African Journal of
Plant Science

  • Abbreviation: Afr. J. Plant Sci.
  • Language: English
  • ISSN: 1996-0824
  • DOI: 10.5897/AJPS
  • Start Year: 2007
  • Published Articles: 718

Full Length Research Paper

Isolation of total RNA from ripe and unripe soursop (Annona muricata L.) fruit

Isabella Montenegro Brasil1*, Maria de Lourdes Oliveira Otoch2, José Hélio Costa3, Geraldo Arraes Maia1, Maria da Guia Silva Lima4, Birgit Arnholdt-Schmitt6 and Dirce Fernandes de Melo4
  1Fruit and Vegetable Laboratory/Food Technology Department, Federal University of Ceará, Fortaleza, Ceará, Brazil. 2Biology Department/University of Ceará State, Fortaleza, Ceará, Brazil. 3Researcher of PRODOC/CAPES/ Biochemistry and Molecular Biology Department, Federal University of Ceará, Fortaleza, Ceará, Brazil. 4Bioenergetics Laboratory/Biochemistry and Molecular Biology Department, Federal University of Ceará, Fortaleza, Ceará, Brazil. 5Biocenter Klein Flottbek, Institute of Botany (AMP II), University of Hamburg, Germany.
Email: [email protected]

  •  Accepted: 16 September 2008
  •  Published: 30 September 2008

Abstract

Soursop fruit tissue is known by its acidic pH and high levels of polysaccharides, polyphenolics and secondary metabolites. These conditions are recognized to interfere unfavorably with conventional methodologies for RNA isolation. We describe here a rapid and simple method for the isolation of total RNA from soursop fruit. RNA was extracted in less than 4 h through a combination of SDS/potassium acetate precipitation and selective binding on a silica-gel-based membrane (Qiagen) through microspin speed technology. In comparison to other methods applied for RNA extraction from soursop fruit, our protocol improved substantially RNA quality as well as RNA yield. The isolated RNA served as a robust template for RT-PCR analysis. Comparable RNA quality and yield per dry weight were obtained from unripe and ripe fruits. This makes the method appropriate to being used in studies on differential gene expression in post-harvest behavior.

 

Key words: Annona muricata L., RNA isolation, tropical fruit.