Shallot latent virus,a new carlavirus

A new carlavirus, apparently omnipresent in shallot (Allium ascalonicum) without causing symptoms, is described as shallot latent virus. It has also been detected in naturally infected onion (A. cepa) and leek (A. porrum). The virus was easily transmissible in sap and could be transferred withMyzus ascalonicus. Infection after inoculation was symptomless in onion, leek,A. fistulosum andA. jailae. Chenopodium album, C. amaranticolor andC. quinoa reacted with local lesions.A. neapolitanum andA. schoenoprasum and 20 other plant species were immune. Dilution end-point was 10⁴–10⁵, thermal inactivation at ca. 80°C and ageing in vitro 8–11 days. Purification by molecular sieving on Sephadex G-200 followed by equilibrium density-gradient centrifuging in CsCl was successful. Sedimentation coefficient was 147.5 S, bouyant density 1.313 g/cm³ and molecular mass of protein subunits 23 200 dalton. With the antiserum (titre 1024) distant serological relationships to some carlaviruses were determined. No inclusion bodies were observed with the light microscope. With the electron microscope a high concentration of straight or slightly curved particles with a normal length of 650 to 652 nm could easily be detected in crude sap and in purified preparations.     

Partial purification of a virus associated with a Spanish isolate of citrus ringspot

A citrus ringspot isolate from Star Ruby grapefruit (RS-SR) was mechanically transmitted to Chenopodium quinoa. RS-SR was partially purified by differential centrifugation, fractionation in a sucrose gradient, and agarose gel electrophoresis of selected fractions. Infectivity of concentrated extracts on C. quinoa was lost in individual fractions of the gradient, but it was recovered by combining a top and a bottom component. Both components contained a 48-kDa protein not found in similar preparations from healthy plants. After further purification the 48 kDa protein was detected at the top edge of the agarose gel. In the initial experiments a 38-kDa protein was found in the same fractions that later contained the 48-kDa protein. An antiserum obtained to the 38 kDa protein reacted in Western blots with both the 38- and the 48-kDa proteins, whereas another antiserum raised to the Florida isolate CRSV-4 (also containing a 48-kDa protein) did not react with the 38-kDa protein, indicating that the latter was probably a degradation product of the 48-kDa protein. Filamentous flexous particles were observed by serologically specific electron microscopy in crude extracts from RS-SR-infected C. quinoa plants. These results indicate that RS-SR is associated with a two-component virus similar to those associated with several psorosis and ringspot isolates, and serologically related to CRSV-4.     

Spinach latent virus,a new ilarvirus seed-borne in Spinacia oleracea

During 1977 and 1978 an apparently new virus was isolated from samples of 12 out of 142 lots of spinach seed from a number of countries which did not produce symptoms in spinach. In one sample seed infection was over 50%. The virus was later found to be identical with a virus encoded GE36, earlier presumed to have been isolated from apple and pear (Van der Meer, 1968; Maat and Vink, 1971). It has now been further characterized and named spinach latent virus.Seventeen out of 36 plant species tested were susceptible, most without producing symptoms.Myzus persicae did not transmit the virus, but rates of seed transmission in artificial hosts were high: 53% inCelosia cristata, over 90% inChenopodium quinoa, 30% inNicotiana rustica, 72% inN. tabacum Samsun, 90% in White Burley and 94% in Xanthi. Infection is in the embryo. The virus can also be detected in dry seeds.Purified virus contained three components sedimenting at 87, 98 and 108 S. In the electron microscope the virus particles were irregularly spherical and c. 27 nm in diameter. They were hard to detect in crude plant sap. Some particles were bacilliform. The buoyant density in cesium sulphate was 1.269×10³ kg.m^–3. Five RNA components were detected having relative molecular masses of 1.30, 1.18, 0.91, 0.35 and 0.27×10⁶, respectively. To induce infection the three largest components are required plus the two smallest components or the coat protein. Molecular mass of the coat protein subunit was 28000. These characters are typical of ilarviruses.The virus proved weakly immunogenic. An antiserum with a titre of 64 was produced. No serological differences could be found between the type isolate from spinach and GE36. The virus did not react with antisera to any of 36 spherical viruses and alfalfa mosaic virus.Samenvatting In 1977 kon uit 5 van de 43 en in 1978 uit 7 van de 99 getoetste partijen spinaziezaad afkomstig uit een aantal landen een klaarblijkelijk nieuw virus worden geïsoleerd zonder dat in de uit zulk zaad opgekweekte planten afwijkingen konden worden geconstateerd. Eén zaadherkomst was voor meer dan 50% geïnfecteerd. Het virus werd aangetoond in zaad van in totaal 12 verschillende spinazierassen. Tevens werd in enkele zaadmonsters komkommermozaïekvirus en in één het tabaksratelvirus aangetroffen.Biologisch, biofysisch en serologisch onderzoek heeft aangetoond dat het virus identiek is aan een eerder onder de code-naam GE36 beschreven virus, dat naar eerst werd vermoed, via toetsplanten was geïsoleerd uit appel en peer. Dit virus is nu vooral aan de hand van het spinazie-isolaat Sp20-9 uitvoerig gekarakteriseerd en beschreven als hetlatente spinazievirus (spinach latent virus). Zeventien van de 36 getoetste plantesoorten bleken vatbaar, de meeste echter symptoomloos. InChenopodium amaranticolor ontstonden karakteristieke, voornamelijk droge puntlesies en op de primaire bladeren vanPhaseolus vulgaris Bataaf opvallende necrotische lokale lesies, terwijl inC. quinoa de systemische reactie meer opviel dan de lokale. Ook bieteblad reageerde soms met lokale symptomen.De houdbaarheid van het infectievermogen in uitgeperst plantesap was bij verdunning 10³–10⁴, bij verhitting 60–65°C, en bij bewaring 4–5 dagen en éénmaal zelfs langer dan 13 dagen.MetMyzus persicae kon het virus niet op non-persistente wijze worden overgebracht. Zaadoverdracht werd ook aangetoond bij vier van de zes hierop onderzochte kunstmatige waardplanten. BijC. quinoa enN. tabacum White Burley en Xanthi bedroeg dit percentage zaadoverdracht zelfs meer dan 90. GE36 ging minder gemakkelijk over met zaad.Zuivering door butanol-klaring, differnetiële en dichtheidsgradiëntcentrifugering leverde aanvankelijk nog met celbestanddelen verontreinigd virus op. Afzondering van de viruszone, concentrering en verdere dichtheidsgradiëntcentrifugering verschafte echter zeer zuiver virus. Dit virus sedimenteerde in een suikergradiënt in twee zones en bij lage concentratie in drie zones. De drie componenten hadden sedimentatiecoëfficienten van 87, 98 en 108 S. De zweefdichtheid van het virus in een cesiumsulfaatgradiënt bedroeg 1,269×10³ kg. m^–3.Bij polyacrylamidegel-elektroforese ontstonden vijf nucleïnezuurbanden, elk bestaand uit RNA met een relatieve moleculaire massa van respectievelijk 1,30, 1,18, 0,91, 0,35 en 0,27 × 10⁶. De moleculaire massa van het eiwit bedroeg 28000. Intacte deeltjes verplaatsten zich in 2,5% polyacrylamide-gel als twee banden.Met de elektronenmicroscoop konden virusdeeltjes met moeite in ruw sap worden waargenomen, wel echter iets beter na fixatie en gemakkelijk in gezuiverde preparaten. De deeltjes waren onregelmatig van vorm, c. 27 nm in diameter, en soms bacilvormig.Bij infectieproeven met de verschillende groepen RNA-componenten en viruseiwit verkregen na SDS-afbraak bleek dat voor infectie de componenten 1, 2 en 3 nodig zijn tezamen met de componenten 4 en 5 òf het eiwit.Het virus bleek slechts zwak immunogeen (antiserumtiter 64) en in agargel vormden zich tenminste twee lijnen, indien de agar was bereid in een fysiologische zoutoplossing, doch slechts één lijn indien de agar was bereid in 0,05 M fosfaat-citroenzuurbuffer pH 7. Het bleek serologisch identiek aan GE36 en was niet verwant aan 36 verschillende bolvormige plantevirussen en aan luzernemozaïekvirus.De relatieve moleculaire massa's van de RNA-componenten, de uniforme zweefdichtheid in cesiumsulfaat, de aantoonbaarheid van meer dan één component in polyacrylamide-gel en bij suikergradiëntcentrifugering, het tripartite genoom en de eiwitafhankelijkheid ervan voor infectie, alsmede de onregelmatige deeltjesvorm en de kleine aantallen bacilvormige deeltjes rechtvaardigen plaatsing van het virus in de ilarvirusgroep.     

Parsley latent virus,a new and prevalent seed-transmitted,but possibly harmless virus of Petroselinum crispum

Parsley latent virus, a hitherto undescribed virus, was isolated from 38 out of 54 samples of seed of parsley (Petroselinum crispum) of 17 out of 24 cultivars and from all five European countries tested, but not from some samples from the USA. It could easily be detected in seedlings and also in seeds germinated on moist filter paper, but not in dry seeds or in seeds soaked in water. Strawberry latent ringspot virus was detected in five samples. The parsley virus is symptomless in parsley and caused latent systemic infection inGomphrena globosa, three cultivars ofSpinacia oleracea and weak and often transient systemic symptoms inChenopodium amaranticolor, C. giganteum, C. glaucum andC. quinoa, but did not infect any other species out of all 32 species of seven plant families tested in total.The virus could easily be transmitted mechanically but not by seven aphid species in the non-persistent manner. Dilution end-point was between 100 and 1000, thermal inactivation between 55 and 60°C and ageing in vitro between 7 and 10 days.Purification yielded a single infectious component. The particles were spherical, ca. 27 nm in diameter, with a sedimentation coefficient of 127.5 S, a buoyant density of 1.449 g/ml, an RNA content of 36% and one type of protein with a relative molecular mass of 22×10³. Purificition without Triton and urea resulted in preparations with aggregates each consisting of 12 particles in icosahedral array.The virus differs from all viruses described so far and did not show clear serological affinity with antisera to any of 34 widely differing viruses tested. It does not seem of direct practical importance and may be easily overlooked.Samenvatting In zaailingen van peterselie (Petroselinum crispum) werd een nog niet eerder beschreven virus aangetroffen. Het virus kon niet worden aangetoond door toetsing van droge of in water geweekte zaden opChenopodium quinoa maar wel in op filtreerpapier gekiemde zaden en vooral in zaailingen. Het werd aangetroffen in 38 van de 54 getoetste herkomsten, in 17 van de 24 getoetste rassen en in zaad vermeerderd in alle zes hierop onderzochte Europese landen maar niet in enkele zaadmonsters uit de USA. In sommige monsters bevatten nagenoeg alle zaden het virus. In vijf herkomsten werd eveneens het nog niet eerder in peterselie gerapporteerde latente aardbeikringvlekkenvirus geconstateerd. Dit virus kan bij toetsing gemakkelijk worden herkend door systemische symptomen inC. amaranticolor en komkommer.In geïnfecteerde peterselieplanten zijn geen afwijkingen waargenomen. Het virus kon niet op non-persistente wijze worden overgebracht met zeven bladluissoorten maar wel gemakkelijk met sap. Van 32 getoetste plantesoorten van zeven families, waaronder vier schermbloemigen, kon het virus slechts worden overgebracht op vierChenopodium-soorten,Gomphrena globosa en alle drie getoetste spinazierassen. AllenC. quinoa (Fig. 1),C. giganteum, C. glaucum en soms ookC. amaranticolor (Fig. 2) reageerden met vaak voorbijgaande systemische symptomen. Een lokalelesietoetsplant werd niet gevonden. Zaadovergang bijC. quinoa kon niet worden aangetoond.Voor de houdbaarheid van het infectievermogen werden de volgende waarden gevonden: verdunningseindpunt 100–1000, thermaal inactiveringspunt 55–60°C en houdbaarheid in vitro 7–10 dagen.Zuivering door homogenisatie in fosfaatcitroenzuurbuffer, behandeling met Triton X-100 en ureum en differentiële en daarna dichtheidsgradiëntultracentrifugering leverde preparaten op met uniforme deeltjes van ca. 27 nm diameter (Fig. 3B), een sedimentatiecoëfficiënt van 127,5 S, een zweefdichtheid van 1,449 g/ml, een RNA-gehalte van 36% en een relatieve moleculaire massa van de eiwitondereenheid van 22×10³. Bij zuivering zonder toepassing van Triton en ureum werd een extra zone verkregen met aggregaten van 12 deeltjes in icosaëdrische rangschikking (Fig. 4). In ruw plantesap waren slechts met grote moeite enkele deeltjes met behulp van de elektronenmicroscoop te vinden.Het virus reageerde niet met antisera tegen 33 bolvormige virussen en luzernemozaïekvirus (Tabel 1). Of de zwakke reactie verkregen met één antiserum tegen het tomate-aspermievirus een verre serologische verwantschap inhoudt, dan wel het gevolg is van een verontreiniging, werd niet vastgesteld.Het virus wordt beschouwd als een geheel nieuw virus waarvoor de naamlatent peterselievirus wordt voorgesteld. Het lijkt door zijn beperkte waardplantenreeks en symptoomloosheid in de vatbaar bevonden soorten, behalve in enkele als toetsplant te gebruikenChenopodium-soorten, nauwelijks van praktische betekenis.     

6种柑橘类植物对柑橘衰退病毒分离株TR-L514变异的影响

 柑橘衰退病毒(Citrus tristeza virus,CTV)存在复杂的株系分化现象,运用弱毒株交叉保护防治柑橘衰退病时需了解不同类型柑橘对CTV构成的影响。作者将CTV分离株TR-L514嫁接接种到6类柑橘植物上获得了18个亚分离株,并对这18个亚分离株和TR-L514进行了指示植物鉴定,p25基因的限制性片段长度多态性(RFLP)和单链构象多态性(SSCP)分析,p23基因的序列比较。结果表明,CTV株系对不同柑橘类植株的适应性存在差异。TR-L514嫁接到不同柑橘类植株其构成会发生变化,在甜橙上可以同时检测出p25//HinfⅠ RFLP第1和6组群,并具有比在其它4类柑橘上更为复杂的SSCP谱型构成,因此甜橙可能更适宜于CTV的增殖。TR-L514及18个亚分离株的p23基因序列差异可能与不同类柑橘植株的适应性有关。     

Potato latent virus: a proposed new species in the genus Carlavirus

A previously undescribed carlavirus, potato latent virus (PotLV), was found infecting the potato cultivar Red La Soda imported from the USA. The particles were filamentous and slightly curved, with modal lengths of 530 and 670 nm. The 11 kDa protein encoded downstream from the coat protein contained a 'zinc-finger' motif characteristic of carlaviruses, and RT–PCR using a carlavirus-specific primer gave a PCR product of 857 bp. Antibodies produced to PotLV did not detect other carlaviruses when used in ELISA and the coat-protein nucleic acid sequence of PotLV showed < 67% similarity with the other carlaviruses tested. The closest similarity was with the Andean strain of potato virus S. Unusually for a carlavirus, PotLV systemically infected Nicotiana bigelovii , N. glutinosa , N. rustica , N. tabacum and Physalis floridana .     

Nigerian tobacco latent virus: a new Tobamovirus from tobacco in Nigeria

Field-grown tobacco plants in Nigeria showing chlorotic mottle and marginal veinbanding on the leaves apparently contained several viruses. One of them proved to be a new Tobamovirus for which we suggest the name Nigerian tobacco latent virus (NTLV), because it did not produce systemic symptoms on various cultivars of Nicotiana tabacum. Sequence analyses of the coat and movement protein genes and their translation products, as well as serological studies, revealed that NTLV is only distantly related to known Tobamoviruses from which it also differs in host range and symptomatology. Its closest relationship was found to Tobacco mild green mosaic virus (TMGMV). The percentages of amino acid sequence identity amounted to 73% for the coat proteins and to 64% for the movement proteins of the two viruses. The total sequence of 1415 nucleotides analysed share 63% identity with the corresponding region of TMGMV. In the immunoelectron microscopical decoration test using antisera at a dilution of 1 : 50, reactions of NTLV were observed only with its own antiserum and one out of two antisera to TMGMV. An antiserum to NTLV diluted 1 : 2 failed to react with TMGMV. NTLV induces the formation of characteristic inclusions in infected cells.     

Effects of artichoke latent virus and broad bean wilt virus isolate FA on components of growth and yield of globe artichoke

The effects of single and mixed artificial infections by artichoke latent virus (ALV) and broad bean wilt virus isolate FA (BBWV-FA) on components of growth and yield of three globe artichoke lines were evaluated in an aphid-free glasshouse. Virus infections reduced plant vigour (length of leaves and height of the ramifications) and dry weight, increased dry matter percentage, lengthened harvest period, and induced significant losses in the number of harvested buds, which resulted from reduced production of ramifications. However, the magnitude of affected parameters depended on inherent line differences and the type of virus infection. The greatest reductions occurred with the ALV + BBWV-FA infection. In single infections ALV led to the more severe effects, especially on components of yield. The level of ALV coat protein, as determined by ELISA, indicated differences in titre between plants of three lines. Can line, which exhibited greater yield reduction than Cada or CBT lines, had the higher ALV virus titre. Conversely Cari was less susceptible to BBWV-FA infection than Cada. The detrimental effects of virus infection emphasize the need for virus control, and differences in responses among lines indicate the possibility of control by plant breeding.     

Separation and interference of strains from a citrus tristeza virus isolate evidenced by biological activity and double-stranded RNA (dsRNA) analysis

Separation of strains of citrus tristeza virus (CTV), differentiated by their double-stranded RNA (dsRNA) profiles, was obtained by graft-inoculating citron plants from a Mexican lime that had been recently aphid- or graft-inoculated with a mild CTV isolate (T-385). Up to 24 sub-isolates with differing dsRNA profiles were obtained from the aphid-inoculated lime. Some of these sub-isolates induced stronger symptoms in several citrus species than the original T-385 isolate. One sub-isolate, T-385-33, was mild in Mexican lime, but induced stem pitting on sweet orange. Inoculation of this isolate on Mexican lime, sour orange and Eureka lemon induced mild or no symptoms when inoculum was taken from citron, but very severe symptoms when the inoculum was from sweet orange. Mexican lime and sweet orange plants co-inoculated with T-385-33 from sweet orange in combination with the other 23 sub-isolates showed mild symptoms. The results obtained suggest that there is natural cross-protection among sub-isolates in the original T-385 isolate.     

Lonicera latent virus,a new carlavirus serologically related to poplar mosaic virus: some properties and inactivation in vivo by heat treatment

A virus with elongate particles (656 nm) was isolated from severalLonicera species. This virus, apparently belonging to the carlavirus group, is serologically distantly related to shallot latent virus and closely related to poplar mosaic virus. The inability to infect poplar and two other hosts of poplar mosaic virus characterizes the virus fromLonicera as a new virus which was namedLonicera latent virus.The virus was easily sap-transmissible but was not transmitted byMyzus persicae.Dilution end-point was about 10^–3, thermal inactivation between 65°C and 80°C and ageing in vitro 1–6 days.Heat treatment, combined with tip-rooting appeared to be a good method to eliminate the virus from severalLonicera species and cultivars.Samenvatting In verschillende soorten en cultivars van het geslachtLonicera (kamperfoelie) blijkt een virus voor te komen dat gemakkelijk door sapinoculatie kan worden overgebracht op kruidachtige planten.Een tegen gezuiverd virus bereid antiserum had een titer van ca. 4096. Er kon mee worden aangetoond dat het virus van kamperfoelie serologisch nauw verwant is met populieremozaïekvirus (Tabel 1). Het virus van kamperfoelie is echter niet in staat om populier,Phaseolus vulgaris Bataaf enVigna sinensis te infecteren en wordt mede daarom als een afzonderlijk virus beschouwd. Het wordt aangeduid als latent kamperfoelievirus (Lonicera latent virus) en behoort evenals populieremozaïekvirus tot de carlavirusgroep (aardappelvirus-S-groep).Het virus blijkt vrij gemakkelijk te kunnen worden geëlimineerd door besmette kamperfoelieplanten gedurende ongeveer zes weken een warmtebehandeling (37°C) te geven en daarna de uiterste toppen (1 cm) te stekken. Van verschillende cultivars werd op deze wijze virusvrij uitgangsmateriaal verkregen.     

Colletotrichum gloeosporioides, a new causal agent of citrus post-bloom fruit drop

Citrus post-bloom fruit drop (caused by Colletotrichum acutatum) frequently occurs in the southwestern region of São Paulo State, Brazil. A survey of Colletotrichum isolates associated with symptoms of post-bloom fruit drop in São Paulo State showed C. gloeosporioides in addition to C. acutatum. The objectives of this study were to confirm the identification of C. gloeosporioides isolated from symptomatic citrus flowers, to test the pathogenicity of C. gloeosporioides isolates, to compare the development of disease caused by C. gloeosporioides and C. acutatum, and to determine the frequency of C. gloeosporioides in a sample of isolates obtained from symptomatic flowers in different regions of São Paulo State. Through the use of species-specific primers by PCR, 17.3% of 139 isolates were C. gloeosporioides, and the remaining 82.7% were C. acutatum. The pathogenicity tests, carried out in 3-year old potted plants of sweet oranges indicated that both species caused typical symptoms of the disease including blossom blight and persistent calyces. Incubation periods (3.5 and 3.9 days, respectively, for C. acutatum and C. gloeosporioides) and fruit sets (6.7 and 8.5%, respectively for C. acutatum and C. gloeosporioides) were similar for both species. The incidences of blossom blight and persistent calyces were higher on plants inoculated with C. acutatum than in those inoculated with C. gloeosporioides. Conidial germination was similar for both species under different temperatures and wetness periods. Under optimal conditions, appressorium formation and melanisation were higher for C. gloeosporioides than for C. acutatum. These results indicated that Colletotrichum gloeosporioides is a new causal agent of post-bloom fruit drop.     

Research on olive leaf spot,olive knot and verticillium wilt of olive in Turkey1

In Turkey, studies on olive pathology have concerned spore release, overwintering and control of Spilocaea oleagina, control of Pseudomonas syringae subsp. savastanoi by antibiotic drench, and isolation and host range of strains of Verticillium dahliae from olive.     

Isolation of Olive latent virus 1 from Tulip in Toyama Prefecture

A virus whose coat protein gene had a high sequence homology with the coat protein gene of Olive latent virus 1 was isolated from diseased tulip in Toyama Prefecture. Received 1 June 2001/ Accepted in revised form 1 August 2001     

Characterization of a vector-non-transmissible isolate of Tomato spotted wilt virus

The characteristics of a thrips‐non‐transmissible isolate of Tomato spotted wilt virus (TSWV), designated TSWV‐M, were compared with those of a thrips‐transmissible isolate, designated TSWV‐T. TSWV‐M showed a narrower host range than TSWV‐T. Adult thrips failed to transmit TSWV‐M, although the vector acquired the virus during the larval stages. TSWV‐M was detected by RT‐PCR in adult thrips bodies, but not in thrips heads, suggesting that loss of thrips transmissibility was the result of the absence of virus in adult thrips salivary glands. Whereas N (nucleoprotein), NSs (non‐structural protein) and G_C (the C‐terminal portion of the glycoprotein precursor protein) were present in similar amounts in leaf tissue from TSWV‐M‐ or TSWV‐T‐infected plants, G_N (the N‐terminal portion of the glycoprotein precursor protein) was present at much lower amounts in TSWV‐M‐ than in TSWV‐T‐infected plants. SDS‐PAGE and immunoblotting analysis of TSWV‐M and TSWV‐T virion preparations with G_N‐ and G_C‐specific antibodies revealed similar amounts of the G_N and G_C glycoproteins in TSWV‐T virions, but lower amounts of G_N than G_C in TSWV‐M virions. This resulted in a statistically significant reduction in the G_N/G_C ratio in TSWV‐M virions. In affinoblots, the G_C and G_N glycoproteins of TSWV‐M exhibited weak binding with lectins showing affinity for N‐linked oligosaccharide structures. Sequence analysis of M RNA (medium segment of the TSMV genome) revealed no deletions or frameshift mutations in the G_N/G_C precursor of TSWV‐M. However, five amino acid changes were detected in the G_N/G_C precursor. A single, relatively conservative amino acid substitution (V→I) was observed in the NSm protein. Sequence analysis of S RNA (small portion of the TSMV genome) revealed a large intergenic region with no changes in the N protein and with three amino acid changes in the NSs protein.     

Biological, serological and molecular characterization of a cucumber mosaic virus isolate from India

A. virus causing mosaic and leaf deformation of Physalis minima has been identified as an isolate of cucumber mosaic virus (CMV) on the basis of its transmission by aphids in a non-persistent manner, polyhedral particles of 29 nm diameter, molecular weight of coat protein subunits us 24-5 kDa. serological relationship with a CMV isolate and a tripartite single-stranded RNA genome with a subgenomic RNA4- Furthermore. cDNA representing coat protein gene was synthesized and cloned. Complete nucleotide sequences (890 nt) were obtained which showed a coat protein gene open reading frame of 657 residues. THE nucleotide sequences provided the 218 amino ACID sequences of the coat protein. Nucleotide as well as amino acid sequences revealed more than 90% identity with the CMV subgroup I strains.     

辣椒轻斑驳病毒湖南分离物的全基因序列测定及结构分析

采自湖南地区的辣椒轻斑驳病毒Pepper mild mottle virus(PMMoV)样品经单斑分离后,根据已经报道的PMMoV序列基因保守区设计6对简并引物,采用片段重叠法和RACE方法扩增、克隆获得一个全长为6 356bp的湖南分离物(PMMoV-HN1,登录号:KP345899)全基因组序列,编码4个蛋白,分别为126kD蛋白(70~3 423nt)、183kD蛋白(70~4 908nt)、28kD蛋白(4 909~5 682nt)和17.5kD蛋白(5 685~6 158nt),5′-非编码区(5′-UTR)和3′-非编码区(3′-UTR)分别含有69和198个碱基,其中5′-UTR存在一个序列为m7G5′pppG的甲基化核苷酸帽子结构。一致性分析发现PMMoV-HN1与PMMoV其他分离物的核酸一致性为94%~99%,编码的氨基酸一致性为94%~99%。全基因组序列系统进化分析表明PMMoV-HN1分离物与中国首次报道的PMMoV-CN分离物亲缘关系最近。本研究是国内报道的第二例PMMoV全基因组序列。     

Further characterization of celery latent virus

Celery latent virus has been isolated from plants of celeriac (Apium graveolens var.rapaceum).Chenopodium amaranticolor andC. quinoa are good assay plants. Celery (A. graveolens var.dulce) and celeriac (13 cultivars tested) did not react with visible symptoms.Fourteen new artificial hosts were found. New systemic symptomless hosts areAnthriscus cerefolium, Nicotiana megalosiphon, Pisum sativum, Spinacia oleracea andTrifolium incarnatum. Systemic symptoms were caused in the pea cultivars Cicero and Dark Skin Perfection.Five aphid species, includingCavariella aegopodii, four of which not tested with the virus before, were unable to transmit the virus.Seed transmission was confirmed in celeriac (up to 34%) and inC. quinoa (up to 67%) and detected for the first time inAmaranthus caudatus. Detection is easier in seedlings and germinated seeds than in dry seeds.Results of purification were erratic but best at high pH (8 or 9). Sedimentation coefficient was 161 S.An antiserum reacted with purified virus in micro-precipitin tests (titer 256) but, especially at high salt concentrations also in agar gel (titer 64), presumably because of easy degradation of virus protein. Reactions in agar gel also occurred with crude extracts from virus-infectedC. quinoa and celeriac.With the electron microscope flexuous virus particles were found in low concentration in crude sap and in high concentration in purified preparations. Particle measurements revealed an average length of 885 nm (in one preparation 940 nm).Light microscopy did not show inclusion bodies in epidermal strips nor did electron microscopy of ultrathin sections reveal pinwheels and other structures typical of the potyvirus group. The virus evidently belongs to a new morphological group, possibly together with some other viruses hitherto insufficiently studied.The virus seems of potential economic importance only, but it is advised to use virus-free mother plants of celery for seed production.participating in the investigations from August 20th to December 19th 1972, supported by a fellowship of the Fundacion Juan March, Madrid, Spain.     

一个马铃薯Y病毒山东分离物的分离与鉴定

 从具有典型花叶症状的马铃薯叶片中分离到马铃薯Y病毒(Potato virus Y,PVY)(本文称PVY-SD-TA分离物),扩繁后,提纯病毒,电镜下可观察到700~900 nm×11 nm的病毒粒体,病组织超薄切片观察可见风轮状的内含体结构,寄主反应特性研究表明其能侵染2科13种植物。SDS-PAGE电泳检测病毒编码的外壳蛋白亚基的分子量为33 kDa。以PVY-SD-TA基因组RNA为模板,应用RT-PCR方法和特异引物合成了外壳蛋白基因。对cDNA全序列分析表明,PVY-SD-TA CP基因核苷酸序列与N株系的同源性为96%,与GenBank中登录序列号为AJ390306的O株系分离物的同源性最高,为99%;与国内不同学者报道的PVY中国流行株的同源性分别为96%,97%和98%。通过以上生物学特性和分子水平的研究将PVY-SD-TA鉴定为普通株系(PVY^O株系)。