Plant Molecular Biology Reporter

, Volume 16, Issue 1, pp 49–59 | Cite as

DNA Isolation and AFLP™ Genetic Fingerprinting of shape Theobroma cacao (L.)

  • Matthew D. Perry
  • Michael R. Davey
  • J. Brian Power
  • Kenneth C. Lowe
  • H. Frances J. Bligh
  • Paul S. Roach
  • Chris Jones
Article

Abstract

Accurate identification of parental plants and their hybrids is essential for an effective breeding programme. Traditional classification of cocoa varieties relies on the characterisation of agricultural traits at plant maturity. A rapid and reliable method is described, based on genotypic analysis. An efficient DNA isolation procedure was developed, yielding unsheared DNA of high purity. Two genetic fingerprinting techniques, RAPD and AFLP™, were evaluated for their suitability in distinguishing cocoa varieties. RAPD analysis was unsatisfactory due to the low frequency of polymorphisms and poor reproducibility. AFLP™ was reliable in distinguishing phenotypically identical, known varieties of cocoa. Importantly, AFLP™ also revealed intra- and inter-varietal variation.

Abbreviations: AFLP™, amplified fragment length polymorphism; APS, ammonium persulphate; CTAB, hexadecyltrimethylammonium bromide; DEB, DNA extraction buffer; f.wt., fresh weight; NEB, nuclei extraction buffer; PMSF, phenylmethanesulphonyl fluoride; RAPD, random amplified polymorphic DNA; T4 PNK, Bacteriophage T4 polynucleotide kinase; Taq, Thermus aquaticus; TBE, tris-borate-EDTA; TEMED, NNN′N′ tetramethylethylenediamine.

amplified fragment length polymorphism cocoa RAPD woody plant 

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Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Matthew D. Perry
    • 1
  • Michael R. Davey
    • 1
  • J. Brian Power
    • 1
  • Kenneth C. Lowe
    • 1
  • H. Frances J. Bligh
    • 1
  • Paul S. Roach
    • 2
  • Chris Jones
    • 2
  1. 1.Plant Science Division, School of Biological SciencesUniversity of NottinghamNG7 2RDUK
  2. 2.School of Biomedical Sciences, Queens Medical CentreUniversity of NottinghamNG7 2UHUK

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