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PCR Detection of Banana Bunchy Top Virus (BBTV) at Tissue Culture Level for the Production of Virus-free Planting Materials

Author Affiliations

  • 1Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore, 641-003, INDIA
  • 2 Molecular Cell Physiology Laboratory, Department of Applied Bioresources, Faculty of Agriculture, 3-5-7 Tarumi, Ehime University, Matsuyama, 790-8566, JAPAN
  • 3 Department of Horticulture, Sunchon National University, 413 Jungangno, Suncheon, Jeonnam 540-742, REPUBLIC OF KOREA

Int. Res. J. Biological Sci., Volume 2, Issue (6), Pages 22-26, June,10 (2013)


Banana bunchy top disease, caused by Banana bunchy top virus (BBTV), is a serious disease that affects the productivity of banana to a greater extent. The production of virus-free planting material is an efficient method in controlling this disease and increase productivity. In vitro propagation, especially meristem tip culture, has played a key role for obtaining a large number of virus free, homogenous planting materials in plantains and bananas (Musa spp.). In our previous study we have demonstrated that direct regeneration from male floral meristems is a rapid and simple method for clonal and mass propagation of Musa spp. cultivars ‘Virupakshi’ and ‘Sirumalai’ (AAB). Detection of BBTV by polymerase chain reaction (PCR) is very reliable and economically important for testing micropropagated banana plantlets for the absence of virus before release to farmers. In this study we have developed a protocol for early diagnosis of BBTV at tissue culture level itself through optimizing the Polymerase Chain Reaction (PCR) method. We designed the viral gene specific primers [coat protein (CP) gene and nuclear shuttle protein (NSP) gene] and standardize PCR conditions for detection of presence of BBTV. All the regenerated plantlets were tested for BBTV presence with standardized PCR and found to be completely free of virus. PCR tests are more sensitive than ELISA tests and are better to be suited to plant tissue with lower number of pathogens, such as in tissue culture conditions.


  1. Dale J.L., Banana bunchy top: an economically important tropical plant virus disease, Advances in Virus Research, 33, 325-333 (1987)
  2. Hafner G.J., Harding R.M. and Dale J.M, Movement and transmission of banana bunchy top virus DNA component one in bananas, Journal of General Virology, 76, 2279-2285, (1995)
  3. Burns T.M., Harding R.M. and Dale J. L., The genome organization of banana bunchy top virus: analysis of six ssDNA components, Journal of General Virology, 76, 1471-1482 (1995)
  4. Harding R.M., Burns T.M., Hafner G.J., Dietzgen R. and Dale J. L., Nucleotide sequence of one segment of the banana bunchy top genome contains a putative replicase gene, Journal of General Virology, 74, 323-328, (1993)
  5. Wanitchakorn R., Harding R.M. and Dale J. L., Banana bunchy top virus DNA-3 encodes the viral coat protein, Archives of Virology, 142, 1673-1680 (1997)
  6. Wanitchakorn R., Hafner G.J., Harding R.M. and Dale J.L., Functional analysis of proteins encoded by banana bunchy top virus DNA-4–6, Journal of General Virology, 81,299–306 (2000)
  7. Thomas J.E., Geering A.D.W., Dahal G., Lockhart B.E.L. and Thottappilly G., Virus and virus-like diseases of major crops in developing countries, in Banana and plaintain.Edited by G. Loebenstein & G. Thottappilly. Dordrecht: Kluwer Academic Publishers, 477–496, (2003)
  8. Novak F.J., Afza R., van Duren M., Perea-Dallos M., Conger B.V. and Xiaolang T., Somatic embryogenesis and plant regeneration in suspension cultures of dessert (AA and AAA) and cooking (ABB) banana (Musa spp.), Biotechnology,7, 154–159 (1989)
  9. Ganapathi T.R., Suprasanna P., Bapat V.A., Kulkarni V.M. and Rao P.S., Somatic embryogenesis and plant regeneration from male flower buds in banana, Current Science, 76, 1228-1231 (1999)
  10. Kulkarni V. M., Varshney L.R., Bapat V.A. and Rao P.S., Somatic embryogenesis and plant regeneration in a seeded banana [Ensete superbum (Roxb.) Cheesman], Current Science, 83, 939-941 (2002)
  11. Assani A., Haicour R., Wenzel G., Cote F., Bakry F., Foroughi-Wehr F., Ducreux G.M., Aguillar E.A. and Grapin A., Plant regeneration from protoplasts of dessert banana cv. Grandnaine (Musa spp., Cavendish sub-group AAA) via somatic embryogenesis, Plant Cell Reports, 2, 482-488 (2001)
  12. Shelake R. M., Angappan K. and Murugan M., An efficient protocol for large-scale plantlet production from male floral meristems of Musa spp. cultivars Virupakshi and Sirumalai, In Vitro Cellular & Developmental Biology – Plant, 47,611-617 (2011)
  13. Wu R. Y. and Su H. J., Production of Monoclonal Antibodies Against Banana Bunchy Top Virus and Their Use in Enzyme-Linked Immunosorbent Assay, Journal of Phytopathology, 128, 203–208 (1990)
  14. Dietzgen R.G. and Thomas J.E., Properties of virus-like particles associated with banana bunchy top disease in Hawaii, Indonesia and Tonga, Australasian Plant Pathology, 20, 161-165 (1991)
  15. Geering A.D.W. and Thomas J.E., A comparison of four serological tests for the detection of banana bunchy top virus in banana, Australian Journal of Agricultural Research, 47, 403–412 (1996)
  16. Xie W.S. and Hu J.S., Molecular cloning, sequence analysis and detection of banana bunchy top virus in Hawaii, Phytopathology 85, 339–347 (1994)
  17. Hafner G.J., Harding R.M. and Dale J.M., A DNA primer associated with banana bunchy top virus, Journal of General Virology, 78, 479–486 (1997)
  18. Chandrasekar A., Kalaiponmani K., Elayabalan S., Kumar K.K., Angappan K. and Balasubramanian P., Screening of banana bunchy top virus through multiplex PCR approach, Archives of Phytopathology and Plant Protection, 44,1920-1925 (2011)
  19. Harding R.M., Sadik A.S., Bahieldin A. and Dale J.L., A sensitive detection of banana bunchy top nanovirus using molecular genetic approaches. Arabian Journal of Biotechnology, 3, 103-114 (2000)
  20. Galal A.M., Use of Polymerase Chain Reaction for Detecting Banana Bunchy Top Nanovirus, 6, 53-56, (2007)
  21. Mansoor S., Qazi J., Amin I., Khatri A., Khan I.A., Raza S., Zafar Y. and Briddon R.W., A PCR-based method, with internal control, for the detection of Banana bunchy top virus in banana, Molecular Biotechnology,30, 167-169 (2005)