• Plant Biotechnology Reports
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• 제5권3호
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• pp.255-263
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• 2011

Codonopsis lanceolata (Campanulaceae) has been used in traditional medicines, as its roots contain several kinds of triterpenoid saponin with high medicinal values. In this work, we induced transgenic hairy roots of C. lanceolata and analyzed triterpenoid saponins from the hairy roots and hairy root-derived transgenic plants. Hairy roots were obtained from leaf explants by the transformation of Agrobacterium rhizogenes R1000. Transgenic hairy root lines were confirmed by the transcriptional activities of rolA, B, C, and D genes by RT-PCR. Transgenic root lines actively proliferated on hormone-free medium but not in nontransformed roots. Hairy roots contained richer triterpenoids (lancemaside A, foetidissimoside A, and aster saponin Hb) than nontransformed roots. Transgenic plants were regenerated from the hairy roots via somatic embryogenesis. They showed phenotypic alterations such as shortened shoots and an increased number of axillary buds and adventitious roots. The transgenic plants also contained higher triterpenoid levels than wild-type plants. These results suggest that hairy roots and transgenic plants of C. lanceolata could be used as medicinal materials for the production of triterpene saponins.

• 식물조직배양학회지
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• 제27권6호
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• pp.485-489
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• 2000

생장이 우수한 인삼모상근 세포주 (KGHR-1, KGHR-5, KGHR-8) 및 6년생 인삼근의 부위별로 ginsenoside 양상 및 생성특성을 조사하였다. 인삼모상근 및 6년생 인상근에서 ginsenoslde-Rb$_1$, Rb$_2$, Rc, Rd, Re, Rf, Rg$_1$, Rg$_2$을 확인하였으며, 인삼모상근 세포주간 및 인삼근 부위별로 ginsenoside의 함량은 큰 차이를 나타내었다. 8종류의 ginsenoside함량이 가장 높은 인삼모상근은 KGHR-1 세포주로 17.42 mg/g dry wt와 함량을 나타내었다. 모상근세포주 KGHR-1은 ginsenoside-Rd, Rg$_1$을, KGHR-5는 ginsenoside-Rb$_1$, Rg$_1$을, 그리고 KGHR-8은 ginsenoside-Rd, Re을 상대적으로 많이 생성하는 특징을 지니고 있으며, ginsenoside-Rf의 생성은 매우 낮았다. 6년생 인삼근의 부위별 ginsenoside의 함량은 주근, 지근, 세근순으로 많았으며, 주근에서 ginsenoside-Rc의 생성은 ginsenoside의 50.99%로써 모상근 세포주의 4.90~6.89%보다 매우 높았다. 6년생 인삼근의 총 ginsenoside에 대한 ginsenoside-Rg$_1$의 비율은 3.43~14.18% 수준으로 주근, 지근, 세근순으로 급격히 감소하였으며, 모상관의 17.14~24.43%와 비교할 때 매우 낮은 수준을 나타내었다. 따라서 인삼모상근 배양을 통하여 특정 ginsenosides생산이 가능하리라 생각된다.

• Journal of Plant Biology
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• 제29권4호
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• pp.275-283
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• 1986

Inoculation of carrot discs with Agrobacterium rhizogenes harboring Ri plasmid resulted in transformation of cells, as revealed by the tumors and hairy roots arising from them. Measurements of IAA content using HPLC indicate that it is higher in tumors and inoculated tissues than in uninoculated tissue. A lot of meristemoids and vessel elements formed I tumor tissue and the hairy roots differentiated from meristemoids. IAA content in tumor tissues is decreased with hairy root and vessel elements formation from them. Formation of wound callus in uninoculated tissues resulted on wound healing but no formation of vessel elements and hairy roots. Tumor tissues show continuous growing on hormone free medium.

• KSBB Journal
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• 제17권6호
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• pp.525-530
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• 2002

멸종위기의 자생식물인 미치광이풀로부터 중요 의약품인 tropane alkaloid를 대량 생산하기 위한 연구의 일환으로 모상근의 최적 배양조건 및 XAD를 이용한 물질 생산 최적 조건을 확립하였다. 모상근은 표면 살균된 지하경을 1.0 mg/1 IBA가 첨가된 B5배지에 배양하여 유도하였으며, 다양한 배양조건에서 생장된 모상근의 hyoscyamine과 scopolamine 함량은 HPLC로 분석하였다. 모상근의 생장에 적합한 배지로는 SH배지로 구명되었고, tropane alkaloid 생산을 위한 최적 배지로는 l/2B5배지로 나타났다. 모상근 생장에 가장 좋은 생장조절제로는 NAA로 나타났으며, 물질생산에는 생장조절제가 첨가되지 않은 배지에서 좋았다. 모상근 생장에 적합한 탄소원으로는 3% sucrose였으며, 물질생산에는 5% scurose가 가장 좋았다. 모상근의 생장에 가장 양호한 접종농도는 0.5 g 였으며, 물질생산에는 1 g 처리구에서 좋았다. XAD 처리는 tropans alkaloid 생산성을 증가시켰으며, 배지내 물질방출을 유도하였으며, 총생산 물질의 50-80%를 회수하는 것으로 나타났다. 본 연구 결과는 미치광이풀 모상근 배양을 통한 tropane alkaloid 생산을 위한 생물공정 발전에 기여할 것이다.

• Applied Biological Chemistry
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• 제37권4호
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• pp.237-242
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• 1994

유동층 생물반응기에서의 anthocyanin 생산을 위하여 당근의 모상근배양을 검토하여 보았다. 이 생물반응기에서의 모상근의 성장은 2.5배 증가되었지만 anthocyanin 생산은 낮았다. 그러나 유동층 생물반응기에서의 anthocyanin 생산은 fungal elicitor의 처리에 의해 2.3배 향상되었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2002년도 제9차 국제심포지움 및 추계정기학술발표회
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• pp.23-23
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• 2002

We established hairy root cultures of F. esculentum transformed with A. rhizogenes for in vitro rutin production. Additionally, we describe the effects of different media and plant growth regulators on growth and rutin biosynthesis in buckwheat hairy root cultures. Excised leaves of P. tinctorium from 10-day-old seedlings were used as the explant material for co-cultivation with A. rhizogenes 15834. The hairy culture of Fagopyrum esculentum Moench. was established by infecting leaf explants with Agrobacterium rhizogenes 15834. About four to five weeks after co-cultivation with A. rhizogenes, 10 hairy roots were excised from the necrotic explant tissues. After repeated transfer to fresh medium for three months, ten clones were transferred to MS liquid culture medium. The growth and rutin production of each clone differently response to the MS liquid medium. Among these clones, H8, which had exhibited good growth rate and one of the highest rutin productivity, was selected for the following experimment.(중략)

• 한국자원식물학회지
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• 제12권1호
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• pp.1-9
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• 1999

인삼 뿌리 절편으로 부터 모상근 유기를 위한 최적 조건을 확립하고자 Agrobacterium rhirogenes와 인삼 뿌리 절편의 항생제 내성 조사 및 최적의 모상근 유도 배지를 조사하기 위하여 수행하였다. NaOCl로 인삼 뿌리를 멸균하였을 때, 오염 정도가 감소하면서 조직의 손상이 일어나지 않는 NaOCl의 농도는 7% NaOCl에서 15-20분, 9% NaOCl에서 5분으로 나타났다. 인삼근은 년수가 증가할수록 오염 정도가 심하였으며, 특히 6년근중 표피가 있는 처리구는 오염 정도가 매우 높았다. Agrobacterium의 성장억제를 위한 항생제는 tetracycline이 가장 효과적이었으며, 30mg/L 이상의 농도에서 균의 성장이 억제되었다. 하지만 30mg/L tetracycline에서 인삼 조직이 고사하였으며, cefotaxime(500mg/L), carbenicillin(500mg/L)에서 균의 성장을 완전히 억제하였으며, 조직의 손상이 일어나지 않았다. 3년근 인삼에서 모상근 유도을 위한 배지로는 1/2MS 배지에 500mg/L의 cefotaxime이 첨가된 배지가 가장 좋았으며, 인삼 뿌리 절편에 Agrobacterium을 발라주는 것 보다는 균과 공동배양할때가 절편이 좋았다. Agrobacterium접종 2주 후부터 callus가 유기되기 시작한 후, 다시 2주 후에 모상근이 유도되었다. 유도된 hairy roots는 PCR에 의하여 rol C유전자를 조사함으로서 형질전환체임을 확인하였다.

• Journal of Photoscience
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• 제8권1호
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• pp.19-22
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• 2001

It was reported that Red ginseng contains specific ginsenoside-Rg$_2$,-Rg$_3$,-Rh$_1$and -Rh$_2$, which show various pharmacological effects. However, production of these specific ginsenosides from Red ginseng is not commercially applicable because of high cost of the raw material, roots. This work was carried out to examine the production of Red ginseng specific ginsenosides from Agrobacterium-transformed hairy roots. Hairy roots were induced from 3 year-old root segment of Korean ginseng (Panax ginseng C.A. Meyer) after infection with Agrobacterium rhizogenes A4. Among many lines of hairybroots, KGHR-8A was selected. Steam heat treatment of hairy roots was resulted in the changes of ginsenoside composition. Eleven ginsenosides were detected in heat-treated hairy roots but eight in freeze dried hairy roots. In heat treated hairy root, content of ginsenoside-Rb$_1$,Rb$_2$,Rc, Rd, Re, Rf, and Rg$_1$were decreased compared to those of freeze dried hairy roots. However, heat treatment strongly enhanced the amount of Red ginseng specific ginsenogides (ginsenoside-Rg$_2$,-Rg$_3$,-Rh$_1$and -Rh$_2$). Amounts of ginsenoside-Rg$_3$,-Rh$_1$and -Rh$_2$ in heat-treated hairy roots were 2.58, 3.62 and 1.08 mg/g dry wt, respectively, but these were detected as trace amount in hairy roots without heat treatment. Optimum condition of heat treatment for the production of Red ginseng specific ginsenoside was 2 h at 105$^{\circ}C$. This result represents that Red ginseng specific ginsenoside can be producted from hairy roots by steam heat treatment.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.105-109
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• 2006

A transformed hairy root clone of Scutellaria baicalensis was established following infection with Agrobacterium rhizogenes ATCC15834. Three root clones of S baicalensis were selected by growth habit and baicalin content. The most active strain-the SR-03 clone-was examined for its growth and baicalin content under various culture conditions. The root growth and baicalin content were maximized in a Schenk and Hildebrandt medium supplemented with 4 and 6% sucrose, respectively. The accumulation of baicalin in transformed hairy roots was enhanced through exposure to various elicitors. Elicitation was attained by the addition of methyl jasmonate, salicylic acid, and various concentrations of fungal cell wall elicitors to the medium. The accumulation of baicalin in the elicited cultures ranged from 10.5 to 18.3 mg/g dry weight of the roots, which was 1.5- to 3-fold the amount attained in controls.

• 한국미생물·생명공학회지
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• 제22권4호
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• pp.395-400
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• 1994

In order to improve anthocyanin production, effects of fungal elicitation in hairy root culture were investigated. fungal elicitor prepared from Fusarium moniliforme was the best in enhancement of anthocyanin production among the eight fungal elicitors tested. The optimum treat-ment time and concentration of treated elicitor for anthocyanin production were 12 hours and 3.28 mg carbohydrates per liter medium. Also, fungal elicitor was treated to hairy root culture in flat-bottomed fluidized-bed bioreactor. The anthocyanin production of elicited culture was enhanced 227% than non-treated.