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Phytoplasma pp 233–244Cite as

T-RFLP for Detection and Identification of Phytoplasmas in Plants

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 938))

Abstract

Conventionally, detection of phytoplasmas has been performed by PCR of the 16S rRNA gene, followed by either RFLP or DNA sequencing to determine the phytoplasma 16Sr group. This chapter demonstrates the technique of terminal restriction fragment length polymorphism (T-RFLP), a fingerprinting technique which combines both detection and identification in a single method, with the added benefit of inbuilt controls which removes the risk of false negative results and in addition highlights potential false positive results.

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References

  1. Smart CD et al (1996) Phytoplasma-specific PCR primers based on sequences of the 16S–23S rRNA spacer region. Appl Environ Microbiol 62:2988–2993

    PubMed  CAS  Google Scholar 

  2. Duduk B, Paltrinieri S, Lee IM, Bertaccini A (2012) Nested PCR and RFLP analysis based on the 16S rRNA gene. In: Dickinson M, Hodgetts J (eds) Methods in molecular biology. Springer, New York

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  3. Harrison NA, Womack M, Carpio ML (2002) Detection and characterization of a lethal yellowing (16SrIV) group phytoplasma in Canary Island date palms affected by lethal decline in Texas. Plant Dis 86:676–681

    Article  CAS  Google Scholar 

  4. Anthony RM, Brown TJ, French GL (2000) Rapid diagnosis of bacteremia by universal amplification of 23S ribosomal DNA followed by hybridisation to an oligonucleotide array. J Clin Microbiol 38:781–788

    PubMed  CAS  Google Scholar 

  5. Liu W-T et al (1997) Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA. Appl Environ Microbiol 63:4516–4522

    PubMed  CAS  Google Scholar 

  6. Hodgetts J et al (2007) Use of terminal restriction fragment length polymorphism (T-RFLP) for identification of phytoplasmas in plants. Plant Pathol 56:357–365

    Article  CAS  Google Scholar 

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Acknowledgement

This work was completed as part of a Defra Plant Health Fellowship for JH. We would also like to thank all individuals and organizations that have provided us with phytoplasma DNA/infected plant samples to enable us to validate the assay.

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Correspondence to Jennifer Hodgetts .

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Hodgetts, J., Dickinson, M. (2013). T-RFLP for Detection and Identification of Phytoplasmas in Plants. In: Dickinson, M., Hodgetts, J. (eds) Phytoplasma. Methods in Molecular Biology, vol 938. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-089-2_20

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  • DOI: https://doi.org/10.1007/978-1-62703-089-2_20

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-088-5

  • Online ISBN: 978-1-62703-089-2

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