Cucumber mosaic virus 2b-Deficient Mutant Causes Limited, Asymptomatic Infection of Bell Pepper

Cucumber mosaic virus Fast New York strain (CMV-Fny) containing a mutated 2b protein (CMV-FnyΔ2b) was evaluated for the ability to infect ‘Calwonder’ bell pepper (Capsicum annuum) plants in comparative tests with the parent virus, CMV-Fny. Plants inoculated with CMV-FnyΔ2b did not develop local or systemic symptoms of infection, whereas CMV-Fny-infected plants developed systemic chlorosis by 7 days post inoculation (dpi), followed by mosaic and leaf deformation. Virus accumulation, determined by enzyme-linked immunosorbent assay (ELISA), revealed that CMV-FnyΔ2b accumulated in inoculated Calwonder leaves and inconsistently infected some noninoculated leaves at a low titer but was not detected in the youngest, noninoculated leaves. Immuno-tissue blot tests did not detect CMV-FnyΔ2b in the stems of infected plants, whereas CMV-Fny accumulated throughout the length of the stems of inoculated plants. In two experiments, protoplasts were isolated from Calwonder leaves, inoculated with viral RNAs of CMV-Fny or CMV-FnyΔ2b, and tested by ELISA for infection. In both experiments, less CMV-FnyΔ2b than CMV-Fny accumulated in protoplasts. These results suggest that the CMV 2b protein is needed for systemic infection of Calwonder pepper plants and for accumulation of the virus in inoculated protoplasts.

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Multigenic System Controlling Viral Systemic Infection Determined by the Interactions Between Cucumber mosaic virus Genes and Quantitative Trait Loci of Soybean Cultivars

Phytopathology ◽

10.1094/phyto-06-10-0154 ◽

2011 ◽

Vol 101 (5) ◽

pp. 575-582 ◽

Cited By ~ 5

Author(s):

Shizen Ohnishi ◽

Issei Echizenya ◽

Eri Yoshimoto ◽

Kim Boumin ◽

Tsuyoshi Inukai ◽

...

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Quantitative Trait ◽

Single Gene ◽

Systemic Infection ◽

Northern Hybridization ◽

Enzyme Linked Immunosorbent Assay ◽

Cdna Clones ◽

Long Distance ◽

Chimeric Rnas

Soybean ‘Harosoy’ is resistant to Cucumber mosaic virus soybean strain C (CMV-SC) and susceptible to CMV-S strain D (CMV-SD). Using enzyme-linked immunosorbent assay and Northern hybridization, we characterized the Harosoy resistance and found that CMV-SC did not spread systemically but was restricted to the inoculated leaves in Harosoy. Harosoy resistance was not controlled by either a dominant or recessive single gene. To dissect this system controlling long-distance movement of CMV in soybean, we constructed infectious cDNA clones of CMV-SC and CMV-SD. Using these constructs and the chimeric RNAs, we demonstrated that two viral components were required for systemic infection by the virus. The region including the entire 2b gene and the 5′ region of RNA3 (mainly the 5′ untranslated region) together were required. By quantitative trait locus (QTL) analysis using an F2 population and the F3 families derived from Harosoy and susceptible ‘Nemashirazu’, we also showed that at least three QTLs affected systemic infection of CMV in soybean. Our study on Harosoy resistance to CMV-SC revealed an interesting mechanism, in which multiple host and viral genes coordinately controlled viral systemic infection.

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First Report of Cucumber mosaic virus in Teucrium fruticans

Plant Disease ◽

10.1094/pdis.2003.87.2.200c ◽

2003 ◽

Vol 87 (2) ◽

pp. 200-200 ◽

Cited By ~ 1

Author(s):

L. Cardin ◽

A. Poupet ◽

J. P. Onesto

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Chenopodium Quinoa ◽

Mediterranean Area ◽

Uranyl Acetate ◽

Enzyme Linked Immunosorbent Assay ◽

Public Gardens ◽

Serotype 1 ◽

Systemic Symptoms ◽

Antigen Antibody

Teucrium fruticans (shrubby germander), family Lamiaceae, is a hardy shrub. Being drought tolerant, it is widespread in the Mediterranean area. Because it is readily propagated through cuttings, it is also planted in hedges. In 1997 and 2000, respectively, yellow chlorotic areas were observed on the foliage of T. fruticans in Saint Jean Cap Ferrat (France) and San Remo (Italy). These symptoms were distinct from those produced by a rust that frequently affects T. fruticans in these areas. Viruses from both locations were identified as Cucumber mosaic virus (CMV) based on the following: (i) symptoms after mechanical inoculation of Nicotiana tabacum cv. Xanthi nc, N. tabacum cv. Samsum, Chenopodium quinoa, C. amaranticolor, Vigna unguiculata cv. Black, and Cucumis sativus cv. Poinsett; (ii) the morphology of particles observed in electron microscopy of uranyl acetate stained leaf dips from tobacco; and (iii) positive result from leaves of diseased T. fruticans and mechanically inoculated host plants cited above based on enzyme-linked immunosorbent assay (ELISA) using CMV antisera. On tobacco cv. Xanthi nc, the French (F) and Italian (I) isolates first induced essentially necrotic rings on the inoculated leaves followed by the same systemic symptoms as described above. The two isolates were cloned from local lesions after two successive inoculations in V. unguiculata cv. Black, multiplied in tobacco, purified with the citrate-chloroform method, and stabilized with formaldehyde (1). The serotype determination was made by double immunodiffusion in agar gel with the CMV-D and CMV-To strains and homologous antisera (1,2). The formation of spurs and antigen-antibody lines indicated that both isolates belonged to the ToRS serotype (1). Thirty plants of T. fruticans cv. Azureum, first tested negative for CMV using ELISA, were mechanically inoculated with the F isolate (25 plants) and the CMV-D strain (five plants) and cultivated in a hydroponic system. Three months later, plants inoculated with the F isolate were positive for CMV using ELISA and displayed clear symptoms with chlorotic spots, which were sometimes ring-shaped. As plants mature, symptoms tend to disappear on young shoots. For the CMV-D strain, three plants of five were ELISA positive, but did not show any typical symptoms. This report demonstrates the infection of T. fruticans by CMV and the symptom induction by some CMV isolates. In September 2002, two CMV isolates were collected from T. fruticans in public gardens in Menton (France) and Genoa (Italy). These new isolates have the same characteristics as those described in this report. References: (1) J. C. Devergne and L. Cardin. Ann. Phytopathol. 7:225, 1975. (2) M. H. V. van Regenmortel. Adv. Virus Res. 12:207, 1966.

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Susceptibility of Vegetatively Propagated Petunia to Tobamovirus Infection and Its Possible Control

HortScience ◽

10.21273/hortsci.34.2.292 ◽

1999 ◽

Vol 34 (2) ◽

pp. 292-293 ◽

Cited By ~ 2

Author(s):

J. Cohen ◽

Noga Sikron ◽

S. Shuval ◽

A. Gera

Keyword(s):

Mosaic Virus ◽

Immunosorbent Assay ◽

Phosphate Buffer ◽

Sodium Phosphate ◽

Tomato Mosaic Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Post Inoculation ◽

Sodium Phosphate Buffer ◽

Local Lesions ◽

Systemic Symptoms

In this study, 18 Petunia ×hybrida Hort. Volm.-Andr. cultivars were mechanically inoculated with the tobamoviruses tobacco mosaic (TMV) or tomato mosaic virus (ToMV) (20 μg·L-1 in 0.05 m sodium phosphate buffer). One and 2 weeks post-inoculation (PI), inoculated and noninoculated upper leaves were harvested and assayed for TMV infection using enzyme-linked immunosorbent assay (ELISA). Local lesions developed on inoculated leaves of 16 cultivars 3-5 days PI. A total of 11 and 16 of the cultivars developed systemic symptoms characteristic of tobamovirus infection 2 weeks after inoculation with TMV and ToMV, respectively. All cultivars were positive in ELISA tests. Large amounts of virus were recovered from the upper, noninoculated leaves of all cultivars, including symptomless plants. Up to 95% infection by TMV occurred when a sterilized knife was passed through an infected shoot of petunia prior to its being used to remove cuttings from healthy petunia plants. Heat sterilization of knives and/or treatment with 2.8 g·L-1 sodium troclosene was very effective in controlling TMV transmission.

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Topical Application of Double-Stranded RNA Targeting 2b and CP Genes of Cucumber mosaic virus Protects Plants against Local and Systemic Viral Infection

Plants ◽

10.3390/plants10050963 ◽

2021 ◽

Vol 10 (5) ◽

pp. 963

Author(s):

Maria C. Holeva ◽

Athanasios Sklavounos ◽

Rajendran Rajeswaran ◽

Mikhail M. Pooggin ◽

Andreas E. Voloudakis

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Topical Application ◽

Plant Virus ◽

Plant Protection ◽

Post Inoculation ◽

Tobacco Plants ◽

Double Stranded Rna

Cucumber mosaic virus (CMV) is a destructive plant virus with worldwide distribution and the broadest host range of any known plant virus, as well as a model plant virus for understanding plant–virus interactions. Since the discovery of RNA interference (RNAi) as a major antiviral defense, RNAi-based technologies have been developed for plant protection against viral diseases. In plants and animals, a key trigger of RNAi is double-stranded RNA (dsRNA) processed by Dicer and Dicer-like (DCL) family proteins in small interfering RNAs (siRNAs). In the present study, dsRNAs for coat protein (CP) and 2b genes of CMV were produced in vitro and in vivo and applied onto tobacco plants representing a systemic solanaceous host as well as on a local host plant Chenopodium quinoa. Both dsRNA treatments protected plants from local and systemic infection with CMV, but not against infection with unrelated viruses, confirming sequence specificity of antiviral RNAi. Antiviral RNAi was effective when dsRNAs were applied simultaneously with or four days prior to CMV inoculation, but not four days post inoculation. In vivo-produced dsRNAs were more effective than the in vitro-produced; in treatments with in vivo dsRNAs, dsRNA-CP was more effective than dsRNA-2b, while the effects were opposite with in vitro dsRNAs. Illumina sequencing of small RNAs from in vivo dsRNA-CP treated and non-treated tobacco plants revealed that interference with CMV infection in systemic leaves coincides with strongly reduced accumulation of virus-derived 21- and 22-nucleotide (nt) siRNAs, likely generated by tobacco DCL4 and DCL2, respectively. While the 21-nt class of viral siRNAs was predominant in non-treated plants, 21-nt and 22-nt classes accumulated at almost equal (but low) levels in dsRNA treated plants, suggesting that dsRNA treatment may boost DCL2 activity. Taken together, our findings confirm the efficacy of topical application of dsRNA for plant protection against viruses and shed more light on the mechanism of antiviral RNAi.

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Lower temperature influences Cauliflower mosaic virus systemic infection

Journal of General Plant Pathology ◽

10.1007/s10327-021-01003-y ◽

2021 ◽

Author(s):

Roberto Alers-Velazquez ◽

Sushant Khandekar ◽

Clare Muller ◽

Jennifer Boldt ◽

Scott Leisner

Keyword(s):

Nucleic Acid ◽

Actin Cytoskeleton ◽

Mosaic Virus ◽

Virus Particle ◽

Systemic Infection ◽

Cauliflower Mosaic Virus ◽

Particle Release ◽

Systemic Symptoms ◽

Lower Temperature ◽

Temperature Exposure

AbstractLower temperatures delayed development of systemic symptoms by Cauliflower mosaic virus (CaMV) in two different plant hosts. However, lower temperature exposure increased CaMV nucleic acid levels in leaves of systemically-infected turnips. Furthermore, lower temperature altered the formation of aggregates formed by the CaMV major inclusion body (IB) protein, P6. Finally, lower temperature altered the architecture of the actin cytoskeleton. These data may suggest that lower temperatures alter the actin cytoskeleton, facilitating the formation of larger IBs that hold on to their internal virions more strongly than small ones, impairing virus particle release and causing a delay in systemic infection.

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Epidemic of Potato virus Y and Cucumber mosaic virus in Henan Province Tobacco

Plant Disease ◽

10.1094/pdis.2001.85.4.447c ◽

2001 ◽

Vol 85 (4) ◽

pp. 447-447 ◽

Cited By ~ 1

Author(s):

X. D. Li ◽

Y. Q. Li ◽

H. G. Wang

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Potato Virus ◽

Potato Virus Y ◽

Early Stage ◽

Potato Virus X ◽

Henan Province ◽

Enzyme Linked Immunosorbent Assay ◽

Resistant Varieties ◽

Vein Necrosis

Flue-cured tobacco is an important crop in Henan Province, China. During the 2000 growing season, many tobacco plants showed various degrees of mottling, mosaic, vein clearing, or vein necrosis in most of the counties. Some plants even died at an early stage of growth. A survey was conducted in May-June in several tobacco-growing counties, and the incidence of symptomatic plants in individual fields ranged from 10 to 85%. The most widely planted tobacco varieties, NC89, K326, and K346, were highly susceptible. Symptomatic plants were collected from Jiaxian and Xiangcheng counties and samples were tested by enzyme-linked immunosorbent assay for Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Potato virus Y (PVY), and Potato virus X (PVX). Of 65 samples tested, 21 were positive for only PVY, 16 positive for only CMV, one each was positive for only TMV or PVX. Nineteen samples were doubly infected with various combinations of these viruses and six were infected with combinations of three viruses. The causal agent(s) in the remaining sample could not be determined. In total, CMV was detected in 40 samples, PVY in 38, PVX in 10, and TMV in 7 samples. TMV and CMV used to be the most important viruses and PVY occurred only rarely. But PVY has become prevalent in Henan and in neighboring Shandong province (2). CMV and TMV were reported to be the most prevalent viruses in Shanxi (1) and Fujian Provinces (3). Because resistant varieties are not available, and mixed infections are more common, the results presented here explain why huge damage is occurring in tobacco crops in recent years. Some varieties are partially resistant to TMV and CMV but the varieties commonly grown are highly susceptible to PVY. Therefore, breeding for resistance to viruses, especially to PVY, is urgent to control the occurrence of tobacco viral diseases. References: (1) J. L. Cheng et al. Acta Tabacaria Sin. 4:43, 1998. (2) J. B. Wang et al. Chinese Tobacco Sci. 1:26, 1998. (3) L. H. Xie et al. Acta Tabacaria Sin. 2:25, 1994.

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Allamanda Mosaic Caused by Cucumber mosaic virus in Taiwan

Plant Disease ◽

10.1094/pd-89-0529b ◽

2005 ◽

Vol 89 (5) ◽

pp. 529-529 ◽

Cited By ~ 1

Author(s):

Y. K. Chen ◽

C. C. Yang ◽

H. T. Hsu

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Chenopodium Quinoa ◽

Rabbit Antiserum ◽

Nucleotide Sequence Analysis ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Indicator Plants

Allamanda (Allamanda cathartica L., family Apocynaceae) is native to Brazil and is a popular perennial shrub or vine ornamental in Taiwan. Plants showing severe mosaic, rugosity, and leaf distortion symptoms on leaves are common in commercial nurseries and private gardens. Examination of crude sap prepared from symptomatic leaves using an electron microscope revealed the presence of spherical virus particles with a diameter of approximately 28 nm. The virus was mechanically transmitted to indicator plants and induced symptoms similar to those incited by Cucumber mosaic virus (CMV). The virus caused local lesions on inoculated leaves of Chenopodium quinoa and C. amaranticolor and systemic mosaic in Cucumis sativus, Lycopersicon esculentum, Nicotiana benthamiana, N. glutinosa, N. rustica, and N. tabacum. On N. tabacum, necrotic ringspots developed on inoculated leaves followed by systemic mosaic. Tests of leaf sap extracted from naturally infected allamanda and inoculated indicator plants using enzyme-linked immunosorbent assay were positive to rabbit antiserum prepared to CMV. Viral coat protein on transblots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis reacted with CMV subgroup I specific monoclonal antibodies (2). With primers specific to the 3′-half of RNA 3 (1), amplicons of an expected size (1,115 bp) were obtained in reverse transcription-polymerase chain reaction (RT-PCR) using total RNA extracted from infected allamanda and N. benthamiana. The amplified fragment (EMBL Accession No. AJ871492) was cloned and sequenced. It encompasses the 3′ part of the intergenic region of RNA 3 (158 nt), CP ORF (657 nt), and 3′ NTR (300 nt) showing 91.8–98.9% and 71.4–72.8% identities to those of CMV in subgroups I and II, respectively. Results of MspI-digested restriction fragment length polymorphism patterns of the RT-PCR fragment and the nucleotide sequence analysis indicate that the CMV isolate from allamanda belongs to subgroup IB, which is predominant on the island. To our knowledge, CMV is the only reported virus that infects allamanda and was first detected in Brazil (3), and this is the first report of CMV infection in allamanda plants occurring in Taiwan. References: (1) Y. K. Chen et al. Arch. Virol. 146:1631, 2001. (2) H. T. Hsu et al. Phytopathology 90:615, 2000. (3) E. W. Kitajima. Acta. Hortic. 234:451, 1988.

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First Report of Cucumber mosaic virus Subgroup II Infecting Lycopersicon esculentum in India

Plant Disease ◽

10.1094/pd-90-1457b ◽

2006 ◽

Vol 90 (11) ◽

pp. 1457-1457 ◽

Cited By ~ 15

Author(s):

N. Sudhakar ◽

D. Nagendra-Prasad ◽

N. Mohan ◽

K. Murugesan

Keyword(s):

Coat Protein ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Tamil Nadu ◽

Indian Subcontinent ◽

Enzyme Linked Immunosorbent Assay ◽

Infected Leaf ◽

Polymorphism Analysis ◽

First Report ◽

Subgroup Ii

During a survey in January 2006 near Salem in Tamil Nadu (south India), Cucumber mosaic virus was observed infecting tomatoes with an incidence of more than 70%. Plants exhibiting severe mosaic, leaf puckering, and stunted growth were collected, and the virus was identified using diagnostic hosts, evaluation of physical properties of the virus, compound enzyme-linked immunosorbent assay (ELISA) (ELISA Lab, Washington State University, Prosser), reverse-transcription polymerase chain reaction (RT-PCR), and restriction fragment length polymorphism analysis (DSMZ, S. Winter, Germany). To determine the specific CMV subgroup, total RNA was extracted from 50 infected leaf samples using the RNeasy plant RNA isolation kit (Qiagen, Hilden, Germany) and tested for the presence of the complete CMV coat protein gene using specific primers as described by Rizos et al. (1). A fragment of the coat protein was amplified and subsequently digested with MspI to reveal a pattern of two fragments (336 and 538 bp), indicating CMV subgroup II. No evidence of mixed infection with CMV subgroup I was obtained when CMV isolates representing subgroups I (PV-0419) and II (PV-0420), available at the DSMZ Plant Virus Collection, were used as controls. Only CMV subgroup I has been found to predominantly infect tomato in the Indian subcontinent, although Verma et al. (2) identified CMV subgroup II infecting Pelargonium spp., an ornamental plant. To our knowledge, this is the first report of CMV subgroup II infecting tomato crops in India. References: (1) H. Rizos et al. J. Gen. Virol. 73:2099, 1992. (2) N. Verma et al. J. Biol. Sci. 31:47, 2006.

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Cucumber Mosaic Virus, Tobacco Streak Virus, and Cucumber Mosaic Virus Satellite RNA Associated with Mosaic and Ringspot Symptoms in Ajuga reptans in Ohio

Plant Disease ◽

10.1094/pdis.1997.81.10.1214d ◽

1997 ◽

Vol 81 (10) ◽

pp. 1214-1214 ◽

Cited By ~ 1

Author(s):

J. R. Fisher ◽

S. T. Nameth

Keyword(s):

United States ◽

Molecular Weight ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

The United States ◽

Negative Control ◽

Enzyme Linked Immunosorbent Assay ◽

Low Molecular Weight ◽

Streak Virus ◽

Ajuga Reptans

Creeping bugleweed (Ajuga reptans L.) is a perennial ornamental commonly grown as a ground cover in temperate climates. Commercial samples of the A. reptans cultivars Royalty, var. Atropurpurea Bronze, Bronze Beauty, and Burgundy Glow showing mosaic and ringspot symptoms were tested for the presence of virus infection by direct antibody sandwich enzyme-linked immunosorbent assay (ELISA) and viral-associated double-stranded (ds) RNA analysis. Cucumber mosaic cucumovirus (CMV) was detected by ELISA and dsRNA analysis in symptomatic samples of all cultivars tested. ELISA values were considered positive if the absorbance values were twice the negative control. Negative control values were established with asymptomatic tissue of the cv. Bronze Beauty. Tobacco streak ilarvirus (TSV) was detected only by ELISA in symptomatic samples of all cultivars except Royalty. No dsRNA suggestive of TSV was detected. Alfalfa mosaic virus (AMV) was detected by ELISA and dsRNA analysis in symptomatic samples of all cultivars tested except Royalty and var. Atropurpurea Bronze. dsRNA analysis also indicated the presence of a low molecular weight, possible satellite (sat) RNA associated with all symptomatic and asymptomatic Royalty and var. Atropurpurea Bronze plants tested. Northern (RNA) blot analysis with a digoxigenin-labeled full-length clone of the (S) CARNA-5 (-) CMV satRNA (ATCC no. 45124) confirmed that the low molecular weight RNA associated with the Royalty and var. Atropurpurea Bronze cultivars was indeed CMV satRNA. Only AMV has been previously reported in A. reptans in the United States (1). This is the first report of CMV and its satRNA, as well as TSV, in A. reptans in the United States. Reference: (1) W. T. Schroeder and R. Provvidenti. Plant Dis. Rep. 56:285, 1972.

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Dalmatian Toadflax (Linaria dalmatica): New Host for Cucumber Mosaic Virus

Weed Technology ◽

10.1614/wt-06-005.1 ◽

2007 ◽

Vol 21 (1) ◽

pp. 41-44 ◽

Cited By ~ 1

Author(s):

Courtney L. Pariera Dinkins ◽

Sue K. Brumfield ◽

Robert K. D. Peterson ◽

William E. Grey ◽

Sharlene E. Sing

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Aphid Transmission ◽

Negative Control ◽

Enzyme Linked Immunosorbent Assay ◽

New Host ◽

Tobacco Plants ◽

Linaria Dalmatica ◽

Mechanical Methods ◽

Dalmatian Toadflax

To date, there have been no reports of Dalmatian toadflax serving as a host for cucumber mosaic virus (CMV). Infestations of Dalmatian toadflax may serve as a reservoir of CMV, thereby facilitating aphid transmission of CMV to both agricultural crops and native plants. The goal of this study was to determine whether Dalmatian toadflax is a host for CMV. Dalmatian toadflax seedlings were randomly assigned to two treatments (18 replicates/treatment): no inoculation (control) and inoculation with CMV (Fast New York strain). The Dalmatian toadflax seedlings were inoculated by standard mechanical methods and tested for the presence of CMV using enzyme-linked immunosorbent assay (ELISA). Ten of the 18 CMV-inoculated toadflax plants tested positive for the virus; 6 of the 18 displayed systemic mosaic chlorosis and leaf curling. All control plants tested negative. Transmission electron microscopy obtained from CMV-positive plants confirmed the presence of CMV based on physical properties. To verify CMV infestation, tobacco plants were assigned to the following treatments (six replicates/treatment): no inoculation (control), CMV-negative (control) inoculation, and a CMV-positive inoculation. Plants were inoculated by standard methods. Five of the 6 tobacco plants treated with the CMV-positive inoculum tested positive for CMV using ELISA. All control plants tested negative for the virus.

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