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Research Article

Identification of a Novel DFR-A Mutant Allele Determining the Bulb Color Difference between Red and Yellow Onions (Allium cepa L.)

Bongju Kim1, Youngcho Cho2, Sunggil Kim1,*
Plant Breeding and Biotechnology 2017;5(1):45-53.
Published online: March 1, 2017

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*Corresponding author: Sunggil Kim, dronion@jnu.ac.kr, Tel: +82-62-530-2061, Fax: +82-62-530-2069
• Received: January 4, 2017   • Revised: January 26, 2017   • Accepted: January 26, 2017

Copyright © 2017 The Korean Society of Breeding Science

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Identification of a Novel DFR-A Mutant Allele Determining the Bulb Color Difference between Red and Yellow Onions (Allium cepa L.)
Plant Breed. Biotech.. 2017;5(1):45-53.   Published online March 1, 2017
DOI: https://doi.org/10.9787/PBB.2017.5.1.45
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Identification of a Novel DFR-A Mutant Allele Determining the Bulb Color Difference between Red and Yellow Onions (Allium cepa L.)
Plant Breed. Biotech.. 2017;5(1):45-53.   Published online March 1, 2017
DOI: https://doi.org/10.9787/PBB.2017.5.1.45
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Identification of a Novel DFR-A Mutant Allele Determining the Bulb Color Difference between Red and Yellow Onions (Allium cepa L.)
Image Image Image Image Image
Fig. 1 The onion bulb of the F1 hybrid between a yellow cultivar, Santero, and a yellow breeding line, OT803.
Fig. 2 Organization of the inactive mutant DFR-A alleles isolated from onions. Arrow-shaped boxes indicate coding regions and the 5′-to-3′ direction. The empty and gray boxes in the coding regions indicate exons and introns, respectively. The vertical filled arrows on the exons indicate the positions of sequence changes. The nucleotide sequences on the vertical empty arrow indicate the normal sequences of the functional alleles. The dotted lines in the promoter region indicate the deleted sequences. The dotted line outside the DFR-ADEL allele indicate entire deletion of the genic region. Nucleotide sequences and names on the inverted triangles indicate inserted sequences and transposable elements, respectively. Lengths of transposable elements are shown in parenthesis.
Fig. 3 Alignment of partial nucleotide sequences of the DFR-AR4 and DFR-APS2 alleles. The vertical filled arrow indicates the positon of single base-pair deletion. The rectangular box indicates the position of the premature stop codon.
Fig. 4 RT-PCR amplification of the DFR-A gene. 1–5: individuals of OT803 containing the homozygous DFR-APS2 allele. 6–10: individuals of H6 containing the homozygous DFR-AR1 allele. CHS-A: chalcone synthase-A, F3H: flavanone 3-hydroxylase
Fig. 5 Flowchart showing the process for identification of 16 DFR-A alleles. The SNPs are shown with numbers indicating the distances from the start codon of the DFR-A gene. Four active and 12 inactive DFR-A alleles are shown in filled and empty oval-shaped circles, respectively.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Identification of a Novel DFR-A Mutant Allele Determining the Bulb Color Difference between Red and Yellow Onions (Allium cepa L.)

Primer sequences used in this study.

Name Sequence (5′ to 3′) Application Reference
DFR-F ATGCCAGTGGAGTGCATGTTGAATGGT DFR-A allele identification Kim et al. (2005b)
DFR-R TGGGTAGCGATTGGTTCATTCTCTTCA DFR-A allele identification and RT-PCR Kim et al. (2005b)
DFR-LR3 TTGCAAACTCCCATGCAGCTTTCTCTG DFR-A allele identification Song et al. (2014)
DFR-LR4 TTAACCATCTGCCCCACAAT DFR-A allele identification This study
DFR-LF5 GAGGAGCACCAAAAGCCCGAATACGAT RT-PCR Song et al. (2014)
Tubulin-F CTGGGAGCTTTACTGCCTTG RT-PCR Song et al. (2014)
Tubulin-R CAAGGGACCTCCTGCAAATA RT-PCR Song et al. (2014)
CHS-F GAGGGTCCAGCCACGGTGTTAGCTG RT-PCR Kim et al. (2004b)
CHS-R ATCAATGGCCACACTCCTAAGCACC RT-PCR Kim et al. (2004b)
F3H-F GAAGACGAGCGTCCCAAGATTGCAT RT-PCR Kim et al. (2004b)
F3H-R TCCACCATCTTTAGTCGCTTGCAGACC RT-PCR Kim et al. (2004b)

Composition of DFR-A and ANS alleles of parental lines (Santero and OT803) and the F1 hybrid. The active DFR-A and ANS alleles are shown in boldfaces. The DFR-AR4-like allele is designated as DFR-APS2 allele.

Population Genotype
DFR-A ANS
Santero DFR-AR3/DFR-AR3 ANSPS/ANSS188L
OT803 Mixed with homozygous and heterozygous DFR-APS2 and DFR-ATTA ANSh1/ANSh1
F1 hybrid DFR-APS2/DFR-AR3 ANSh1/ANSS188L
Table 1 Primer sequences used in this study.
Table 2 Composition of DFR-A and ANS alleles of parental lines (Santero and OT803) and the F1 hybrid. The active DFR-A and ANS alleles are shown in boldfaces. The DFR-AR4-like allele is designated as DFR-APS2 allele.

Table 1

Table 2

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