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Research Article

Kompetitive Allele-Specific PCR Marker Development and Quantitative Trait Locus Mapping for Bakanae Disease Resistance in Korean Japonica Rice Varieties

Kyeong-Seong Cheon1, Young-Min Jeong2, Youn-Young Lee2, Jun Oh1, Do-Yu Kang1, Hyoja Oh1, Song Lim Kim1, Nyunhee Kim1, Eungyeong Lee1, Jeongho Baek1, Inchan Choi1, Kyung-Hwan Kim1, Yong Jae Won3, In Sun Yoon1, Young-il Cho2, Jung-Heon Han1, Hyeonso Ji1,*
Plant Breeding and Biotechnology 2019;7(3):208-219.
Published online: September 1, 2019

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*Hyeonso Ji, jhs77@korea.kr, Tel: +82-63-238-4657, Fax: +82-63-238-4654
• Received: August 2, 2019   • Revised: August 12, 2019   • Accepted: August 12, 2019

Copyright © 2019 The Korean Society of Breeding Science

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Kompetitive Allele-Specific PCR Marker Development and Quantitative Trait Locus Mapping for Bakanae Disease Resistance in Korean Japonica Rice Varieties
Plant Breed. Biotech.. 2019;7(3):208-219.   Published online September 1, 2019
DOI: https://doi.org/10.9787/PBB.2019.7.3.208
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Kompetitive Allele-Specific PCR Marker Development and Quantitative Trait Locus Mapping for Bakanae Disease Resistance in Korean Japonica Rice Varieties
Plant Breed. Biotech.. 2019;7(3):208-219.   Published online September 1, 2019
DOI: https://doi.org/10.9787/PBB.2019.7.3.208
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Kompetitive Allele-Specific PCR Marker Development and Quantitative Trait Locus Mapping for Bakanae Disease Resistance in Korean Japonica Rice Varieties
Image Image Image Image
Fig. 1 Construction of a physical map of japonica genome with 771 KASP markers which include 400 markers from Cheon et al. (2018), and 371 new KASP markers. Numbers on the left side and letters on the right side of each chromosome (Chr) indicate the physical location (bp) and the name of each marker, respectively. Black and red letters on the right side represent KASP markers developed in previous and present studies, respectively.
Fig. 2 Phenotypic differences of parental varieties and F3 families to BD. (A) Plants at 4 weeks after inoculation with BD show the different responses to BD. (B) Distribution of the mortality rate of the F3 families from Junam/Nampyeong and Saenuri/Nampyeong. Solid inverse triangles indicate the average mortality values of the three parents.
Fig. 3 A genetic map of KASP markers identified in the F2 families of Junam/Nampyeong (A), Saenuri/Nampyeong (B), and position of QTLs for BD resistance. The numbers on the left side and letters on the right side of each chromosome (Chr) indicate the genetic distance of each marker from the first marker at the top of each chromosome and the name of each marker, respectively. The genetic distance (cM) was calculated by the Kosambi function. Genetic maps of 12 chromosomes are presented in Fig. S1 and in the previous study (Cheon et al. 2018). The QTL interval at 95% probability is indicated by the filled black box.
Fig. 4 Detection of indel regions and development of selection markers for BD resistance. (A) PCR amplification of Os01g0601625 and Os01g0601675 regions. (B) Positions of Os01g0601625 and Os01g0601675 regions in the Nipponbare reference genome (IRGSP-1.0) and each indel identified in the Os01g0601625 and Os01g0601675 regions on chromosome 1. The numbers in the upper line indicate the position on chromosome 1 of the Nipponbare reference genome. Gray areas and red boxes in the lower line indicate the predicted gene regions and indel positions, respectively. (C, D) Comparison with Nipponbare (Nip) and Nampyeong (Nam) sequences of Os01g0601625 (C) and Os01g0601675 (D) regions. Arrows indicate the direction and position of primers, and dots indicate the indel regions in each gene. (E, F) PCR band patterns of 1625IND (E) and 1675IND (F) in 27 Korean japonica varieties: 1, Dongjin; 2, DongjinAD; 3, Nampyeong; 4, Hwayeong; 5, Junam; 6, Hopum; 7, Anmi; 8, Ilmi; 9, Hiami; 10, Samgwang; 11, Ungwang; 12, Seolhyangchal; 13, Chilbo; 14, Saenuri; 15, Odae; 16, Joun; 17, Dongan; 18, Geumo; 19, Hwaseong; 20, Nagdong; 21, Giho; 22, Paldal; 23, Jinheung, 24, Jodongji; 25, Damageum; 26, Jungseangeunbangju; 27, Jujido; M, 1 kb Plus DNA ladder.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Kompetitive Allele-Specific PCR Marker Development and Quantitative Trait Locus Mapping for Bakanae Disease Resistance in Korean Japonica Rice Varieties

Summary of KASP assays from the resequencing data of 13 Korean japonica varieties.

Assay (n) Polyz) (n, %) Monoy) (n, %) Nox) (n, %) Reference
13 Korean japonica rice varieties 1st 506 400 (79.1) 89 (17.6) 17 (3.4) Cheon et al. (2018)
2nd 504 371 (73.6) 126 (25.0) 7 (1.4) In this study
Total 1010 771 (76.3) 215 (21.3) 24 (2.4)

z)Polymorphism.

y)Monomorphism.

x)No amplification.

Summary of the 771 annotated KASP assay sites.

Regions No. %
Intergenic 600 77.8
 Genic
  5′ UTR 11 1.4
  CDS
   NSz) 31 4.0
   SYy) 17 2.2
  Intron 89 11.5
  3′ UTR 21 2.7
  Exons 2 0.3

z)Non-synonymous.

y)Synonymous.

Identification of two QTLs related to BD resistance.

Cross Combinationz) QTL name Chry) Location (cM) Closest marker QTL intervalx) (cM) LOD Additive Dominant R2 Reference
J/N qFfR1-1 1 98.9 KJ01_075 95.8–100.7 21.4 19.45 −7.44 0.499 qBK1 (Hur et al. 2015)
qBK1.1 (Fiayz et al. 2016)
qFfR1 (Ji et al. 2018)
S/N qFfR6 6 61.7 KJ06_055 57.1–62.7 5.99 9.44 0.74 0.124
KJ06_059

z)J/N and S/N indicate Junam/Nampyeong and Saenuri/Nampyeong, respectively.

y)Chromosome number.

x)Interval at 95% probability.

Table 1 Summary of KASP assays from the resequencing data of 13 Korean japonica varieties.

Polymorphism.

Monomorphism.

No amplification.

Table 2 Summary of the 771 annotated KASP assay sites.

Non-synonymous.

Synonymous.

Table 3 Identification of two QTLs related to BD resistance.

J/N and S/N indicate Junam/Nampyeong and Saenuri/Nampyeong, respectively.

Chromosome number.

Interval at 95% probability.

Table 1

Table 2

Table 3

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