Title: Real-Time PCR Genotyping using Taqman Probes to Detect High Oleic Acid Peanuts Authors
Submitted to: American Peanut Research and Education Society Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: June 1, 2009
Publication Date: September 25, 2009
Citation: Barkley, N.L., Chamberlin, K.D., Wang, M.L., Pittman, R.N. 2010. Real-Time PCR Genotyping using Taqman Probes to Detect High Oleic Acid Peanuts. American Peanut Research and Education Society Abstracts. Online DOI 10.1007/s11032-009-9338-z 25:541-548. Technical Abstract: Oleic acid, a monounsaturated, omega-9 fatty acid is an important agronomic trait in peanut cultivars because it provides increased shelf life, improved flavor, enhanced fatty acid composition, and a beneficial effect on human health. Currently, most high oleic peanuts confer limited resistance to disease, which makes them highly vulnerable to infection such as TSWV. In an attempt to increase genetic diversity specifically disease resistance of high oleic acid lines, crosses between lines containing high oleic to linoleic ratios (high O/L), wild species, and cultivated botanical varieties (Arachis hypogaea ssp. hypogaea var. hirsuta or peruviana) were prepared. The main bottleneck of breeding research is rapid detection of the trait(s) of interest. Therefore, a Real-Time PCR assay was developed to identify the high oleic trait in the parents and the resulting progenies. This test utilizes Taqman probes to detect the presence of an indel (insertion/deletion) that causes a frameshift which ultimately alters the gene product. This procedure can distinguish all F435 derived material, which is recognized as containing high levels of oleic acid, from normal oleic lines. Moreover, this assay differentiates hybrid F1’s from the selfed progeny, and furthermore, can be employed from either seed or leaf material. Overall, the Real-Time PCR test facilitates the identification of progeny carrying the high oleic trait, and thus, undesirable non-high oleic lines in the segregating population can be rapidly eliminated.