DNA ISOLATION PROCEDURE FOR LESQUERELLA SP.

Authors: Kaufman, Benjamin; Richards, Stacy; DIERIG, DAVID

Submitted to: Association for the Advancement of Industrial Crops Conference
Publication Type: Abstract Only
Publication Acceptance Date: 6/2/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: A prerequisite for molecular genetic studies is, by definition, the ability to isolate DNA. In plant species with high contents of polysaccharide and polyphenolic compounds, this basic requirement becomes a challenge. In these species, the carbohydrates coprecipitate with the nucleic acids, forming large pellets in which the DNA is trapped in gum and is no longer retrievable. A species that exemplifies the problem is Lesquerella fendleri, a new industrial crop that has evoked interest for its unique oil and hydroxy fatty acid (lesquerolic acid). The difficulty in isolating DNA from this species impeded applying new methods of molecular breeding to the improvement process. We are describing a novel DNA isolation method developed by the New Crops group at U.S. Water Conservation Labortaory in Phoenix. In this procedure, after the cell walls are broken down, the cell nuclei are being separated from the cytoplasmatic and intracellular fluids by differential centrifugation in a viscous medium; this preparatory step separates between the cellular compartments that contain the desired DNA and the problematic carbohydrates. The DNA is then isolated from the nuclei without interference. High quality DNA was obtained and used successfully in various analyses.