Agrobacterium rhizogense에 의해 형질 전환된 인삼의 모상근 배양에서 Ginsenoside의 생산

고경수(Kyung-Soo Ko),허인옥(In-Ok Heo),고정삼(Jeong-Sam Koh),이윤진(Woon-Jin Lee) 한국생물공학회 1990 KSBB Journal Vol.5 No.3

Agrobacterium rhizogenes 를 이용한 Atropa belladonna 의 형질전환과 Tropane alkaloid 성분에 관한 연구

한태원(Tae Wan Han),허인옥(In Ok Heo),김창민(Chang Min Kim),고경수(Kyung Soo Ko) 한국생약학회 1992 생약학회지 Vol.23 No.1

Agrobacterium rhizogenes를 이용한 까마중의 형질전환과 모상근 배양

고경수(Kyung Soo Ko),허인옥(In Ok Heo),양관팔(Kwan Pal Yang),이운진(Woon Jin Lee),김창민(Chang Min Kim),조필형(Pill Hyeong Jo) 한국생약학회 1991 생약학회지 Vol.22 No.1

The study aimed to confirm transformation, morphology, steroid alkaloidal TLC pattern and growth rate of hairy roots. The Solanum nigrum plantlets were inoculated with Agrobacterium rhixogenes strain 15834. Hairy roots were induced by plasmid. Agropine and mannopine were detected in the hairy roots. The organization of hairy roots on the transectional morphology was undifferentiated. Culture on the medium containing hormone(IBA 2, kinetin 0.1 ㎎/ℓ) altered hairy roots into callus. The growth rate of hairy roots on the NN30 liquid medium was 52 times heavier than in the original state and this was higher than on the other media. The results of TLC analysis indicated that the hairy roots produced steroidal alkaloids resembling those of normal roots.

해녀콩의 유식물 (幼植物) 자엽내 BAρNAase 의 성질

고석찬(Suck Chan Koh),허인옥(In Ok Heo),권영명(Young Myung Kwon) 한국식물학회 1994 Journal of Plant Biology Vol.37 No.2

N-α-benzoyl-DL-arginine ρ-nitroanilide hydrolase (BAρNAase) was partially purified and characterized from the cotyledons of Canavalia lineata. The overall purification and final recovery were 77.5-fold and 7%, respectively. The apparent mol wt of the native enzyme was estimated as 200 kD from the gel filtration and this enzyme was revealed as one of the gelatin-hydrolyzed proteases by activity staining with gelatin-containing gel. The optimum pH for the hydrolysis of N-α-benzoyl-DL-arginine ρ-nitroanilide (BAρNAase) was about 9.5. This enzyme catalyzed the hydolysis of BAρNA with Vmax of 15.5 unit/mg and Km value of 1.6 mM which were different from them of bovine-pancreatic trypsin. The catalytic activity of this enzyme was strongly inhibited by phenylmethanesulfonyl fluoride (PMSF) but insensitive to aprotinin, indicating that it was a serine proteinase but not a trypsin-like one. And this enzyme was slightly stimulated by Ca^2+ and mg^2+ but remarkably inhibited by Mn^2+, HG^2+ and Zn^2+.

녹차(綠茶) 개발에 관한 연구 : 조기 육묘법을 중심으로 early propagation

허인옥 濟州大學校 基礎科學硏究所 2001 基礎科學硏究 Vol.14 No.1

This study was conducted to development of early propagation method on Thea sinensis L. The results obtained are summerized as follow: 1) Auxin treatment was effective both the softwood and hardwood, especially was effective in treatment of IBA 40000 ppm. 2) Everage of new shoot length on the 29th of Jun to the hardwood was 6.4 em and to the softwood was 1.0 cm. 3) In survial ratio, the softwood was 96%, while the hardwood was 83%. 4) In march, the polyethylene cover over the hardwood can be shorten propatation period.

Agrobacterium rhizogenes에 의해 유도된 Atropa belladonna 毛狀根의 배양과 Tropane Alkaloid의 成分에 관한 硏究

許仁玉,韓太完 제주대학교 1993 논문집 Vol.37 No.-

. The study was aimed to confirm transformation, tropane alkaloid production and growth rate of hairy roots. Hairy roots were induced by inoculation of stem of sterile plants of Atropa belladonna with Agrobacterium rhizogenes strain 15834, A4, R1000. Agropine and Mannopine were detected in the hairy roots. The growth rate of hairy roots according to medium and sucrose concentration was higher in hairy roots induced by strain 15834 than other strains. especially, the hairy roots induced by strain 15834 produced 113.5g ger 1 in NN 30 liquid medium cultured for 40 days. Besides, the production of tropane alkaloid was 0.086%~0.628% per dry weight according to strain and produced 0.628% in hairy roots induced by R1000.

溫州蜜柑(Cirus unshiu Marc.) 葉內 Abscisic Acid 含量의 李節的 變化와 器官別 脫離에 관한 硏究

許仁玉,申昌勳 제주대학교 1991 논문집 Vol.33 No.-

溫州蜜柑(Citrus unshiu Marc.) 葉內 Abscisic acid(ABA) 含量의 季節的 變化 및 器官別 脫離에 따른 ABA 含量의 差를 HPLC로 分析한 結果는 다음과 같다. 1. 葉內 ABA 含量의 季節的 變化는 1~4月에 비교적 낮았으며, 7月에 가장 높은 반면 8月에 급격히 減少한 후, 12月까지 완만하게 增加하였다. 2. 꽃의 發達에 따른 ABA 含量의 變化는 開花후 增加하는 傾向을 나타냈고, 특히 脫離되는 時期에 가장 많았다. 3. 잎과 과실의 脫離에 따른 ABA 含量은 脫離直前의 離層部位에서 가장 많았다. The present study was conducted to investigate seasonal changes of ABA content in the leaves and differences of ABA content according to abscission in organs of Citrus unshiu Marc. ABA content was determined by HPLC. 1. Seasonal changes of ABA content in liaves were relatively low from January to April and were the highest in July. On the other hand, ABA content was decreased rapidly in August and then increased gradually until December. 2. The content of ABA during development of flower increased after blooming and was especially the highest in abscission stage. 3. The content of ABA according to abscission of leaves and fruits were the highest in abscission zone before abscission.

Atropa belladonna 모상근의 생장량과 tropane alkaloid 생산에 관한 연구

허인옥,오순자,한태완,김철수 濟州大學校 基礎科學硏究所 1996 基礎科學硏究 Vol.9 No.1

Agrobacterium rhizogenes strain R1000에 의해 유도된 Atropa belladonna 모상근을 배지별, NAA, IBA, IAA, 2,4-D를 농도별(0.01-100 mg/l)로 첨가한 NN액체배지에서 생장량과 tropane alkaloid 함량을 HPLC로 이용하여 정량하였다. 모상근 배양 30일째의 각 auxin의 농도별에 따른 생장율은 고농도보다 저농도일때 대체적으로 양호하였으나, 0.1mg/1 2,4-D농도에서는 callus화 하였고 10mg/l 2,4-D, 1mg/l IBA일때 2.96g, 2.36g으로 대조구보다 2.3배의 생장량을 보였다. 농도별에 따른 Tropane alkaloid 함량은 atropine이 대조구 0.141%, 원식물체가 0.059%로 나타났으며, 0.01mg/l NAA와 0.1mg/l IAA에서 배양한 모상근이 각각 1.378%, 1.814%의 생산을 보여 대조구와 원식물체보다 9-30배의 생산을 보였다. 또한 scopolamine 함량은 대조구와 원식물체는 각각 0.109%. 0.050%에 비해 0.01mg/l NAA에서 배양한 모상근에서 0.505%로 대조구 원식물체보다 4-10배의 생산량을 보였다. The study aimed to measure growth rate of Atropa belladonna hairy root cultured under medium, various auxin(NAA, IBA, IAA, 2.4-D) and auxin concentration(0.01-100mg/l) and investigated tropane alkaloid content using HPLC. The growth rate of hairy root cultured for 30 days in MS and NN liquid medium were higher in NN medium as 3.51g than MS medium, growth rate of belladonna hairy root cultured in NN medium added various auxin and auxin concentration was the most effective in low concentration than high concentration, especially, was best in the medium supplemented with 0.01mg/l 2,4-D. Atropine content of hairy root cultured in 0.01mg/l and 0.1mg/l IAA produced 9-30 times than control and normal plant. scopolamine measured each 0.109%, 0.050% in control and normal plant, hairy root cultured in 0.01mg/l NAA produced 4-10 times than control and normal plant.

차나무 callus 의 catechin류 함량분석

오순자,허인옥,한태완 濟州大學校 基礎科學硏究所 1996 基礎科學硏究 Vol.9 No.2

본 연구는 차나무의 신초를 이용하여 기내에서 여러가지 배양조건에 따른 캘러스의 catechin류 함량을 분석하여 생산성이 높은 배양조건을 찾고자 실시하였다. 신초배양시 캘러스는 0.5mg/L NAA와 1mg/L BA 조합 처리구에서 가장 잘 유도되었다. 이 조건에서 유도된 캘러스를 가지고 thiamin-HCI, PVP, taxifolin과 quercetin을 농도별로 처리하여 4주간 배양하여 생장 및 catechin류 함량을 조사하였다. 캘러스의 생장은 30mg/L thiamin-HCL, 200mg/L PVP, 1 mM taxifolin, 0.01 mM quercetin처리시 대조구에 비해 2~3배의 생장를 보였다. 차나무 잎의 주성분인 catechin류를 분석하고자 신초와 여러조건에서 배양된 캘러스를 가지고 분석한 결과를 보면, 캘러스에서 높은 함량을 보였다. 또한 신초에서의 catechin류는 EGC, EGCG, ECG등은 검출되었으나, EC는 검출되지 않았다. 그러나 배양된 캘러스에서는 4가지 성분이 모두 검출되었다. 각각의 성분들의 최적 배양조건을 보면, EGC는 500mg/L PVP, EC는 100mM quercetin, EGCG는 10 mg/L thiamin-HCI, ECG는 10mM taxifolin처리구에서 가장 높은 함량을 보였다. This study aimed to measure the growth of callus cultured under MS medium with thiamin-HCI as a vitamine. PVP as the antioxidant. taxifolin and quercetin as precursors of catechins and to select the clone producing catechins highly. Callus induction from Thea sinensis young leaf was great in the MS medium with 0.5mg/L NAA and 1 mg/L BA. When components of callus were compared with those of young leaf. Four kinds of catechins(EC, EGC, EGCG, ECG) were detected in cultured callus whereas EC not in young leaf. Catechins were detected most in the medium with 10mg/L thiamin-HCI. Expecially, EC was most as a 6.39mg/g. The content of catechins in PVP treatment was not different each other but was higher in 100 mg/L and 200 mg/L PVP than in control. When the callus was cultured in the medium with taxifolin and quercetin the caffeine content was lower than in the medium with not them. Also, the content of catechins in taxifolin and quercetin treatment was not different each other but it was highest in 10mM taxifolin and 100mM quercetin treatment.

삼보감(Citrus sulcata Hort. et Takahashi) 유실물체에서 유도된 캘러스의 체세포배 형성과 식물체 재분화

박수영,허인옥,부지현,한태완,송관필 濟州大學校 基礎科學硏究所 1998 基礎科學硏究 Vol.11 No.1

삼보감에 있어서 기내 체세표배 발생을 통한 다량증식을 도모하기 위해서 캘러스 증식 및 체세포배 발생, 배로부터 식물체 재분화에 미치는 배지 및 Polyamine, 생장조절제의 효과를 구명하기 위해 실시하였다. 삼보감 과실에서 채집된 종자를 호르몬이 첨가되지 않은 MS배지에서 무균발아시켰다. 발아된 유식물체의 줄기에서 캘러스를 유도하기 위해 NAA와 BA가 첨가된 MT 배지에서 배양하였고 5mg/L 2,4-D와 1mg/L BA가 첨가된 배지에서 계대배양하였다. 캘러스를 증식시키기 위한 배양조건 설정으로 배지별과 Polyamine(Spermidine, Spermine, Putrecine) 농도별(0 - 1mM)에 따른 생장량을 좌하였다.배지별에 따른 생장율은 MT배지에서 0.968g(fr wt)으로 가장 높았고, Polyamine의 영향은 0.01 mM Putrecine 처리구에서 0.78g으로 가장 높게 나왔다. 배발생 캘러스는 0.1mg/L NAA와 0.5mg/L BA가 첨가된 MT배지에서 유도하였으며 유되된 체세포배를 식물체로 재분화시키기 위해서 0.1mg/L NAA와 1mg /L BA가 첨가 된 MT배지에서 배양하였다. This study was performed to investigate the culture condition induction of somatic embryo and plant regeneration in callus induced from Citrus sulcata leaf and stem as a basic research for breeding of new plant. The seeds of Sambokam were germinated in hormone free MS medium under sterile condition. Callus induced form stem and leaf geminated young plant in MT supplemented with NAA plus BA and subcultured in MT supplemented with 5 ㎎/ L 2,4-D and 1 ㎎/L BA. As a investigation of culture condition for callus proliferation, growth rate of callus were investigated in various medium and polyamine concentrations. The effect of medium was most effective in MT medium as 0.968g and polyamine was most effective of in 0.01mM putrecine among various concentration. Formation of embryogenic callus induced from MT medium containing 0.1 ㎎/L and 0.5 ㎎/L BA. The geminated embryos developed to complete plantlet when cultured on MT medium supplemented with 0.1 ㎎/L NAA and 1㎎/L BA.