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Phytoplasma Plasmid DNA Extraction

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Phytoplasma

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

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Abstract

Phytoplasma plasmids have generally been detected from DNA extracted from plants and insects using methods designed for the purification of total phytoplasma DNA. Methods include extraction from tissues that are high in phytoplasma titre, such as the phloem of plants, with the use of CsCl–bisbenzimide gradients that exploit the low G+C content of phytoplasma DNA. Many of the methods employed for phytoplasma purification have been described elsewhere in this book. Here we describe in detail two methods that are specifically aimed at isolating plasmid DNA.

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References

  1. Hinnebusch J, Tilly K (1993) Linear plasmids and chromosomes in bacteria. Mol Microbiol 10:917–922

    Article  PubMed  CAS  Google Scholar 

  2. Davis MJ et al (1988) Cloning of chromosomal and extrachromosomal DNA of the mycoplasmalike organism that causes maize bushy stunt disease. Mol Plant Microbe Interact 1:295–302

    Article  Google Scholar 

  3. Tran-Nguyen LTT, Gibb KS (2006) Extrachromosomal DNA isolated from tomato big bud and Candidatus Phytoplasma australiense phytoplasma strains. Plasmid 56:153–166

    Article  PubMed  CAS  Google Scholar 

  4. Namba S, Oshima K, Gibb K (2005) Phytoplasma genomics. In: Blanchard A, Browning GS (eds) Mycoplasmas: molecular biology, pathogenicity and strategies for control. Horizon Bioscience, Norfolk, pp 97–133

    Google Scholar 

  5. Panopoulos NJ, Peet RC (1985) The molecular genetics of plant pathogenic bacteria and their plasmids. Annu Rev Phytopathol 23:381–419

    Article  CAS  Google Scholar 

  6. Vivian A, Murillo J, Jackson RW (2001) The roles of plasmids in phytopathogenic bacteria: mobile arsenals? Microbiology 147:763–780

    PubMed  CAS  Google Scholar 

  7. Toruño TY et al (2010) Phytoplasma PMU1 exists as linear chromosomal and circular extrachromosomal elements and has enhanced expression in insect vectors compared with plant hosts. Mol Microbiol 77:1406–1415

    Article  PubMed  Google Scholar 

  8. Nishigawa H et al (2002) Evidence of intermolecular recombination between extrachromosomal DNAs in phytoplasma: a trigger for the biological diversity of phytoplasma? Microbiology 148:1389–1396

    PubMed  CAS  Google Scholar 

  9. Niemark H, Kirkpatrick BC (1993) Isolation and characterization of full-length chromosomes from non-culturable plant-pathogenic Mycoplasma-like organisms. Mol Microbiol 7:21–28

    Article  Google Scholar 

  10. Lee I-M, Davis RE (1983) Phloem-limited prokaryotes in sieve elements isolated by enzyme treatment of diseased plant tissues. Phytopathology 73:1540–1543

    Article  Google Scholar 

  11. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  12. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21

    Article  CAS  Google Scholar 

  13. Liefting LW et al (2006) Comparative analysis of the plasmids from two isolates of “Candidatus Phytoplasma australiense”. Plasmid 56:138–144

    Article  PubMed  CAS  Google Scholar 

  14. Harrison NA et al (1991) Molecular cloning and detection of chromosomal and extrachromosomal DNA of mycoplasmalike organisms associated with witches’-broom disease of pigeon pea in Florida. Mol Plant Microbe Interact 4:300–307

    Article  CAS  Google Scholar 

  15. Kollar A et al (1990) Isolation of the DNA of various plant pathogenic mycoplasmalike organisms from infected plants. Phytopathology 80:233–237

    Article  CAS  Google Scholar 

  16. Bai X et al (2006) Living with genome instability: the adaptation of phytoplasmas to diverse environments of their insect and plant hosts. J Bacteriol 188:3682–3696

    Article  PubMed  CAS  Google Scholar 

  17. Denes AS, Sinha RC (1991) Extrachromosomal DNA elements of plant pathogenic mycoplasmalike organisms. Can J Plant Pathol 13:26–32

    Article  CAS  Google Scholar 

  18. Kuske CR, Kirkpatrick BC (1990) Identification and characterization of plasmids from the western aster yellows mycoplasmalike organism. J Bacteriol 172:1628–1633

    PubMed  CAS  Google Scholar 

  19. Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Article  PubMed  CAS  Google Scholar 

  20. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbour Press, New York

    Google Scholar 

  21. Ausubel FM et al (eds) (1992) Current protocols in molecular biology. Wiley, New York

    Google Scholar 

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Correspondence to Mark T. Andersen .

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Andersen, M.T., Liefting, L.W. (2013). Phytoplasma Plasmid DNA Extraction. 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_31

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  • DOI: https://doi.org/10.1007/978-1-62703-089-2_31

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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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