• Molecules and Cells
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• 제25권2호
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• pp.231-241
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• 2008

Indole glucosinolates (IG) play important roles in plant defense, plant-insect interactions, and stress responses in plants. In an attempt to metabolically engineer the IG pathway flux in Chinese cabbage, three important Arabidopsis cDNAs, CYP79B2, CYP79B3, and CYP83B1, were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B3 or CYP83B1 did not affect IG accumulation levels, and overexpression of CYP79B2 or CYP79B3 prevented the transformed callus from being regenerated, displaying the phenotype of indole-3-acetic acid (IAA) overproduction. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, the transformed calli were regenerated into whole plants that accumulated higher levels of glucobrassicin, 4-hydroxy glucobrassicin, and 4-methoxy glucobrassicin than wild-type controls. This result suggests that the flux in Chinese cabbage is predominantly channeled into IAA biosynthesis so that coordinate expression of the two consecutive enzymes is needed to divert the flux into IG biosynthesis. With regard to IG accumulation, overexpression of all three cDNAs was no better than overexpression of the two cDNAs. The content of neoglucobrassicin remained unchanged in all transgenic plants. Although glucobrassicin was most directly affected by overexpression of the transgenes, elevated levels of the parent IG, glucobrassicin, were not always accompanied by increases in 4-hydroxy and 4-methoxy glucobrassicin. However, one transgenic line producing about 8-fold increased glucobrassicin also accumulated at least 2.5 fold more 4-hydroxy and 4-methoxy glucobrassicin. This implies that a large glucobrassicin pool exceeding some threshold level drives the flux into the side chain modification pathway. Aliphatic glucosinolate content was not affected in any of the transgenic plants.

• BMB Reports
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• 제41권6호
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• pp.472-478
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• 2008

Three Arabidopsis cDNAs, MAM1, CYP79F1, and CYP83A1, required for aliphatic glucosinolate biosynthesis were introduced into Chinese cabbage by Agrobacterium tumefaciens-mediated transformation. The transgenic lines overexpressing MAM1 or CYP83A1 showed wild-type phenotypes. However, all the lines overexpressing CYP79F1 displayed phenotypes different from wild type with respect to the stem thickness as well as leaf width and shape. Glucosinolate contents of the transgenic plants were compared with those of wild type. In the MAM1 line M1-1, accumulation of aliphatic glucosinolates gluconapin and glucobrassicanapin significantly increased. In the CYP83A1 line A1-1, all the aliphatic glucosinolate levels were increased, and the levels of gluconapin and glucobrassicanapin were elevated by 4.5 and 2 fold, respectively. The three CYP79F1 transgenic lines exhibited dissimilar glucosinolate profiles. The F1-1 line accumulated higher levels of gluconapoleiferin, glucobrassicin, and 4-methoxy glucobrassicin. However, F1-2 and F1-3 lines demonstrated a decrease in the levels of gluconapin and glucobrassicanapin and an increased level of 4-hydroxy glucobrassicin.

• 원예과학기술지
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• 제30권6호
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• pp.743-750
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• 2012

본 연구는 UPLC를 이용하여 국내 브로콜리 육성계통 95점의 화구 내 desulfo-glucosinolate를 표준물질과 비교하여 정량 분석하고 그 함량변이를 평가하여, 궁극적으로 고기능성 국내 브로콜리 품종 육성의 기초자료로 이용하고자 수행되었다. 브로콜리 화구에서 9종의 표준물질과 비교하여 progoitrin, glucoraphanin, sinigrin, gluconapin, glucobrassicanapin 및 glucobrassicin 등 6종의 glucosinolate가 동정되었다. 전체 브로콜리 육성계통 95점에 대해서 총 glucosinolate 함량은 $4.2-29.0{\mu}mol{\cdot}g^{-1}$ DW의 변이를 보였다. 총 glucosinolate 중 glucoraphanin의 함량은 $1.6-13.9{\mu}mol{\cdot}g^{-1}$ DW로 가장 높은 비율을 차지하였고 progoitrin은 제한적인 13개의 계통에서만 특이적으로 함유되어 있었으며 glucoraphanin의 함량과는 부의 상관관계를 보였다. 동정된 glucosinolate 중 glucobrassicanapin, glucoraphanin, glucobrassicin 및 gluconapin의 4종 물질이 전체 브로콜리 계통의 함량변이에 크게 영향하였고 이들 4종 물질 상호 간에는 높은 정의 상관관계가 있음이 확인되었다. 본 연구결과에서 선발된 브로콜리 유망계통에 대해서는, 향후 재배 작형 및 연차간 물질함량의 안정성 평가와 유전자원 간 추가적인 비교평가 연구를 통하여 고기능성 육종소재로의 개발이 가능할 것으로 판단된다.

• 원예과학기술지
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• 제32권5호
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• pp.730-738
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• 2014

The objective of this study was to evaluate glucosinolate (GSL) profiles and variation of total and individual GSLs concentrations within seedling (0-14 days) and growth stages (0-15 weeks) of Korean Chinese cabbage (Brassica rapa L. ssp. pekinensis). Ten GSLs (progoitrin, glucoraphanin, glucoalyssin, gluconapin, glucobrassicanapin, 4-hydroxyglucobrassin, glucobrassicin, 4-methoxyglucobrassicin, neoglucobrassicin and gluconasturtiin) were identified from Korean Chinese cabbage. In general, total GSL content significantly decreased during seedling (from 92.89 to $35.26{\mu}mol{\cdot}g^{-1}$ DW) and g rowth stages ( from 74.11 to $1.97{\mu}mol{\cdot}g^{-1}$ DW). Gluconapin was the highest in seeds and in the germination period ($73.1{\mu}mol{\cdot}g^{-1}$ DW) and declined gradually from 73 to 15% during seedling stages. The level of the major aliphatic GSLs, gluconapin and progoitrin, tended to decrease sharply, whereas levels of indolic GSLs (4-methoxyglucobrassicin, glucobrassicin) and aromatic GSLs (gluconasturtiin) were found to increase generally at the beginning of growth stages.

• 농업과학연구
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• 제38권2호
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• pp.285-294
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• 2011

Glucosinolate (GSL) contents were investigated (i) at 1~7 days after sowing (DAS) in seed sprouts and (ii) at 3-7 weeks after sowing for the time-course. Moreover, (iii) They were compared with five different cultivars of rocket salad (Eruca sativa). Seventeen GSLs were separated by HPLC analysis, and 10 GSLs among them were identified as glucoraphanin, sinigrin, glucoalyssin, diglucothiobeinin, glucobrassicanapin, glucoerucin, glucobrassicin, dimeric, 4-mercaptobutyl GSL, 4-methoxy glucobrassicin, gluconasturttin by using LC-APCI-MS analysis, but 7 compounds were not identified. (i) The total GSL content in seed sprouts initially increased up to 3 DAS and then decreased according to their seedling growth. In particular, glucoraphanin known as a strong anti-cancer reagent was found the highest level (5.05 ${\mu}mol/g$ dry wt.) at 3 DAS. The most abundant GSL was glucoerucin ranged from 26.0~49.6 ${\mu}mol/g$ dry wt. (ii) In the time-course, the total GSL contents increased dramatically from 3-week (5.91 ${\mu}mol/g$ dry wt.) to 7-week after sowing (32.2 ${\mu}mol/g$ dry wt.). The major GSLs were glucoraphanin, glucoerucin and 4-methoxy glucobrassicin. (iii) By comparing GSL contents with five different cultivars, the total GSL contents increased from 4-week to 6-week after sowing, regardless of cultivar. In 4-week-old, the order with the total GSL content was "Rucola" > "Rocket Herbs" ${\geqq}$ "Odyssey" > "Takii" > "Herb", but in 6-week-old it is changed as "Takii" > "Herb" > "Odyssey" > "Rucola" > "Rocket Herbs" even there was almost no significantly difference between them.

• 농업과학연구
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• 제44권2호
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• pp.261-271
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• 2017

The aim of this study was to investigate the amount of glucosinolates (GSLs) in kale sprouts (Brassica oleracea L. var. acephala) ('TBC') according to different concentrations of sulfur ions in sprout's nutrient solutions (0.0, 0.5, 1.0, and 2.0 mM) and to different light sources [Fluorescent lamp, Red, Blue, and Mix (R+B) LED]. Kale sprouts were cultivated in a growth chamber for 13 days in sulfur solutions. Kale sprouts were treated with fluorescent lamp and LED light sources for 5 days, from eight days after sowing to harvest. Amount of seven types of GSLs (progoitrin, sinigrin, 4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, gluconasturtiin, and neoglucobrassicin) were measured in kale sprouts after harvest. The total GSL content was influenced by different sulfur solution concentration, and it was the highest at S 0.5 mM ($172.54{\mu}mol{\cdot}g^{-1}DW$) and the lowest at S 2.0 mM ($163.09{\mu}mol{\cdot}g^{-1}DW$). The GSL content was influenced by different light source, and it was the highest with Red LED ($159.23{\mu}mol{\cdot}g^{-1}DW$) and the lowest with Blue LED ($147.57{\mu}mol{\cdot}g^{-1}DW$). As the sulfur solution concentration increased under all light source, progoitrin and sinigrin contents tended to decrease while glucobrassicin content showed an upward tendency for all of the light sources. The content of glucobrassicin was higher than that of progitrin when treated with sulfur solutions for all LED light sources. Sinigrin, which has excellent anti-cancer effects, showed the highest rate (92.2%) among all the GSLs, under all of the light sources.

• 한국식품영양학회지
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• 제25권2호
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• pp.259-265
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• 2012

본 연구에서는 배추와 양배추 에탄올 추출물의 총 폴리페놀 함량 및 총 플라보노이드 함량을 측정하고, glucosinolates 함량을 HPLC로 분석하였다. 또한, MTT assay를 통한 암세포 증식 억제 활성을 측정하였다. 배추와 양배추 에탄올 추출물의 항산화물질로 알려진 총 폴리페놀 함량은 각각 308.48 및 344.75 ${\mu}g$ GAE/g dry weight으로 나타났으며, 총 플라보노이드 함량은 각각 5.33 및 5.95 ${\mu}g$ QE/g dry weight으로 나타났다. 배추 추출물에서는 progoitrin, glucoalyssin, gluconapin, glucobrassicanapin, glucobrassicin, 4-methoxyglucobrassicin의 총 6개 glucosinolates를 확인하였다. 양배추 추출물에서는 glucoraphanin, sinigrin, glucobrassicin 및 4-methoxyglucobrassicin의 총 4개 glucosinolates를 확인하였다. 배추와 양배추 에탄올 추출물이 AGS 인체 위암세포주, HepG2 인체 간암세포주, LNCaP 인체 전립선암 세포주 증식에 미치는 영향을 MTT assay를 통해 알아보았다. 배추와 양배추 추출물의 농도가 증가함에 따라 농도 의존적으로 암세포 증식 억제 효능이 증가하였다. 또한, 배추와 양배추 추출물을 암세포에 처리하고 배양하는 시간이 24시간에서 48시간으로 길어질수록 암세포 성장 억제 효능도 증가하였다. 배추와 양배추의 추출과정 중에 생성된 glucosinolate 가수분해 산물과 폴리페놀, 플라보노이드 등의 생리활성 물질들이 암세포 성장 억제에 직접적인 영향을 주었을 것으로 사료된다. 한국인들이 자주 섭취하고 있는 배추와 양배추에는 주요 생리활성 물질인 글루코시놀레이트뿐 아니라 폴리페놀, 플라보노이드가 함유되어 있으며, 이들 추출물들은 암세포 증식 억제 효능이 있음을 보여주었다.

• 한국식품과학회지
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• 제52권6호
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• pp.587-594
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• 2020

본 연구에서는 부위별 배추와 배추김치에 함유되어 있는 글루코시놀레이트류의 조성을 UPLC-MS/MS를 이용하여 탐색하고, 인돌류 대사체를 MS/MS를 이용하여 분석한 결과, 글루코시놀레이트류는 음이온 모드([M-H]-)에서 검출되었으며, glucobrassicanapin (m/z 386), glucoalyssin (m/z 450), glucobrassicin (m/z 447), 4-methoxyglucobrassicin (m/z 477), neoglucobrassicin (m/z 477)는 MS scan 모드에서, gluconapin (m/z 372→97), progoitrin (m/z 388→97), glucoiberin (m/z 422→97), 4-methoxyglucobrassicin (m/z 477→97), neoglucobrassicin (m/z 477→447)는 MS/MS MRM 모드에서 검출되었다. Glucobrassicin과 같은 인돌기 함유 글루코시놀레이트류들이 대사되어 생성되는 인돌류 대사체로는 ascorbigen (m/z 306→130)과 I3A (m/z 146→118)가 MS/MS MRM ([M+H]+) 모드에서 검출되었다. Ascorbigen은 NKC보다 IPC와 OPC에 유의적으로 많이 함유되어 있었다. I3A는 NKC에 가장 많이 함유되어 있었으나 시료 간 유의적인 차이는 없었다. 본 연구를 통하여 배추와 배추김치에 존재하는 글루코시놀레이트류와 그 대사체인 인돌류 물질을 UPLC-MS/MS를 이용하여 분석할 수 있었다.

• 농업과학연구
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• 제45권1호
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• pp.1-8
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• 2018

Glucosinolates are one of the important plant secondary metabolites that are produced mainly in Brassicaceae plants. The compounds are primarily involved in defense responses to biotic and abiotic resistance in plants and play important biological roles during plant growth and development. In this study, the glucosinolate profiles in leaves of two different Brassica oleracea populations were compared using high-performance liquid chromatography (HPLC). The nine major glucosinolates compounds in cabbage leaves were identified as belonging to the aliphatic and indolic groups. Among them, sinigrin, which belongs to the aliphatic group, was recorded to be 41% whereas glucobrassicin and 4-methoxyglucobrassicin, which belong to the indolic group, were recorded to be 53.8%. In addition, we performed a genetic analysis to identify regions of the genome regulating glucosinolates biosynthesis in the $F_3$ population of Brassica oleracea. A total of 9 glucosinolates were used for the quantitative trait locus (QTL) analysis. Out of 9, a total of 3 QTLs were identified and they were associated with sinigrin, glucobrassicin, and 4-methoxyglucobrassicin synthesis located in Chromosome 1 and Chromosome 8, respectively. The results of this study will provide valuable information for the breeding of cabbage containing high glucosinolate content, and our next target is to develop component-specific and tightly linked markers for various glucosinolates.

• 원예과학기술지
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• 제33권2호
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• pp.177-185
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• 2015

케일 내 유용성분인 GSL 함량은 작물의 재배방법과 기후조건, 영양상태, 유전적 특성 등 재배 환경에 따라 영향을 받을 수 있으므로 본 연구에서는 생장단계에 따른 케일(Brassica oleracea var. acephala) 내 GSL 함량을 조사하였다. 케일 품종은 'Manchoo Collard', 'TBC' 두 품종으로 재배기간은 2012년 2월 20일부터 동년 7월 3일까지 133일이었으며 수확시기는 파종 후 35, 63, 91, 105, 119, 133일(days after sowing, DAS)이었다. 'TBC'에서는 총 10종의 GSL(progoitrin, sinigrin, glucoalyssin, gluconapin, glucoiberverin, 4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, gluconasturtiin, neoglucobrassicin)가 분리 및 동정되었으나 'Manchoo Collard'에서는 glucoiberverin를 제외한 9종의 GSL가 분리 및 동정되었다. 'Manchoo Collard'의 GSL 함량은 133DAS에서(평균 $6.12{\mu}mol{\cdot}g^{-1}$ DW)가장 높았고, 35DAS($1.16{\mu}mol{\cdot}g^{-1}$ DW)에서 가장 낮았다. 'TBC'의 GSL 함량은 91DAS(평균 $13.41{\mu}mol{\cdot}g^{-1}$ DW)에서 가장 높았고, 35DAS($0.31{\mu}mol{\cdot}g^{-1}$ DW)에서 가장 낮았다. 총 GSL 함량 중 sinigrin이 'Manchoo Collard'(133DAS)에서 57%, 'TBC'(91DAS)에서 44%로 가장 높았다. 항암효과가 뛰어난 crambene, allyl isothiocynate, indole-3-cabinol, phenethyl isothiocyanate의 전구체인 progoitrin, sinigrin, glucobrassicin, gluconasturtiin은 'Manchoo Collard'와 'TBC'의 총 GSL 함량 중 각각 94, 78%로 나타나 두 품종 모두 항암 효과를 가질 것으로 기대된다.