KR101975797B1 - 고추의 웅성불임 회복과 관련된 핵산 분자, 프라이머 세트, 및 그의 용도 - Google Patents
고추의 웅성불임 회복과 관련된 핵산 분자, 프라이머 세트, 및 그의 용도 Download PDFInfo
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- KR101975797B1 KR101975797B1 KR1020140095013A KR20140095013A KR101975797B1 KR 101975797 B1 KR101975797 B1 KR 101975797B1 KR 1020140095013 A KR1020140095013 A KR 1020140095013A KR 20140095013 A KR20140095013 A KR 20140095013A KR 101975797 B1 KR101975797 B1 KR 101975797B1
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Abstract
Description
도 2는 AFLP 분자표지와 G05G1 및 OPP13 분자표지에 대한 연관지도를 나타낸다.
도 3은 고추 회복 유전자 연관 분자표지를 선별된 재조합 개체에 적용한 결과를 나타낸다.
도 4는 고추 회복 유전자 연관 분자표지를 이용하여 그 유전자형을 분석한 결과를 나타낸다.
도 5는 토마토 유전체, 고추 scaffold 서열, AFLP 유전자 지도, BAC 말단 서열 유래 분자표지들에 대한 연간 지도 간의 상관 관계를 나타내는 모식도이다.
도 6은 고추 회복 유전자와 공동분리되는 DNA 구역을 결정하기 위해 수행한 염색체 워킹 과정에 대한 모식도이다.
도 7은 3개의 서로 다른 고추 계통에서 얻은 4개의 PPR6 유사 유전자의 단백질 서열을 비교한 결과를 나타낸다.
도 8은 부강 C와 부강 A에서 분석된 전사체의 read들을 PPR6 유전자 상에 배열한 결과를 나타낸다.
도 9는 RfRf 또는 rfrf 유전자형을 갖는 청양 F2 개체의 여러 조직에서 RT-PCR 분석을 수행한 결과를 나타낸다.
분자표지명 | 프라이머 조합 |
E32/M44 | EcoRI+AAC 및 MseI+ATC |
E33/M36 | EcoRI+AAG 및 MseI+ACC |
E41/M62 | EcoRI+AGG 및 MseI+CTT |
E48/M50 | EcoRI+CAC 및 MseI+CAT |
E49/M40 | EcoRI+CAG 및 MseI+AGC |
E51/M36 | EcoRI+CCA 및 MseI+ACC |
E54/M36 | EcoRI+CCT 및 MseI+ACC |
E59/M59 | EcoRI+CTA 및 MseI+CTA |
분자표지명 | 프라이머 서열 (서열번호) |
제한 효소 | PCR시 Tm 값 |
유전적 거리 |
ABHD 1.5-SCAR | TGGTGCCAATGCAATGTGGC (10) |
|
56.0 |
6/1068 |
GCTTCTCAGGTGTCTGTGGC (11) | ||||
CRF-SCAR | ATTTTCAGATTGTGGCGACG (12) |
|
57.0 |
1/1068 |
CGACCATCACGACGAGG (13) | ||||
4940 CAPS | CCCAGCCCAGGCTTACTTAAC (14) |
RsaI
|
56.0 |
1/1068 |
CTGAAGGTGTTGTCGCCTTGC (15) | ||||
3336-last2 SCAR | CATCGAACTGATACGGAAGGAC (16) |
|
55.0 |
0/1068 |
TAACACTACTTGGGGAAAGCG (17) | ||||
0.9-10K CAPS | GCCTAATCTAACAGGTGACTC (18) |
Bcl1
|
51.0 |
0/1068 |
GGATAAGTAAGGGTGGAAAAGG (19) | ||||
120K F2R3 SCAR | TAAAAAAATAGCGGGGGGTAAG (20) |
|
53.0 |
0/1068 |
CCACTTGTCTTGAAGCACTC (21) | ||||
PPR6 Co1Mod1 CAPS | CCACCCCAAACGTAAGGGAT (22) |
Bgl2
|
56.0 |
0/1068 |
CAACTTAGCCAATACAATCCCAC (23) | ||||
PPR12 SCAR | CAAGGTGTTGTCTATGTGGG (24) |
|
49.5 |
3/1068 |
CATCAACGTGATTAGTGGATAC (25) | ||||
415 SP6-3 SNP |
CACTAAATCTCCCGCTATGAGT (26) |
|
55.0 |
3/1068 |
GGAGTGGGGTAAGAAAAAGATTTACG (27) | ||||
4162-2 SCAR | GCAGTTCGGTTTTACGGAGTTAC (28) |
|
56.0 |
3/1068 |
CCATTGGACAAAAGGGGATC (29) | ||||
1.85Mb HRM | GACATGCAAGGTAAGGCTGC (30) |
|
53.0 |
12/1068 |
CACAAATTCTGGCTATCGGTC (31) |
분자표지 |
분자표지에 따른 유전자형 |
식물의 웅성불임 회복 유전자형 |
판별 성공률 (%) |
|
RfRf | rfrf | |||
OPP13-CAPS |
OPP-haplotype 1 | 6 | - | 27.5 |
OPP-haplotype 2 | 1 | 8 | ||
OPP-haplotype 3 | 14 | 22 | ||
PR-CAPS |
PR-haplotype 1 | 8 | 9 | 33.3 |
PR-haplotype 2 | 9 | 9 | ||
PR-haplotype 3 | 4 | 12 | ||
CRF-SCAR |
RfRf 또는 Rfrf | 19 | 3 | 90.2 |
rfrf | 2 | 27 | ||
4940 CAPS |
RfRf | 19 | 2 | 92.2 |
rfrf | 1 | 17 | ||
증폭 안 됨 | 1 | 11 | ||
3336-last2 SCAR | RfRf 또는 Rfrf | 19 | 3 | 90.2 |
rfrf | 2 | 27 | ||
PPR6 Co1Mod1 CAPS |
RfRf | 19 | 1 | 92.2 |
Rfrf | - | 1 | ||
rfrf | 2 | 28 | ||
4162-2 SCAR |
RfRf 또는 Rfrf | 19 | 22 | 52.9 |
rfrf | 2 | 8 | ||
PPR12 SCAR |
RfRf 또는 Rfrf | 19 | 28 | 41.2 |
rfrf | 2 | 2 | ||
총 계통수 | 21 | 30 |
분자표지 |
분자표지에 따른 유전자형 |
식물의 웅성불임 회복 유전자형 |
판별 성공률 (%) |
||
RfRf | rfrf | Rfrf | |||
OPP13-CAPS |
OPP-haplotype 1 | 6 | 0 | 1 | 24.0 |
OPP-haplotype 2 | 0 | 0 | 0 | ||
OPP-haplotype 3 | 8 | 6 | 13 | ||
Heterotype | 0 | 6 | 0 | ||
CRF-SCAR |
RfRf 또는 Rfrf | 15 | 8 | 2 | 70.0 |
rfrf | 9 | 4 | 12 | ||
4940 CAPS |
RfRf 또는 Rfrf | 14 | 6 | 1 | 66.0 |
rfrf | 10 | 6 | 13 | ||
3336-last2 SCAR | RfRf 또는 Rfrf | 21 | 12 | 1 | 92.0 |
rfrf | 3 | 0 | 13 | ||
120kb F2R3 SCAR | RfRf 또는 Rfrf | 21 | 12 | 1 | 92.0 |
rfrf | 3 | 0 | 13 | ||
PPR6 Co1Mod1 CAPS |
RfRf | 19 | 0 | 1 | 88.0 |
Rfrf | 2 | 12 | - | ||
rfrf | 3 | - | 13 | ||
415 SP6-3 SCAR | RfRf 또는 Rfrf | 16 | 12 | 11 | 62.0 |
rfrf | 8 | 0 | 3 | ||
총 계통수 | 24 | 12 | 14 |
Contig명 | 길이 | BLASTX 추정 단백질 기능 | 공동분리 DNA 구역에서의 위치 | 상동성 (%) | Contig 내 최장 orf의 길이 | ||
시작점 | 끝점 | BAC 클론 DNA 서열 기준 | 전사체 유래 contig 기준 | ||||
contig_135691 | 498 | NAC domain-containing protein | 107390 | 107887 | 98.8 | 213 | 213 |
contig_90919 |
718 |
P-selectin-glycoprotein |
168207 | 168412 | 100 | 365 |
362 |
168619 | 168682 | 98.65 | |||||
170818 | 171265 | 96.65 | |||||
contig_56827 | 499 | transposable element | 173685 | 173187 | 100 | 120 | 120 |
contig_133674 | 429 | transposable element | 175961 | 176389 | 99.77 | - | - |
contig_130916 |
637 |
unknown |
404350 | 404727 | 100 | 228 |
228 |
405587 | 405744 | 100 | |||||
445115 | 445204 | 100 | |||||
contig_115973 | 393 | Protein binding protein | 405115 | 404723 | 100 | 260 | 266 |
contig_137829 | 479 | unknown | 406210 | 406689 | 99.38 | 244 | 222 |
contig_132622 | 666 | unknown | 409366 | 408702 | 99.85 | 330 | 337 |
contig_36222 | 738 | transposable element | 410038 | 410775 | 99.86 | 297 | 297 |
contig_133779 | 383 | transposable element | 412183 | 411801 | 100 | 351 | 357 |
contig_84137 | 584 | transposable element | 414086 | 413503 | 99.66 | 168 | 168 |
contig_81669 |
2108 |
NBS-LRR root knot nematode resistance |
442820 | 441345 | 100 | 444 | 444 |
440995 | 440901 | 98.11 | |||||
440797 | 440645 | 100 | |||||
440486 | 440366 | 100 | |||||
440286 | 440024 | 99.27 | |||||
contig_112857 | 1298 | NBS-LRR root knot nematode resistance | 443460 | 444757 | 100 | 606 | 606 |
contig_64819 | 436 | disease resistance RPP13-like | 446306 | 445873 | 99.31 | 114 | 114 |
contig_90053 | 364 | dihydroorotase | 447536 | 447173 | 100 | 129 | 129 |
contig_110311 |
726 |
NBS-LRR root knot nematode resistance |
450064 | 450157 | 95.74 | 357 |
357 |
450634 | 451263 | 99.37 | |||||
contig_107563 |
986 |
TOM40-1 |
456816 | 457344 | 99.05 | 237 |
207 |
457339 | 457700 | 98.08 | |||||
contig_116115 | 441 | PPR1 | 459290 | 458850 | 100 | 441 | 435 |
contig_112011 | 678 | transposable element | 806082 | 805524 | 98.21 | 453 | 575 |
Claims (11)
- 삭제
- 삭제
- 삭제
- 서열번호 9의 뉴클레오티드 서열로 이루어지는, 웅성불임 회복 특성을 갖는 고추에 특이적인 핵산 분자.
- 서열번호 9의 뉴클레오티드 서열로부터 제작되는, 고추의 웅성불임 회복 특성을 예측하기 위한 프라이머 세트.
- 청구항 5에 있어서, 상기 프라이머 세트는 서열번호 14 및 15의 올리고뉴클레오티드로 이루어지는 프라이머 세트; 서열번호 16 및 17의 올리고뉴클레오티드로 이루어지는 프라이머 세트; 서열번호 20 및 21의 올리고뉴클레오티드로 이루어지는 프라이머 세트; 및 서열번호 22 및 23의 올리고뉴클레오티드로 이루어지는 프라이머 세트로 구성된 군으로부터 선택되는 하나 이상의 프라이머 세트를 포함하는 것인 프라이머 세트.
- 청구항 5에 있어서, 상기 프라이머 세트는 서열번호 14 및 15의 올리고뉴클레오티드로 이루어지는 프라이머 세트; 서열번호 16 및 17의 올리고뉴클레오티드로 이루어지는 프라이머 세트; 서열번호 20 및 21의 올리고뉴클레오티드로 이루어지는 프라이머 세트; 및 서열번호 22 및 23의 올리고뉴클레오티드로 이루어지는 프라이머 세트를 포함하는 것인 프라이머 세트.
- 청구항 5의 프라이머 세트를 포함하는, 고추의 웅성불임 회복 특성을 예측하기 위한 키트.
- 고추로부터 유래된 게놈 DNA를 주형으로 하고 청구항 5의 프라이머 세트를 프라이머로 사용하여 핵산을 증폭하는 단계; 및
얻어진 증폭 산물로부터 상기 고추가 웅성불임 회복 특성을 갖는지를 결정하는 단계를 포함하는, 고추의 웅성불임 회복 특성을 예측하는 방법. - 청구항 9에 있어서, 상기 프라이머 세트가 서열번호 14 및 15의 올리고뉴클레오티드로 이루어지는 프라이머 세트 또는 서열번호 22 및 23의 올리고뉴클레오티드로 이루어지는 프라이머 세트를 포함하는 경우, 얻어진 증폭 산물을 제한 효소로 절단하는 단계를 더 포함하는 것인 방법.
- 청구항 10에 있어서, 상기 제한 효소는 AluI, BglⅡ, BsaWI, BsoBI, BsrGI, BstYI, HindⅢ, Hpy188I, Hpy188Ⅲ, MspI, RsaI, TaqI, TatI, XbaI, 또는 XhoI인 것인 방법.
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