DNA Isolation and AFLP™ Genetic Fingerprinting of shape Theobroma cacao (L.)
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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.
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- Bachmann K (1994) Molecular markers in plant ecology. New Phytol 126: 403-418.Google Scholar
- Bennett MD and Leitch IJ (1995) Nuclear DNA amounts in Angiosperms. Ann Bot 76: 113-176.Google Scholar
- Couch JA and Fritz PJ (1990) Isolation of DNA from plants high in phenolics. Plant Mol Biol Rep 8: 8-12.Google Scholar
- Figueira A, Janick J and Goldsbrough P (1992) Genome size and DNA polymorphism in Theobroma cacao. J Am Soc Hort Sci 117: 673-677.Google Scholar
- Gawel NJ and Jarret RL (1991) A modified CTAB DNA extraction procedure for Musaand Ipomoea.Plant Mol Biol Rep 9: 262-266.Google Scholar
- Greathouse DC, Laetsch WM and Phinney BO (1971) The shoot-growth rhythm of a tropical tree, Theobroma cacao. Am J Bot 58: 281-286.Google Scholar
- Henry RJ, Ko HL and Weining S (1997) Identification of cereals using DNA-based technology. Cereal F W 42: 26-29.Google Scholar
- Hunter JR (1990) The status of Cacao (Theobroma cacao,Sterculiaceae) in the western hemisphere. Econ Bot 44: 425-439.Google Scholar
- Jofuku KD and Goldberg RA (1988) Analysis of plant gene structure, In: Shaw CH (ed.), Plant Molecular Biology - A Practical Approach, pp. 37-66. IRL Press, Oxford.Google Scholar
- Lanaud C, Risterucci AM, N'Goran AKJ, Clement D, Flament MH, Laurent V, Falque M (1995) A genetic linkage map of Theobroma cacaoL. Theor Appl Genet 91: 987-993.Google Scholar
- Laurent V, Risterucci, AM and Lanaud C (1993) Variability for nuclear ribosomal genes within Theobroma cacao. Hered 71: 96-103.Google Scholar
- Patwary MU, MacKay RM, van der Meer JP (1993) Revealing genetic markers in Gelidium vagum(Rhodophyta) through the random amplified polymorphic DNA (RAPD) technique. J Phycol 29: 216-222.Google Scholar
- Penner GA, Bush A, Wise R, Kim W, Domier L, Kasha K, Laroche A, Scoles G, Molnar SJ and Fedak G (1993) Reproducibility of Random Amplified Polymorphic DNA (RAPD) analysis among laboratories. PCR Meth Appl 2: 341-345.Google Scholar
- Ronning CM, Schnell RJ and Kuhn DN (1995) Inheritance of Random Amplified Polymorphic DNA (RAPD) markers in Theobroma cacao. J Am Soc Hort Sci 120: 681-686.Google Scholar
- Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, Frijters A, Pot J, Peleman L, Kuiper M and Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23: 4407-4414.Google Scholar
- Weising K, Nybom H, Wolff K and Meyer W (1995) Evaluation of fragment patterns, In: DNA fingerprinting in plants and fungi, pp. 136-155. CRC Press, London.Google Scholar
- Wilde J, Waugh R and Powell W (1992) Genetic fingerprinting of Theobromaclones using randomly amplified polymorphic DNA markers. Theor Appl Genet 83: 871-877.Google Scholar
- Williams JGK, Kubelik AR, Livark KJ, Rafalski JA and Tingey JV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18: 6531-6535.Google Scholar
- Wood GAR (1985) History and development. In Wood GAR and Lass RA (eds), Cocoa (4th edn.), pp. 1-10. Longman, London.Google Scholar