초록 ▼

본 연구는 고랭지 환경에 적합한 고품질의 고랭지양파를 육성하기 위하여 2004년 5월 부터 2007년 5월까지 3년간 고령지농업연구소와 건국대학교가 공동으로 다음과 같이 수행하였다.
1. 고랭지양파 우량 웅성불임계통 육성
고랭지 환경에 적응하는 우수한 웅성불임계통을 육성하기 위하여 웅성불임 계통을 육성하기 위하여 B/C 조합의 후대에서 임성을 확인하여 웅성불임 계통을 육성하고, B/B 및 B/C의 교배조합을 통하여 새로운 웅성불임 계통을 육성하였다. 웅성불임 유지친으로 확인된 것은 A line 과 교배를 통하여 A, B

본 연구는 고랭지 환경에 적합한 고품질의 고랭지양파를 육성하기 위하여 2004년 5월 부터 2007년 5월까지 3년간 고령지농업연구소와 건국대학교가 공동으로 다음과 같이 수행하였다.
1. 고랭지양파 우량 웅성불임계통 육성
고랭지 환경에 적응하는 우수한 웅성불임계통을 육성하기 위하여 웅성불임 계통을 육성하기 위하여 B/C 조합의 후대에서 임성을 확인하여 웅성불임 계통을 육성하고, B/B 및 B/C의 교배조합을 통하여 새로운 웅성불임 계통을 육성하였다. 웅성불임 유지친으로 확인된 것은 A line 과 교배를 통하여 A, B line을 육성하였다. 다양한 웅성불임친을 육성하기 위하여 표지인자를 통하여 세포질이 N으로 확인된 개체 및 계통에 대하여 우수한 품질친을 교배하여(B/C) 및 B/B를 통하여 새로운 웅성불임 계통을 육성하고자 시도하였다. 이와 같은 방법으로 숙기 및 구형 등 다른 여러 가지 계통의 웅성불임 유지친 계통을 육성하였고 금후 세대진전을 통하여 고정한 다음 $F_1$ 품종육성의 재료로 사용할 것이다.
2. F1 조합능력 성능검정
고랭지양파 F1 품종을 육성하기 위하여 만추황 및 도입된 웅성불임 계통을 이용하였고, 품질친으로 육성된 다양한 계통을 가지고 F1 성능검정을 실시하였다. 1년차 성능검정에서 우수한 조합은 2차 년도에 생산력 검정을 실시하였고, 생산력검정에서 도 우수한 조합은 지역적응성을 검정하였다. 3년간 약 150조합을 작성하여 성능검정을 실시하였다 성능검정은 숙기, 내병성, 수량 및 품질친등을 조사하였다.
3. 고랭지양파 우량계통 지역적응 시험
F1 품종을 육성하기 위하여 조합성능검정 및 생산력 검정에서 우수한 조합은 다양한 고랭지 환경에 적응성을 알아보고자 실시하였다. 1년차에 6조합, 2년차에 2조합, 3년차에 5조합 등 총 13조합을 공시하여 '04년에는 4개 지역, '05~'06년에는 3개지역에서 실시하였다. 그 지역 및 해발을 보면 대관령(해발 800m), 진부(600m), 인제(500m), 홍천 내면(500m)이다. 이 중 인제는 2004년에만 실시하였다.
4. 생명공학기법을 이용한 웅성불임 관련 DNA 표지인자 개발 및 실용화연구
본 연구는 양파의 웅성불임을 조절하는 것으로 알려진 세포질 웅성불임 인자와 연관된 DNA 표지인자를 실제 육종에 이용하기 위한 실용화 연구, 그리고 웅성불임인자를 회복시키는 핵내 웅성불임회복 유전자와 연관된 DNA표지인자의 개발로 구성 되어 있다. 또한 DNA 표지인자를 이용해서 웅성불임친을 유지할 수 있는 유지친의 조기육성과 핵내 웅성불임회복 유전자형별 집단을 육성하고자 하였다.
먼저 세포질 웅성불임 인자와 연관된 DNA 표지인자 실용화 연구에서는 실시간 유전자 증폭기술을 이용한 high through-put 기술 개발 하고 다양한 계통의 세포질 웅성불임인자를 동시에 판별하기 위한 multiplex PCR-RFLP 및 Real-time PCR의 검출한계 설정하였다. 세포질 웅성불임인자를 이용한 유지친 육성을 위해 우수 고정계통내의 N- 세포질 선발 및 A line 이용한 임성확인용 교배조합 작성하고 후대 유전분석을 실시하였다. 둘째 핵내 웅성불임회복 유전자와 연관된 DNA표지인자의 개발하기 위해 핵내 웅성불임 유전자형 확인 및 DNA표지인자 개발 용 집단육성과 웅성불임회복 유전자 연관 SSR(simple sequence repeats) 표지인자 개발을 실시하였다.
5. 고랭지 양파의 품종 및 계통간 저장성 및 기능성 비교
고랭지 양파의 고기능성 성분(황화합물(ACSOs) 비교분석하기 위하여 황화합물의 추출 및 분석조건 확립하였으며 품종 및 계통간 황화합물의 정성 및 정량분석하였다. 저장성이 좋은 고랭지 양파의 저장성 관련 하여 양파성분 조성과 저장성과의 상관관계 구명하고 저장기간 중 생리적 특성을 연구하였다. 고랭지양파 저장성이 기능성이 우수한 계통 선발효율을 높이기 위하여 기능성 및 저장성 평가기법 개발하였으며 외관 특성 및 간단한 분석에 의해 기능성 및 저장성 평가기법 개발하였다.

Abstract ▼

1. Research Results
가. Breeding of promising male sterile lines in long-day onion
In first year, a total of 175 combination was made for identifying of maintainer line(B line) using test cross. Among them, 22 lines were identified as new B line. The selected B line was selfed for multiplicatio

1. Research Results
가. Breeding of promising male sterile lines in long-day onion
In first year, a total of 175 combination was made for identifying of maintainer line(B line) using test cross. Among them, 22 lines were identified as new B line. The selected B line was selfed for multiplication and crossed with male sterile line(A line) for introducing S cytoplasmic factor to breed new A line.
For the breeding of new B line , the 32 combination cross between B line and B line were made. 'Exhibition' and 'Sapporoki' were used for the cross between maintainer line and restore fertility line for introducing high yield to breeding lines. In second year, some promising B lines were selected in combination between B lines. The lodging date of cross between W202B and W404B-2 was July 25th and that of cross between W205B and W4166B-5 was Aug. 10th. Th bulb weight of cross between 2002-213B and W202B/W404B, W205B and 44166B-2 was 183g, 164g, respectively. The bulb color was yellowish brown and bulb shape ratio was from 0.88 to 1.14 that meant circular or oval shape. The number of growing points was below 2.0 and the distance between two growing points was below 1.4cm that could be used for single center breeding program. The number of rings was above 8.0 and total sugar contents was highest in cross between W202B and (W404B cross with 2002-213B) as ${11.3^{\circ}}$Brix.
In third year, 30 male sterile lines were selected as promising breeding lines. Especially, the cross between 44166B and hybrid flamenco had high disease resistance and higher yield, 181g and high purity among the population. Two red male sterile lines which had anthocyanin were selected and further works will be made, fertility confirmation and selection.
나. Examination of F1 hybrid combining ability
The fifty, fifty and sixty crosses were made for F1 hybrid combining ability in 2004, 2005 and 2006 year, respectively. In first year, four F1 hybrid cross lines 'Manchuhwang and Wolryun', 'W910 and Kamui', 'W205 and jp6' and 'W910 and Kamui' were selected for promising new $F_1$ hybrid lines. In second year, four F1 hybrid corss lines 'NY36-A and Wolryun', In third year, Manchuhwang and RADAR, Manchuhwang and tuskimikari, W910A and Kamui F4, W202A/Wolf F4, were selected. In third year, selected four promising F1 hybrid lines were tested for regional adaptation. In the lodging date, the cross between W404A and Swift was June 18th and crosses 'NY36-A and Topone', 'Manchuhwang and jp-4' and '44165 and Chunsim' were August 2nd. When we analyzed the bulb weight, the corss between 'Manchuhwang and jp-6' was shown in highest figure, 130.2g and bulb weight of 8 combination were higher than that of Higuma, control variety. The sugar content and dry matter(%) of 'Higuma' was 7.10 and 11.94, respectively but the higher sugar content and dry matter(%) of the cross between W910A and Kamu was shown in 9.44 and 15.69, respectively. The range of number of growing points was 1.3~2.3(ea/bulb) among the breeding lines and this range was similar to that of 'Higuma' and 'Kamu'.
The sprouting ratio of 'Higuma', 'Wolf' and 'Kamui' was 69.7, 40.0 and 13.0%, respectively. On the other hand, the storage ability of new F1 breeding lines was higher than that of control varieties, especially the sprouting ratio was 0% in the cross between W205 and jp-4.
다. Regional adaptation experiment of promising breeding lines
The six, two and five crosses were tested for regional adaptation at three highland areas in 2004, 2005 and 2006 year, respectively. The bulbing time was earliest in 'Inje' area and latest in 'Dargwallyeong' area. The highest average bulb weight was shown in 'Jinbu' area and the lowest average bulb weight was shown in 'Daegwallyeong' area. That was assumed that short growing period because of high altitude resulted in poor yield in 'Daegwalllyeong'. Bulb weight of hybrid line, NIHA-GA-1, was 156.4g and that of 'Higuma' was 142.8g in average at 'Jinbu' area. There were same trend in 'Hongcheon' and 'Inje' area, the bulb weight of NIHA-GA-1 was 173.8g and 102.5g in 'Hongchoen' and 'Inje' area , respectively. The bulb weight of other lines were fluctuating depending on the area and year. After regional adaptation experiment in three year and three highland areas, the average yield of NIHA-GA-1 had 3,585kg/10a and that of 'Higuma' was 3,457kg/10a, respectively. The sprouting ratio of NIHA-GA-1 was 37.8% and that of 'Higuma' was 49.8%. In conclusion, NIHA-GA-1 was applied for new cultivar enrollment as 'Daegwanhwang'. In the future, another promising $F_1$ hybrid lines will be selected through regional adaptation experiment and be applied for new F1 hybrid cultivars.
라. Application of DNA markers of CMS factors in cytoplasm and to develop of DNA markers linked to fertility restore gene in nucleus
1) Application of DNA markers linked to CMS factors
We have developed a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) marker that can identify male-fertile (N) and male-sterile (S) cytoplasm in onions. The PCR-RFLP marker is retained in a chloroplast gene psbA amplicon. Digesting the amplicons from different cytoplasm-containing varieties with the MspI restriction enzymes hows differences at this recognition sequence (CCGG). Although the N-cytoplasm has one viable MspI site, the S-cytoplasm plants retain a point mutation(CTGG), and thus no MspI site. Moreover, this marker can discriminate between N- and S-cytoplasms in the same DNA sample mixture at a ratio of up to 10-fold, indicating its high diagnostic precision. Furthermore, we assessed the validity of the SNP detected in psbA gene for high-through put discrimination of CMS factors using Real-time PCR and a TaqMan probe assay. For the identification of CMS type in onion, two different DNA markers, cob and orf501A, have been used for 60 breeding lines. Among the these breeding lines, 7 lines were shown in CMS-T.
2) Development of DNA markers linked to restore fertility gene
For the making population of restore fertility, we selected N-cytoplam plants from "Manchuhwang(open-pollinated cultivar)" which was bred by NIHA using SCAR marker. When CMS-factor of 150 plants from 'Manchuhwang' was analyzed, 124 plants had N-cytoplasm. N-cytoplasm plants were crossed with male sterile inbred line(W202A, bred by Wisconsin Univ.). Total 66 crosses were accomplished, but 34 crosses could be analyzed for the nuclear restore allele. Among 34 combination, the offsprings of one combination showed all male sterility and this lines thought be the maintainer line in the 'Manchuhwang'. This maintainer line was just available with materials for $F_1$ hybrid seed production without fixing of characters. To develop DNA markers linked to fertility restore gene in onion, we investigated DNA polymorphism between two different DNA bulks that is dominant and recessive plants in 'Manchuhwang', respectively using onion SSR(simple sequence repeats) based on GeneBank (Http://www.ncbi.nlm.nih.gov). Among the SSR in onion, ACM103 which has TCC motif in genome was shown in polymorphism in those two DNA bulks. Dominant and recessive fertility restore allele plants has 150bp and 180bp PCR products, respectively PCR products were sequenced and analyzed multiple sequence alignment using Vector NTI(ver. 10). As a results, there is three major tandem repeats(TCC) missing in dominant fertility restore line(RfRf). This SSR marker linked to fertility restore gene in onion can be used for direct selection of male sterile line with DNA marker related to the cytoplasmic male sterile factors.
마. Comparison of storage ability and functionality among the long-day onion
When we analyzed the sulfide compounds, especially ACSOs(S-alk(en)yl-L-cysteine sulfoxides) compounds, in long-day onion, MCSO (methyl-cysteine sulfoxide) concentration of Wolf-$F_3$ line was highest figure, 0.83mg/L and the highest concentration of PrenCSO(1-propenyl-cysteine sulfoxide) was shown in JP-22 as 1.62mg/g. The total contents of ACSOs(S-alk(en)yl-L-cysteine sulfoxides) compounds of onion cultivated in different area was analyzed. When 'Higuma' cultivar was grown in
'Daegwallyeong' area, the highest contents of ACSOs was appeared and vise versa results in 'Hongcheon'. But the highest contents of ACSOs was shown in 'Hongcheon' area in 'Manchuhwang/Wolf' breeding line.
The onion extracts that was extracted by MCW(methanol:Chloroform:Water) solution had low anti-oxidant ability. When $IC_{50}$ was investigated among the breeding lines, the highest of $IC_{50}$ was shown in 'Manchuhwang/JP_selfing_4' and HDH-3, and the lowest of IC50 was shown in 'Red Beauty'. In soluble solid contents(SSC), 'Kamui', 'Amposta', 'HDH-3' and 'Manchuhwang' had slightly higher contents than that of other breeding lines. When we compared the concentration of pyruvic acid in onion bulb, breeding lines such as '5226', '5690' and '5241' were higher than that of other breeding lines and '5564' and '5606' were lowest, 2.80mM/gFW. The correlation between the concentration of pyruvic acid and ACSOs, and the concentration of ACSOs and the ability of anti-oxidant was not significant in onion bulb. But when the correlation between SSC and rot ratio was shown, r=0.2802.

목차 Contents

• 표지...1
• 제 출 문...3
• 요 약 문...5
• SUMMARY...13
• 목 차...20
• 제 1 장 연구개발과제의 개요...22
• 제1절 연구개발의 목적...22
• 제2절 연구개발 필요성...23
• 제3절 연구범위...24
• 제 2 장 국내외 기술개발 현황...26
• 제1절 국내기술개발 현황...26
• 제2절 국외기술개발 현황...27
• 제 3 장 연구개발수행 내용 및 결과...28
• 제1절 고랭지양파 우량 웅성불임계통 육성...28
• 1.서 설...28
• 2.재료 및 방법...28
• 3.결과 및 고찰...31
• 4.종합결과...41
• 제2절 F1 조합능력 성능검정...42
• 1.서 설...42
• 2.재료 및 방법...43
• 3.결과 및 고찰...44
• 4.종합결과...55
• 제3절 고랭지양파 우량계통 지역적응 시험...56
• 1.서 설...56
• 2.재료 및 방법...57
• 3.결과 및 고찰...59
• 4.종합결과...70
• 제 4절 생명공학기법을 이용한 웅성불임 관련 DNA 표지인자 개발 및 실용화연구...73
• 1.서 설...73
• 2.재료 및 방법...75
• 3.결과 및 고찰...79
• 4.종합결과...92
• 제5절 고랭지 양파의 품종 및 계통간 저장성 및 기능성 비교...93
• 1.서 설...93
• 2.재료 및 방법...96
• <시험1>양파의 황화합물 분석조건 확립 및 분획 분리...96
• <시험2>양파의 품종 및 계통간 황화합물의 정량 분석과 항산화 활성 조사...104
• <시험3>고랭지 양파의 품종 및 계통간 저장성 조사...106
• 3.결과 및 고찰...108
• <시험1>양파의 황화합물 분석조건 확립 및 분획 분리...108
• <시험2>양파의 품종 및 계통간 황화합물의 정량 분석과 항산화 활성 조사...112
• <시험3>고랭지 양파의 품종 및 계통간 저장성 조사...117
• 4.종합결과...122
• 제 4 장 목표달성도 및 관련분야에의 기여도...124
• 제 5 장 연구개발결과의 활용계획...127
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보...129
• 제 7장 참고문헌...130