Background: Cultivated peanut (
Arachis hypogaea
L.) is a major oilseed cropworldwide. Fatty acid composition of
peanut oil may affect the flavor and shelf life of the resulting food products. Oleic acid and linoleic acid are
the major fatty acids of peanut oil. The conversion from oleic acid to linoleic acid is controlled by the Δ12 fatty
acid desaturase (FAD) encoded by
AhFAD2A and
AhFAD2B, two homoeologous genes from A and B
subgenomes, respectively. One nucleotide substitution (G:C → A:T) of
AhFAD2A and an “A” insertion of
AhFAD2B resulted in high-oleic acid phenotype. Detection of
AhFAD2 mutation had been achieved by cleaved
amplified polymorphic sequence (CAPS), real-time polymerase chain reaction (qRT-PCR) and allele-specific
PCR (AS-PCR). However, a low cost, high throughput and high specific method is still required to detect
AhFAD2 genotype of large number of seeds. Kompetitive allele specific PCR (KASP) can detect both alleles in a
single reaction. The aim of this work is to develop KASP for detection
AhFAD2 genotype of large number of
breeding materials.
Results: Here,we developed a KASPmethod to detect the genotypes of progenies between high oleic acid peanut
and common peanut. Validationwas carried out by CAPS analysis. The results fromKASP assay and CAPS analysis
were consistent. The genotype of 18 out of 179 BC
4F
2 seeds was aabb.
Conclusions: Due to high accuracy, time saving, high throughput feature and low cost, KASP is more suitable for
determining
AhFAD2 genotype than other methods.
