• 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.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.91-98
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• 2005

The hyoscyamine 6${\beta}$-hydroxylase (h6h) gene was introduced into the genome of Scopolia parviflora through the Agrobacterium rhizogenes binary vector system. The enzyme was expressed ally and tissue specific selectively in roots, resulting in five transgenic hairy root lines. The presence of the h6h gene in kanamycin-resistant hairy roots and its overexpression were confirmed by polymerase chain reaction (PCR), Northern blotting, and Western blotting, respectively. In the transgenic hairy root lines which constitutively expressed the H6H enzyme, hyoscyamine and scopolamine accumulated in high concentration. Among the transgenic hairy root lines that expressed the H6H enzyme, only two were more productive. The levels of tropane alkaloids in transgenic hairy root varied greatly: The best transgenic line (#5) contained 8.12 mg of scopolamine per g dry weight, which produced the compound three times more than wild-type root. These results suggest a possibility of improving the yield of tropane alkaloids in hairy root lines by genetic and metabolic engineering.

• 한국약용작물학회지
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• 제12권2호
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• pp.123-128
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• 2004

Hyoscyamine and scopolamine are two most common tropane alkaloids found in the Solanaceae. The pEB expression vector carrying Nspmt gene was transformed to Agrobacterium rhizogenes. The growth of transgenic hairy roots were approximately to 80% of the wild type root. Transgenic hairy roots are less developed on only their branch roots than those of the wild type root. The extracts from Nspmt transgenic hairy root lines, 3 and 5 contained between 3.52 and 4.23 mg/g dry weight as hyoscyamine and did between 5.23 to 6.40 mg/g dry weight as scopolamine. These results showed that the overexpression of the pmt gene enhanced tropane alkaloids production of S. parviflora transformed roots and this improvement affected both hyoscyamine and scopolamine production.

• 한국약용작물학회지
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• 제22권1호
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• pp.1-7
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• 2014

Codonopsis lanceolata (Campanulaceae) has been used in traditional medicines, as its roots contain several kinds of 3,28-bidesmosidic triterpenoid saponin with high medicinal values. In this study, we induced hairy root-derived transgenic plants of C. lanceolata and analyzed triterpenoid saponins from the leaf, stem and root. Transgenic plants were regenerated from the hairy roots via somatic embryogenesis. The saponins are lancemaside A, B and E, foetidissimoside A, and aster saponin Hb. Transgenic plants contained richer triterpenoids saponin than wild-type plants. Major saponin lancemaside A was the most abundant saponin in the stem from transgenic-plant, $4.76mg{\cdot}1^{-1}dry$ stem. These results suggest that transgenic plants of C. lanceolata could be used as medicinal materials for the production of triterpene saponins.

• 식물조직배양학회지
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• 제25권6호
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• pp.525-530
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• 1998

인삼의 뿌리절편, 줄기, 엽병에 Agrobacterium을 접종하여 고생장 모상근을 선발하고자 수행하였다. 인삼 뿌리 절편에서 A. rhizogenes R1000과 A. rhizogenes $A_4$에 의하여 모상근이 유도되었으며, 인삼의 줄기 및 엽병에서도 모상근이 유도되었다. 유도된 모상근은 rolC및 virC DNA절편의 확인으로 형질전환체임을 확인하였으며, 균이 제거된 모상근을 선발하였다. 선발된 모상근 300 세포주중 성장과 형태적 특징이 뚜렷한 11 모상근을 선발하였다. 이들 모상근은 주근이 비후된 것, 주근이 가는 것, 측근의 돌기가 많은 것, 측근의 생장이 높은 것 등의 특징을 나타내었다. 이들 11 모상근중 주근이 가늘고 측근의 생장이 왕성한 형태가 모상근의 생장이 좋았으며, 주근이 비후되고 측근의 돌기가 많은 형태는 생장이 낮았다. 본 연구에서 인삼 뿌리절편으로부터 고생장 모상근인 KGHR-1, KGHR-5, KGHR-8을 선발하였다.

• 한국약용작물학회:학술대회논문집
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• 한국약용작물학회 2009년도 심포지엄 및 춘계학술발표회
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• pp.257-258
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• 2009

• KSBB Journal
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• 제21권2호
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• pp.123-128
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• 2006

황기의 형질전환된 모상근으로부터 약리물질인 astragalosides I, II, 그리고 III을 효율적으로 생산하기 위하여 다양한 물리화학적 조건들 즉, 배지, 초기 당농도, 교반속도, 초기 접종농도, 광조사, 그리고 chitosan 처리에 한 elicitation 효과를 조사 하였다. 선발된 AG-04 clone의 성장과 astragalosides의 함량은 4종의 배지들 중 SH배지에서 가장 좋았으며, 배지에 첨가하는 초기 당 농도는 3%와 6%에서 각기 건물중과 astragalosides의 함량이 가장 높게 나타났다. 또한, 초기 접종량은 50 ml 배지에 500 mg(FRW)씩을 주입하는게 가장 효과적이었으며, 18시간 광조사가 되는 Shaking incubator에서 90 rpm과 120으로 교반시켰을 때 모상근의 성장과 astargalosides의 함량이 높게 나타났다. 배양 2주 후 chitosan의 처리는 모상근의 성장에는 크게 영향을 미치지 않았으나, 100 mg/L 처리구에서 대조구의 약 2.1배의 astragalosides 함량 증가를 가져왔다. 이와같은 연구결과는 황기의 대량배양을 통한 saponins 생산 연구에 효율적으로 활용될 수 있을 것으로 본다.

• The Plant Pathology Journal
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• 제35권6호
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• pp.692-697
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• 2019

Agrobacterium rhizogenes-mediated transformation of sugar beet hairy roots expressing single-chain variable fragment (scFv) was exploited to evaluate the efficacy of four antibody-based constructs for interfering with the Beet necrotic yellow vein virus infection. The scFv specific to a major coat protein of virus, p21, was targeted to various cellular compartments including the cytosol (pIC and pICC constructs), apoplast (pIA), and mitochondrion (pIM). After mechanical virus inoculation, most of the hairy root clones expressing scFv in the cytosol displayed low virus titers while the majority of transgenic hairy root clones accumulated antibody in outer membrane of mitochondria or apoplast were infected. This hairy root system provided an efficient and rapid approach to initially investigating root disease resistance like rhizomania prior to transform whole recalcitrant plants such as sugar beet.

• 한국자원식물학회지
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• 제11권2호
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• pp.176-176
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• 1998

This study was carried out to obtain the information for growth characteristics of crown gall tumor and hairy root transformed by Agrobacterium spp,. on the media with phytohormones, casein hydrolysate and activated charoal. Crown gall tumors and hairly roots were formed respectively on potato tuber discs infected by tumerfaciens A ch 5 and A.rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. PCR analysis of Rol C and Vir C gene fragments confirmed that crown gall root was prompted on the medium containing 2,4-D 2mg/l with casein hydrolysate lg/l. The survival ration of crown gall tumor callus derived from potato increased on medium containing the activated charcoal 0.5∼0.2mg/l because of the prevention, on the other hand, hairly roots were necrosis on the same medium. Callus derived from hairly root were excellently grown for a short time by suspension culture on liquid medium containing 2.4-d 2mg/L and casein hydrolysate lg/l.

• 한국자원식물학회지
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• 제11권2호
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• pp.179-187
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• 1998

This study was carried out to obtain the information for growth characteristics of crown gall tumor and hairy root transformed by Agrobacterium spp,. on the media with phytohormones, casein hydrolysate and activated charoal. Crown gall tumors and hairly roots were formed respectively on potato tuber discs infected by tumerfaciens A ch 5 and A.rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. PCR analysis of Rol C and Vir C gene fragments confirmed that crown gall root was prompted on the medium containing 2,4-D 2mg/l with casein hydrolysate lg/l. The survival ration of crown gall tumor callus derived from potato increased on medium containing the activated charcoal 0.5∼0.2mg/l because of the prevention, on the other hand, hairly roots were necrosis on the same medium. Callus derived from hairly root were excellently grown for a short time by suspension culture on liquid medium containing 2.4-d 2mg/L and casein hydrolysate lg/l.