초록 ▼

Ⅳ. 연구개발결과
○ 콩 분자육종플랫폼 구축을 위해 대풍 등 34품종의 전장유전체 정보를 해독하였다. 이를 통해 Variation Block(VB) 기반의 Indel 마커(4bp 이상) 17,127 개를 개발하였다. 또한 유용 QTL 탐색을 위한 황금/대풍 F9세대 RILs 620계통을 육성하였다.
○ 콩 분자육종플랫폼 활용에서는 수량 관련 유용 VB 탐색, 종자크기 및 이소플라본 함량 관련 QTL 탐색 등 VB 기반의 질적형질 탐색 방법(variation block association study, VBAS)을 구축하고

Ⅳ. 연구개발결과
○ 콩 분자육종플랫폼 구축을 위해 대풍 등 34품종의 전장유전체 정보를 해독하였다. 이를 통해 Variation Block(VB) 기반의 Indel 마커(4bp 이상) 17,127 개를 개발하였다. 또한 유용 QTL 탐색을 위한 황금/대풍 F9세대 RILs 620계통을 육성하였다.
○ 콩 분자육종플랫폼 활용에서는 수량 관련 유용 VB 탐색, 종자크기 및 이소플라본 함량 관련 QTL 탐색 등 VB 기반의 질적형질 탐색 방법(variation block association study, VBAS)을 구축하고 대풍의 제색 개량 등 맞춤형 품종개발에 활용하였다.
○ 품종 인식을 위한 바코드시스템 개발에서는 VB 기반의 Indel 마커 202개를 활용한 바코드시스템과 이를 구동하는데 필요한 프로그램을(MyCrops_soybean) 개발하고 산업체에 기술이전하였다.
○ 대풍 및 황금 콩에서의 종자 발달단계에 따른 전사체 발현비교를 RNA-Seq을 통해 다양한 novel transcript 와 alternative splicing 발굴하였다. 황금콩에서 대풍 종실 외관 특성 개량을 위한 유용 유전자 탐침의 주요한 후보군으로서 DHN 유전자의 과발현을 동정 하였으며, 유전자의 기능 분석(종실외관개선, 재해내성 및 생산성 증대)은 형질전환 효모균주 및 벼 형질전환체에서 확인하였다.
○ 종자발달단계별 RNA-Seq을 통한 이소플라본 합성 유전자의 발현 변화를 측정하였으며, 그에 따른 품종간의 이소플라본 성분 분석을 수행하였다. 이소플라본의 항산화 효과 및 매커니즘 분석은 효모 균주를 통해 실시하였다.
○ 황금 콩에서의 저온스트레스 하에서 단백질체 분석을 통한 다양한 스트레스 반응유용 유단백질의 발현변화를 확인하였다.
○ Vegetative stage에서의 (V1) 품종간(신팔달 2호, 신기, 백운, 대풍, 황금)의 유전자 발현분석을 RNA-Seq을 통해 수행 완료하였으며, 동일한 조건에서의 품종 간의 단백질체 분석을 LC-ESI-MS/MS를 통해 수행하였다. 전사체 및 단백질체 결과의 비교 분석을 통한 품종별 특이 유전자의 발현을 동정하였다.
○ 초다수성 집단을 이용한 Sequencing based genetic map의 작성 및 장려품종들의 염기서열 변이 정보의 확보 및 수량 관련 QTL 및 유전자의 확인을 위해 풍산나물콩, SS2-2, SS404-T5-76, 대원콩, 신팔달콩2호 등 국내 우량콩 품종 resequencing 수행하였다. 대원콩xS404-T5-76(high-yield line)의 유전자 지도 작성를 위한 F6세대 recombinant inbred line (RIL) 육성하였다.
○ 초다수성 콩 계통의 육성을 위해 교배조합별 SSD에 의한 F5세대 RIL 육성하였다. 양질 다수성 콩 품종 육성 연구에서는 우량 콩 계통육성, 양질 다수성 계통 생산력검정시험 및 지역검정실험을 통해 우량 다수성 나물용 콩 품종을 기술이전하였다.
○ 최종 선별된 콩 품종 또는 유효성분의 생리활성(암예방/체중조절/피부미용/심혈관질환 예방 효능) 관련 분자표적 단백질 규명연구에서 콩 아이소플라본인 daidzein의 대사체인 7,3’,4’-THIF의 미백 효능 관련 작용기전 및 분자표적을 규명한 결과, 7,3’,4’-THIF를 세포주에 처리시, AKT/mTOR/GSK3beta signal은 활성화되고 MKK3/6/p38/MSK 및 PKA signal은 억제되는 것으로 확인하였다. 7,3’,4’-THIF가 alpha-MSH와 경쟁적으로 MC1R과 결합한다는 것을 computer modeling을 이용하여 규명하였다.

Abstract ▼

In contrast with wild species, cultivated crop genomes consist of reshuffled recombination blocks, which occurred by crossing and selection processes. Accordingly, recombination block-based genomics analysis can be an effective approach for the screening of target loci for agricultural traits. We pr

In contrast with wild species, cultivated crop genomes consist of reshuffled recombination blocks, which occurred by crossing and selection processes. Accordingly, recombination block-based genomics analysis can be an effective approach for the screening of target loci for agricultural traits. We propose the variation block method, which is a three-step process for recombination block detection and comparison. The first step is to detect variations by comparing the short-read DNA sequences of the cultivar to the reference genome of the target crop. Next, sequence blocks with variation patterns are examined and defined. The boundaries between the variation-containing sequence blocks are regarded as recombination sites. All the assumed recombination sites in the cultivar set are used to split the genomes, and the resulting sequence regions are termed variation blocks. Finally, the genomes are compared using the variation blocks. The variation block method identified recurring recombination blocks accurately and successfully represented block-level diversities in the publicly available genomes of 31 soybean and 23 rice accessions. The practicality of this approach was demonstrated by the identification of a putative locus determining soybean hilum color. We suggest that the variation block method is an efficient genomics method for the recombination block-level comparison of crop genomes. We expect that this method will facilitate the development of crop genomics by bringing genomics technologies to the field of crop breeding.
Gene expression analysis of Daepoong (DP) and Hwanggeum (HG) during the seed development stage revealed that no gene expression was constitutively up- or downregulated during the ripening period, (i.e., from flowering to a seed size of 10.0 mm); the exception being the DHN genes. However, there was a clear difference when gene expression patterns of the first and second half of the seed development stage were compared. Gene expression was upregulated in HG with compared to DP. Yeast transformants expressing DHN (CaDHN) showed higher abiotic stress resistance and alcohol fermentation capacity (particularly at low temperature) than the wild-type yeast strain containing a control vector. We also confirmed the increased improvement of and rice grain yield in rice transformants expressing DHN. These results suggest the potential of the DHN gene as a useful genetic resource for improving the and productivity of soybeans. Isoflavone biosynthetic genes dependent on the seed development stage were overexpressed in DP compared to HG, and the isoflavone content was proportional to the gene expression level. We also confirmed the antioxidant effect of isoflavone in the yeast system, and present the antioxidative mechanism of isoflavone through gene expression analysis using a microarray. Proteomic analysis using HG under low temperature stress revealed the overexpression of various cell rescue systems (e.g., protein synthesis and degradation, antioxidant system, electron donor system). Gene expression analysis under using RNA-Seq showed a marked difference between varieties and various novel genes were accordingly verified. Pearson correlation analysis showed that phylogenetic classification of strains is possible through gene expression analysis. Furthermore, proteomic analysis under the same conditions revealed a clear difference in gene expression between different varieties. These gene expression results obtained through transcriptomic and proteomic analyses provide significant information indicating that the physiological characteristics of each strain under normal growth conditions can be compared at the molecular level. In the future, having completed a comparative gene expression analysis of different varieties with respect to their protective capacity against stress types, the stress adaptation ability of each strain will be systemized. This will enable the development of protocols to establish a platform for developing strains with multiple stress tolerance and enhanced productivity.
The objective of this project was to develop a soybean cultivar and promising inbred lines with high-quality and high-yielding potential through genomics-assisted breeding. Firstly, we performed replicated yield trials in different locations during three years using six superior recombinant inbred lines (RILs) derived from different cross combinations for registration of a new soybean variety. The SS0404-T5-63 line used for soybean sprouts, which were derived from a cross of Pungsanamulkong x SS2-2 showed higher yield and more resistant to lodging and bacterial pustule than Pungsanamulkong as a control in three locations, Suwon, Naju, and Jeju. So, we had a technology transfer deal for the variety registration right of SS0404-T5-63 with CJ CheilJedang company. Additional RILs with high-yielding were developed through single seed descendant method and line selection following artificial crossing between the high-yield line and elite Korean cultivars. Meanwhile, five soybean genotypes including the high-yield line SS0404-T5-76 were resequenced to catalog sequence variations among these genotypes. These data will be useful to construct a genetic linkage map for the dissection of genomic elements responsible for high-yielding. We also surveyed genome-wide mutations in dwarf soybean mutant derived from fast neutron bambardment, revealing that loss of peroxidase gene due to a large deletion event is attributed to the dwarfism in soybean. In addition, we developed two sets of near isogenic lines (NILs) carrying high and low protein alleles at the quantitative trait locus for high seed protein on Chr 20 and whole genomes of these NILs were sequenced. The investigation of nucleotide variations between high and low protein lines enabled to predict feasible candidate genes involved in accumulation of seed storage protein.
Soybean isoflavones provide beneficial effects for several of the most common diseases that afflict humans such as cancer. The health benefits of soybean products have been well documented. Growing evidence has shown that soybean consumption may prevent certain cancers, has a beneficial role in chronic renal disease, lowers plasma cholesterol, and decreases the risk of coronary heart disease. The active components of the soy products responsible for these effects have not yet been defined. However, the components of isoflavones, genistein and daidzein, are thought to be responsible for the observed health benefits. The objective of this study was to evaluate the concentration of functional materials in high producible soybean. A total of five varieties, SS0403-T5-38, SS0407-T5-67, SS0408-T5-99, SS0406-T5-1, and SS0404-T5-63, as an experimental group and six varieties, Daewonkong, Danbaekkong, SS2-2, Iksannamulkong, Pungsannamulkong, and Taekwangkong, as a control group were used. The study found that about twice as much isoflavone contents were observed in the control group as compared to the experimental group. FC assay and DPPH assay were performed for the measurement of antioxidant effects and polyphenol contents. SS0408-T5-99 and SS0403-T5-38 had the highest values of total phenol content (11.9mg GAE/g dry weight) and antioxidant effect (182.6mg VCEA C/g dry weight). However, there was no significantly difference observed between the experimental and control groups. SS0408-T5-99 known as a normal soybean cultivar and SS0404-T5-63 known as a sprout soybean cultivar had the highest values of isoflavone content, having a threefold increase in isoflavones when they transformed into soybean production. These two varieties revealed higher production yields, 135%~150%, than other varieties tested in three different regions.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 4
• SUMMARY ... 8
• 목차 ... 11
• 제1장 서 론 ... 12
• 제1절 연구개발의 목적 ... 12
• 제2절 연구개발의 필요성 ... 12
• 제2장 국내외 기술개발 현황 ... 16
• 제1절 국내외 연구 현황 ... 16
• 제2절 국내외 연구현황 비교 및 필요 연구 분야 ... 19
• 제3장 연구개발 수행 내용 및 결과 ... 21
• <제1세부과제 : 맞춤형 콩 품종 개발을 위한 유전체기반의 분자육종플랫폼 구축> ... 21
• <제1협동과제 : 콩 전사체 발현 분석을 통한 유용 유전자 탐색> ... 57
• <제2협동과제 : 유전체 정보를 이용한 다수성 양질 고기능성 콩 품종 육성> ... 107
• <제3협동과제 : 다수성 양질 콩의 기능성 규명 및 콩 가공 식품 개발> ... 138
• 제4장 연구개발 목표 달성도 및 대외 기여도 ... 185
• 제1절 목표대비 대외 달성도 ... 185
• 제2절 정량적 성과 ... 189
• 제3절 대외기여도 ... 191
• 제5장 연구개발 결과의 활용계획 ... 192
• 제6장 연구개발 과정에서 수집한 해외과학기술정보 ... 194
• 제7장 기타 중요 변동사항 ... 199
• 제8장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 200
• 제9장 참고문헌 ... 201
• 끝페이지 ... 211