Note: Comparison of antibody- and genome-based diagnostic techniques forSugarcane mosaic virus in sugarcane
- 68 Downloads
Different antibody-based diagnostic techniques such as direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA), electroblot immunoassay, immunosorbent electron microscopy and dot blot immunoassay were compared with a genome-based technique,viz., reverse transcriptase polymerase chain reaction (RT-PCR) againstSugarcane mosaic virus (SCMV) in different sugarcane cultivars expressing predominant and less predominant symptoms as well as in asymptomatic ones. Polyclonal antiserum raised against SCMV antigen purified from sugarcane cv. CoC 671 was used. All of the antibody-based tests reacted positively with plants showing predominant foliar symptoms; however, the sensitivity was not uniform, with samples from the plants showing less predominant symptoms and in asymptomatic plants. The RT-PCR assay method was more sensitive in detecting the virus from plants of less predominant symptoms also. Hence, a combined diagnostic system utilizing antibody-based techniques can be employed for mass screening and in quarantine operations against the virus, but in doubtful cases RT-PCR can be employed. Among the different sugarcane tissues, SCMV was diagnosed in the leaf lamina, bud, leaf sheath and midrib tissues, but root and pith tissues were negative to the virus in RT-PCR assay. In RT-PCR, 870 bp size cDNA was amplified from infected sugarcane with SCMV specific primer.
Key WordsSugarcane mosaic virus diagnosis enzyme linked immunosorbent assay electroblot immunoassay dot blot immunoassay immunosorbent electron microscopy reverse transcriptase polymerase chain reaction
Unable to display preview. Download preview PDF.
- 1.Agnihotri, V.P. (1996) Current sugarcane disease scenario and management strategies.Indian Phytopathol. 49:109–126.Google Scholar
- 2.Anon. (2001) Statistics.Indian Sugar 51:265–284.Google Scholar
- 3.Balamuralikrishnan, M., Sabitha Doraisamy, Ganapathy, T. and Viswanathan, R. (2002) Serological specificity and titre of sugarcane mosaic virus polyclonal antisera raised under varying immunization procedures and bleeding time.J. Plant Dis. Prot. 109:646–654.Google Scholar
- 4.Gallagher, S., Wintson, S.E., Fuller, S.H. and Hurre, J.G.R. (1995) Immunoblotting and immunodetection.in: Ausubel, F., Bernt, R., Kingston, E.E., Mooree, D.D., Seidman, J.D., Smith, J.A.et al. [Eds.] Short Protocols in Molecular Biology. John Wiley and Sons, Inc., New York, NY. pp. 1040–1048.Google Scholar
- 7.Rao, G.P., Jain, R.K. and Varma, A. (1998) Identification of sugarcane mosaic and maize dwarf mosaic potyviruses infecting poaceous crops in India.Indian Phytopathol. 51:10–16.Google Scholar
- 8.Sambrook, J., Fritsch, E.F. and Maniatis, R. (1989) Molecular Cloning. A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.Google Scholar
- 9.Shukla, D.D., Frenkel, M.J., McKern, N.M., Ward, C.W., Jilka, J., Tosic, M.et al. (1992) Present status of the sugarcane mosaic subgroup of potyviruses.in: Bernett, O.W. [Ed.] Potyvirus Taxonomy. Springer, New York, NY. pp. 363–373.Google Scholar
- 10.Shukla, D.D., Tosic, M., Jilka, J., Ford, R.E., Taker, R.W. and Langham, M.A.C. (1989) Taxonomy of poty viruses infecting maize, sorghum and sugarcane in Australia and the United States as determined by reactivities of polyclonal antibodies directed towards virus specific N-termini of coat proteins.Phytopathology 79:223–279.CrossRefGoogle Scholar
- 11.Shukla, D.D., Ward, C.W. and Brunt, A.A. (1994) The sugarcane mosaic virus subgroup.in: The Potyviridae. CAB International, Wallingford, UK. pp. 360–371.Google Scholar
- 12.Viswanathan, R., Balamuralikrishnan, M., Premachandran, M.N. and Tripathi, B.K. (1998) Sugarcane bacilliform virus: Symptoms, detection and distribution in world germplasm collection at Cannanore.Proc. 60th Annual Convention Sugar Technologists’ Association of India (Shimla, India), pp. 17–23.Google Scholar
- 13.Yambao, M.L.N., Cabauatan, P.Q. and Assam, O. (1998) Differentiation of rice tungro spherical virus variants by RT-PCR and RFLP.Int. Rice Res. Newsl. 23(2):22–24.Google Scholar