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유전체재해석(resequencing)에 의한 가야벼의 벼멸구 저항성 유전자 탐색 및 선발마커 개발

지현소1,*, 안억근2, 서보윤1, 강현주1, 이상복2, 현웅조2, 최인찬1, 김경환1, 김송림1, 이승범1, 서석철1, 이강섭1

Detection of Genes Conferring Resistance to the Brown Planthopper (BPH) in Gayabyeo Through Genome Resequencing and Development of Their Selection Markers

Hyeonso Ji1,*, Eokkeun Ahn2, Bo Yoon Seo1, Hyun-Ju Kang1, Sang Bok Lee2, Ung-Jo Hyun2, Inchan Choi1, Kyung-Hwan Kim1, Song Lim Kim1, Seung-Bum Lee1, Seok-Chul Suh1, Gang-Seob Lee1
Korean Journal of Breeding Science 2018;50(2):104-115.
Published online: June 1, 2018

농촌진흥청 국립농업과학원

농촌진흥청 국립식량과학원

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(E-mail: jhs77@korea.kr, Tel: +82-63-238-4657, Fax: +82-63-238-4654)
• Received: January 1, 2018   • Accepted: January 16, 2018

Copyright: © 2018 by 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/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Detection of Genes Conferring Resistance to the Brown Planthopper (BPH) in Gayabyeo Through Genome Resequencing and Development of Their Selection Markers
Korean. J. Breed. Sci.. 2018;50(2):104-115.   Published online June 1, 2018
DOI: https://doi.org/10.9787/KJBS.2018.50.2.104
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Detection of Genes Conferring Resistance to the Brown Planthopper (BPH) in Gayabyeo Through Genome Resequencing and Development of Their Selection Markers
Korean. J. Breed. Sci.. 2018;50(2):104-115.   Published online June 1, 2018
DOI: https://doi.org/10.9787/KJBS.2018.50.2.104
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Detection of Genes Conferring Resistance to the Brown Planthopper (BPH) in Gayabyeo Through Genome Resequencing and Development of Their Selection Markers
Image Image Image Image Image
Fig. 1 Genetic map of developed CAPS markers.
Fig. 2 Read mapping of Gayabyeo genome sequences onto BPH resistance genes. Reference BPH resistance gene sequences were shown at the top row of each panel, and mapped read sequences were shown below the reference sequences. White letters indicate identical nucleotides while red letters indicate different nucleotides. a: Bph3, b: BPH14, c: BPH18, d: BPH26, e: BPH9.
Fig. 3 Detection of InDels between Gayabyeo and Nipponbare reference sequences for development of InDel markes in Bph3 and BPH26 region. a: Bph3 region, b: BPH26 region.
Fig. 4 Genotyping of Gayabyeo and other Korean rice varieties with Bph3IND and BPH26IND markers. a: genotyping Gayabyeo and 34 Korean japonica rice varieties, b: genotyping Gayabyeo and 15 Korean Tongil-type rice varieties.
Fig. 5 Distribution of BPH resistance scores of the 180 F3 families.

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Fig. 4

Fig. 5

Detection of Genes Conferring Resistance to the Brown Planthopper (BPH) in Gayabyeo Through Genome Resequencing and Development of Their Selection Markers

The list of primers for amplification of Bph3 and BPH26 genes of Gayabyeo

   Gene         forward primer         reverse primer gene size (bp)
OsLecRK1 CCCACTCAATTATAGCTACACA TCGACGACAGCAAATCCT 2442
OsLecRK2 ACAAATGCCGTTCTCCAAG AGGATCATGTGGTTGTAGAGG 2436
OsLecRK3 ACGCGGTAGGAAAGACCA AAGCATCAGTGCCCGAACA 2436
BPH26 exon1 TAGCATCAGTCCCTTGCTTGTTTGC ATTGATTTAATTAGCAGACAAGTTG 1089
BPH26 exon2 AACTCTCGTCTCGTCTTAAAATATA AGTAGTAATGTGCGTAGCAATGGAG 624
BPH26 exon3 CTAGTGCCAGTTACTCCGATAAATAT TATGCACTAGCATCACTACA 1944

* Bph3 is composed of three genes that are OsLecRK1, OsLecRK2, and OsLecRK3.

** Sequencing of BPH26 is done on its three exons according to Tamura et al. (2014).

List of developed CAPS markers

no. marker name       forward primer       reverse primer restriction enzyme chromosome position (bp)
1 GT01002 GCTGATGATCGTTTGCCACA TGCGATGTATTTTCGCGTTTT EcoRV 1 1,065,395
2 GT01003 AAATTGGCTGGGGCACTTCT TGCAGGATCAGGAAGAGCTG EcoRI 1 1,600,379
3 GT01007 CAGGGCCGCAGTGTTAATTC AGCGAGACGTTCGTGATGGT EcoRI 1 5,522,341
4 GT01008 AATAAGCGCTTGACGCGAAA CGCTCAACCACCTCGTCATT EcoRV 1 6,037,347
5 GT01011 TCCTGAGCATCCTTGCATGA AAAACGAGAGGGCGCATGT EcoRV 1 9,187,583
6 GT01018 ACTTGGAGGCGCATAACGAA GCATAATGACATGTCGCATTGAA EcoRI 1 23,956,461
7 GTS01007 AAGGGTTGTATGAACCGTCAAC TCCATGGGACACTAAAGAGAGG SalI 1 25,601,011
8 GTS01008 CGTCGAAGAAAGTGACTCAAGA CCTCTTTTGTTGGTATCATTGG NruI 1 26,694,741
9 GTS01009 ACTCCCAAAACCAGCGTAAATA CGTCAGGAATGAATTCGAAACT SacI 1 27,335,280
10 GTS01010 AGAACCCGTGTAGAGCACATTA CAATTGATCTGCTTGCTCTCTG PshBI 1 32,523,034
11 GT02001 GCATATGTTTGACCGTTCGTCTT CGGCGCTACACCATACCAAT EcoRI 2 198,216
12 GTS02001 AACTAGCACGCTTCGTTTTAGC CGGATTGGTGGAATTGAAAA StuI 2 1,330,779
13 GTS02002 CCATTTGGCAATGTTCTCATAC CACCCTTCTATGCTCTCATCCT BamHI 2 4,001,036
14 GTS02003 TGTGTGGTAGAAAATGGACCTG CCATGATTGCCAGTGTTACATC EcoRI 2 5,066,630
15 GTS02004 TACCGATCAGCTGAAAACACAC TCTTGTTCCCTTCCCTTTAACA BamHI 2 6,854,129
16 GTS02006 GTGACCTCCGATCAAATCAACT CTACTGTGGGCAGTGAAGAGGT BamHI 2 8,587,683
17 GTS02007 ATTTCCCTCCTGTTCCTCATTT GGTTGGGTTTTGGTATAGCTGA HindIII 2 9,657,613
18 GTS02008 AGCTGGAAAATGTTGTATGCAG GGGATGAATCAGTAGTTCAGCA HindIII 2 10,645,645
19 GT02014 GTGTTGGCTAGGGCCTTTGA TTCTCTGGTTGGCCTTCAGC EcoRI 2 21,056,727
20 GT02021 CGCCTCTTGAAGACCCAGAG CTGACCGGCATGGGAGTTTA SacI 2 33,887,583
21 GT03001 GCTGTTGCCGCAATATCACA AGGCATTGGCATTGTTAGGC EcoRI 3 855,210
22 GT03003 TGGGGATTGCCAGATGTGAT GCATGGCATTTGAAAGAGTGA PvuII 3 2,538,138
23 GT03004 CAGTGTCTGGGCACAGCAAG GCGTTCCACACTTCCACTCC HindIII 3 3,811,855
24 GT03005 GGAAATACCTGCCCCGTTTT CAGACGATCGGGAATTTCTACG EcoRV 3 4,753,112
25 GT03006 CTCCTCTCGCACGCAGAAGT CCAGCTTCTCCACGGAACAA EcoRV 3 5,592,985
26 GT03009 TCTACCTGCCCAGGAAGCAA TTTCAGGCAAAGCAAAGCAG EcoRV 3 9,205,476
27 GTS03002 AGCATCACGATGATTCTTTTGC TTATTGTGCCCTAGACGAATGA SacI 3 11,547,474
28 GTS03004 TAGGGTTGGCTCTCAATCTCTC ACACCTACAAGGTTGTGACTGC EcoRI 3 13,593,894
29 GTS03008 ATCGCACGGAAGGATAACTG GTTGCTCTAGAACCTCGCAATC SacII 3 19,416,808
30 GTS03009 ATTATCTCACGACCTGGGACTG AGCTAGCTAGGGTTTGTGTTGC EcoRI 3 20,345,341
31 GTS03010 TGAGGAAGAGGAGGAGATTGAG CTGCTTCATTGTGTCTGAGGAC SacI 3 21,733,967
32 GT03021 GAATCTGATTGCCCCCTAGC GGACCCAAATTGAAAGGCAAT EcoRV 3 25,459,877
33 GT03024 TTGGAAGCTTGAAACGATGG TCGCAAGTTCGAAACCCACT EcoRI 3 28,056,489
34 GT03025 TGCGAATACAATTCGGACTGC CGTGCAGTACCCTCCAAACC EcoRV 3 29,184,626
35 GT03030 AATTAAACTCCGCGCCTGGT AAACCATGCATCCGATAGGG HindIII 3 36,245,706
36 GT04004 TGGCGACAAACAGAGCACAC CCTTCTTGCGCAGGACCTC EcoRI 4 4,875,618
37 GT04005 CCAGCCAAACTCAGTTGTCCA CAGACCATACCTCCTTCAGAGCA EcoRI 4 5,924,429
38 GTS041424 ACTGGGAAGAAGGCAAGACA GTGAGCATCCCACTCTCCAT EcoRI 4 6,623,655
39 GTS041472 CCTGAAAGGGCTCAAAAACA TCTTGCAGGTGACAAATCCA PshBI 4 6,914,922
40 GTS041749 TGAACTATTGCGGTTGACCA TGCGGATTGTGTAAGACAGG EcoRV 4 8,481,173
41 GT04014 TGGATCGCTTGATGTGGTTG ATGCCTCAAATACCGCAGCA EcoRI 4 14,685,347
42 GTS043975 TTCGAGGGTTGGTGACTGAT AGAGGGTCCATGGTTTTGTG EcoRV 4 16,607,468
43 GT04020 TCCTTGCAAACGCAAAGTCA GCAACAAAACATTGCGAAGG EcoRI 4 21,083,274
44 GT05001 TTGCAGCTGAAGAAAATGTG TTTGGCACATGAAAATGATG EcoRI 5 602,152
45 GT05003 TGTTGCAGAGTAGCAAACCA TATAAGCTGCAGCCTGGAAT EcoRV 5 2,107,453
46 GTC05003 CCGAAGAGCGACTCCTAGTG AGAGATGCACAGAGGGAGGA PshBI 5 6,043,943
47 GT05013 GAGCATAAGGGGTTTCCCTA TAATGCTGGTGTGGACTCAA EcoRI 5 15,946,504
48 GTC05008 TAATTGCCTCCCAAACAACC TGTCATGCAAAGCAAAAAGG PshBI 5 18,530,587
49 GT05015 CGACGACTATATGGCGGATA AACTCCAATCGGTCATACCA EcoRI 5 20,548,891
50 GT05016 CCGTTTTGTCAGAAACCTGT TAAATCGCGAGACGAATCTT EcoRV 5 22,331,098
51 GTC05010 AGGTGGTGGTGGTTCCATT TCACTGCCGATTTCTCACTG PvuII 5 25,145,098
52 GT05024 TGATCCCCAATGAAAGCTAC TCTTGTACTCCCTTCGTTTCA EcoRI 5 29,110,069
53 GT06013 TCCAACAGCAACAATCTTCA TCTCAAGCTCAATTCCGAGT EcoRI 6 24,403,695
54 GT06015 AGAGCAGAGCTCGTGTTAGTG TGGTGATTTTTGGATGGATT EcoRI 6 26,356,825
55 GT06016 GCGAAGATGCTAGTGGAAAA GTTGCAAAGGATCTGGAAAA EcoRV 6 27,324,979
56 GT06017 GATTCGACGTCTGAAAAGGA ATATGCATGGTTCACTGCAA EcoRI 6 29,168,500
57 GT06019 TGCTCAGCAGAGTCTTGCTAT GCTTGAGACTGGTTCGTGTT SacII 6 31,015,468
58 GT07002 CAAAAGTAACCAATGGCAAGA GCACATTTGAACTGCATCTG EcoRV 7 5,619,862
59 GT07008 TGGAATCAAAGTGAAGGTCAG TGGCAATCTAAAGAGGAGGA EcoRI 7 17,797,502
60 GT07010 TATGGGGACTCCAATAAGCA TACGGTCTCGTCGGTATCAT EcoRV 7 19,489,327
61 GTS07491 AAGGCCAAATGGAGGTTAGC CTGGGAGTCCAGTCATCACA BglII 7 21,220,123
62 GT07013 ATTCAGCCCCTTCTGGATAC ACCTAGGGCAGTCCTTGATT EcoRI 7 22,517,510
63 GTS07984 GCCGCTTTCTAAAAGGATTT GACCGAAATTTTGGCTCAAT EcoRV 7 23,387,924
64 GTS07117 AGCCTGCCACTCTTCCTGTA CGAGGACATGGCATCCTATT EcoRV 7 27,812,959
65 GT07018 TTCAACCTTAGGCACTCGTC TGAGATTCTCCTTGCGTGTT EcoRV 7 28,923,650
66 GTS08007 GATCAGTCGTCCTCGTCCTC GCATTGAGCACATCCATGAC HindIII 8 1,150,395
67 GTS08142 ATGCTCCAATTGAGCTGACC TACTCATGCAGGAACGCATC EcoRI 8 6,123,634
68 GTS08218 AAATGGTGTTTTTCCCGTTG AGGTCGTGGCATGATTTAGC PshBI 8 8,787,812
69 GTS08298 CGATCGGCTCTTACATGACA AGTGGATTTGCAGGCAGCTA EcoRI 8 27,252,203
70 GTS08328 GAGTTACAAACCGCCTTTGC TTGCTAGGGGTCAAGGAAAA BglII 8 28,243,051
71 GTS09004 AGCACCCAGAGCAACAGTTT CCCACTAATTAGCCCATTGC EcoRV 9 620,373
72 GTS09063 GCTTGGATGACAAGCGTTCT AGATCGGCTTCCGAAAGAGT BglII 9 9,835,918
73 GTS09106 ACGCGACTGCTAAGGTTCAT CTCGATCGGTCCCTTTCAT EcoRV 9 11,310,635
74 GTS09188 AGTGGTAGATCCCGTTGAGC AACCGGAGACCTTCAGTGTG EcoRV 9 13,099,244
75 GTS09302 GCATGCATCCGTTGAATAAA GAGATGCTCGTCAGGGACAC HindIII 9 16,712,838
76 GTS09305 ATGGCTTTCGGGCATAGTAA GGATGGCCTATTCAGCAAAA EcoRV 9 21,161,450
77 GTS10005 TCTCAGCCACTCGTTGAATG TCTGGATGGGCTAAATGAGG EcoRV 10 721,349
78 GTS10029 CCTCCCTTCAGTTTGTGAGC CATTGGATCCCCAAAGAAAA EcoRI 10 3,044,726
79 GTS10116 CCCCCTAAGATGGTTTTTGC TGAGAAGTGAAGTGTGAGTGTGG EcoRV 10 4,577,975
80 GTS10199 AGCCTGAGAAGCAGATTGGA TGCAATGACGTGATGGTACA EcoRI 10 6,149,421
81 GTS10204 TTTCACACGGGACATTCAAA TCGACCTTGGACTTCTAGGC StuI 10 9,961,760
82 GTS10214 TAGGATCTCGCCGAACATCT ATGCACCTCCCCAACATATC EcoRV 10 11,290,285
83 GTS10276 TGGTCTTTGAAGCATTCTGG TTCAATATTGGCCGTAAACC EcoRV 10 13,637,268
84 GTS11014 CTTCGCTTCATTTCGAGTTTG CATTTCTGCAGTCGCATTTG EcoRV 11 353,397
85 GTS11016 CGCACGAGATGCTGTCTAAC GAAAAACCCTCTGCCCTCTT NruI 11 1,389,204
86 GTS11086 TGCTGGTGTGCTATTTGAGC CGTGGCCATCCAAAAGTAAT EcoRI 11 17,330,622
87 GTS11088 CGCCTAGGTGCAGAAAAGAG ACACCATGAGCCTCCTCACT BamHI 11 20,838,960
88 GTS11330 TGTCATTGGTTGCCCTGTAA AGCATGGAGGTGGTGCTAAC EcoRV 11 26,534,991
89 GTS11350 AGGAATATGAATCGCGTGTG TTTGGGAGCTATGGCCTATG EcoRI 11 27,197,820
90 GTS120003 GCGTTTTTCCTCCAATTCAA ACCGACAACTTTTGCCCTTT PvuII 12 3,007,338
91 GTS120032 AACGAAGAGGCCAATGGATA GTTGACCATGGGTGCTCACT PvuII 12 3,807,192
92 GTS120139 TGCTTTCAGGAGAGCAGGAT CCACCAGCATCACTCTGCTA EcoRI 12 9,263,934
93 GTS120457 GCATGAGAGACAGCGGAGTT GTGATTTGCCATGCCTTTTT EcoRV 12 13,674,598
94 GTS120652 TGGGCACAACTACAAAGGTG CATTCCCATGTTCCACATCA EcoRI 12 17,438,893
95 GTS120760 CCCAGCCACAAGAAATGAAT TTTTCCTTTTCCCTGCTGTG EcoRI 12 19,070,762
96 GTS120842 AAGCCATCATGTTCCATTCC TTTTGGTACGTCATCCGTGT EcoRI 12 21,522,146
97 GTS120941 CAGGTGGTCCTTTTCAGCAT TAGTGTCATGTGCCCTGAGC EcoRI 12 22,565,939
98 GTS120989 CCTTTGGAGGGCTTGACATA TACCCACTGGAAACGGAAGA EcoRV 12 23,622,982
99 GTS121030 CGTTGCTCCGTTCATCTTCT GCTAGCCCCTTTTCATCTCC EcoRV 12 26,276,873

Primer sequences of Bph3IND and BPH26IND markers.

marker name      forward primer      reverse primer
Bph3IND GCTTCACCTGTGTCCTGTT GTGTATGAGGGTTGTTCTGGAT
BPH26IND GGGCAGCTAAGCGCATGGTT GGGTGATAAATCGAACAGCAGAGG

Identification of QTLs for BPH resistance

QTL name chromosome location (cM) QTL interval* (cM) LOD additive effect dominance effect R2
 qBPH4 4 0 0-6.6 7.22 0.94 -0.20 0.138
 qBPH12 12 51.4 40.8-54.4 6.64 0.87 0.55 0.131

*interval at 99% probability

Table 1 The list of primers for amplification of Bph3 and BPH26 genes of Gayabyeo

* Bph3 is composed of three genes that are OsLecRK1, OsLecRK2, and OsLecRK3.

** Sequencing of BPH26 is done on its three exons according to Tamura et al. (2014).

Table 2 List of developed CAPS markers
Table 3 Primer sequences of Bph3IND and BPH26IND markers.
Table 4 Identification of QTLs for BPH resistance

interval at 99% probability

Table 1

Table 2

Table 3

Table 4

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