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Dual labeled probe based real time PCR method for detection of 16SrXI-B sub-group phytoplasma associated with coconut root wilt disease in India

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Abstract

The Root Wilt Disease (RWD) is a non-lethal debilitating disease of coconut palms in India. Phytoplasma belonging to 16SrXI-B sub-group are associated with the disease. Rapid and specific detection protocol will greatly aid the current RWD management of replanting with disease-free seedlings. Here we report the development of a real-time PCR-based rapid and specific detection method for RWD phytoplasma. The phytoplasma 16S rRNA gene was used for designing primers and probe. The probe was dual-labeled with fluorescent molecule FAM at the 5′ end and quencher TAMRA at the 3’end. The real time assay was optimised for primer/probe, template DNA annealing temperature and cycling. The optimised assay could detect the phytoplasma in 85 % of the samples tested. Amplicon sequence confirmation ensured the fidelity of the assay. The assay developed here is suited for 16SrXI-B sub-group phytoplasmas.

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References

  • Ahrens U, Seemuller E (1992) Detection of DNA of plant pathogenic mycoplasma like organisms by a polymerase chain reaction which amplifies a sequence of the 16S rRNA gene. Phytopathology 82:828–832

    Article  CAS  Google Scholar 

  • Angelini E, Bianchi GL, Filippin L, Morassutti C, Borgo M (2007) A new TaqMan method for the identification of phytoplasmas associated with grapevine yellows by real-time PCR assay. J Microbiol Meths 68:613–622

    Article  CAS  Google Scholar 

  • Cordova C, Oropeza C, Hau P, Harrison N, Rodriguez AC, Narvaez M, Matos GN, Reyes C, Saenz L (2014) A real-time PCR assay for detection of coconut lethal yellowing phytoplasmas of group 16SrIV subgroups A, D And E found in the Americas. J Plant Pathol 96:343–352

    CAS  Google Scholar 

  • Galetto, L. and Marzachi, C. (2010) Real time PCR diagnosis and quantification of phytoplasma. In Phytoplasmas: Genomes, Plant hosts and Vectors (Eds). Phyllis G. Weintraub and Phil Jones. CAB International. Pp 1–18

  • Holland PM, Abramson RD, Watson R, Gelfand DH (1991) Detection of specific polymerase chain reaction product by utilizing the 5' - 3' exonuclease activity of Thermus aquaticus DNA polymerase. Proc Natl Acad Sci 88:7276–7280

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Manimekalai R, Nair S, Soumya VP, Roshna OM, Thomas GV (2011) Real-time PCR technique-based detection of coconut root (wilt) phytoplasma. Curr Sci 101:1209–1213

    CAS  Google Scholar 

  • Manimekalai R, Soumya VP, Nair S, Thomas GV, Baranwal VK (2014) Molecular characterization identifies 16SrXI-B group phytoplasma(‘Candidatus Phytoplasma oryzae’-related strain) associated with root wilt disease of coconut in India. Scientia Hortic 165:288–294

    Article  CAS  Google Scholar 

  • Nikolic P, Mehle N, Gruden K, Ravnikar M, Dermastia M (2010) A panel of real-time PCR assays for specific detection of three phytoplasmas from the apple proliferation group. Mol Cell Probes. doi:10.1016/j.mcp.2010.06.005

    PubMed  Google Scholar 

  • Solomon JJ, Nair CPR, Srinivasan N, Gunasekaran M, Sasikala M (1999) Coconut root (wilt) –the malady and remedy. J Plantation Crops 27:71–92

    Google Scholar 

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Correspondence to R. Manimekalai.

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Nair, S., Manimekalai, R., Soumya, V.P. et al. Dual labeled probe based real time PCR method for detection of 16SrXI-B sub-group phytoplasma associated with coconut root wilt disease in India. Australasian Plant Pathol. 45, 187–189 (2016). https://doi.org/10.1007/s13313-016-0406-7

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  • DOI: https://doi.org/10.1007/s13313-016-0406-7

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