orchids.     

Detection of plum pox virus by isolation of double-stranded ribonucleic acid (dsRNA)

Double-stranded RNA (dsRNA) associated with plum pox virus (PPV) in Nicotiana clevelandii and Prunus domestica has been isolated. While dsRNA was detected in N. clevelandii in considerable amounts by electrophoresis, only small amounts were found in P. domestica. This may be due to viscous substances in the leaves of this woody host. Different PPV strains (NAT - not aphid-transmissible; AT - aphid-transmissible) showed specific patterns in electrophoresis gels. When PPV was assayed in N. clevelandii by dsRNA detection or by standard ELISA or ISEM, all three methods were found to be efficient, with none being superior. ELISA, as a simple and fast routine method, is still the method of choice. DsRNA detection will be suitable for plant disease agents undetectable by ELISA and ISEM.     

Non-structural plum pox potyvirus proteins detected by immunogold labelling

Plum pox potyvirus (PPV) induces in infected Nicotiana clevelandii cells characteristic crystalline inclusions known as nuclear inclusions (NI) when located in the nucleus and as dense material (Dm) when located in the cytoplasm. Crystalline inclusions contain protease (NIa) and RNA-dependent RNA polymerase (NIb) proteins. It is now well established for all potyviruses that cylindrical inclusions contain CI helicase ATPase protein (Martin et al., 1992). The intracellular location of other non-structural PPV proteins remains unknown. Using Escherichia coli expression vectors, specific antibodies were obtained against P1, P3, 6K2 and NIb PPV proteins for which antibodies were not yet available. As expected, NIb antiserum labelled crystalline inclusions. P1, P3 and 6K2 proteins were present in both types of crystalline inclusions found in the nucleus and in the cytoplasm of PPV-infected leaves of N. clevelandii, suggesting that nuclear inclusions and dense material were composed of the same proteins. This composition is discussed.     

山东省大豆花叶病毒原及其生物学的研究

 山东省田间侵染大豆的花叶病毒,粒体线条状,大小714-729×12-13nm,粗提纯液与SMV抗血清呈阳性反应。寄主范围窄,接种5科14种植物只侵染大豆,呈系统花叶症状。体外抗性。钝化温度65-70℃,稀释限点10^-3-10^-4,体外保毒划3-4天。传毒方式:可通过种子、蚜虫和摩擦接种传毒。根据以上特性,侵染大豆的花叶病毒,毒原应为大豆花叶病毒(SMV),属于马铃薯y病毒组(Potyvirus group)的成员。     

The occurrence of three viruses in hop (Humulus lupulus) in China

Forty-two percent of leaf samples taken from hop cv. Qingdao Dahua growing in plantations in northeastern China in 1983-1984 were infected with hop mosaic virus (HMV) and/or hop latent virus (HLV). Mosaic or line-pattern symptoms were seen in some plants but only in spring and it is uncertain if the symptoms were caused by either virus which appeared in our studies to be similar, both serologically and in host range, to HMV and HLV described in Europe and North America. In host range studies, HMV infected Nicotiana clevelandii systemically without inducing symptoms. Occasionally it caused chlorotic spots in the inoculated leaves of Chenopodium quinoa and brown star-point lesions in inoculated leaves of Phaseolus vulgaris cv. Topcrop, The virus was purified by PEG precipitation from extracts of N. clevelandii plants; yields were about 40-120 mg/kg fresh leaves. The virus contained a single protein (estimated molecular weight 34 200) that was estimated to contain 272 amino acids with no methionine, and a single RNA species (estimated molecular weight 295 x 10⁶) that represented c. 6% of the particle weight. HLV sometimes caused inconspicuous chlorotic spots in inoculated leaves of C. murale and local pinpoint lesions in P. vulgaris in winter only. A virus similar to alfalfa mosaic virus was isolated from leaves of hop cv. Golding and caused systemic necrotic symptoms in C. quinoa and C. amaranticolor , and systemic chlorotic spots in hop cv. Styrian.     

Characterization of a strain of clover yellow vein potyvirus infecting borage (Borago officinalis L.) in Spain

A disease of borage ( Borago officinalis ) in Spain, characterized by severe mosaic and deformation of the leaves, was shown to be caused by a potyvirus. The borage-infecting potyvirus was characterized biologically by the symptoms induced in 23 indicator species and was shown to be transmitted experimentally by the aphid Myzus persicae in a non-persistent manner. In order to classify the borage-infecting potyvirus we have cloned and sequenced the entire coat protein gene and 3' non-coding region of the viral RNA. By comparing this nucleotide sequence with those of other members of the Potyviridae , we can identify the Spanish borage-infecting potyvirus as an isolate of clover yellow vein virus (CYVV), a virus so far only known to cause important diseases in forage legumes. This is the first record of CYVV in Spain and of CYVV infecting a natural host of the Boraginaceae.     

Severe mosaic caused by zucchini yellow mosaic virus in cucurbits from Jordan

A virus disease causing severe mosaic in melon (the melon isolate) was identified as a strain of zucchini yellow mosaic virus (ZYMV). Identification was based on host range, aphid transmissibility, electron microscopy, and serological tests. The virus was recovered from all cultivated cucurbits in Jordan and from naturally infected Moluccella laevis. It was seed-transmitted in Ranunculus sardous. Host-range comparison showed that the melon isolate and a French isolate belong to a different biotype group from a Connecticut isolate (ZYMV-CT); this was confirmed by indirect ELISA. During 1987–1988. ZYMV appeared to be the predominant virus affecting cucurbits.     

Identification and partial characterization of a <Emphasis Type="Italic">Carica papaya</Emphasis>-infecting isolate of <Emphasis Type="Italic">Alfalfa mosaic virus</Emphasis> in Brazil

A Carica papaya plant with severe yellow leaf mosaic, leaf distortion, and systemic necrosis was found in the municipality of Piracicaba, state of São Paulo, Brazil. Transmission electron microscopy (TEM) analysis revealed the presence of potyvirus-like particles and bacilliform particles similar to those of the Alfamovirus genus. The potyvirus was identified as Papaya ringspot virus-type P (PRSV-P). Biological, serological, and molecular studies confirmed the bacilliform virus as an isolate of Alfalfa mosaic virus (AMV). Partial nucleotide and amino acid sequences of the coat protein gene of this AMV isolate shared 97–98% identity with the AMV isolates in the GenBank database. This report is the first of the natural infection of papaya plants by AMV.