scholarly journals Biological and serological procedures to detect three nepoviruses in fruit trees

2010 ◽  
Vol 40 (No. 4) ◽  
pp. 121-127
Author(s):  
J. Polák ◽  
M. Chaloupková ◽  
M. Jokeš
Keyword(s):  
Phosphate Buffer ◽  
Leaf Roll ◽  
Chenopodium Quinoa ◽  
Fruit Trees ◽  
Ringspot Virus ◽  
Mechanical Transmission ◽  
Biological Detection ◽  
Cysteine Hydrochloride ◽  
Cherry Leaf Roll Virus ◽  
Veronal Buffer

Cherry leaf roll virus (CLRV), Myrobalan latent ringspot virus (MLRSV) and Strawberry latent ringspot virus (SLRSV) were transferred by budding to woody trees, hybrid Ishtara, peach cv. GF 305 and cv. Lesiberian. Three buffers with antioxidants and stabilisers: 0.01M phosphate with 1% caffeine; 0.007M phosphate-0.01M veronal with 0.01M cysteine hydrochloride and 0.007 EDTA; 0.015M phosphate with 1% nicotine and 0.066M phosphate buffer without additives were compared for their efficiency in mechanical transmission from woody sources to herbaceous hosts (Chenopodium quinoa and C. amaranticolor). 0.007M phosphate-0.01M veronal buffer with 0.01M cysteine hydrochloride, and 0.007 EDTA and 0.015M phosphate buffer with 1% nicotine were found to be the best buffers for the three nepoviruses. Both biological transmission to herbaceous assay hosts and detection by ELISA in the investigated tree are necessary to reliably detect the three nepoviruses. Biological detection is reliable from April to June, and in September and October. ELISA detection is also more difficult in July and August. The suitability of C. quinoa and C. amaranticolor to maintain CLRV, MLRSV and SLRSV was compared. C. amaranticolor plants were found to be more suitable for CLRV and SLRSV, infected plants grow over 6 months after mechanical inoculation by the nepoviruses. C. quinoa plants proved to be most suitable for maintenance of MLRSV, while C. amaranticolor is a symptomless host of MLRSV. Reinoculation with the nepoviruses is recommended in intervals of 4 to 6 months.

scholarly journals Localization of Transmissible and Nontransmissible Viruses in the Vector Nematode Xiphinema americanum

2002 ◽  
Vol 92 (6) ◽  
pp. 646-653 ◽  
Author(s):  
Shouhua Wang ◽  
Rose C. Gergerich ◽  
Sandra L. Wickizer ◽  
Kyung S. Kim
Keyword(s):  
Electron Microscopy ◽  
Leaf Roll ◽  
Transmission Efficiency ◽  
Ringspot Virus ◽  
Nematode Population ◽  
Virus Like Particles ◽  
Food Canal ◽  
Cherry Leaf Roll Virus ◽  
The Status ◽  
Section Electron

The inner lining of the food canal of nematodes that transmit plantinfecting viruses is regarded as the retention region of viruses. To characterize the location of transmissible and nontransmissible viruses in the vector nematode Xiphinema americanum, three nepoviruses, Tobacco ringspot virus (TRSV), Tomato ringspot virus(TomRSV), and Cherry leaf roll virus(CLRV), and one non-nematode-transmissible virus, Squash mosaic virus (SqMV), were evaluated for transmission efficiency and localization sites in the nematode. Transmission trials showed highest transmission efficiency for TomRSV (38% with 1 and 100% with 10 nematodes, respectively), intermediate efficiency for TRSV (27% with 1 and 65% with 10 nematodes, respectively), and no transmission for CLRV and SqMV. Electron microscopy and immunofluorescent labeling revealed that TRSV was primarily localized to the lining of the lumen of the stylet extension and the anterior esophagus, but only rarely in the triradiate lumen. Within a nematode population, particles of TRSV were no longer observed in these three regions 10 weeks after acquisition, and it is assumed that there was gradual and random loss of the virus from these areas. The percentage of nematodes that were labeled by virus-specific immunofluorescent labeling in a population of viruliferous nematodes decreased gradually after TRSV acquisition when the nematodes were placed on a nonhost of the virus, and the loss of immunofluorescent labeling paralleled the decrease in the ability of the nematode population to transmit the virus. TomRSV was localized only in the triradiate lumen based on thin-section electron microscopy. No virus-like particles were observed in any part of the food canal of nematodes that had fed on CLRV-infected plants. Virus-like particles that appeared to be partially degraded were observed only in the triradiate lumen of nematodes that had fed on SqMV-infected plants. These results clarified the status of localization of two nontransmissible viruses in X. americanum and presented evidence that two nematode-transmissible viruses, TRSV and TomRSV, are localized in different regions of the food canal of X. americanum.

scholarly journals First Report of Nasturtium as a Natural Host of Cherry leaf roll virus on Amsterdam Island

Plant Disease ◽  
2010 ◽  
Vol 94 (4) ◽  
pp. 477-477 ◽  
Author(s):  
A. Marais ◽  
C. Faure ◽  
T. Candresse ◽  
M. Hullé
Keyword(s):  
Natural Infection ◽  
Leaf Roll ◽  
Plant Viruses ◽  
Natural Host ◽  
Mechanical Transmission ◽  
Sequence Information ◽  
Coding Region ◽  
Rt Pcr ◽  
First Report ◽  
Cherry Leaf Roll Virus

Cherry leaf roll virus (CLRV) is a well-known virus belonging to the genus Nepovirus, but unlike most members of this genus, it is not known to be transmitted by nematodes but only through seeds and pollen. Since its first description in 1955 on Prunus avium L. in England (1), CLRV has been shown to have a worldwide distribution and a wide natural host range. During a survey of plant viruses in the French sub-Antarctic islands, samples from nasturtium plants (Tropaeolum majus), an introduced plant species, showing symptoms of leaf mosaic, deformation, and veinal necrosis were collected on Amsterdam Island. Upon mechanical transmission with sap extracts, necrotic ringspot and oak-leaf symptoms typical of Nepovirus infection were observed on the leaves of inoculated Nicotiana clevelandii and N. tabacum plants. Inoculation of healthy nasturtium plants resulted in mosaic and pin-point necrosis symptoms. Electron microscopy on negatively stained sap extracts revealed the presence of icosahedral virions, 28 to 30 nm in diameter, in the symptomatic Nicotiana leaves. Amplification by reverse transcription (RT)-PCR with a polyvalent test, which identifies viruses belonging to the family Comoviridae (2), yielded the expected 248-bp fragment. Sequencing of the cloned amplicon showed 80% nucleotide and 90% amino acid identity with a part of the RNA dependent RNA polymerase (RdRp) of CLRV (CAE83562). To confirm the presence of CLRV, an approximate 4.6-kbp cDNA fragment was PCR amplified from double-stranded RNAs purifed from infected Nicotiana plants using the sense primer 5′-GTGGGACTGCCATGCACCTACTC-3′ and an oligo-T25 as antisense primer. This PCR product (GenBank Accession No. GU167974) spans the region between the VPg gene and the polyA tail at the 3′ end of the genome and thus provides approximately 2.8 kb of new internal sequence information on RNA1 of CLRV. The presence of CLRV in the initial nasturtium samples was confirmed with a CLRV-specific RT-PCR assay that amplifies the 3′ non-coding region of the CLRV genome (3). Sequence of the amplified fragment showed it to be identical to the corresponding part of the 3′ non-coding region of 4.6-kbp clone obtained from the CLRV isolate mechanically transmitted to the N. tabacum and N. clevelandii plants. Experimental infection of nasturtium by CLRV has been reported (4), but to the best of our knowledge these results represent the first report of natural infection of T. majus by CLRV. Given its seed transmissible character in many hosts, CLRV likely was introduced in infected seeds of T. majus imported to the remote sub-Antarctic Amsterdam Island. References: (1) R. Cropley. Ann. Appl. Biol. 49:524, 1961. (2) V. Maliogka et al. J. Phytopathol. 152:404, 2004. (3) K. Rebenstorf et al. J. Virol. 80:2453, 2006. (4) K. Schmelzer. Phytopathol. Z. 55:317, 1966.

scholarly journals Detection and phylogeny of viruses in native Albanian olive varieties

2021 ◽  
Vol 60 (1) ◽  
pp. 165-174
Author(s):  
Toufic ELBEAINO ◽  
Magdalena CARA ◽  
Shpend SHAHINI ◽  
Pasko PANDELI
Keyword(s):  
Mosaic Virus ◽  
Leaf Roll ◽  
Olive Tree ◽  
Ringspot Virus ◽  
Arabis Mosaic Virus ◽  
Cherry Leaf Roll Virus ◽  
Leaf Yellowing ◽  
Standard Quality ◽  
Olive Trees ◽  
Olive Varieties

Forty samples representing 14 native Albanian and two foreign olive varieties were collected from an olive varietal collection plot in the Valias region (Tirana, Albania). The samples were assayed by RT-PCR for presence of olive-infecting viruses, including arabis mosaic virus (ArMV), cherry leaf roll virus (CLRV), cucumber mosaic virus (CMV), olive latent ringspot virus (OLRSV), olive latent virus 1 (OLV-1), olive leaf yellowing-associated virus (OLYaV), strawberry latent ringspot virus (SLRSV) and by PCR for the bacterium Xylella fastidiosa (Xf). Ninety-eight percent of the samples were infected with at least one virus. OLYaV was the most prevalent (85% of samples), followed by OLV-1 (50%), OLRSV (48%), CMV (28%), SLRSV (3%) and CLRV (5%), whereas ArMV and Xf were absent. Fifty-five percent of the samples were infected with one virus, 13% with two viruses, 20% with three, and 5% with four. Analyses of the nucleotide sequences of the Albanian virus isolates generally showed low genetic variability, and that most were phylogenetically related to Mediterranean isolates, in particular to those from Greece and Italy. Five olive trees, representing three native cultivars (‘Managiel’, ‘Kalinjot’ and ‘Kushan-Preze’) and one foreign (‘Leccino’), were found to be plants of the Conformitas Agraria Communitatis (“CAC”) category i.e. free of ArMV, CLRV, SLRSV and OLYaV. Only one tree of the native cultivar ‘Ulliri i kuq’ was free of all tested viruses, so this is plant material of the “Virus-tested” category. Olives derived from both categories could be used for propagation of standard quality plant materiel in a future certification programme for olive in Albania. This is the first report of CLRV, OLRSV, CMV and OLV-1 in Albania. The study also reveals the precarious health status of native olive varieties in the Valias varietal collection plot. However, the discovery of six plants representing two certifiable categories is a first step in a future olive tree certification program in the country.

scholarly journals First Report of a Disease of Peony Caused by Alfalfa mosaic virus

Plant Disease ◽  
2003 ◽  
Vol 87 (1) ◽  
pp. 99-99 ◽  
Author(s):  
M. G. Bellardi ◽  
C. Rubies-Autonell ◽  
A. Bianchi
Keyword(s):  
Mosaic Virus ◽  
Leaf Roll ◽  
Tobacco Rattle Virus ◽  
Alfalfa Mosaic Virus ◽  
Ringspot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
First Report ◽  
Systemic Leaf ◽  
Cherry Leaf Roll Virus ◽  
Line Patterns

During the summers of 2001 and 2002, Japanese peony (Paeonia albiflora Pall., synonym P. lactiflora, family Paeoniaceae) plants, cultivated in the Botanical Garden of the University of Parma (Emilia Romagna Region of northern Italy), were found affected by a disease with virus-like symptoms. The oldest leaves showed yellow, mosaic, oak-like arabesques and line-patterns; the remaining leaves and pink flowers were symptomless. A disease of peony, known as peony ring spot disease, has been reported worldwide (Europe, United States, Japan, and New Zeland) for several years and is associated with strains of Tobacco rattle virus (TRV) (1). Electron microscopic observations of peony leaf sap (leaf dip preparations stained with uranyl acetate and phospotungstic acid) did not show the presence of any rod-shaped virus particles, including TRV. Mechanical inoculations of sap from symptomatic leaves caused symptoms typical of Alfalfa mosaic virus (AMV) on Chenopodium amaranticolor Coste & Reyn. (local chlorotic and necrotic lesions and systemic periveinal line-patterns), Ocimum basilicum L. (yellow mosaic), Vigna unguiculata (L.) Walp. (red, local necrotic lesions), and Nicotiana tabacum cv. Samsun (white, necrotic lesions, systemic leaf malformation, and mosaic), and N. glutinosa L. (systemic leaf variegation). Symptomatic leaves of peony and infected herbaceous plants were analyzed for virus presence by protein A sandwich enzyme-linked immunosorbent assay (PAS-ELISA). The polyclonal antisera tested were those to AMV (PVAS 92, American Type Culture Collection, Manassas, VA), AMV-Vinca minor L. (DiSTA collection, Italy), and the nepoviruses Strawberry latent ringspot virus, Tomato ringspot virus, and Cherry leaf roll virus. PAS-ELISA revealed only the presence of AMV. Immunoelectron microscopy and gold-labeled decoration confirmed the identity of the virus. In 2001, five symptomless peony plants (provided by a commercial grower and previously analyzed for AMV and TRV presence) were grafted with shoots from peony showing yellow mosaic on the leaves and maintained in a greenhouse under aphid-proof cage. During the summer of 2002, one of the grafted plants showed a mild mosaic on the leaves; PAS-ELISA revealed this peony was infected by AMV. To our knowledge, this is the first report of AMV in peony. Reference: (1) Chang et al. Ann. Phytopathol. Soc. Jpn. 42:325, 1976.

scholarly journals Virus Surveys in Olive Orchards in Greece Identify Olive Virus T, a Novel Member of the Genus Tepovirus

Pathogens ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 574
Author(s):  
Evanthia Xylogianni ◽  
Paolo Margaria ◽  
Dennis Knierim ◽  
Kyriaki Sareli ◽  
Stephan Winter ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Leaf Roll ◽  
Virus Infections ◽  
Northern Greece ◽  
Coat Protein Region ◽  
Cherry Leaf Roll Virus ◽  
Leaf Yellowing ◽  
Olive Trees ◽  
Olive Varieties ◽  
Viral Sequences

Field surveys were conducted in Greek olive orchards from 2017 to 2020 to collect information on the sanitary status of the trees. Using a high-throughput sequencing approach, viral sequences were identified in total RNA extracts from several trees and assembled to reconstruct the complete genomes of two isolates of a new viral species of the genus Tepovirus (Betaflexiviridae), for which the name olive virus T (OlVT) is proposed. A reverse transcription–polymerase chain reaction assay was developed which detected OlVT in samples collected in olive growing regions in Central and Northern Greece, showing a virus prevalence of 4.4% in the olive trees screened. Sequences of amplified fragments from the movement–coat protein region of OlVT isolates varied from 75.64% to 99.35%. Three olive varieties (Koroneiki, Arbequina and Frantoio) were infected with OlVT via grafting to confirm a graft-transmissible agent, but virus infections remained latent. In addition, cucumber mosaic virus, olive leaf yellowing-associated virus and cherry leaf roll virus were identified.

scholarly journals Detection of Cherry leaf roll virus and Strawberry latent ring spot virus by one-step RT-PCR

2009 ◽  
Vol 45 (No. 4) ◽  
pp. 140-143 ◽  
Author(s):  
S. Kumari
Keyword(s):  
Leaf Roll ◽  
Rt Pcr ◽  
Spot Virus ◽  
Cherry Leaf Roll Virus ◽  
One Step ◽  
Polymerase Chain ◽  
The One ◽  
Ring Spot ◽  
Screen House ◽  
Xiphinema Diversicaudatum

A one-step reverse transcription-polymerase chain reaction (RT-PCR) protocol was developed and used for the detection of <i>Cherry leaf roll virus</i> (CLRV) and <i>Strawberry latent ring spot virus</i> (SLRSV). The protocol was used to test infected screen house plants and also plants from orchards and vineyards where the vector (<i>Xiphinema diversicaudatum</i>) of SLRSV was detected from the soil. The one-step RT-PCR protocol is rapid and sensitive and has the potential to be used for the diagnosis of CLRV and SLRSV in routine diagnostic laboratories.

Serological Comparison of Isolates of Cherry Leaf Roll Virus from Diseased Beech and Birch Trees in a Forest Decline Area in Germany with Other Isolates of the Virus

1990 ◽  
Vol 129 (4) ◽  
pp. 339-344 ◽  
Author(s):  
A. T. Jones ◽  
Renate Koenig ◽  
D.-E. Lesemann ◽  
J. Hamacher ◽  
F. Nienhaus ◽  
...  
Keyword(s):  
Forest Decline ◽  
Leaf Roll ◽  
Cherry Leaf Roll Virus ◽  
Birch Trees
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