Back
 AJPS  Vol.3 No.11 , November 2012
Rolling Circle Amplification Is More Sensitive than PCR and Serology-Based Methods in Detection of Banana streak virus in Musa Germplasm
Abstract: Banana (Musa sp.) is a popular and important crop among many communities in East Africa. Banana production is however threatened by the wide-spread banana streak disease (BSD), caused by Banana streak virus (BSV). The success of BSV management is inherently coupled to the availability of a sensitive indexing method. In this study, the sensitivity of three BSV detection techniques: rolling circle amplification (RCA), immunocapture PCR (with degenerate and Gold finger primers) and standard PCR was compared. A set of 32 BSD-asymptomatic samples were used to compare the techniques. Analysis of variance (ANOVA) for comparison of the four techniques showed that there were significant differences (P < 0.05) among all the means, with RCA and direct PCR having the highest detection mean values. Owing to its fidelity and capacity to circumvent the amplification of the integrated nuclear viral sequences, the RCA technique is recommended for routine indexing of Musa tissues for BSV. This study unveils a more reliable BSV detection method, a need that has remained unaddressed for a long while.
Cite this paper: M. Wambulwa, F. Wachira, L. Karanja and S. Muturi, "Rolling Circle Amplification Is More Sensitive than PCR and Serology-Based Methods in Detection of Banana streak virus in Musa Germplasm," American Journal of Plant Sciences, Vol. 3 No. 11, 2012, pp. 1581-1587. doi: 10.4236/ajps.2012.311191.
References

[1]   J. W. Daniells, A. D. W. Geering, N. J. Brynde and J. E. Thomas, “The Effect of Banana streak virus on the Growth and Yield of Dessert Bananas in Tropical Australia,” Annals of Applied Biology, Vol. 139, No. 1, 2001, pp. 51-60. doi:10.1111/j.1744-7348.2001.tb00130.x

[2]   B. E. L. Lockhart and D. R. Jones, “Banana Streak,” In: D. R. Jones, Ed., Diseases of Banana, Abacá and Ensete, CABI Publishing, Oxon, 1999, pp. 262-274.

[3]   B. E. L. Lockhart, T. C. Ndowora, N. E. Olszewski and G. Dahal, “Studies on Integration of Banana streak badnavirus Sequences in Musa: Identification of Episomally-Expressible Badnaviral Integrants in Musa Genotypes,” In: E. A. Frison and S. E. Sharrock, Eds., Banana streak virus: A Unique Virus—Musa Interaction: Proceedings of a Workshop of the PROMUSA Virology Working Group, Montpellier, 19-21 January 1998, pp. 35-42.

[4]   A. D. W. Geering, J. N. Parry, L. Zhang, N. Olszewski, B. E. L. Lockhart and J. E. Thomas, “Is Banana streak virus Strain OL the Only Activatable Virus Integrant in the Musa Genome?” Proceedings of the 2nd International Conference on Molecular Cellular Biology of Banana, InfoMusa, Vol. 10, No. 2, 2001, pp. 13-20.

[5]   G. Harper, R. Hull, B. E. L. Lockhart and N. Olszewski, “Viral Sequences Integrated into Plant Genomes,” Annual Review of Phytopathology, Vol. 40, 2002, pp. 119-136. doi:10.1146/annurev.phyto.40.120301.105642

[6]   International Network for the Improvement of Banana and Plantain (INIBAP), “Report of the Meeting,” In: E. A. Frison and S. E. Sharrock, Eds., Banana streak virus: A Unique Virus—Musa Interaction: Proceedings of a Workshop of the PROMUSA Virology Working Group, Montpellier, 19-21 January 1997, pp. 7-14.

[7]   G. Thottappilly, G. Dahal and B. E. L. Lockhart, “Studies on a Nigerian Isolate of Banana streak badnavirus. I. Purification and Enzyme-Linked Immunoassay,” Annals of Applied Biology, Vol. 132, No. 2, 1998, pp. 253-261. doi:10.1111/j.1744-7348.1998.tb05201.x

[8]   B. E. L. Lockhart and N. E. Olszewski, “Serological and Genomic Heterogeneity of Banana streak badnavirus: Implications for Virus Detection in Musa Germplasm,” In: J. Ganry, Ed., Breeding Banana and Plantain for Resistance to Diseases and Pests, CIRAD, Montpellier, 1993, pp. 105-130.

[9]   D. Haible, S. Kober and H. Jeske, “Rolling Circle Amplification Revolutionizes Diagnosis and Genomics of Geminiviruses,” Journal of Virological Methods, Vol. 135, No. 1, 2006, pp. 9-16. doi:10.1016/j.jviromet.2006.01.017

[10]   N. J. Gawel and R.L. Jarret, “A Modified CTAB Extraction Procedure for Musa and Ipomoea,” Plant Molecular Biology Reporter, Vol. 9, No. 3, 1991, pp. 262-266. doi:10.1007/BF02672076

[11]   A, P. James, R. J. Geijskes, J. L. Dale and R. M. Harding, “Development of a Novel Rolling—Circle Amplification Technique to Detect Banana streak virus Which Also Discriminates between Integrated and Episomal Virus Sequences,” Plant Disease, Vol. 95, No. 1, 2011, pp. 57-62.

[12]   G. Harper, D. Hart, S. Moult and R. Hull, “Detection of Banana streak virus in Field Samples of Bananas from Uganda,” Annals of Applied Biology, Vol. 141, No. 3, 2002b, pp. 247-257. doi:10.1111/j.1744-7348.2002.tb00216.x

[13]   M. Bousalem, E. J. P. Douzery and S. E. Seal, “Taxonomy, Molecular Phylogeny and Evolution of Plant Reverse Transcribing Viruses (Family Caulimoviridae) Inferred from Full-Length Genome and Reverse Transcrip-tase Sequences,” Archives of Virology, Vol. 153, No. 6, 2008, pp. 1085-1102. doi:10.1007/s00705-008-0095-9

[14]   L. Blanco and M. Salas, “Relating Structure to Function in Phi29 DNA Polymerase,” Journal of Biological Chemistry, Vol. 271, No. 15, 1996, pp. 8509-8512. doi:10.1074/jbc.271.15.8509

[15]   L. Blanco, A. Bernad, J. M. Lazaro, G. Martin, C. Garmendia and M. Salas, “Highly Efficient DNA Synthesis by the Phage Phi29 DNA Polymerase,” Journal of Biological Chemistry, Vol. 264, No. 15, 1989, pp. 8935- 8940.

[16]   B. O. Agindotan, G. Thottappilly, A. Uwaifo and S. Winter, “Production of Monoclonal and Polyclonal Antibodies Against a Nigerian Isolate of Banana streak virus,” African Journal of Biotechnology, Vol. 2, No. 7, 2003, pp. 171-178.

[17]   L. Karanja, A. Wangai, G. Harper and R. S. Pathak, “Molecular Identification of Banana streak virus Isolates in Kenya,” Journal of Phytopathology, Vol. 156, No. 11-12, 2008, pp. 678-686. doi:10.1111/j.1439-0434.2008.01415.x

[18]   G. Dahal, F. Dahal, C. Pasberg-Gauhl, J. D. A. Hughes, G. Thottapilly and B. E. L. Lockhart, “Evaluation of Micropropagated Plantain and Banana (Musa spp.) for Banana streak badnavirus Incidence Under Field and Screen-house Conditions in Nigeria,” Annals Applied Biology, Vol. 134, No. 2, 1999, pp. 181-191. doi:10.1111/j.1744-7348.1999.tb05254.x

 
 
Top