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ARS Home » Research » Publications at this Location » Publication #90928
Title: DNA ISOLATION METHOD FOR HIGH POLYSACCHARIDE LESQUERELLA SPECIES

Author
item Kaufman, Benjamin
item Richards, Stacy
item Dierig, David

Submitted to: Industrial Crops and Products
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/5/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary: A prerequisite for molecular genetic studies is, by definition, the ability to isolate DNA. In Lesquerella fendleri and other plant species with a high carbohydrate content, this basic requirement becomes a challenge. The carbohydrates do not separate from the DNA and together they form large pellets, from which the DNA is no longer retrievable. We are describing a simple DNA isolation method that solves this problem. In one preparatory step, we separated the cellular compartments, which contain the desired DNA, from the problematic carbohydrates. The DNA is then isolated without interference. High quality DNA was obtained this way and used successfully in various analyses. The ability to employ DNA markers in the selection process for the improvement of Lesquerella will greatly facilitate the rate and efficiency of the breeding program. Lesquerella has a long growing season and molecular markers data will permit selection at the seedling stage rather than at maturity. The The expression of many plant traits is affected by the environment; therefore, these traits are unreliable as selection criteria. Molecular markers are not affected by environment; therefore, selection based on the presence or absence of DNA segments of interest is more reliable and efficient. Being able to isolate DNA is the first step in enabling researchers and breeders of Lesquerella to benefit from these advantages.

Technical Abstract: A prerequisite for molecular genetic studies is, by definition, the ability to isolate DNA. In Lesquerella fendleri and other plant species with a high content of polysaccharide, this basic requirement becomes a challenge. The carbohydrates coprecipitate with the nucleic acids forming large pellets, in which the DNA is trapped in gum, and is no longer retrievable. We are describing a simple DNA isolation method that solves this problem. After breaking down the cell walls, the cell nuclei are separated from the cytoplasmatic and intracellular fluids by differential centrifugation in a viscous medium. This preparatory step separates the cellular compartments which 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.