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국내 밀 품종의 오메가5-글리아딘 () 유전자좌 분석

Allelic Analysis of Omega-5 Gliadin (Gli-B1) in Korean Wheat Cultivars

You-Ran Jang1,†, Hye-Rang Beom1,†, Jung-Bong Kim1, Chul-Soo Park2, Yeong-Tae Kim3, Chon-Sik Kang4, Sun-Hyung Lim1, Young-Mi Kim1, Jong-Yeol Lee1,*
The Korean Journal of Breeding Science 2016;48(2):159-167.
Published online: May 31, 2016

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*Corresponding author (jy0820@korea.kr, + 82-63-238-4616, + 82-63-238-4604))

Author Contributions: Y. R. Jang and H.R. Beom contributed eq ually.

• Received: April 8, 2016   • Accepted: April 27, 2016

© The Korean Society of Breeding Science

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  • The ω5-gliadins are the major allergens in wheat-dependent excise-induced anaphylaxis (WDEIA). In this study, SDS-PAGE analysis was used to assign the ω5-gliadins (Gli-B1) alleles in thirty two Korean wheat cultivars, compared with eleven standard wheat cultivars for Gli-B1a~m alleles. These results were reconfirmed with their complementary Glu-B3 low-molecular-weight glutenin subunits alleles tightly linked with Gli-B1 locus revealed with 2-DGE in our previous study. As a result, one Gli-B1b, four Gli-B1d, two Gli-B1f, six Gli-B1m and nineteen Gli-B1h varieties were identified. This is the first report on revealing the Gli-B1 alleles in Korean wheat cultivars and represents valuable basic data on wheat allergy, relationship between gliadin and wheat quality, and development of hypo-allergenic wheat.
밀은 종피가 13-17%, 배가 2-3% 그리고 배유가 81-84%를 차지한다. 주요한 배유의 구성은 전분이 60-75%, 단백질이 6-20%, 수분이 약 10% 그리고 지질이 1.5-2%로 구성된다 (Barak et al. 2015). 배유 저장 단백질 중 글루텐으로 인해 밀은 빵, 라면, 과자와 같은 가공적성을 가진다. 글루텐 단백질 은 알코올 수용액 (aqueous alcohol) 용해도에 따라 녹는 글리 아딘 (gliadin)과 녹지 않는 글루테닌 (glutenin)으로 나뉜다 (Wieser 2007). 글루테닌은 분자량에 따라 고분자 및 저분자 글루테닌 서브유닛 (high/low molecular weight subunits; HMW/LMW-GS)으로 나누어 지고, 각각의 분자량은 70-90KDa, 20-45KDa 사이에 존재하며, 이들간의 이황화 결합 으로 글루텐 거대 중합체 (gluten macro polymer)를 형성한다 (Wasik & Bushuk 1975, Payne 1987). HMW-GS는 1번 염색체 1A, 1B 및 1D의 장완에 위치한 Glu-A1, Glu-B1 및 Glu-D1에 의해 암호화 된다 (Shewry et al. 1995). LMW-GS는 SDS-PAGE의 이동성 (mobility) 에 따라 B-, C- 그리고 D- 그룹 서브유닛으로 나누어지며 (D'Ovidio & Masci 2004), 1번 염색체 단완의 Glu-A3, Glu-B3 및 Glu-D3 에 의해 암호화된다. LMW-GS 유전자를 발현시키는 Glu-3 위치와 글리아딘 유전자 의 일부를 발현시키는 Gli-1 위치는 염색체상에서 매우 밀접하게 연관되어있다 (Gao et al. 2007, Pogna et al. 1990, Ruiz & Carrillo 1993).
글리아딘은 밀 글루텐의 주요 구성 성분이며, 전체 저장단백질 의 40-50% 차지한다 (Anderson et al. 1997). 대부분의 글리아딘 이 단량체로 존재하고 글리아딘은 낮은 pH의 A-PAGE (Acid-PAGE)나 SDS-PAGE의 이동성에 따라 ω5-, ω1,2-, α-/β -, γ-글리아딘 4그룹으로 나뉘며 (Wieser 2007, Banc et al. 2009) 분자량은 30-75kDa정도이고 (Fido et al. 1997) 글루텐 복합체에 점성 및 신장성을 부여한다 (Khatkar et al. 2002, Wieser 2007). γ-, ω-글리아딘은 1번 염색체의 단완에 존재하며 Gli-A1, Gli-B1 및 Gli-D1에 의해 암호화되고, α-/β-글리아딘은 6번 염색체의 단완에 존재하며 Gli-A2, Gli-B2 및 Gli-D2로 암호 화 된다 (Ferranti et al. 2007, Wrigley & Shepherd 1973, Brown & Flavell 1981). 전체 글리아딘 단백질 중 α-/β- 및 γ-글리아딘이 메이져 단백질로 구성되고, 각각 28-33% 및 23-31%를 차지한다. ω-글리아딘은 훨씬 적으며, ω-1,2글리아딘 은 4-7%, ω-5글리아딘은 3-6%를 차지한다 (Wieser & Kieffer 2001). 특히 대부분의 γ-글리아딘과 ω-글리아딘이 Gli-1에 의해 발현되며 Glu-3에 의해 발현되는 저분자 글루테닌 유전자와 염색체 상에 밀접히 연관되어 있다 (Wieser & Kieffer 2001).
ω-글리아딘은 분자량이 39,000-55,000Da 범위에 있고, 아미 노산 조성에 있어서도 다른 글리아딘과 차이가 나는데 특히 이항화 결합을 일으키는 시스테인이나 메티오닌 잔기들이 부족 하다 (S-poor prolamin). 따라서 ω-글리아딘은 이황화결합이나 밀 반죽내에서의 수소결합을 하지 않는다고 알려져 있다 (Tatham & Shewry 2000). ω-1,2 글리아딘은 N-말단 아미노산 서열이 ARE, ARQ 또는 KEL로 시작하며 PQQPFP의 반복서열 을 가지며 (Bouchez-Manhiout et al. 2010), 1번 염색체의 A 및 D 장완에 암호화 되어있다 (DuPont et al. 2004). ω5-글리아 딘은 분자량이 약 50,000 Da으로 약 40,000 Da인 ω1,2-글리아 딘에 비해 A-PAGE나 SDS-PAGE 이동성에서 위에 존재한다 (Wieser 2007). ω-5글리아딘은 N-말단 아미노산 서열이 SRL로 시작되며 (DuPont et al. 2000) FPQQQ와 QQIPQQ의 반복서열 을 가진다 (Battais et al. 2005b, Matsuo et al. 2004, Matsuo et al. 2005). 이들은 1B 염색체의 Gli-B1 위치에서 암호화되며 Glu-B3 저분자 글루테닌과 밀접히 연관되어 있으며, ω5-글리아 딘 또는 Glu-B3 저분자 글루테닌 유전자좌 분석시 이들의 결과를 상호보완적으로 사용 가능하다. 이들 ω5-글리아딘은 심각한 음 식 알러지인 밀 의존성 운동 유발성 과민증 (wheat-dependent exercise-induced anaphylaxis; WDEIA) 의 중심 항원이라고 알려져 있고 (Battais et al. 2005a, Matsuo et al. 2004, Matsuo et al. 2005, Morita et al. 2003, Palosuo et al. 1999, Palosuo et al. 2001), 심한 경우 사망에 이르게 한다. 밀이 들어간 음식을 섭취 후 물리적인 운동을 했을 때 증상이 발생하며 주로 성인들에 서 발병한다고 알려져 있다. WDEIA 환자들의 혈청 immunoglobulin E (IgE)와 결합하는 주된 항원결정기 (predominant epitope)는 ω5-글리아딘 ESTs (Matsuo et al. 2004, Denery-Papini et al. 2011)와 분리 된 유전자 (Matsuo et al. 2005)를 참고로 합성하여 확인 되었으며, 공통적인 IgE 결합 모티프는 QQX1PX2QQ(X1=I,L,F,S또는 Y 이고 X2=Q 또는 E) 으로 확인되었다. 심각한 밀 알러지인 WDEIA의 위험성 을 줄이기 위한 여러 육종적 및 생명공학적 시도가 있었다. Denery-Papini et al. (2007)은 호밀 유래의 1BL/1RS 밀 전좌 (translocation) 계통에서 IgE와의 결합이 낮은 것을 확인하였으 나, 전좌 밀 계통은 일반적으로 가공적성이 낮아 실제로 사용하지 않았다. Waga & Skoczowski (2014)는 전통적인 육종법으로 ω5-글리아딘과 ω1,2-글리아딘이 모두 결손 된 합성 밀 계통을 육성하였으며, 밀 알러지 환자의 혈청을 이용한 효소면역분석법 (Enzyme linked immunosorbent assay, ELISA)에서 보통 빵용 밀보다 30% 감소된 면역반응 실험결과를 보였다. RNAi 기술을 이용하여 ω5- 글리아딘을 저해시킨 미국 밀 품종 Butte 86 형질 전환체 두 계통을 육성하였고 (Altenbach & Allen 2011, Altenbach et al. 2014a, Altenbach et al. 2014b), 11명의 WDEIA 환자의 혈청으로 이차원전기영동을 이용한 immunoblotting을 실시하였다. 11명중 7명의 환자의 혈청에서 이들 두 계통의 ω5-글리아딘과 IgE과의 반응성이 현격히 줄어들 었으나, 낮은 수준에서 다른 글루텐 단백질과 결합하였다. 다른 세 명의 환자 혈청은 ω5-글리아딘이 아닌 HMW-GS, 알파 글리 아딘 또는 비 글루텐 단백질과 반응이 현저히 증가하였다. 국내에 서도 밀 알러지 환자가 증가하고 글루텐 프리 시장이 폭발적으로 증가함에 따라 밀 알러지 또는 셀리악 병의 주요 원인이 되는 ω5-글리아딘, ω1,2-글리아딘, 알파/베타 글리아딘 또는 감마 글리아딘 군 (群)이 결손된 알러지 저감 밀 (Hypo-allergenic wheat) 계통을 찾는 시도가 진행되고 있으며, 본 연구팀에서 일부 글루텐 단백질이 결손된 계통을 찾아 동정 중에 있다.
현재까지 고분자 및 저분자 글루테닌에 대한 유전자좌 및 이들과 밀 품질에 관련 연구는 많은 진전이 있었지만, 이들 보다 복잡하고 어려운 글리아딘은 일부 알러지 및 기본적인 물성 연구 외에는 미미한 실정이다 (Weegels et al. 1996, Metakovsky et al. 1997). 따라서 본 연구에서는 국내 밀 품종의 Gli-1 ω-5글리 아딘 (GliB-1)의 유전자좌 분석을 위해 표준품종들 (standard wheat cultivars)의 글리아딘에 대한 SDS-PAGE 패턴분석을 실 시하여 국산밀의 유전자좌를 명명하였다. 이러한 연구는 고품질 밀 육성을 위한 분자육종적 연구 및 WDEIA를 포함한 밀 알러지 연구의 기반자료로 활용될 가능성이 충분하다고 생각한다.
재료
본 연구에 사용된 재료는 농촌진흥청 국립식량과학원에서 2014년에 수확된 금강 밀 등 32품종의 기본식물 밀가루를 분양 받아 사용하였으며, Gli-B1 유전자좌 판별을 위한 표준품종들(St andard wheat cultivars)은 Liu et al. (2010)과 Lauriere et al. (2007) 그리고 일본 밀 유전자원 데이터 베이스(http://shigen.nig.ac.jp/wheat/komugi/genes/symbolClassListAction.do?geneClassificationId=318) 를 참고하여 선정하였다. Gli-B1a의 Chi nese Spring, Gli-B1b의 Gabo, Gli-B1c의 Siete Cerros 66, Gli-B1d의 Suneca, Gli-B1e의 Apexal, Gli-B1f의 Cappelle-Desprez, Gli-B1g의 Champtal, Gli-B1h의 Rudi, Gli-B1i의 Insignia, Gli-B1l의 Clement 그리고 Gli-B1m의 Norin61은 미국 농업연구청 (National Plant Germplasm System)에서 분양 받아 이용하였다 (Table 1).
Table 1.
Standard wheat cultivars for Gli-B1 allele analysis.
Table 1.
Allele(Glu-B3 locusa) Standard Cultivar
Gli-B1a (Glu-B3a) Chinese Spring
Gli-B1b (Glu-B3b) Gabo
Gli-B1c Siete Cerros 66
Gli-B1d (Glu-B3h) Suneca
Gli-B1e Apexal
Gli-B1f (Glu-B3g) Cappelle-Desprez, Glenlea
Gli-B1g (Glu-B3f) Champtal
Gli-B1h (Glu-B3d) Rudi
Gli-B1i (Glu-B3c) Insignia
Gli-B1l (Glu-B3j) Clement
Gli-B1m (Glu-B3i) Norin61

aComplementary Glu-B3 locus.

글리아딘 추출
글리아딘 추출은 Dziuba et al. (2014)를 참고하였다. 미세하 게 분쇄한 밀 100mg에 0.15M NaCl 수용액 1ml을 혼합하여 25°C 에서 2시간 동안 반응시키고 20°C 15000 x g에서 5분 동안 원심 분리 후 상등액을 제거하였다. 침전물에 70% Ethanol 수용액 1ml을 혼합하여 동일 온도로 12시간 동안 반응시킨 후에 20°C 15000 x g에서 5분 동안 원심 분리하여 상등액을 -20°C에 보관하였다.
SDS-PAGE 분석
-20°C에 보관중인 글리아딘과 sample buffer(50mM Tris-HCl pH6.8, 8% β-mercaptoethanol, 2% SDS, 20% glycerol)를 1:2의 비율로 섞어 제조하고 50ul를 12.5% SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis ) 젤에 로딩하여 70V로 13시간, 110V로 4시간 동안 전기영동을 실시하였다. 전기영동 후 commassie brilliant blue R-250 (Bio-Rad) 용액으로 젤을 4시간 염색 하였고, 탈색용 액(10% glacial acetic acid, 10% 메탄올, 80% 증류수)로 5시간 탈색시켜 형성된 단백질 밴드 패턴을 분석 하였다.
글리아딘 SDS-PAGE 분석에 의한 국내 밀 품종들의 Gli-B1 유전자좌 분석
국내 밀 32품종의 Gli-B1 (ω5-글리아딘) 유전자좌 분석을 위해 글리아딘의 SDS-PAGE를 실시하고 Gli-B1 표준품종들 의 글리아딘 SDS-PAGE 결과와 비교하여 유전자좌를 분석하 였다. 또한 Gli-B1 과 1BS 염색체 상에서 밀접하게 연관되어 있고 유전자좌 분석시 상호 보완적인 관계에 있는 본 연구팀의 Glu-B3 LMW-GS의 이차원 전기영동(two-dimensional gel electrophoresis; 2DGE) 유전자좌 분석 결과 (Lee et al (2016) Allelic analysis of low molecular weight glutenin subunits using 2-DGE in Korean wheat cultivars. Euphytica. 심사중) 를 참고 하였다 (Table 2). 유전자좌 판별을 위한 Gli-B1 표준품종 11품종은 Table 1에 표시하였다 (Liu et al. 2010, Lauriere et al. 2007). 각 글리아딘은 SDS-PAGE 이동성 (mobility)에 따라 ω-5-, ω-1,2-, γ- 그리고 α/β- 글리아딘으로 표시하였다 (Liu et al. 2010, Lauriere et al. 2007).
Table 2.
Allelic compositions of Gli-B1 in Korean wheat cultivars by gliadin SDS-PAGE. The pedigree each cultivar is indicated.
Table 2.
Cultivar Pedigree Glu-B3a Gli-B1
Sukang Suwon266/Asakaje b b
Hanbaek Shann7859/Keumkang//Guamuehill h d
Joeun Eunpa/Suwon242 h d
Jokyung Seri82/Keumkang h d
Keumkang Geuru/Kanto75//Eunpa h d
Jonong Suwon234/SW80199 g f
Sinmichal 1 Alchan//Kanto107/BaiHuo g f
Alchan Suwon210/Tapdong d h
Baekjoong Keumkang/Olgeuru d h
Chungkye Norin4/Sharbatisonora d h
Dabun Suwon234//sw76039/Suwon220/3/Keumkang d h
Eunpa Chukoku81/3/Tob-CNO// Yuksung3/Suwon185 d h
Gobun Eunpa/Tapdong//Eunpa/Shannung6521 d h
Goso Gobun/Ol d h
Jeokjoong Keumkang/Tapdong d h
Jinpoom Geuru/Genaro81 d h
Milsung Sirogane//Noring43/Sonalika d h
Namhae Ol/Calidad d h
Ol Norin72/Norin12 d h
Olgeuru Geuru/Chokwang//Nishigai143 d h
Saeol Sirogane//Norin43/Sonalika d h
Seodun Geuru/Genaro81 d h
Sinmichal Olgeuru//Kanto107/BaiHuo d h
Tapdong Chukoku81//Suwon158/Toropi d h
Uri Geuru/Ol d h
Younbaek Keumkang/Tapdong d h
Dahong Norin72/Wonkwang i m
Anbaek Sae/Geuru ad m
Dajoong SW992114-NM-131-7/Gobun ad m
Geuru Strampelli/69D-3607//Chokwang ad m
Jopoom Kanto75//OR8500494P/Bezostaya ad m
Suan Keumkang/Eunpa//Keumkang ad m

a)Glu-B3 alleles using 2DGE in this group.

Gli-B1a~m 유전자좌에 해당하는 표준품종들은 Liu et al. (2010) 과 Lauriere et al. (2007) 를 참고로 각 유전자좌를 화살표 로 표시하였다 (Fig. 1). 이 각각의 유전자좌 패턴분석결과를 참고로 국내 밀 32품종의 Gli-B1 유전자좌를 분석하였다. 이들의 Gli-B1 유전자좌 SDS-PAGE 패턴에 대해서 설명하면, Gli-B1b 는 Fig. 1과 Fig. 2의 Gabo에서 나타난 것과 같이 ω5-글리아딘 부분에 2개의 밴드가 해당되며 수강이 국내 밀 중 유일하게 해당되었다. Gli-B1d는 Fig. 1과 Fig. 3의 Suneca에서 나타난 것과 같이 3개의 밴드를 확인하였고, 한백, 조은, 조경 그리고 금강이 해당되었다. Gli-B1f는 Fig. 1과 Fig. 2의 Cappelle-Desprez나 Glenlea에 표시한 1개의 밴드를 확인하였 고, 조농과 신미찰1이 해당되었다. Gli-B1h는 Fig. 1과 Fig. 4의 Rudi에서처럼 한 개의 밴드가 나타났으며, 국산 밀 중 가장 많은 19개 품종이 해당되었다. Gli-B1m은 Fig. 1과 Fig. 5의 Norin61에서 나타난 것과 같이 한 개의 밴드이고, 다홍, 안백, 다중, 그루, 조품 그리고 수안 6품종이 해당되었다. 이들 유전자 좌 중 Gli-B1h와 Gli-B1m의 밴드 패턴은 비슷해 보이나 Gli-B1m 의 밴드가 조금 더 높음을 Fig. 1에서 알 수 있었고, 이들 유전자좌 의 최종 판정은 상호보완적인 Glu-B3 LMW-GS의 SDS-PAGE 및 2-DGE 결과를 종합적으로 고려하여 판단하였다 (아래의 내용 참조).
Fig. 1.
SDS-PAGE analysis of gliadin fractions of the standard wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to individual Gli-B1 alleles.
KJBS-48-2-159_F1.gif
Fig. 2.
SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1b and Gli-B1f.
KJBS-48-2-159_F2.gif
Fig. 3.
SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1d.
KJBS-48-2-159_F3.gif
Fig. 4.
SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1h.
KJBS-48-2-159_F4.gif
Fig. 5.
SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1m.
KJBS-48-2-159_F5.gif
국내 밀 32 품종 중 Gli-B1b 1개, Gli-B1d 4개, Gli-B1f 2개, Gli-B1m 6개 그리고 Gli-B1h는 19개의 빈도를 보였다 일반적으 로 알려진 Gli-B1a~m의 11개 유전자좌 중 5개의 낮은 다형성을 보였고, 이들 중에서도 Gli-B1h는 32개 중 19개로 59.4%의 빈도를 보였다. Gli-B1과 1B 염색체 상에서 상호보완적으로 밀접하게 연관되어 있는 Glu-B3의 저분자 글루테닌 유전자좌 분석 결과와 정확히 일치함을 알 수가 있었다 (Table 1, 2). 일반적으로 Glu-B3 저분자 글루테닌은 빵 가공적성에 많은 영향 을 미친다고 알려져 있으며, Glu-B3 j, h, b, g의 순으로 빵 품질에 영향을 미친다고 알려져 있다. 하지만 글루테닌 분획의 SDS-PAGE 분석 시 Glu-B3 저분자 글루테닌은 글리아딘 단백 질의 혼입과 다른 저분자 글루테닌 단백질과의 중첩으로 해석에 상당히 어려움을 겪는데, 상호보완적인 Gli-B1 유전자좌 분석으 로 이런 어려움을 상당 부분 해결할 수 있으며 정확도를 높일 수 있다. 특히 Glu-B3f와 g는 글루테닌 분획의 SDS-PAGE 분석 시 구분이 불가능하다고 보고되고 있는데 (Gupta et al. 1994, Branlard et al. 2003, Wrigley et al. 2006, Liu et al. 2010), Fig. 1에서 나타난 것과 같이 Gli-B1 SDS-PAGE 분석에서 Gli-B1f (Cappelle-Desprez)와 g (Champtal)처럼 쉽게 구분 된 다. 이와 유사한 예로 Glu-B3b와 g는 글루테닌 분획의 SDS-PAGE 상에서의 구별이 어려우나 글리아딘 분획의 SDS-PAGE 분석 시 Fig. 1의 Gli-B1b (Gabo)와 f (Cappelle-Desprez)로 구분이 가능하며 확실한 구별법은 pH3-11의 글루테닌 분획의 2DGE 에서 Glu-B3g는 Glu-B3b와 는 달리 pH6 근처의 고분자 글루테닌과 글리아딘 사이에 분명한 스팟을 하나 가지고 있다 (Data not shown; Lee et al (2016) Allelic analysis of low molecular weight glutenin subunits using 2-DGE in Korean wheat cultivars. Euphytica. 심사중; Liu et al. 2010). 반대의 경우로 Fig. 1의 Gli-B1h (Rudi)와 m (Norin61)은 글리아딘 분획의 SDS-PAGE 분석 시 구분이 어려우나 글루테닌 분획의 SDS-PAGE에서 Glu-B3d와 i는 쉽게 구별이 가능하다고 알려져 있다 (Liu et al. 2010, Park et al. 2011, Shin et al. 2012). 또한 글루테닌 분획의 Glu-B3 LMW-GS 2-DGE 분석에서는 d와 i의 패턴은 동일하며 이들 d와 i 그리고 ad는 쉽게 구별이 가능하다 (Data not shown; Lee et al (2016) Allelic analysis of low molecular weight glutenin subunits using 2-DGE in Korean wheat cultivars. Euphytica. 심사중; Liu et al. 2010). 상기의 결과들을 종합해 보면 Glu-B3 저분자 글루테닌 유전자좌 분석 시에는 Gli-B1 유전자좌 분석이 필수 불가결하며 반대의 경우도 마찬 가지이다.
본 연구에서 WDEIA의 중심 항원인 ω5-글리아딘 (Gli-B1)의 유전자좌를 분석하기 위해 표준 품종을 이용하여 최초로 국내 밀 32품종의 유전자좌를 명명하였고, 빵 가공적성에 많은 영향을 미치며 상호보완적인 Glu-B3 저분자 글루테닌의 유전자좌를 확인하는 보조적인 수단으로 가능성을 확인하였다. 본 연구는 밀 알러지 및 글리아딘과 밀 품질과의 관계 그리고 알러지 저감 밀 계통 육성에 기반이 되는 연구로 중요한 의미를 가진다.
오메가 (ω)5-글리아딘은 밀 의존성 운동 유발성 과민증 (wheat-dependent exercise-induced anaphylaxis; WDEIA)의 중심 항원이다. 본 연구에서는 국내 밀 품종의 ω5-글리아딘 (Gli-B1)의 유전자좌를 명명하기 위해 Gli-B1(a~m) 11 유전자좌 의 표준품종 글리아딘에 대한 SDS-PAGE 패턴 비교 분석을 통해 유전자좌를 명명하였고, 보조적으로 1B 염색체에 밀접히 연관되어 있는 본 연구팀의 Glu-B3 저분자 글루테닌 유전자좌를 상호보완적으로 활용하여 재확인하였다. 그 결과 Gli-B1b은 1 품종, Gli-B1d는 4 품종, Gli-B1f 는 2품종 Gli-B1m 은 6품종 나머지 19품종은 Gli-B1h를 확인하였다. 본 실험은 국내 밀 품종 의 ω5-글리아딘 에 대한 최초의 연구이며 밀 알러지 및 글리아딘 과 밀 품질과의 관계 그리고 알러지 저감 밀 계통 육성에 기반이 되는 중요한 자료로 가치가 있다.
본 연구는 농촌진흥청 국립농업과학원 농업과학기술 연구개 발사업 (과제번호: PJ010116) 및 농촌진흥청 국제공동연구사업 (과제번호: PJ012096) 지원에 의해 이루어진 것임.
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Allelic Analysis of Omega-5 Gliadin (Gli-B1) in Korean Wheat Cultivars
Korean. J. Breed. Sci.. 2016;48(2):159-167.   Published online June 30, 2016
DOI: https://doi.org/10.9787/KJBS.2016.48.2.159
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Allelic Analysis of Omega-5 Gliadin (Gli-B1) in Korean Wheat Cultivars
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DOI: https://doi.org/10.9787/KJBS.2016.48.2.159
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Allelic Analysis of Omega-5 Gliadin (Gli-B1) in Korean Wheat Cultivars
Image Image Image Image Image
Fig. 1. SDS-PAGE analysis of gliadin fractions of the standard wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to individual Gli-B1 alleles.
Fig. 2. SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1b and Gli-B1f.
Fig. 3. SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1d.
Fig. 4. SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1h.
Fig. 5. SDS-PAGE analysis of gliadin fractions of the Korea wheat cultivars for Gli-B1 locus. The arrow indicates the band corresponding to the Gli-B1m.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Allelic Analysis of Omega-5 Gliadin (Gli-B1) in Korean Wheat Cultivars

Standard wheat cultivars for Gli-B1 allele analysis.

Allele(Glu-B3 locusa) Standard Cultivar
Gli-B1a (Glu-B3a) Chinese Spring
Gli-B1b (Glu-B3b) Gabo
Gli-B1c Siete Cerros 66
Gli-B1d (Glu-B3h) Suneca
Gli-B1e Apexal
Gli-B1f (Glu-B3g) Cappelle-Desprez, Glenlea
Gli-B1g (Glu-B3f) Champtal
Gli-B1h (Glu-B3d) Rudi
Gli-B1i (Glu-B3c) Insignia
Gli-B1l (Glu-B3j) Clement
Gli-B1m (Glu-B3i) Norin61

Complementary Glu-B3 locus.

Allelic compositions of Gli-B1 in Korean wheat cultivars by gliadin SDS-PAGE. The pedigree each cultivar is indicated.

Cultivar Pedigree Glu-B3a Gli-B1
Sukang Suwon266/Asakaje b b
Hanbaek Shann7859/Keumkang//Guamuehill h d
Joeun Eunpa/Suwon242 h d
Jokyung Seri82/Keumkang h d
Keumkang Geuru/Kanto75//Eunpa h d
Jonong Suwon234/SW80199 g f
Sinmichal 1 Alchan//Kanto107/BaiHuo g f
Alchan Suwon210/Tapdong d h
Baekjoong Keumkang/Olgeuru d h
Chungkye Norin4/Sharbatisonora d h
Dabun Suwon234//sw76039/Suwon220/3/Keumkang d h
Eunpa Chukoku81/3/Tob-CNO// Yuksung3/Suwon185 d h
Gobun Eunpa/Tapdong//Eunpa/Shannung6521 d h
Goso Gobun/Ol d h
Jeokjoong Keumkang/Tapdong d h
Jinpoom Geuru/Genaro81 d h
Milsung Sirogane//Noring43/Sonalika d h
Namhae Ol/Calidad d h
Ol Norin72/Norin12 d h
Olgeuru Geuru/Chokwang//Nishigai143 d h
Saeol Sirogane//Norin43/Sonalika d h
Seodun Geuru/Genaro81 d h
Sinmichal Olgeuru//Kanto107/BaiHuo d h
Tapdong Chukoku81//Suwon158/Toropi d h
Uri Geuru/Ol d h
Younbaek Keumkang/Tapdong d h
Dahong Norin72/Wonkwang i m
Anbaek Sae/Geuru ad m
Dajoong SW992114-NM-131-7/Gobun ad m
Geuru Strampelli/69D-3607//Chokwang ad m
Jopoom Kanto75//OR8500494P/Bezostaya ad m
Suan Keumkang/Eunpa//Keumkang ad m

Glu-B3 alleles using 2DGE in this group.

Table 1. Standard wheat cultivars for Gli-B1 allele analysis.

Complementary Glu-B3 locus.

Table 2. Allelic compositions of Gli-B1 in Korean wheat cultivars by gliadin SDS-PAGE. The pedigree each cultivar is indicated.

Glu-B3 alleles using 2DGE in this group.

Table 1.

Table 2.

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