Cytological, Histological and Molecular Characterization of Two Isolates of Cucumber Mosaic Virus (CMV) in Egypt

Elsayed E. Wagih, Mohamed M. Zalat, Maha Adel Kawanna


Two isolates of Cucumber mosaic virus (CMV), CMV-wild tobacco (from Alexandria governorate) and CMV-cucumber (from Kafr El-Sheikh governorate) were investigated in this study. Cytological studies on epidermal strips of Nicotiana glutinosa leaves separately infected with each isolate revealed the presence of viral crystalline inclusion bodies within the infected cells. Electron microscopy of ultrathin sections of CMV infected N. glutinosa leaves showed significant alterations in the shape and internal structure of chloroplasts. The cell wall had serrated edges in infected cells but was more severe in cells infected with CMV-wild tobacco isolate compared to those infected with CMV-cucumber isolate. CMV-cucumber isolate was partially purified from systemically infected leaves of N. glutinosa. The ratio A260/ 280 was 1.0 and the concentration of the virus in the preparation was estimated using an extinction coefficient of E260nm0.1%, 1cm = 5. Yield of purified virus was about 2.8 mg/100 g fresh weight of infected N. glutinosa leaves. Electron microscopy of the purified preparation of CMV showed the presence of numerous spherical particles with a mean particle diameter of 28 nm. Amplified real-time reverse transcription-polymerase chain reaction (qRT-PCR) product of coat protein gene of each isolate was purified and sequenced. Sequences of both isolates had been submitted to GenBank Database and ware assigned accession number (LT669766) for CMV-cucumber isolate and (LT706517) for CMV-wild tobacco isolate. The sequences were edited using Chromas Pro. Version 1.34 software and compared with previously subgrouping of 27 isolates of the virus retrieved from the GenBank database. Both CMV-wild tobacco and CMV-cucumber isolates were closely related to the isolate with the accession number AJ585086 with a similarity of 97.07% and 98.54%, respectively, suggesting that the two isolates belong to subgroup II. According to the available literature, this is the first report in Egypt where CMV isolates belonging to subgroup II have been obtained


CMV; Cytological studies; Histological studies; Purification; Phylogenetic analysis


Aglave, B. A., M. Krishnareddy, F. S. Patil and M. S. Andhale. 2007. Molecular identification of a virus causing banana chlorosis disease from Marathwada region. International Journal of Biotechnology and Biochemistry, 3: 13-24.

Ashby, J. W. 1977. Infection of karaka (Corynocarpus laevigatus J. R. & G. Forst.) by cucumber mosaic virus. New Zealand Journal of Agricultural Research, 20: 533-34.

Daniels, J. 1992. Characterization of Cucumber Mosaic Virus Isolates from California. Plant disease, 76: 1245-50.

Dheepa, R. and S. Paranjothi. 2010. Transmission of Cucumber Mosaic Virus (CMV) infecting banana by aphid and mechanical methods. Emirates Journal of Food and Agriculture, 22: 117-29.

Dietzgen, R. G., B. Callaghan, C. M. Higgins, R. G. Birch, K. Chen and Z. Xu. 2001. Differentiation of peanut seedborne potyviruses and cucumoviruses by RT-PCR. Plant disease, 85: 989-92. PMid:30823115

El-Baz, R. M. 2004. Physicochemical, physiological and histopathological studies on cucumber mosaic virus, Helwan University.

El-Borollosy, A. M. and H. M. A. Waziri. 2013. Molecular characterization of a cucumber mosaic cucumovirus isolated from lettuce in Egypt. Annals of Agricultural Sciences, 58: 105-09.

Francki, R. I. B. 1985. The Plant Viruses: Polyhedral virions with tripartite genomes. Plenum Press: New York.

Gerola, F. M., M. Bassi and G. Belli. 1965. Some observations on the shape and localization of different viruses in experimentally infected plants, and on the fine structure of the host cells. II. nicotiana glutinosa systemically infected with cucumber mosaic virus, strain y. Caryologia, 18: 567-97.

Gibbs, A. J. and B. D. Harrison. 1970. Cucumber Mosaic Virus. pp. 14.

Honda, Y. and C. Matsui. 1974. Electron microscopy of cucumber mosaic virus-infected tobacco leaves showing mosaic symptoms. Phytopathology, 64: 534-39.

Hull, R. 2002. Purification and Composition of Plant Viruses Matthews' Plant Virology. Elsevier Academic Press. London, UK. pp. 75-107.

Kimura, M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16: 111-20. PMid:7463489

Martelli, G. P. and M. Russo. 1984. Use of thin sectioning for visualization and identification of plant viruses. Methods in Virology, 8: 143-224.

Medina, V., G. Rodrigo, T. Tian, M. Juarez, V. V. Dolja, M. Angeles Achon and B. W. Falk. 2003. Comparative cytopathology of Crinivirus infections in different plant hosts. Annals of Applied Biology, 143: 99-110.

Megahed, A. A. 2008. Effect of antiviral proteins produced by bacterial and fungal isolates on some viruses infecting vegetable crops, Ain Shams University.

Megahed, A. A., K. A. E. Dougdoug, B. A. Othman, S. M. Lashin, M. A. Ibrahim and A. R. Sofy. 2012. A new Egyptian satellite strain of cucumber mosaic cucumovirus. International Journal of Virology, 8: 240-57.

Mochizuki, T. and S. T. Ohki. 2011. Single amino acid substitutions at residue 129 in the coat protein of cucumber mosaic virus affect symptom expression and thylakoid structure. Archives of virology, 156: 881-86. PMid:21221672

Mourembou, G., J. Rathored, J. B. Lekana-Douki, A. Ndjoyi-Mbiguino, F. Fenollar, C. Michelle, P. E. Fournier, D. Raoult and J. C. Lagier. 2016. Noncontiguous finished genome sequence and description of Kallipyga gabonensis sp. nov. New Microbes and New Infections, 9: 15-23. PMid:26862430 PMCid:PMC4706609

Noordam, D. 1973. Identification of plant viruses: Methods and experiments. Center for Agriculture Publishing and Documentation: Wageningen, The Netherlands.

Nour El-Din, H., R. Salem and K. Abdalla. 2013. Cloning and characterization of cucumber mosaic virus coat protein gene from infected banana plants in Egypt. Egyptian Journal of Genetics and Cytology, 42: 151-61.

Owen, J., M. Shintaku, P. Aeschleman, S. B. Tahar and P. Palukaitis. 1990. Nucleotide sequence and evolutionary relationships of cucumber mosaic virus (CMV) strains: CMV RNA 3. Journal of General Virology, 71: 2243-49. PMid:2230731

Rabie, M., C. Ratti, M. Calassanzio, E. A. Aleem and F. A. Fattouh. 2017. Phylogeny of Egyptian isolates of Cucumber mosaic virus (CMV) and Tomato mosaic virus (ToMV) infecting Solanum lycopersicum. European Journal of Plant Pathology, 149: 219-25.

Roossinck, M. J. 2002. Evolutionary history of cucumber mosaic virus deduced by phylogenetic analyses. Journal of Virology, 76: 3382-87. PMid:11884564 PMCid:PMC136033

Sastry, K. S., B. Mandal, J. Hammond, S. W. Scott and R. W. Briddon. 2019. Encyclopedia of plant viruses and viroids. Springer: India.

Seng, P., M. Sarih, C. Socolovschi, N. Boudebouch, M. Hassar, P. Parola, D. Raoult and P. Brouqui. 2009. Detection of anaplasmataceae in ticks collected in Morocco. Clinical Microbiology and Infection, 15: 86-87. PMid:19438648

Shevchenko, T. P., O. V. Tymchyshyn, E. AlDalain, A. S. Bysov, I. G. Budzanivska, O. V. Shevchenko and V. P. Polishchuk. 2015. The first evidence of subgroup IB isolates of Cucumber mosaic virus in Ukraine. Biopolymers and Cell, 31: 57-62.

Sialer, M. M. F., F. Cillo, L. Barbarossa and D. Gallitelli. 1999. Differentiation of cucumber mosaic virus subgroups by RT-PCR RFLP. Journal of Plant Pathology, 81: 145-48.

Sofy, A. R. and A. M. Soliman. 2011. Molecular identification of a Cucumber mosaic virus subgroup I Egyptian isolate from geranium based on bioinformatics analysis of CP gene sequence. Egyptian Journal of Virology, 8: 178-94.

Sulistyowati, E., N. Mitter, S. Bastiaan-Net, M. J. Roossinck and R. G. Dietzgen. 2004. Host range, symptom expression and RNA 3 sequence analyses of six Australian strains of Cucumber mosaic virus. Australasian Plant Pathology, 33: 505-12.

Suzuki, M., S. Kuwata, J. Kataoka, C. Masuta, N. Nitta and Y. Takanami. 1991. Functional analysis of deletion mutants of cucumber mosaic virus RNA3 using an in vitro transcription system. Virology, 183: 106-13.

Taha, M. A. T. 2010. Biological control of Cucumber mosaic virus (Cucumovirus) by certain local streptomycetal isolates, Ain Shams University.

Takeshita, M., N. Shigemune, K. Kikuhara, N. Furuya and Y. Takanami. 2004. Spatial analysis for exclusive interactions between subgroups I and II of Cucumber mosaic virus in cowpea. Virology, 328: 45-51. PMid:15380357

Tamura, K., G. Stecher, D. Peterson, A. Filipski and S. Kumar. 2013. MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30: 2725-29. PMid:24132122 PMCid:PMC3840312

Thompson, J. D., D. G. Higgins and T. J. Gibson. 1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22: 4673-80. PMid:7984417 PMCid:PMC308517

Tidona, C. A. and G. Darai. 2001. The Springer Index of Viruses. Springer-Verlag: Berlin Heidelberg. PMid:11807240

Tomlinson, J. A., A. L. C. Elizabeth, M. Faithfull and M. J. W. Webb. 1973. Purification and serology of the W strain of cucumber mosaic virus. Annals of Applied Biology, 74: 181-89.

Verma, N., B. K. Mahinghara, R. Ram and A. A. Zaidi. 2006. Coat protein sequence shows that Cucumber mosaic virus isolate from geraniums (Pelargonium spp.) belongs to subgroup II. Journal of Biosciences, 31: 47-54. PMid:16595874

Wagih, E. E., M. M. Zalat and M. A. Kawanna. 2021. Partial Characterisation of Two Isolates of Cucumber mosaic virus (CMV). Scientific Journal of Agricultural Sciences, 3.

Zambolim, E. M., M. I. T. Assis, L. Zambolim, J. A. Venturia and M. G. de Carvalho. 1994. Natural infection of banana cultivar Prata (AAB) by cucumber mosaic virus in the state of Minas Gerais, Brazil. Fitopatologia Brasileira, 19: 483-84.

Zitter, T. A. and J. F. Murphy. 2009. Cucumber mosaic virus. Plant Health Instructor.

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DOI: 10.33687/phytopath.010.01.3502


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