Abstract
Cucumber mosaic virus (CMV) causing mosaic, leaf distortion and stunting of vanilla(Vanilla planifolia Andrews) in India was characterized on the basis of biological and coat protein (CP) nucleotide sequence properties. In mechanical inoculation tests, the virus was found to infect members of Chenopodiaceae, Cucurbitaceae, Fabaceae and Solanaceae.Nicotiana benthamiana was found to be a suitable host for the propagation of CMV. The virus was purified from inoculatedN. benthamiana plants and negatively stained purified preparations contained isometric particles of about 28 nm in diameter. The molecular weight of the viral coat protein subunits was found to be 25.0 kDa. Polyclonal antiserum was produced in New Zealand white rabbit, immunoglobulin G (IgG) was purified and conjugated with alkaline phosphatase enzyme. Double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) method was standardized for the detection of CMV infection in vanilla plants. CP gene of the virus was amplified using reverse transcriptase-polymerase chain reaction (RT-PCR), cloned and sequenced. Sequenced region contained a single open reading frame of 657 nucleotides potentially coding for 218 amino acids. Sequence analyses with other CMV isolates revealed the greatest identity with black pepper isolate of CMV (99%) and the phylogram clearly showed that CMV infecting vanilla belongs to subgroup IB. This is the first report of occurrence of CMV onV. planifolia from India.
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Abbreviations
- CMV:
-
Cucumber mosaic virus
- CP:
-
coat protein
- DAC-ELISA:
-
direct antigen coated-enzyme linked immunosorbent assay
- DAS:
-
double antibody sandwich
- EBIA:
-
electroblot immuno assay
- IgG:
-
immunoglobulin G
- RT-PCR:
-
reverse transcriptase-polymerase chain reaction
References
Altschul S F, Madden T L, Schäffer A A, Zhang J, Zhang Z, Miller W and Lipman D J 1997 Gapped BLAST and PSI-BLAST: a new generation of protein database search Programs;Nucleic Acids Res. 25 3389–3402|
Avrameas S 1969 Coupling of enzymes to proteins with glutaraldehyde and use of conjugate for the detection of antigens and antibodies;Immunochemistry 6 43–52
Benson D A, Boguski M S, Lipman D J, Ostell J, Ouellette B F, Rapp B A and Wheeler D L 1999 GenBank;Nucleic Acids Res. 27 12–17
Bhai R S, Joseph Thomas, Potty S N, Geetha L and Solomon J J 2003 Mosaic disease of vanilla(Vanilla planifolia Andrews) the first report from India;J. Spices Aromatic Crops 12 80–82
Bhat A I, Faisal T H, Madhubala R, Hareesh P S and Pant R P 2004a Purification, production of antiserum and development of enzyme linked immunosorbent assay-based diagnosis forCucumber mosaic virus infecting black pepper(Piper nigrum L.);J. Spices Aromatic Crops 13 16–21
Bhat A I, Venugopal M N, Pant R P and Bhai R S 2004b Occurrence and distribution of viral diseases on vanilla in India;J. Spices Aromatic Crops 13 143–148
Clark M F, Lister R M and Bar-Joseph M1986 ELISA Techniques;Methods Enzymol. 118 742–766
Farreyrol K, Pearson M N, Grisoni M and Leclercq Le Quillec F 2001 Severe stunting ofVanilla tahitensis in French Polynesia caused byCucumber mosaic virus (CMV) and the detection of the virus inV. fragrans in Reunion Island;Plant Pathol. 50 414–415
Hsu H T, Barzuan L, Hsu Y H, Bliss W and Perry K L 2000 Identification and subgrouping ofCucumber mosaic virus with mouse monoclonal antibodies;Phytopathology 90 615–620
Laemmli U K 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4;Nature (London) 227 680–685
Lot H, Marrou J, Quiot J B and Eswan C 1972Cucumber mosaic virus CMI/AAB;Descriptions Plant Viruses No. 213
O’Donell I J, Shukla D D and Gough K H 1982 Electroblot immunoassay of virus infected plant sap — A powerful technique for detecting plant viruses;J. Virol. Methods 4 19–26
Page RDM 1996 TREEVIEW: an application to display phylogenetic trees on personal computers;Comput. Appl. Biosci. 12 357–358
Palukaitis P, Roossinck M J, Dietzgen R G and Francki RIB 1992Cucumber mosaic virus;Adv. Virus Res. 41 281–348
Pappu S S, Brand R, Pappu H R, Rybicki E, Gough K H, Frenkel M J and Niblett C L 1993 A polymerase chain reaction method adapted for selective cloning of 3′ non translated regions of potyviruses: application todasheen mosaic virus;J. Virol. Methods 41 9–20
Rhoads D D and Roufa D S 1985 Emetine resistance of Chinese hamster cells: structure of wild type and mutant ribosomal protein S14 messenger RNA species;Mol. Cell. Biol. 5 1655–1659
Roossinck M J 1999 Cucumoviruses (Bromoviridae)-general features; inEncyclopedia of Virology, 2nd edition (eds) L Granoof and R G Webster (SanDiego: Academic Press) pp 315–320
Roossinck M J 2002 Evolutionary history of cucumber mosaic virus deduced by phylogentic analysis;J. Virol. 76 3382–3387
Roossinck M J, Zhang L and Hellwald K-H 1999 Rearrangements in the 5′ nontranslated region and phylogenetic analyses ofCucumber mosaic virus RNA 3 indicate radial evolution of three subgroups;J. Virol. 73 6752–6758
Sambrook J and Russell D W 2001Molecular cloning: A laboratory manual (New York: Cold Spring Harbor laboratory Press)
Sarma Y R, Kiranmani G, Sreenivasulu P, Anandaraj M, Hema M, Venkatramana M, Murthy A K and Reddy D V R 2001 Partial characterization and identification of a virus associated with stunt disease of black pepper(Piper nigrum) in South India;Curr. Sci. 80 459–462
Sudharshan M R, Radhika N S and Prakash K V 2003 Vanilla viral diseases in Karnataka;Spice India 16 28–29
Srivastava A and Raj S K 2004 High molecular similarity among Indian isolates of Cucumber mosaic virus suggests a common origin;Curr. Sci. 87 1126–1131
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Madhubala, R., Bhadramurthy, V., Bhat, A.I. et al. Occurrence ofCucumber mosaic virus on vanilla (Vanilla planifolia Andrews) in India. J. Biosci. 30, 339–350 (2005). https://doi.org/10.1007/BF02703671
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DOI: https://doi.org/10.1007/BF02703671