An effective method for isolating high quality RNA from spathe tissue of Anthurium andraeanum Hort. and other members of the Araceae family

Authors

  • Vern E. Collette Ag. Research Grasslands, Tennent Drive, Private Bag 11008, Palmerston North, New Zealand
  • Paula E. Jameson Massey University, Private Bag 11-222, Palmerston North, New Zealand
  • Kathy E. Schwinn New Zealand Institute for Crop & Food Research Limited, Priv ate Bag 11-600, Palmerston North, New Zealand
  • Chris S. Winefield Lincoln University, PO Box 84, Canterbury, New Zealand
  • Pathmanathan Umaharan Department of Life Sciences, University of the West Indies, St. Augustine, Trinidad and Tobago

Keywords:

Anthurium, RNA extraction, spathe, Spathyphyllum, Zantedeschia

Abstract

The abaxial and adaxial surfaces of Anthurium andraeanum Hort. (anthurium) spathe have marked cuticular thickening. In addition, the spathe tissue has high levels of polyphenols, evidenced by rapid browning upon injury. Both of these features hinder the extraction of high-quality RNA. To overcome these difficulties, an effective reproducible extraction method was developed using polyvinylpolypyrrolidone (PVPP) and 2?-mercaptoethanol. PVPP binds phenolics while 2?-mercaptoethanol prevents oxidation of phenolics. Both of these chemicals acting in concert possibly accounted for the marked improvement in the yield and quality of isolated RNA from anthurium spathe. Using this approach we obtained high quality RNA suitable for cDNA library construction, RT-PCR and northern analysis. The method was also effective with other members of the Araceae family.

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Issue

Volume 85 Number 3

Section

Research Papers