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Jie, Eun-Yee,Kim, Suk-Weon,Jang, Hye-Rim,In, Dong-Su,Liu, Jang-Ryol The Korean Society of Plant Biotechnology 2011 식물생명공학회지 Vol.38 No.1
This study describes the effect of myo-inositol on sustained cell division and plant regeneration from cotyledon-derived protoplast of cabbage (Brassica oleracea var. capitata). Freshly isolated protoplasts were cultured in modified Murashige and Skoog (MS) medium removed ammonia ions and containing $0.4\;mg\;l^{-1}$ thiamine HCl, $100\;mg\;l^{-1}$ myo-inositol, $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and several concentrations of myo-inositol (2, 4, 6, 8, 10% (w/v)) as an osmotic stabilizer. After 3 weeks of culture in the dark at $25^{\circ}C$, the plating efficiency of cabbage protoplasts reached to $22.5{\pm}2.9%$ when cultured in modified MS medium supplemented with $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and 8% (w/v) of myo-inositol at a density of $2{\times}10^5$ protoplasts/ml. Rapidly growing cell colonies after 3 weeks of culture were transferred to the same culture medium removed osmoticum. To induce shoot regeneration from calluses, calluses with about 2 mm in diameter were transferred to the MS medium containing $2\;mgl^{-1}$ BA and $0.5\;mgl^{-1}$ NAA. After further three weeks of incubation onto the medium in the light, green shoots were formed on the surface of calluses at a frequency of 30%. Upon transfer to half-strength MS basal medium, roots were formed onto the bottom of regenerated shoots without auxin treatments. These regenerated plantlets were successfully acclimatized to soil transfer, grown to normal mature plants. The cabbage protoplast culture system established in this study could be applied for production of somatic hybrids or cybrids by asymmetric protoplast fusion and mass proliferation of elite somatic clones of cabbage.
주수지,최승희,Jie Eun Yee,이옥란,Kim Suk Weon 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.6
Phytosulfokines (PSK) belong to a new class of peptide growth factors involved in intercellular signaling in plants. In this study, we examined the efect of PSK on cell division and subsequent adventitious shoot formation in protoplast-derived callus culture of Nicotiana benthamiana. Protoplast cultures and protoplast-derived calluses of N. benthamiana were treated with diferent concentrations of PSK (0, 0.01, 0.03, 0.1, 0.3, and 1 µM). Cell division efciency was the highest at 32% in the 0.1 µM PSK treatment, which was about two times higher than that of control treatment. PSK concentrations<0.1 µM were more efective in inducing cell division in the protoplasts than those>0.3 µM. In contrast, higher concentrations of PSK were more efective in inducing adventitious shoot formation in protoplast-derived calluses. The frequency of adventitious shoot formation was the highest at 8.5% when the calluses were treated with 1 µM PSK. qRT-PCR analysis showed an increase in the expression of genes involved in cell division and diferentiation: G1/S specifc cyclin (CYCD3-1), cytokinin dependent kinase (CDK), and WUSCHEL (WUS) at 0.1 µM PSK treatment. These results indicate that PSK plays a signifcant role in promoting cell division and adventitious shoot formation in N. benthamiana. The results obtained in this study could be applied to promote cell division and plant regeneration in protoplast cultures of diverse recalcitrant crop plant species.
Kwon, Yong-Kook,Jie, Eun Yee,Sartie, Alieu,Kim, Dong Jin,Liu, Jang Ryol,Min, Byung Whan,Kim, Suk Weon Elsevier 2015 Food chemistry Vol.166 No.-
<P><B>Abstract</B></P> <P>To determine whether or not FT-IR spectroscopy could be used for taxonomic and metabolic discrimination of African yam lines, tuber samples from African and Asian yam species were subjected to FT-IR. Most remarkable spectral differences between African and Asian yams were found in the 1750–1700cm<SUP>−1</SUP> region, polysaccharide (1200–900cm<SUP>−1</SUP>) and protein/amide I and II (1700–1500cm<SUP>−1</SUP>) regions of FT-IR spectra. A hierarchical dendrogram based on partial least square-discriminant analysis (PLS-DA) of FT-IR data from 7 African yam species show phylogenetic relationship. In addition, the content of dioscin, a steroidal saponin found in yam tuber, was predicted using a PLS regression model with regression coefficient <I>R</I> <SUP>2</SUP> <I>= </I>0.7208 indicated that prediction model had average accuracy. Thus, considering these results we suggest that FT-IR combined with multivariate analysis could be applied as a novel tool for metabolic evaluation and high-throughput screening of African yam lines with higher content of dioscin.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Rapid metabolic discrimination of African yams was possible by FT-IR spectroscopy. </LI> <LI> Hierarchical dendrogram from African yams shows probable taxonomic relationship. </LI> <LI> Prediction for dioscin content from FT-IR spectra shows an average accuracy. </LI> <LI> FT-IR could be applied as a novel tool for metabolic evaluation of yam tubers. </LI> </UL> </P>
FT-IR 스펙트럼 데이터의 다변량 통계분석을 이용한 고기능성 아프리칸 얌 식별 및 기능성 성분 함량 예측 모델링
송승엽(Seung Yeob Song),지은이(Eun Yee Jie),안명숙(Myung Suk Ahn),김동진(Dong Jin Kim),김인중(In-Jung Kim),김석원(Suk Weon Kim) 한국원예학회 2014 원예과학기술지 Vol.32 No.1
본 연구에서는 UV-VIS spectrophotometer를 이용한 total carotenoids, flavonoids, phenolics 함량 데이터와 FT-IR 스펙트럼 데이터를 다변량통계분석법을 통하여 기능성 성분 함량이 높은 아프리칸 얌 고속 선발 시스템을 구축하였다. 62개 아프리칸 얌의 total carotenoids 함량은 0.01-0.91μg・g<sup>-1</sup> dry wt 나타냈다. Total flavonoids와 phenolics 함량은 12.9-229.0μg・g-1 dry wt와 0.29-5.2mg・g-1 dry wt로 각각 나타났다. 아프리칸 얌은 FT-IR 스펙트럼상의 1700-1500, 1500-1300, 1100-950cm<sup>-1</sup> 부위에서 중요한 스펙트럼 변화가 나타났다. 이 부위는 각각 amide I과 II을 포함하는 아미노산 및 단백질계열의 화합물, phosphodiester group을 포함한 핵산 및 인지질 그리고 단당류나 복합 다당류를 포함하는 carbohydrates 계열의 화합물들의 질적, 양적 정보를 반영하는 부위이다. PCA 분석과 PLS-DA 분석에서 62개 아프리칸 얌은 유연성이 높은 종으로 3개의 그룹을 형성하였다. 아프리칸 얌의 FT-IR 스펙트럼 데이터와 UV-VIS spectrophotometer 을 이용한 total carotenoids, flavonoids, phenolics 함량 데이터 간에 PLS regression 분석하였다. Total carotenoids, flavonoids, phenolics 함량 성분의 실측 값과 예측 값간에 상관계수(R²)가 각각 0.83, 0.86, 0.72로 나타났다. 이 결과, 아프리칸 얌으로부터 FT-IR 스펙트럼을 이용한 total carotenoids, flavonoids, phenolics 함량 예측이 가능하였다. 본 연구에서 확립된 대사체 수준에서 아프리칸 얌의 유용 기능성 성분 함량 예측 모델링을 통해 품종, 계통의 신속한 선발 수단으로 활용이 가능할 것으로 예상된다. We established a high throughput screening system of African yam tuber lines which contain high contents of total carotenoids, flavonoids, and phenolic compounds using ultraviolet-visible (UV-VIS) spectroscopy and Fourier transform infrared (FT-IR) spectroscopy in combination with multivariate analysis. The total carotenoids contents from 62 African yam tubers varied from 0.01 to 0.91 μg・g<sup>-1</sup> dry weight (wt). The total flavonoids and phenolic compounds also varied from 12.9 to 229 μg・g<sup>-1</sup>and from 0.29 to 5.2 mg・g<sup>-1</sup> dry wt. FT-IR spectra confirmed typical spectral differences between the frequency regions of 1,700-1,500, 1,500-1,300 and 1,100-950 cm<sup>-1</sup>, respectively. These spectral regions were reflecting the quantitative and qualitative variations of amide I, II from amino acids and proteins (1,700-1,500 cm<sup>-1</sup>), phosphodiester groups from nucleic acid and phospholipid (1,500-1,300 cm<sup>-1</sup>) and carbohydrate compounds (1,100-950 cm<sup>-1</sup>). Principal component analysis (PCA) and subsequent partial least squarediscriminant analysis (PLS-DA) were able to discriminate the 62 African yam tuber lines into three separate clusters corresponding to their taxonomic relationship. The quantitative prediction modeling of total carotenoids, flavonoids, and phenolic compounds from African yam tuber lines were established using partial least square regression algorithm from FT-IR spectra. The regression coefficients (R²) between predicted values and estimated values of total carotenoids, flavonoids and phenolic compounds were 0.83, 0.86, and 0.72, respectively. These results showed that quantitative predictions of total carotenoids, flavonoids, and phenolic compounds were possible from FT-IR spectra of African yam tuber lines with higher accuracy. Therefore we suggested that quantitative prediction system established in this study could be applied as a rapid selection tool for high yielding African yam lines.
Kim Young-Cheon,안우석,차아라,Jie Eun Yee,Kim Suk Weon,Hwang Byung-Ho,이상협 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.1
Broccoli, a Brassica crop, is an important vegetative crop because of its nutritional components and beneficial phytochemi- cals. Glucoraphanin (GR), a major glucosinolate (GSL) in broccoli, is converted by hydrolyzation of the endogenous enzyme, myrosinase, into sulforaphane (SR), which protects the body against a variety of chronic diseases. Despite their economic importance, biotechnological approaches for increasing GR content in Brassica species are still limited. The main objective of this study was to develop a GR-rich broccoli cultivar using the CRISPR/Cas9-mediated DNA-free genome-editing technique. It is considered that MYB28 is one of the key genes involved in the accumulation of GSL levels in broccoli. Furthermore, with increased GSL levels by introgression of MYB28 from wild species, B. villosa showed a 9 bp deletion in exon 3, leading to one amino acid substitution and the deletion of three amino acids. Therefore, we considered the 9 bp deletion to be the most significant change in GR-rich broccoli and conducted Cas9 protein and single-guide RNA transfection into broccoli protoplasts for editing the flanking sequence of the 9 bp deleted MYB28 gene. Finally, increased GR content was observed in broccoli regenerated from protoplasts with specifically edited MYB28.
쥐에서 dimethylhydrazine 처리와 식이지방 종류에 따라 HMG CoA reductase 활성도와 bile acid 및 중성 sterol 배설에 미치는 영향
김영현,박현서,지은이 한국지질학회 1999 韓國脂質學會誌 Vol.9 No.2
The study was designed to observe the effect of 1,2-dimethylhydrazine (DMH) and the type of dietary fat on plasma lipids, hepatic microsomal level of HMG-CoA reductase, membrane fluidity, fecal excretion of bile acids and neutral sterols in rats. Male Sprague Dawley rats, at 7 wks of age, were divided into 2 groups (control and DMH), and each group was subdivided into corn oil and perilla oil groups, and fed experimental diet for 20 wks. Each rat of DMH group was intramuscularly injected with DMH (total dose of 180 ㎎/㎏). Regardless of the type of dietary fat, DMH treatment had shown significant reduction in plasma cholesterol and TG levels. Perilla oil rich in n-3 α-linolenic acid significantly reduced plasma cholesterol level compared with corn oil rich in n-6 linoleic acid, but plasma TG level was not significantly reduced by perilla oil. There was no significant effect on hepatic microsomal level of cholesterol and HMG CoA reductase activity by DMH and dietary oil. Microsomal PL content was significantly increased by perilla oil, which resulted in the reduction of cholesterol/phospholipid (Chol/PL) ratio and increased membrane fluidity. DMH treatment significantly increased the excretion of secondary bile acid (doxycholic and lithocholic acid), but perilla oil significantly reduced its excretion. DMH treatment had shown the trends of increasing the excretion of neutral sterols, but perilla oil significantly decreased its excretion in DMH group. In conclusion, plasma cholesterol and TG levels were reduced by DMH treatment which might be from the increased requirement of nutrients for tumor growth. However, perilla oil could reduce fecal excretion of secondary bile acid (colon cancer promotor) which was increased by DMH, and also reduced plasma cholesterol level by increasing hepatic microsomal fluidity. Therefore, it could be recommended to use more perilla oil in meal preparation to prevent cardiovascular disease and colon cancer.