보고서 정보
주관연구기관 |
서울대학교 Seoul National University |
연구책임자 |
이석하
|
참여연구자 |
하정민
,
김문영
,
이영호
,
양슈에페이
,
윤민영
,
장현주
,
이재언
,
이은수
,
박세원
,
정준희
,
조은비
,
권하경
,
조강흠
,
최재아
,
김진숙
,
이금순
|
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2021-02 |
과제시작연도 |
2020 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
연구관리전문기관 |
농촌진흥청 Rural Development Administration |
등록번호 |
TRKO202100009787 |
과제고유번호 |
1395063039 |
사업명 |
차세대바이오그린21(R&D) |
DB 구축일자 |
2021-09-18
|
키워드 |
녹두.고품질표준유전체.후성유전인자.복총상화서.동시등숙.mungbean.high-quality reference genome sequence.epigenetic factors.synchronous pod maturity.compound raceme inflorescence.
|
초록
▼
1. 선화녹두 x IT208075 RIL 집단에서 genotype-by-sequencing을 이용한 SNP 토대 유전자연관 지도 작성
- 1,799개의 SNP 마커를 이용하여 11개 염색체로 이루어진 선 화녹두 x IT208075 집단의 genetic map을 작성함음.
2. 선화녹두 x IT208075 RIL 집단에서 복총상화서 조절 유전자좌 동정
- 복총상화서는 단일유전자에 의해 결정되는 질적형질이며 이 유전자좌를 Comraceme 으로 명명함
- Comraceme 유전자좌는 4번 염색체상에 마커들 chr4
1. 선화녹두 x IT208075 RIL 집단에서 genotype-by-sequencing을 이용한 SNP 토대 유전자연관 지도 작성
- 1,799개의 SNP 마커를 이용하여 11개 염색체로 이루어진 선 화녹두 x IT208075 집단의 genetic map을 작성함음.
2. 선화녹두 x IT208075 RIL 집단에서 복총상화서 조절 유전자좌 동정
- 복총상화서는 단일유전자에 의해 결정되는 질적형질이며 이 유전자좌를 Comraceme 으로 명명함
- Comraceme 유전자좌는 4번 염색체상에 마커들 chr4_26997427과 chr4_27545988사이에 위치하고 헤테로인 계통들이 복총상화서를 나타내는 것으로 보아 복총상화서가 단총상화서에 대해 우성임을 알 수 있음
- 염기서열변이와 유전자발현 분석을 통해 B3 Trasncription family gene의 homologue 유전자인 Vradi04g00002481가 Comraceme 후보유전자로 동정함
3. 선화녹두 x IT208075 RIL 집단에서 주요 농업형질 연관 QTL 동정
- 주요 농업 형질 표현형 조사는 식물의 구조와 관련된 마디(node) 수, 가지(branch) 수, 꽃대 수(total peduncles), 곁가지의 꽃대 수(axillary peduncle)와 수확량과 관련하여 꼬투리(total pods) 수, 개체당 꼬투리(podsper plant) 수를 조사하였고, 개화기(first flowering time), 성숙기(maturity time) 등 8개 형질에 대해 총 14개의 QTL들을 동정함
4. 녹두 집단의 다양성 분석
- 23개국으로부터 수집한 276개 녹두 수집종에 대해 GBS을 수행하고 분석된 SNP로 PCA 분석한 결과, 집단 구성과 위도와 상당한 연관 관계가 있음을 보여주었음.
5. 선화녹두 x 경기재래5호 집단에서 주요 농업형질 QTL 동정
- 선화녹두 x 경기재래5호 RIL 집단에 대해 약 9000개의 마커로 이루어진 초고밀도 유전자지도를 이용하여 키, 개화시, 가지수, 마디수, 동시등숙에 연관된 총 9개의 양적형질 유전자좌를 동정하였다
6. 녹두 동시등숙과 연관된 후성유전인자 탐색
- DNA 메틸화 정도가 동시등숙성과 통계적으로 유의미하게 다른 DNA 영역 (Differentially Methylated Region,이하 DMR)을 유형별로 조사하고 DNA 염기서열변이와 독립적이면서 동시등숙성에 관여하는 pure genic DMR 탐색함
7. 첫수확이 녹두 동시등숙에 미치는 영향
- 꼬투리를 제거 하지 않은 대조군과의 비교를 통해 처리전과 후, 그리고 선화녹두와 경기재래5호간 dfferentially expressed genes (DEGs) 분석하고 sucrose 함량 차이 비교 분석함
(출처 : 요약서 3p)
Abstract
▼
□ Purpose&Contents
< Research purposes>
Identification of genetic factors associated with agronomic traits through genetic and epigenetical approaches and trait improvement inmungbean
< Research contents >
◯ Identification of QTLs and DNA markers linked to agronomic traits in mungbean us
□ Purpose&Contents
< Research purposes>
Identification of genetic factors associated with agronomic traits through genetic and epigenetical approaches and trait improvement inmungbean
< Research contents >
◯ Identification of QTLs and DNA markers linked to agronomic traits in mungbean using high-density linkage maps
◯ Detection of differences in penotypes/genotypes/transcriptome/methylome between mappingparents in mungbean
◯ Genetic mapping of loci controlling inflorescence, synchronous maturity and branching
◯ Detection of epigenetic factors associated with agronomic traits in mungbean
□ Results
1. Construction of a genetic map using the VC1973A x IT208075 RIL population: 4.3 Gb raw data from 31 million reads were generated from IT208075 and a total of 672 million GBS reads of 235 RILs were mapped to the reference genome. Of the 4,177 SNP markers available, the genetic map contained 1,799 SNPs over 11 chromosomes (Vr01 to Vr11) and spanned 1,353.5 cM with a mean marker interval of 0.82 cM by the ML method (Supplementary Table 4). The number of SNPs mapped to each chromosome ranged between 54 (Vr07) and 357 (Vr02) with a mean of 164.
2. Identification of a locus controlling compound raceme inflorescence: Using a RIL population derived from VC1973A, an elite cultivar with a compound raceme type, and IT208075, a natural mutant with a simple raceme type, a single locus that determined the inflorescence type was identified based on 1:1 segregation ratio in the F8 generation, and designated Comraceme. Linkage map analysis showed Comraceme was located on chromosome 4 within a marker interval spanning 520 kb and containing 64 genes. RILs carrying heterozygous fragments around Comraceme produced compound racemes, indicating this form was dominant to the simple raceme type. Expression of Vradi04g00002481, a development-related gene encoding a B3 transcriptional factor was significantly lower at the early vegetativestage and higher at the early reproductive stage, in IT208075 than inVC1973A. Vradi04g00002481 was therefore likely to determine inflorescence type in mungbean.
3. Identification of QTLs linked with agronomic traits using a VC1973C x IT208075 RIL population: We penotyped eight agronomic traits including node number, branch number, total number of peduncles, axillary peduncle number, total pods, pods per plant, first flowering time and maturity time for two years. In total, 14 QTLs associated with the traits were identified on chromosome 4, 8 and 9. The QTL for node number, branch number, total number of peduncles, total pods, pods per plant and first flowering time on chromosome 4 was co-located with the locus Comraceme.
4. Molecular diversity among Asian mungbean accessions: The new mungbean genome assembly was used as a reference to assess genetic variation among 276 accessions, including 233 cultivars, 42 wild mungbean accessions, and one Vigna glabrescens accession (outgroup), collected from 23 countries. Raw sequence reads were generated via genotyping-by-sequencing (GBS) at a sequencing depth of 4.3–13X,depending on the accession. Principal component analysis (PCA) of selected mungbean cultivars with reliable origin data showed somecorrelation between population group membership and geographical origin.
5. Identification of QTLs associated with agronomic traits in a VC1973C x V2984 RIL population: QTLs regulating plant height, node number, branch number, flower initiation, and synchronous pod maturity were identified based on the genetic map constructed using resequencing data. QTLs for plant height were detected on Vr04 (Height4-1) and Vr05 (Height5-1), which explained 23.9% and 6.2% of the variance in plant height, respectively (percentage of variance explained, PVE). QTLs for flower initiation were identified on Vr04 (FI4-1) and Vr09 (FI9-1), with 24.1 and 6.4 PVE, respectively. One QTL affecting branch number was detected on Vr03 (Branch3-1), with 11.1 PVE. QTLs affecting node number were identified on Vr04 (Node4-1) and Vr11 (Node11-1), with 20.0 and 6.3 PVE, respectively. Two QTLsfor SPM were detected on Vr04 (SPM4-1) and Vr07 (SPM7-1), with 10.3 and 6.8 PVE, respectively. On Vr04, Height4-1, Node4-1 and SPM4-1 were identified within the same marker interval and FI4-1 was adjacent to them, sharing one genetic marker (29,615,281 bp)
6. Identification of epigenetic variation associated with synchronous pod maturity in mungbean: To elucidate the epigenetic influences on synchronous pod maturity (SPM) in mungbean, we determined the genome-wide DNA methylation profiles of eight RILs and their parental genotypes, and compared DNA methylation profiles between high SPM and low SPM RILs, thus revealing differentially methylated regions (DMRs). A total of 3, 18, and 28 pure DMRs, defined as regions showing no significant correlation between nucleotide sequence variation and methylation level, were identified in CpG, CHG, and CHH contexts, respectively.
7. Transcriptomic and biochemical analyses of the accumulation of sucrose in mungbean leaves after pod removal: we examined changes in sucrose levels and transcriptome in leaf (source) tissues after pod (sink) removal using two genotypes, VC1973A and V2984; VC1973A had higher synchronicity in pod maturity than V2984. After pod removal, much higher number of pods were produced in V2984 than VC1973A. The sucrose content of leaf tissues significantly decreased in V2984 because it continued to utilize assimilates from leaves for producing new pods, but significantly increased in VC1973A because of the loss of sink. Transcriptome analysis revealed that the number of differentially expressed genes was approximately 4-fold higher in VC1973A than in those of V2984 after pod removal. The expression of two paralogous genes (Vradi01g05010 and Vradi10g08240), encoding beta-glucosidase enzymes, significantly decreased in VC1973A after pod removal and was significantly lower in depodded VC1973A than depodded V2984, indicating these two genes may participate in sucrose utilization for seed development by regulating the level of glucose.
□ Expected Contribution
Development of DNA markers associated with useful traits using the mungbean reference genome sequence improved in both quailty and quantity and the ultrahigh-density genetic linkage map
○ Advancement of genomics-assisted breeding for mungbean improvement through identification of genes controlling agronomic traits based on QTLs linked to those traits
○ Mungbean is lacking a lot of trait improvement and breeding relative to soybean. Mungbean reference genome sequence with high-quality and high-level completion would be a powerful tools for genetic and molecular studies of many agronomic traits and breeding.
○ Suggestion of novel index to evaluate non-synchronous pod maturity in mungbean and improvement of synchronous pod maturity trait for high-yielding and labour-saving
(출처 : SUMMARY 7p)
목차 Contents
- 표지 ... 1
- 제출문 ... 2
- 보고서 요약서 ... 3
- 국 문 요 약 문 ... 5
- Summary ... 7
- 목차 ... 10
- 제 1 장 연구 개발 과제의 개요 ... 11
- 제 1 절 연구 개발 목적 ... 11
- 제 2 절 연구 개발의 필요성 ... 11
- 제 2 장 연구 수행 내용 및 결과 ... 16
- 제 3 장 목표달성도 및 관련분야 기여도 ... 65
- 제 1 절 목표대비 달성도 ... 65
- 제 2 절 정량적 성과(논문게재, 특허출원, 기타)를 기술 ... 66
- 제 4 장 연구 결과의 활용 계획 ... 68
- 제 5 장 연구 개발 결과의 보안 등급 ... 69
- 제 6 장 연구시설·장비종합정보시스템에 등록한 연구시설·장비 현황 ... 70
- 제 7 장 연구개발과제의 대표적 연구실적 ... 71
- 제 8 장 기타사항 ... 72
- 제 9 장 참고문헌 ... 73
- 끝페이지 ... 75
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