• KSBB Journal
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• 제11권5호
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• pp.600-605
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• 1996

Taxus brevifolia와 Taxus cuspidata 현탁세포배 양에서 taxa!의 생산에 가장 적합한 초기 접종농도 를 결정하기 위해 접종농도를 세포생체중량으로 2.5, 5 5, 7.5, 10 g/flask가 되도록 변화시켜 세포의 생장 과 taxol의 생산을 측정하였다. Taxus brevifolia의 경우 초기 접종농도가 높을수록 lag phase가 감소되 는 경향을 보였으며, 두 가지 세포주(Taxus bre­1 vifolia와 Taxus cuspidata)에서 모두 초기 접종농도 를 5 g/flask가 taxol의 생산에 최척 임을 확인할 수 있었다. Taxol 생산에 가장 척합한 초기 접종농도로 생장배지와 생산배지에 세포를 접종시켰을 때 생장 배지에서는 taxal이 검출되지 않았지만 6% 자당을 이용한 생산배지에서는 9일째 taxol이 최대로 생산 되었다가 이후 다시 줄어드는 경향을 보였다.

• 식물조직배양학회지
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• 제27권5호
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• pp.401-405
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• 2000

Influence of fructose addition to the cultivation medium on the production of taxanes and the growth of callus culture of Taxus baccata was studied. The cultures showed an ability to adjust to the substitution of some of the sucrose in the media by fructose and the fresh biomass accumulation was higher on the media containing different concentrations of fructose during the second cultivation period.

• Journal of Plant Biotechnology
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• 제2권1호
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• pp.29-33
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• 2000

To develop an efficient propagation method for yew tree, zygotic embryos were cultured under various conditions. When dissected embryos were cultured on GA$_3$ containing media, the highest germination frequency was observed on WPM medium contaning 1.0 mg/L GA$_3$. For germination of the embryos, two different conditions were compared; culturing embryos with endosperm (Method I), and 2) culturing embryos only (Method II). Maximum germination was achieved in 0.5 mg/L GA$_3$ when embryos with endosperm were cultured on the media. Of the media tested, White and WPM medium were the most suitable on germination of embryos. The abnormality of yew embryos found was observed when it cultured on GA$_3$ or culture media. About 40% of the precociously germinated embryos could be developed into full seedlings. Seedlings contained taxol in high quantity (535 $\mu\textrm{g}$/g dry weight). In vitro techniques will be sewed as a useful tool for the development of transformed root cultures and biosynthesis studies.

• 생약학회지
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• 제27권3호
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• pp.262-266
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• 1996

Undifferentiated callus were induced from the young leaves of Taxus cuspidata by treatment with various combinations of plant growth hormones. The effects of light and culturing temperature on production of baccatin III, the precursor of taxol, were studied. The contents of baccatin III in the cultured callus were analysed by HPLC. The illumination of fluorescence and administration of isoleucine, one of the possible substrates in biosynthesis of terpenoids, to the culturing media increased the production of baccetin III.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권5호
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• pp.350-354
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• 2000

The growth promotion of a Taxus brevifolia cell suspension culture was investigated using conditioning factors. The conditioning factors produced and secreted from cultured cells usually stimulate cell division and the production of secondary metabolites. Therefore, the effective incubation time for the optimal secretion of conditioning factors was firstly determined for the promotion of cell growth. Conditioned media obtained by cultivating for 2 and 5 days showed the promotion of initial cell growth during the early cell growth period. However, the positive effect of the conditioning factors on the initial cell growth did not continue because of the depletion of the medium nutrients. Accordingly, the addition of a carbon source to the conditioned medium prolonged the positive effect on the cell growth. The addition of sucrose to the conditioned medium resulted in the maximum cell density being reached 4 days earlier compared to the control group and an increased substrate yield.

• BMB Reports
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• 제38권6호
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• pp.668-675
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• 2005

A full-length cDNA encoding taxadiene synthase (designated as TmTXS), which catalyzes the first committed step in the Taxol biosynthetic pathway, was isolated from young leaves of Taxus media by rapid amplification of cDNA ends (RACE). The full-length cDNA of TmTXS had a 2586 bp open reading frame (ORF) encoding a protein of 862 amino acid residues. The deduced protein had isoelectric point (pI) of 5.32 and a calculated molecular weight of about 98 kDa, similar to previously cloned diterpene cyclases from other Taxus species such as T. brevifolia and T. chinenisis. Sequence comparison analysis showed that TmTXS had high similarity with other members of terpene synthase family of plant origin. Tissue expression pattern analysis revealed that TmTXS expressed strongly in leaves, weak in stems and no expression could be detected in fruits. This is the first report on the mRNA expression profile of genes encoding key enzymes involved in Taxol biosynthetic pathway in different tissues of Taxus plants. Phylogenetic tree analysis showed that TmTXS had closest relationship with taxadiene synthase from T. baccata followed by those from T. chinenisis and T. brevifolia. Expression profiles revealed by RT-PCR under different chemical elicitor treatments such as methyl jasmonate (MJ), silver nitrate (SN) and ammonium ceric sulphate (ACS) were also compared for the first time, and the results revealed that expression of TmTXS was all induced by the tested three treatments and the induction effect by MJ was the strongest, implying that TmTXS was high elicitor responsive.

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.213-219
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• 2004

Zygotic embryos were excised from immature and mature seeds of the Himalayan yew, Taxus wallichiana. The embryos germinated precociously when kept in darkness for 5 weeks and developed into full seedlings within 10-12 weeks. The highest rate of embryo germination ($81\%$) was obtained in modified Lloyd & McCown' s woody plant medium containing macro and micronutrients at half strength supplemented with $1\%$ activated charcoal, which supported both the best embryonic growth ($43\%$) and seedling development ($32\%$). However, the supplementation of basal media with kinetin, thidiazuron, 6-benzyl aminopurine or $GA_3$ had no effect on the germination of the embryos. The embryos derived from immature seeds germinated but the frequency of embryonic growth was better in mature seeds. Stratification of seeds effected precocious germination of embryos. Seeds kept at $4^{\circ}C$ for 1 week germinated earlier and at a higher frequency irrespective of the stage of seed maturity, while the germination rate declined with prolonged cold treatment for 1 month at that same temperature. Analysis of taxanes in germinating seedlings revealed that root tissues contained high levels of taxol, 10-deacetyl-baccatin ill and baccatin ill as compared to shoots. Thus embryo culture technique appears to overcome the lengthy dormancy requirement of T. wallichiana seeds.

• 한국산림과학회지
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• 제70권1호
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• pp.1-6
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• 1985

주목(朱木)(Taxus cuspidata Sieb. et Zucc)의 휴안지삽수(休眼枝揷穗)를 1982년(年) 4월(月) 20일(日)에 온실내(温室內)와 포지조건(圃地條件)에서 삽목(揷木), 당년(當年) 10월(月) 31일(日)에 발근개체(發根個體)를 가지고 평균발근율(平均發根率)을 계산(計算)하여 통계분석(統計分析)하였다. 본(本) 실험(實驗)의 목적(目的)은 원예적(園藝的)으로 가치(價値)를 평가(評價)받고 있음에 착안하여 가장 합리적(合理的)인 삽목조건(揷木條件)을 찾아내는 데 있다. 결과(結果)를 요약(要約)하면 다음과 같다. 1) 온실내삽목(温室內揷木)은 포지삽목(圃地揷木)에 비해 평균발근율(平均發根率)이 높았다. 이것은 처리별(處理別)의 차이(差異)에 불구하고 같은 경향(傾向)을 나타내었다. 2) IBA처리구(處理區)는 무처리구(無處理區)에 비해 더 높은 평균발근율(平均發根率)을 보였다. 온실내삽목(温室內揷木)의 경우 IBA처리구(處理區)는 평균발근율(平均發根率) 86%, 무처리구(無處理區) 23%, 포지삽목(圃地揷木)인 경우 IBA처리구(處理區) 53%, 무처리구(無處理區) 11%이다. 이것은 모두 삽수장(揷穗長) 20cm, 그리고 사토배지(砂土培地)에 삽목(揷木)되었을 경우이다. 3) 삽목배지(揷木培地)로서 사토(砂土), 양토(壤土), 갈색토(褐色土) 중(中) 온실내삽목(温室內揷木)이나 포지삽목(圃地揷木) 공히 사토구(砂土區)가 가장 높은 발근율(發根率)을 보였다. 4) 삽수장(揷穗長)이 발근(發根)에 마치는 영향을 분석(分析)한 바 15~20cm가 발근(發根)에 적당하고 25cm 이상은 발근율(發根率)이 떨어지는 것으로 나타났다. 5) 고찰(顧察)에 의한 발근양식(發根樣式)은 삽수하단절단면(揷穗下端切斷面)에 켈루스세포괴(細胞塊)가 생기고 그 안에 근기(根基)가 만들어져 근계형성(根系形成)에 관여하는 경우와 삽수(揷穗)의 경부(莖部)에 측근(側根)모양으로 나타나는 불정근(不定根)의 경우이다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1995년도 식물학심포지움 식물로부터 유용 2차대사산물의 생산 PRODUCTION OF USEFUL SECONDARY METABOLITES FROM PLANTS
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• pp.40-56
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• 1995

During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

• Journal of the Korean Wood Science and Technology
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• 제27권3호
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• pp.91-98
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• 1999

우리나라에 식재된 외래주목의 taxol 및 전구체인 10-deacetyl baccatin III(10-DAB III)의 함랑을 조사하기 위해 천리포수목원에 식재된 외래주목의 잎과 줄기로부터 추출하여 HPLC로 정량분석하였다. 외래주목의 잎과 줄기의 taxol 함량은 종간 및 종내, 그리고 부위에 따라 상당한 변이를 보였다. Taxol 함량은 T. X media cv. Vcrmulcun, T. baccata cv. Washingtonii, T. X cuspidata cv. Nana을 제외하고 잎에서 놓았으며, 전구체인 10-deacetyl baccatin III(10-DAB III)는 모든 종의 잎에서 높았다. 그 중 4월에 채집한 T. cuspidata cv. Nigra의 잎의 taxol 함량은 건물중 당 1,027 ${\mu}g$, 10-DAB III 의 함량은 건물중 당 267 ${\mu}g$로 보고된 태평양주목의 함량보다 월등히 높았다. 이종의 부위별 tax이 함량은 생장점 및 자방 조직보다 잎에서 높았으며, 이 결과들은 조직 및 세포배양의 재료로 널리 이용할 수 있을 것이다.