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Korean. J. Breed. Sci. : Korean Journal of Breeding Science

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슈퍼홍미 쌀품종 탁시폴린(Taxifolin) 성분의 환경변이

Environmental Variation of Taxifolin Content in a Rice Variety ‘Superhongmi’

Korean Journal of Breeding Science 2018;50(1):45-49.
Published online: February 28, 2018

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*Corresponding Author (ryusn@knou.ac.kr, +82-2-3668-4630, +82-2-3673-2381)
• Received: January 11, 2018   • Accepted: January 16, 2018

© Korean Society of Breeding Science. All rights reserved.

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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  • This study was conducted to obtain general information for rice breeding materials high in taxifolin content. The taxifolin was isolated from the rice variety, ‘Superhongmi’, and was identified by means of spectral methods and quantitative analysis using a high performance liquid chromatography (HPLC). The taxifolin content of the ‘Superhongmi’ variety was ranged between 32mg and 68mg per 100g seed that differs depending on cultivated regions. The taxifolin content was lower grown at mid north than those at south region. A large amount of taxifolin was found in the aleurone layer of seed as well as in the node of spike of the ‘Superhongmi’. The weight of 1,000 grains was ranged between 23.5g and 27.8g that varied depending on cultivated regions. This variety showed heading in the 6th of September, and its fertility ratio was about 83% in sejong region. These results suggest that the taxifolin-rich rice variety, ‘Superhongmi’, is a good source of functional and stable food.
슈퍼홍미는 전통적인 교배육종을 통해 총 폴리페놀 함량을 높인 천립중이 무거운 만생종으로서 기능성 식품소재로서 유망 한 기능성 쌀품종이다(Ryu 2015). 페놀성 화합물은 식물계에 널리 분포되어 존재하는 2차 대사산물 중 하나로 다양한 구조와 분자량을 가지며 phenolic hydroxyl기가 단백질과 같은 거대분 자와 결합하여 항산화, 항암 및 항균 등의 생리기능을 가지는 것으로 알려져 있다(Choi et al. 2003). 플라보노이드를 포함한 폴리페놀성 화합물은 유리 라디칼을 제거하여 산화를 억제하고 활성산소의 소거 및 산화적 스트레스를 막아 항암, 항균, 항알러 지, 노화 등을 예방하거나 지연시키는 효과를 가진다(Hertog et al. 1992). 플라보이드는 플라본(flavon)을 기본구조로 가지는 식물 색소로 페놀화합물과 함께 식물체 전반에 함유되어 있으며 항산화 활성에 직접적인 영향을 주는 인자로 알려져 있다. 최근 농진청에서 개발한 중만생 적갈색 신품종 홍진주벼는 기능성물 질인 페놀릭산과 플라보노이드 함량이 기존의 유색미 품종보다 많이 함유되어 있는 것으로 보고되었다(Yang et al. 2011).
미강은 우리나라의 중요 식량자원인 벼를 도정할 때의 부산물 로서 현미의 도정 과정상에서 약 7% 정도의 비중으로 얻어지고 있으며, 국내에서는 연간 약 60 만톤 정도의 미강이 발생하는 것으로 알려져 있다. 미강의 조성은 벼의 품종, 도정 방법 등에 따라 다르나 수분함량 14%를 기준으로 할 때 조단백질 11~ 17%, 조지방질 15~20%, 조섬유질 7~11%, 조회분 7~10%, 탄수화물 34~52%인 것으로 보고 되고 있다. 우리나라에서는 미강의 약 30%가량이 미강유 생산에 활용되고 있으며, 전 세계 적으로는 미강의 약 5%정도만이 미강유 생산에 이용되고 있는 것으로 알려져 있다. 특히, 식품 가공기술의 발달로 식품의 기호 성 증진을 위해 곡류의 도정 과정이 과거보다 더 정교해짐에 따라 미강의 발생량은 계속 증가할 것으로 예측되고 있어, 미강을 이용한 새로운 식품의 연구개발이 지속적으로 이루어질 것으로 판단된다.
현미 상태의 유색미는 식이섬유를 다량 함유하고 있으며 각종 단백질, 불포화지방산 및 비타민, 무기질 등이 풍부하여 영양학 적으로 우수한 식품이다. 이런 유색미는 적갈색, 흑자색 등의 다양한 천연색소를 함유하는데 일반적으로 흑자색계 쌀에는 안토시아닌계 색소가 다량 함유되어 있고 적갈색계 쌀에는 탄닌 계 색소가 함유되어 있는 것으로 알려져 있다. 안토시아닌계 색소는 높은 항산화 기능뿐만 아니라 돌연변이 억제효과 등의 높은 기능성으로 인체 면역능을 향상시키며 노화방지, 질방예방 등에 효과가 있어 기능성 식품소재로서 이용가치가 높은 것으로 알려져 있다. 탄닌계 색소 역시 유해성 중금속을 제거하고 변이원 생성을 억제시키는 것으로 알려져 있다(Nam et al. 2002, Oh et al. 1996, Ryu 2014).
당뇨병은 유전적 또는 후천적인 원인에 의해 혈당 조절 호르몬 인 인슐린의 분비량이 부족하거나 인슐린의 기능이 정상적으로 이루어지지 않아 당 대사에 이상이 발생하여 고혈당을 유발하는 대사성 질환이다. 당뇨병은 크게 제1형과 제2형으로 분류된다. 제1형(인슐린 의존성) 당뇨는 유전적인 소인으로 인해 췌장의 베타세포가 파괴되어 인슐린 분비가 저하되어 인슐린 치료가 절대적으로 필요한 반면, 제2형 당뇨는 유전적인 소인보다 고열 량, 고지방 식사, 운동부족, 스트레스 등 후천적인 요인으로 인해 인슐린 분비장애, 인슐린 저항성이 발생하는 경우이다. 제2형 당뇨는 중년 이후에 발병율이 높으며, 국내의 당뇨 환자의 90% 이상이 제2형 당뇨병으로 보고되어 있다. 국내 당뇨병 환자는 약 500만 명 정도로 예상되고 있으며, 당뇨병은 5대 사망원인 중 하나로 암, 뇌혈관질환, 심장질환 다음으로 사망률이 높아 사회적으로 문제가 되고 있다. 2014년 우리나라 당뇨병 유병률 이 13.7%로 조사됐다(Diabets Fact Sheet in Korea 2016). 당뇨 병은 지속적인 고혈당 증상으로 인해 심장질환, 신장질환, 시각 장애 및 신경 장애와 같은 다양한 합병증을 동반하는 것을 특징으 로 하는데, 이러한 합병증이 심각한 문제를 야기하며, 환자들의 삶의 질을 극도로 저하시킨다. 특히 대표적으로 순환기계의 질환 에서 심근경색이나 뇌경색 등의 대혈관장애 등은 당뇨병 상태뿐만 아니라 당뇨병 발병 이전 단계인 내당능장애 단계에서부터 발병 이 증가하는 것으로 나타나 이에 대한 예방 방법 또한 필수적으로 필요한 상태이다(O'Brien & Granner 1996, Reaven 1988). 따라 서 당뇨병의 치료 및 합병증을 예방하기 위해 혈당을 지속적으로 관리하여야 하며, 약물요법과 함께 식사요법, 운동요법 등 생활 습관에서부터 철저한 관리체계가 필요하며 가장 바람직한 방법 은 생활습관과 식습관 조절을 통해서 당뇨병 발병 이전 단계로부 터 혈당을 관리하는 예방 차원에서의 치유가 가장 바람직하다 (Park et al. 1996).
흑자색미에 함유된 안토시아닌 색소의 80% 정도는 cyanidin- 3-glucoside(C3G)이며, 그 외 peonidin-3-glucoside(P3G), malvidin, pelargonidin 및 selphinidin 등을 함유하고 있다(Ryu et al. 2006). 안토시아닌은 항산화활성, 항염효과(Wang H, et al. 2002), 항암 효과, 항아토피효과, 항당뇨효과 및 심혈관계 질병 등의 예방과 치료효과 등이 밝혀진 성분이다(Ryu 2014, Hong et al. 1997). 반면 갈색유색미에는 흑자색미에 없는 탁시폴린(taxifolin) 성분이 있는 것으로 밝혀졌다(Kwon et al. 2017). 슈퍼홍미 추출물은 in vitro 실험에서 항산화활성, 항당뇨활성, 혈당상승억제작용이 높 은 것으로 알려졌다. 또한 슈퍼홍미의 탁시폴린 성분은 약효가 우수한 당뇨병 예방 또는 치료용 약학 조성물로도 밝혀졌다(Kwon et al. 2017, Xiao sun et al. 2014).
본 연구는 쌀에 함유된 혈당저하 성분인 탁시폴린 성분의 품종간 차이, 재배지역과 식물체 및 종실의 부위별 함량변이를 조사하여 육종의 소재로서 기본정보를 얻고자 수행하였다.
1. 재료
슈퍼홍미(국립종자원 출원2015-339호, 2015.03.31.) 벼 품종 은 총 폴리페놀 함량을 높인 기능성 쌀 품종이다(Ryu 2015). 기능 성 성분 추출을 위해 슈퍼홍미 현미 100g에 1L의 MeOH(0.1% trifluou acetic acid)을 혼합한 후 40℃에서 2시간 추출하였다. 추출액을 원심분리기에서 원심분리한 후(8,000rpm, 4℃, 30분) 상등액을 Whatman No. 1 필터로 여과하였다. 여과액을 감압농 축하여 용매를 모두 제거한 후 동결건조기에서 동결건조하여 분석하였다. 탁시폴린 성분의 화학구조는 Fig. 1과 같으며 분석 조건은 Table 1과 같다.
Fig. 1.
Structure of taxifolin component of rice variety Superhongmi.
KJBS-50-45_F1.jpg
Table 1
Analysis condition of taxifolin component in rice variety Superhongmi.
Table 1
Mobile phase A : DW   B : 60% MeOH (pH 2.4)
Detector Agilent 1260 DAD
Flow 1.0 ㎖/min
Column TOSOH TSK-GEL C18
탁시폴린의 표준물질을 각각 25 mg 정평하여 MeOH 25 ml에 용해시킨 용액을 stock solution으로 해서 이를 일정량씩 취한후 각각에 MeOH을 가해 5, 10, 20, 50 mg/ml가 되게 하여 조제하였 다. solution 용액을 각각 10 ㎕ 씩 취하여 chromatogram을 얻고 각각의 chromatogram으로부터 peak area를 구하여 검량선을 작성하였다.
갈색쌀의 탁시폴린 함량차이를 구명하기 위하여 적진주벼, 홍진주벼, 건강홍미, 슈퍼홍미와 슈퍼홍미 유래 계통 5개를 분석 재료로 사용하였다.
슈퍼홍미는 5개 재배지역 여주(경기도 여주시농업기술센터 시험포장), 세종(세종시 전의면 고등리 시험포장), 당진(당진시 우강면 송산리 138 시험포장), 김제(김제시 대창자연미 영농법 인 시험포장), 진도(전남 진도군농업기술센터 시험포장)에서 농 촌진흥청 표준재배법에 의해 재배하여 taxifolin 함량을 분석하 였다. 식물체를 이삭, 줄기, 잎, 뿌리로 구분하여 분석재료로 하였다. 또한, 종자를 배, 배유, 호분층으로 구분하여 탁시폴린 함량을 분석하였다.
1. 슈퍼홍미 Taxifolin성분의 정량분석
슈퍼홍미 추출물의 지표성분인 Taxifolin을 HPLC를 이용한 정량분석방법을 확립하기 위해 Table 1과 같은 방법으로 분석하 였다. Detector로는 UV만 사용하여 분석하였다. Taxifolin 성분 의 retention time은 30분대 임을 알 수 있었다(Fig. 2).
Fig. 2.
HPLC chromatogram for MeOH extract from Superhongmi rice variety. Column : Tosoh TSK-Gel C18.
KJBS-50-45_F2.jpg
확인된 taxifolin 성분의 함량을 결정하기 위하여 표준검량선 을 작성하여 Fig. 3에 나타내었다. 슈퍼홍미 추출물의 taxifolin 성분 농도의 범위를 5~50 ㎍/㎖로 하여 정량곡선을 측정하였다. 정량의 범위는 성분의 농도에 직선성을 나타내는 농도로 표시하 였으며, 면적비를 이용한 검량선으로 상관계수 R2를 구하여 양호 한 직선성을 확인한 결과 5~50㎍/㎖ 구간에서 R2=0.999로 매우 유의적인 상관성을 확인하였다(Fig. 3).
Fig. 3.
Calibration curves for standard materials of taxifolin component.
KJBS-50-45_F3.jpg
슈퍼홍미 추출물을 통해서 taxifolin 함량을 HPLC를 통해 분석 한 결과 슈퍼홍미 현미 100 g에 68 mg 함유된 것으로 확인되었다.
2. 유색미 Taxifolin 함량의 품종 및 식물체 부위별 함량차이
갈색쌀에 함유된 탁시폴린 함량은 Table 2에서와 같다. 현미 100 g에 함유된 양은 0에서 78 mg으로 품종간 차이가 컸다. 슈퍼홍미 유래 계통내에서도 26 mg에서 78 mg의 함량 변이가 있어 고 탁시폴린 품종육성에 중요한 소재가 될 것으로 사료된다. 또한, 식물체 부위와 종실의 부위별 탁시폴린 함량을 Table 3에 서 보면, 종실 뿐만 아니라 이삭에도 다량 함유되어 있어 향후 식물체를 이용한 새로운 부가가치 창출을 위한 소재로의 이용도 가능 할 것으로 보인다(Table 3). 탁시폴린 성분은 종실의 호분층 에 대부분 함유 되어 있어 도정후의 부산물로서 쌀겨의 새로운 용도가 개발될 것으로 보인다.
Table 2
Difference of taxifolin content in brown rice varieties. (mg/100g seed)
Table 2
Varieties and strains Content of taxifolin

Superhongmi 67.7±5.48
〃   Strains 15074 26.3±3.14
〃   Strains 15075 60.3±2.49
〃   Strains 15189 35.7±4.45
〃   Strains 15190 24.7±4.32
〃   Strains 17079 77.6±6.49
Jeokjinju ND*
Hongjinju ND*
Keonganghongmi ND*

*ND: Not Detected

Table 3
Difference of taxifolin contents in plant and seed of rice variety Superhongmi.
Table 3
Parts Contents of taxifolin (mg/100g seed)
Plant node of spike 143.5±8.25
leaf -
root -
Seed embryo -
endosperm -
aleurone layer 106.6±5.32
3. 재배지역에 따른 슈퍼홍미의 작물학적 특성과 taxifolin 함량변이
슈퍼홍미의 재배지역에 따른 작물학적 특성과 탁시폴린 함량 차이를 Table 4에서 보면, 세종지역에서의 출수기는 9월 6일로 서 만생종 품종이다. 이미 보고된 슈퍼홍미의 작물학적 특성과 유사한 결과이다(Ryu 2015). 만생종인 슈퍼홍미를 중북부지역 인 여주에서 재배할 때 출수기가 9일 늦은 9월 15일 이었으며 임실율도 77%로 낮고, 탁시폴린 함량도 다른 지역에 비해 그 함량이 낮아 재배지역으로 부적합 하였다. 반면, 세종 이남지역 인 당진, 김제, 진도지역에서는 함량 차이가 크지 않았다. 그러나 남부 진도지역에서 재배할 때 임실율이 다소 높았다.
Table 4
Difference of major agronomnic traits and seed taxifolin content depending on cultivated region in rice variety Superghongmi.
Table 4
Cultivated region Transplanting date Heading date 1,000 grain weight (g) Ratio of fertility (%) Contents of taxifolin (mg/100g seed)
Yeoju May 30 Sep. 15 23.5±2.89bz 76.6±3.45b 32.72±5.32b
Sejeong May 30 Sep. 6 26.8±2.23a 83.4±4.14a 67.7±5.48a
Dangjin May 30 Sep. 6 26.5±3.14a 82.4±3.15a 63.4±4.32a
Kimje May 30 Sep. 7 27.3±2.89a 83.5±2.65a 65.4±5.11a
Jindo June 5 Sep. 9 27.8±3.16a 86.6±3.19a 68.3±3.39a

zValues are means±standard deviation(n=3). Means in the same column not sharing a common superscript are significantly different at p<0.05.

본 연구는 쌀에 함유된 혈당을 저하하는 탁시폴린 성분에 대한 육종소재로서 기본정보를 얻고자 수행하였다. 슈퍼홍미 벼 품종으로부터 탁시폴린 성분을 분리하고 분광학적 방법에 의해 확인하였다. 또한 HPLC로 검량선을 작성하고 정량분석을 하였다. 다양한 유색미 품종 중에서 슈퍼홍미와 슈퍼홍미 유래 계통에서만 탁시폴린 함량이 확인되었다. 슈퍼홍미의 탁시폴린 함량은 재배지역에 따라서 쌀 100 g에 32 mg에서 68 mg의 함량변이를 보였다. 이들 성분의 함량은 중북부지역에서 재배 되었을 때보다 남부지역에서 재배되었을 때 높게 나타났다. 천립 중은 재배지역에 따라서 23.5 g에서 27.8 g의 변이를 보였다.
탁시폴린 성분은 슈퍼홍미의 이삭뿐만 아니라 종자의 호분층 에도 다량 함유되어 있어 이들 식물체를 기능성 소재로 이용할 수 있을 것으로 사료된다. 따라서 탁시폴린 성분이 다량 함유된 슈퍼홍미는 식량으로서뿐만 아니라 기능성 식품소재로서 높은 가치가 있을 것으로 기대된다.
이 논문은 2017년도 한국방송통신대학교 학술연구비 지원을 받아 작성된 것임.
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Environmental Variation of Taxifolin Content in a Rice Variety ‘Superhongmi’
Korean. J. Breed. Sci.. 2018;50(1):45-49.   Published online March 1, 2018
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Environmental Variation of Taxifolin Content in a Rice Variety ‘Superhongmi’
Korean. J. Breed. Sci.. 2018;50(1):45-49.   Published online March 1, 2018
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Environmental Variation of Taxifolin Content in a Rice Variety ‘Superhongmi’
Image Image Image
Fig. 1. Structure of taxifolin component of rice variety Superhongmi.
Fig. 2. HPLC chromatogram for MeOH extract from Superhongmi rice variety. Column : Tosoh TSK-Gel C18.
Fig. 3. Calibration curves for standard materials of taxifolin component.
Environmental Variation of Taxifolin Content in a Rice Variety ‘Superhongmi’

Analysis condition of taxifolin component in rice variety Superhongmi.

Mobile phase A : DW   B : 60% MeOH (pH 2.4)
Detector Agilent 1260 DAD
Flow 1.0 ㎖/min
Column TOSOH TSK-GEL C18

Difference of taxifolin content in brown rice varieties. (mg/100g seed)

Varieties and strains Content of taxifolin

Superhongmi 67.7±5.48
〃   Strains 15074 26.3±3.14
〃   Strains 15075 60.3±2.49
〃   Strains 15189 35.7±4.45
〃   Strains 15190 24.7±4.32
〃   Strains 17079 77.6±6.49
Jeokjinju ND*
Hongjinju ND*
Keonganghongmi ND*

*ND: Not Detected

Difference of taxifolin contents in plant and seed of rice variety Superhongmi.

Parts Contents of taxifolin (mg/100g seed)
Plant node of spike 143.5±8.25
leaf -
root -
Seed embryo -
endosperm -
aleurone layer 106.6±5.32

Difference of major agronomnic traits and seed taxifolin content depending on cultivated region in rice variety Superghongmi.

Cultivated region Transplanting date Heading date 1,000 grain weight (g) Ratio of fertility (%) Contents of taxifolin (mg/100g seed)
Yeoju May 30 Sep. 15 23.5±2.89bz 76.6±3.45b 32.72±5.32b
Sejeong May 30 Sep. 6 26.8±2.23a 83.4±4.14a 67.7±5.48a
Dangjin May 30 Sep. 6 26.5±3.14a 82.4±3.15a 63.4±4.32a
Kimje May 30 Sep. 7 27.3±2.89a 83.5±2.65a 65.4±5.11a
Jindo June 5 Sep. 9 27.8±3.16a 86.6±3.19a 68.3±3.39a

zValues are means±standard deviation(n=3). Means in the same column not sharing a common superscript are significantly different at p<0.05.

Table 1 Analysis condition of taxifolin component in rice variety Superhongmi.
Table 2 Difference of taxifolin content in brown rice varieties. (mg/100g seed)

ND: Not Detected

Table 3 Difference of taxifolin contents in plant and seed of rice variety Superhongmi.
Table 4 Difference of major agronomnic traits and seed taxifolin content depending on cultivated region in rice variety Superghongmi.

Values are means±standard deviation(n=3). Means in the same column not sharing a common superscript are significantly different at p<0.05.