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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 25, No. 1, 2017, pp. 9-12
Bioline Code: ej17002
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 25, No. 1, 2017, pp. 9-12

 en Development and application of KASP marker for high throughput detection of AhFAD2 mutation in peanut
Zhao, Shuzhen; Li, Aiqin; Li, Changsheng; Xia, Han; Zhao, Chuanzhi; Zhang, Ye; Hou, Lei & Wang, Xingjun

Abstract

Background: Cultivated peanut ( Arachis hypogaea check for this species in other resources 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 BC4F2 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.

Keywords
Arachis hypogaea; Fatty acid desaturase; High oleic acid; Kompetitive allele specific PCR; Linoleic acid; Marker assisted selection; Molecular breeding; Mutation in peanut; Nucleotide substitution; Oilseed crop; Peanut oil

 
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