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CROP BREEDING & GENETICS

Two Alleles of ahFAD2B Control the High Oleic Acid Trait in Cultivated Peanut

^a Dep. of Horticulture, Univ. of Georgia Tifton Campus, Tifton GA 31793
^b Research Geneticist, USDA-ARS, P.O. Box 748, Tifton, GA 31793. Funding was provided by the Georgia Seed Development Commission and the Univ. of Georgia Research Foundation Cultivar Development Program

^* Corresponding author (pozias{at}uga.edu).

A high oleic:linoleic acid ratio (O/L) in peanut (Arachis hypogaea L.) seeds is controlled primarily by two recessive genes, ahFAD2A and ahFAD2B (ol₁ and ol₂). Marker-assisted breeding for high O/L could become routine provided that user-friendly and economical markers could be developed that would target mutant alleles underlying the trait. Previously, a mutant allele of ahFAD2A was characterized as having a 448G>A transition, and two ahFAD2B alleles were found with either a nonsense mutation caused by a single nucleotide insertion (441_442insA) (high O/L line F435) or a transposable element insertion (Flavorunner 458). A cleaved amplified polymorphic sequence (CAPS) marker targeting the 441_442insA site in ahFAD2B was designed. ‘Georgia-02C’ and ‘Georgia Hi-O/L’, derivatives of irradiation-induced high O/L mutants, showed CAPS marker and sequence data diagnostic for the genetic mutations in ahFAD2 found in the spontaneous mutant line F435. In addition, conflicting reports regarding the organization of ahFAD2 genes and ahFAD2B mutations were reexamined. Our data clearly show that molecular markers can be used to screen for known ahFAD2 mutations and that only two mutant ahFAD2B alleles are present in the U.S. high O/L cultivars. Genetic markers based on these mutations have utility for detecting seed mixtures and heterozygotes, and can accelerate introduction of the high O/L trait into peanut breeding lines.

Abbreviations: CAPS, cleaved amplified polymorphic sequence • CTAB, hexadecyltrimethylammonium bromide • MITE, miniature-inverted-repeat-transposable element • NMD, nonsense-mediated mRNA decay • O/L, oleic:linoleic acid ratio • PCR, polymerase chain reaction • RT-PCR, reverse transcription polymerase chain reaction