초록 ▼

▶ 연구목적 :
○ 두과작물(녹두, 팥, 동부, 강낭콩 등)유전자원 자원선발, DNA 분석 및 유용유전자 탐색 활용기반구축
○ 두과작물(녹두, 팥, 동부, 강낭콩 등)의 자원별로 Phenol계 및 Flavonoid계 고 기능
성 성분에 대한 프로파일을 구축 및 고 성분 함유 자원 선발
○ 두과작물(녹두, 팥, 동부, 강낭콩 등) 추출물을 동물에 투여 한 후 지질대사, 당대사, 항염증, 항산화, 골대사 조절 효과 구명 및 자원산업화 기반구축
▶ 주요연구내용 :
○ 1세부과제명 : 두과작물 유용유전자 탐

▶ 연구목적 :
○ 두과작물(녹두, 팥, 동부, 강낭콩 등)유전자원 자원선발, DNA 분석 및 유용유전자 탐색 활용기반구축
○ 두과작물(녹두, 팥, 동부, 강낭콩 등)의 자원별로 Phenol계 및 Flavonoid계 고 기능
성 성분에 대한 프로파일을 구축 및 고 성분 함유 자원 선발
○ 두과작물(녹두, 팥, 동부, 강낭콩 등) 추출물을 동물에 투여 한 후 지질대사, 당대사, 항염증, 항산화, 골대사 조절 효과 구명 및 자원산업화 기반구축
▶ 주요연구내용 :
○ 1세부과제명 : 두과작물 유용유전자 탐색을 위한 SNP마커개발 및 DNA프로파일 대량구축
- 녹두 핵심자원 선발 및 SNP 정보 확보
‣ 두과작물 유용유전자 탐색을 위한 DNA 마커개발 및 DNA프로파일 대량구축
‣ 녹두 유전자원 증식 및 기초특성 평가, 유전적 다양성 분석, 특성평가 DB 구축
- 녹두의 DNA마커개발 및 Mapping 집단의 유전자형 분석
‣ 두과작물 유용유전자 탐색을 위한 DNA 마커개발 및 DNA프로파일 대량구축
- 팥의 핵심자원 선발, DNA마커개발 및 DNA프로파일
‣ 팥의 작물학적 기초특성평가, 분자마커를 이용한 다양성분석 및 선발
- 강낭콩의 핵심자원 선발, DNA마커개발 및 DNA프로파일
‣ 두과작물 유용유전자 탐색을 위한 DNA 마커개발 및 DNA프로파일 대량구축
‣ 강낭콩의 작물학적 기초특성평가, 분자마커를 이용한 다양성분석 및 선발
○ 1협동과제명 : 소면적 두류작물의 특성평가 및 유전적 다양성 분석
- 녹두의 작물학적 기초 특성 평가와 분자마커를 이용한 다양성 분석및 자원 선발
‣ 보존 두류유전자원 현황분석.
‣ 녹두 유전자원 기초특성 평가 검토 및 자원선발
- 팥의 작물학적 기초특성 평가와 분자마커를 이용한 다양성 분석 및 자원선발
‣ 자원증식 및 기초특성 조사
‣ 팥 자원프로파일링 및 데이터 입력
- 강낭콩의 작물학적 기초특성 평가와 분자마커를 이용한 다양성 분석및 자원 선발
‣ 강낭콩자원증식 및 기초특성 조사
‣ 자원선발 및 데이터 입력
○ 2협동과제명 : 두과작물 유전자원의 기능성물질 분석, chemical profile 및 산업화 가능 자원선발
- 녹두 유전자원의 기능성 물질 분석
‣ 녹두 유전자원에서 기능성 물질 분석 및 chemical profile작성 및 자원선발
- 팥 유전자원의 기능성 성분 분석
‣ 팥 유전자원에서 기능성 물질 분석 및 chemical profile작성 및 자원선발
- 강낭콩 유전자원의 기능성 성분 분석
‣ 강낭콩에서 기능성 물질 분석 및 Chemical profile작성 및 자원선발
○ 3협동과제명 : 두과작물의 항염증, 항산화, 지질 및 당대사, 골대사조절 능력 평가를 통한 기능성 성분의 작용기전 규명
- 골다공증 동물 실험을 위한 식이제조 및 동물모델 제작
‣ 난소 절제술을 통해 골다공증 동물 모델 제작 및 식이 제조
- 골다공증 동물모델에서 두과작물 (대두, 녹두, 동부, 팥)의 섭취가 골대사에 미치는 영향 연구
‣ 난소 절제술을 통해 폐경 후 골다공증 동물모델을 확립하여 각종 두과작물 (대두, 녹두, 동부, 팥)을 섭취시킨 후 골대사 관련지표 측정을 통하여 두과작물이 폐경 후 골다공증 예방에 미치는 영향 및 작용기전을 살펴봄
- 골다공증 동물모델에서 두과작물 (대두, 녹두, 동부, 팥)의 섭취가 지질 대사 및 항산화상태에 미치는 영향 연구
‣ 난소 절제술을 통해 골다공증 연구를 위한 동물모델 제작 후 AIN-93M 식이를 기본으로 녹두, 팥, 동부, 강낭콩이 함유된 식이와 대조식이를 제조하여 5주간 섭취시킨 후 혈중 지질과 골밀도와의 상관성을 살펴봤으며, 항염 인자 발현을 측정하여 두과작물이 골다공증 대사 및 항염 효과에 미치는 영향을 살펴봄.

Abstract ▼

1. Legume genetic resources conservation and characterization
Legume PGR conserved in NAC has been characterized about 10-18 of morphological and basic agronomic traits on mung bean, adzuki bean and kidney bean.
2. M ungbean
In growth habitat, indeterminate type in mung bean occupied at 60%

1. Legume genetic resources conservation and characterization
Legume PGR conserved in NAC has been characterized about 10-18 of morphological and basic agronomic traits on mung bean, adzuki bean and kidney bean.
2. M ungbean
In growth habitat, indeterminate type in mung bean occupied at 60% and days from planting to maturity of mung bean were ranged from 57 to 118 days with an average of 78 days.
a. Genetic marker development
Mungbean (Vigna radiata (L.) Wilczek) is a unique species in its ability to fix atmospheric nitrogen, with early maturity, and relatively good drought resistance. We used 454 sequencing technology for transcriptome sequencing. A total of 150 159 and 142,993 reads produced 5 254 and 6 374 large contigs (≥500 bp) with an average length of 833 and 853 for Sunhwa and Jangan, respectively. Functional annotation to known sequences yielded 41.34% and 41.74% unigenes for Jangan and Sunhwa. A higher number of simple sequence repeat (SSR) motifs was identified in Jangan (1 630) compared with that of Sunhwa (1 334). A similar SSR distribution pattern was observed in both varieties. A total of 8 249 single nucleotide polymorphisms (SNPs) and indels with 2 098 high-confidence candidates were identified in the two mungbean varieties.
The average distance between individual SNPs was ∼860 bp. Our report demonstrates the utility of transcriptomic data for implementing a functional annotation and development of genetic markers.
b. Coreset for genetic diversity analysis development
With the rapid increase in the number of germplasm collections, many genebanks face problems of redundant resources. To cope with this problem, the development of an allele-mining set is especially important. Our study support the efficiency of PowerCore in the development of coreset from 705 collected accessions using AFLP markers. The result demonstrated that the higher allele (fragment) capturing efficiency of PowerCore than other strategies (Distance Based, Stratified Random, and Random) tested at any level of sample size (5%, 10%, 15%, 20% of total accessions) selected. Highly significant correlation (r=0.96) was observed between allele frequency distribution of entire collections and that of core set developed by PowerCore.
c. Genetic diversity analysis
Fifteen simple sequence repeat (SSR) markers were used to analyze the population structure of 55 mungbean accessions (34 from South Asia, 20 from West Asia, 1 sample from East Asia). A total of 56 alleles were detected, with an average of 3.73 per locus.
The mean of major allele frequency, expected heterozygosity and polymorphic information content for 15 SSR loci were 0.72, 0.07 and 0.33 respectively. The mean of genetic diversity and polymorphic information content were almost similar for South Asian and West Asian accessions (genetic diversity 0.35 and polymorphic information content 0.29).
Model-based structure analysis revealed the presence of three clusters based on genetic distance. 47 accessions (85.56%) showed membership with the clusters and 8 accessions (14.54%) were categorized as admixture.
The genetic diversity and population structure of 66 and 65 mungbean accessions collected from different part of the world were also analyzed using 15 simple sequence repeat (SSR) markers. All of the results could be useful for future germplasm preservation, conservation and miprovements such as marker assisted selection (MAS) based breeding programs of mungbean.
d. Chemical compound analysis
In the analysis of chemical compound, the average total concentrations of phenolic compound of mungbeans in China (2148.3 ㎍ g^-1), India (2602.5 ㎍ g^-1), Iran (2196.5 ㎍ g^-1), Japan (2506.9 ㎍ g^-1), Korea (2476.1 ㎍ g^-1), Nepal (2374.2 ㎍ g^-1), Pakistan (2425.0 ㎍ g^-1), Philippines (2520.9 ㎍ g^-1), Taiwan (2232.9 ㎍ g^-1), Thailand (2224.7 ㎍ g^-1) and Uzbekistan (2089.1 ㎍ g^-1) showed difference by their origins. In the comparison of two phenolic compound group, flavonoid group in all countries showed higher level than that of phenoilc acids and others. The average percentage of the flavonoids group about 80% and the phenolic acids group composed about 14% in total phenolic compounds concentration. The average concentration of total tocopherols were 111.5 ㎍ g^-1 in China, 109.4 ㎍ g^-1 in India, 115.2 ㎍ g^-1 in Iran, 100.4 ㎍ g^-1 in Japan, 102.8 ㎍ g^-1 in Korean mung bean, 107.7 ㎍ g^-1 in Nepal, 108.9 ㎍ g^-1 in Pakistan, 107.6 ㎍ g^-1 in Philippines, 114.7 ㎍ g^-1 in Taiwan, 107.9 ㎍ g^-1 in Thailand and 101.9 ㎍ g^-1 in Uzbekistan. And the average concentration of γ -tocopherol showed differences between the eleven origins (76.4 ㎍ g^-1 in China, 74.4 ㎍ g^-1 in India, 78.0 ㎍ g^-1 in Iran, 67.2 ㎍ g^-1 in Japan, 68.6 ㎍ g^-1 in Korean mung bean, 72.8 ㎍ g^-1 in Nepal, 73.4 ㎍ g^-1 in Pakistan, 72.7 ㎍ g^-1 in Philippines, 79.4 ㎍ g^-1 in Taiwan, 73.3 ㎍ g^-1 in Thailand and 66.9 ㎍ g^-1 in Uzbekistan).
3. Red bean (A dzuki bean)
In growth habitat, the ratio of determiate and indeterminate type in adzuki bean was
almost similar. Days from planting to maturity of adzuki bean were ranged from 83(K153498) to 118 (K043808, K018714) days, with an average of 101.2 days.
a. Genetic diversity analysis Red bean, also known as azuki bean [Vignaangularis (Willd.) Ohwi & Ohashi], belongs to a group of legumes (family Fabaceae). In total, 178 red bean accessions were taken to analyze the genetic diversity, population structure, and gene flow using 36 polymorphic SSR markers selected from (Hanetal.,2005). In total, 431 alleles were detected, with an average of 11 alleles per locus among 178 tested red bean accessions. Forty-six specific alleles were identified with 20 loci. Locus CEDG090 had the highest number (22) of alleles, whereas only two alleles were observed at loci CEDG144 and CEDC018. The proportion of different alleles for microsatellite loci was analyzed using a microsatellite toolkit. In locus CEDG029, one allele is shared in all three groups of varieties and species, and three alleles are shared between the wild ancestors and cultivated varieties, while in locus CEDG090, one allele is shared in all three groups and 12 alleles are shared between the wild ancestors and cultivated varieties. Our findings will help explain the genetic relationships and population structure of the red bean in Korea and will be useful for designing effective breeding programs and broadening the genetic base of commercial varieties.
b. Chemical compound analysis
One hundred and fifty adzuki bean samples were collected from gene bank of RDA (Rural Development Administration, Suwon, Korea) and they were divided into various types by their origins and seed size. Between two origins, the average total phenolic compound concentrations of Japanese (2432.4 ㎍ g^-1) adzuki beans shown higher than Korean (2255.5㎍ g^-1). The mean of the total phenolic compounds in small size adzuki bean in Japan was 2506.6 ㎍ g^-1 and 2458.8 ㎍ g^-1 in Korean adzuki bean seeds. In the large size of adzuki beans, the mean of total phenolic compounds was 1315.1 ㎍ g^-1 in Japanese adzuki bean seeds and 1231.9 ㎍ g-1 in Korean adzuki bean germplasm. The average total phenolic compounds in medium seeds revealed 2368.6 ㎍ g^-1 in Japan adzuki beans and 1397.0 ㎍ g^-1 in Korean germplasm. In the small seed, the total phenolic compound concentrations of adzuki bean varied from 523.8 ㎍ g^-1 to 6596.9 ㎍ g^-1 in Japanese and from 374.5 ㎍ g^-1 to 6569.2 ㎍ g^-1 in Korean. Among the four tocopherols, δ-tocopherol (75.8 ㎍ g^-1) had the highest levels while, β-tocopherol (29.7 ㎍ g^-1) showed the lowest levels in whole adzuki bean varieties and the mean of total tocopherols was 192.9 ㎍ g^-1.
4. K idney bean (common bean)
In growth habitat, determinate type in kidnety bean was about 66%. Days from planting to maturity of kidney bean were ranged from 83 to 123 days, with an average of 98.8 days. Days from planting to maturity of kidney beans originated from Korea with an average 92.1 days were 6.7days earlier than that from Bulgaria with an average 98.8days.
a. Genetic diversity analysis
In total, 96 common bean accessions from Korea, Bulgaria, Ukraine, Russia, Mexico;
TJ, Rajikistan, Uzbekistan were take to analyze the genetic diversity, population structure, and gene flow using 26 polumorphic SSR markers selecter from (Yu et al.
2000; Guerra‐ Sanz, J. 2004; M., F. Pedraza, et al. 2003). In total, 108 alleles were detected, with an average of 4.2 alleles per locus among 96 tested common bean accessions. Seventeen specific alleles were identified with 7 loci. Locus BMD-31 had the highest number (14) of alleles, whereas only two alleles were observed at locus AZ301561.1, Bng01-R2, BMd-32, BMd-19, AF402604.1, BH126808.1, BMb469, AY007525.1, BMb519, BMb611, BMd-7, BMb266 . The average gene diversity and PIC values ranged from a low of 0.0799 (BMb519) and a high of 0.8368 (BMd-31), and ranged from a minimum of 0.0587 (BH126808.1) to maximum of 0.819 (BMd-31), respectively. A model-based structure analysis revealed the presence of five populations. However, 31 accessions (32.3%) in the sample were categorized as having admixed ancestry. These results could be used for broadening the genetic base of commercially grown varieties.
b. Chemical compound analysis
In 139 kidney beans, homogentisic acid was not detected among 26 phenolic compounds (1. Gallic acid, 2. Pyrogallol, 3. Homogentisic acid, 4. Protocatechuic acid, 5. Gentisic acid, 6. β-Resorculic acid, 7. Chlorogenic acid, 8. Vanillic acid, 9. Caffeic acid, 10. Syringic acid, 11. Vanillin, 12. p-Coumaric acid, 13. Salicylic acid, 14. Ferulic acid, 15. Veratric acid, 16. m-Coumaric acid, 17. Rutin, 18. o-Coumaric acid, 19. Hesperedin, 20. Myricetin, 21. Resveratrol, 22. Quercetin, 23. Naringenin, 24. Kaempferol, 25. Formononetin, 26. Biochanin A). 106 kidney bean was the highest concentrations(2048.85μg/g) of phenolic compounds in all kidney bean germplasms. On the other hand, 139 kidney bean(203.47μg/g) had the lowest concentration of phenolic compounds. Specially, 40 kidney beans(1296.06μg/g) revealed the highest concentrations of anthocyanins.
5. Effect of legumes on the lipid profiles, antioxidant statuses, and inflammation statuses and bone mineral density and bone biomarkers in the osteoporotic rat model
a. Effects on the lipid profiles, antioxidant statuses, and inflammation statuses
Forty-week-old male Sprague-Dawley rats (n=12) were fed a high-fat diet (45 kcal% energy as fat) for eight weeks to induce hyperlipidemia and were then randomly divided into two groups (n=6 each) that were fed diets containing white rice (WR, control) or adzuki beans (AB) for the next five weeks. Blood lipid profiles, hepatic antioxidant parameters, and inflammation-related parameters were measured in all of the groups. The final body weights, liver weights, epididymal, mesenteric, and retrotoneal fat weights were significantly decreased in AB group than WR group. The concentrations of serum triglycerides, total and HDL-cholesterol were significantly lower in the AB groupcompared to those of the WR group. Serum insulin level was lower in AB group than WR group. Hepatic catalase, SOD, and GSH activities, as well as TBARS levels were not significantly different between groups. Levels of TNF-α, IL-6, and CRP, were not significantly different between groups. IkB-α was expressed in the liver cytosolic fraction, and nuclear translocation of NF-$_{\kappa}B$ (p65) into the liver nucleus was blocked in all groups. In conclusion, adzuki beans may prevent CVD by reducing body weight, body fat, plasma triglycerides, and total cholesterol concentrations in hyperlipidemic rats.
b. Osteoporotic rat model and effect of legumes on bone mineral density and bone biomarkers
Ten weeks old female Sprague-Dawley rats were either sham-operated (Sham, n=9) or surgically ovariectomized. After recovery from the surgery, the rats were fed a AIN-93M diet (OVX, n-9) or AIN-93M containing soybean (OVX-S, n-=9), mung bean (OVX-M, n=9), cowpea (OVX-C, n=9), or adzuki beans (OVX-A, n=9) for 12 weeks. BMDs of the right femur, tibia and lumbar spine were measured at the end of the dietary experiment using dual-energy X-ray absorptiometry (DEXA). The concentrations of serum or urinary calcium, phosphate, alkaline phosphatase (ALP), osteocalcin, and deoxypridinoline (DPD) were measured. The total amounts of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), osteoprotegerin (OPG), and receptor activator of nuclear factor κB ligand (RANKL) protein from tibia and femur of the rats were assessed by western blot analysis.
The OVX and all bean groups had a significantly higher final body weight, and subcutaneous fat weight, had a significantly lower uterus weight than in the Sham group, but nor significantly different among the OVX and all bean groups Serum calcium was significantly lower in the OVX and all bean groups compared to theSham group. Serum phosphorus in the OVX -M group was significantly lower than in the Sham group. Serum A LP in the OVX group was significantly higher than Sham, OVX -S, and OVX -M groups. Serum osteocalcin was significantly higher in the OVX -S, OVX -M, and OVX -A groups than in the Sham and OVX groups. Urinary calcium was no significant differences between the Sham group and OVX group, but the OVX group was significantly higher than OVX -A group. Urinary DPD excretion was significantly higher in the OVX group compared to the Sham group and not significantly different among the OVX and all bean groups. The BMD of the femur in the OVX group was significantly lower than in the Sham, OVX -S, and OVX -M groups. The Sham and all bean groups had a significantly lower the expressions of TNF-α and IL-6, had a significantly higher the expressions of OPG/RANKL ratio than in the OVX group. However, the expression of IL-1β did not significantly differ among the OVX and all bean groups.
c. Consumption of legumes improves certain bone markers
Female Sprague-Dawley rats were either sham operated (sham; n = 7) or surgically
ovariectomized and then fed a regular AIN-93M diet (OVX; n = 7) or AIN-93M containing soybean (n = 7), mung bean (n = 7), cowpea (n = 7), or adzuki beans (n = 7) for 10 weeks. No bean consumption significantly altered the body, subcutaneous fat, or uterus weight; however, consumption significantly increased the serum calcium/phosphorous ratio and decreased urinary calcium excretion compared with those of the OVX group. Serum concentration of 17β-estradiol was significantly lower in the OVX group compared with that of the sham group and was lowest in the group fed OVX diet containing soybean. Serum osteocalcin concentration was significantly higher in all OVX rats given a diet with beans compared with the same diet without, but urinary deoxypyridinoline excretion was lowest in the group fed OVX diet containing cowpea.
There were no significant differences in bone mineral density or bone mineral content of the right femur, tibia, or lumbar spine or in the trabecular bone volume of the tibia among the diet groups. In conclusion, the consumption of soybean, mung bean, cowpea, and adzuki bean in OVX rats improved osteocalcin, but only those fed cowpea showed decreased bone resorption biomarker, suggesting that cowpea may have the most protective effect on bone in OVX rats.
e. Effect of Dietary Legumes on Bone-Specific Gene Expression
Forty-two Sprague-Dawley rats were divided into six groups during the 12 week study: rats that underwent sham operations (Sham), ovariectomized rats fed an AIN-93M diet (OVX), and ovariectomized rats fed an AIN-93M diet with soybeans (OVX-S), with mung beans (OVX-M), with cowpeas (OVX-C) and with azuki beans (OVX-A).
Consumption of legumes significantly increased BMD of the spine and femur and bone volume of the femur compared to the OVX. Serum calcium and phosphate ratio, osteocalcin, expression of osteoprotegerin (OPG), and the receptor activator of nuclear factor κB ligand (RANKL) ratio increased significantly, while urinary excretion of calcium and deoxypyridinoline and expression of TNF-α and IL-6 were significantly reduced in OVX rats fed legumes, compared to OVX rats that were not fed legumes.
f. Supplementation of Korean fermented soy paste doenjang reduces visceral fat in overweight subjects with mutant uncoupling protein-1 allele The antiobesity effect of doenjang, a Korean fermented soy paste is different between the mutant and the wild-type alleles of a polymorphism upstream of the uncoupling protein-1 (UCP-1) gene in overweight subjects. In our randomized, double-blind, placebo-controlled trial, a total of 51 subjects with a body mass index of 23 kg/m(2) or greater and a waist-to-hip ratio of 0.90 or greater for men or 0.85 or greater for women were randomly assigned to take 9.9 g/d of either a placebo or doenjang for 12 weeks.
The relative frequency of the mutant G allele of the UCP-1 polymorphism was 0.60 in the placebo group and 0.62 in the doenjang group. Supplementation of doenjang had no significant effect on the visceral fat area compared with that of the placebo group, but there was a significantly reduced amount of visceral fat in subjects with the G allele of UCP-1 polymorphism. Doenjang supplementation was found to significantly increase the free fatty acid concentration in subjects with both the A allele and the G allele. There was a significant association between visceral fat and age in study subjects with both the wild-type and mutant alleles of the UCP-1 gene. Doenjang supplementation significantly reduced visceral fat and increased the free fatty acid concentrations in subjects with the G allele of the UCP-1 polymorphism, which suggests that doenjang may be related to increased free fatty acid levels caused by elevated lipolysis in these subjects.

목차 Contents

• 제출문 ... 1
• 요약문 ... 2
• SUMMARY ... 10
• 제1장 서론 ... 17
• 1. 연구개발의 목적 ... 17
• 2. 연구개발 대상의 기술의 경제적.산업적 중요성 및 필요성 ... 17
• 3. 연구내용의 범위 ... 20
• 제2장 국내외 기술개발 현황 ... 21
• 제1절 연구개발대상 기술의 국내.외 현황 ... 21
• 제2절. 녹두 ... 27
• 제3절. 팥 ... 77
• 제4절. 강낭콩 ... 100
• 제5절. 두과작물의 항염증, 항산화, 지질 및 당대사, 골대사 조절 능력 평가를 통한 기능성 성분의 작용기전 규명 ... 115
• 제4장 연구개발목표 달성도 및 대외기여도 ... 124
• 제1절 목표대비 달성도 ... 124
• 제2절 정량적 성과 ... 125
• 제5장 연구개발결과의 활용계획 ... 129
• 제6장 중요 변동사항 ... 129
• 제7장 참고문헌 ... 129
• 주요 결과 요약서 ... 138