• Journal of Plant Biology
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• 제36권3호
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• pp.225-231
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• 1993

Transformed hairy roots of Scopolia parviflora, producing tropane alkaloids and native to Korea, were obtained following infection of rhizome segments with Agrobacterium rhizogenes A4. Each root tip induced from inoculum sites excised and cultured in MS agar or liquid medium. About seventy of hairy root clones were established. Among them, several fast growing hairy root clones were examined for alkaliod content. Two dimensional TLC analysis showed that the tropane alkaloid pattern of hairy root was more complicated than that in the rhizome of mother plant. On the other hand, some hairy root clones did not produce scopolamine and hyoscyamine. In HPLC analysis, some hairy root clones yield higher levels of scopolamine and hyoscyamine than those of mother plant rhizome which used for infection. Scopolamine and hyoscyamine were identified by comparison of their retention times and of their spectra data with those of authentic compounds.

• KSBB Journal
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• 제13권2호
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• pp.155-161
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• 1998

The hairy roots of Crotalaria sessiliflora were induced from the tissue segments infected with Agrobacterium rhizogenes ATCC 15834. The induced hairy roots were subjected to paper electrophoresis fro the detection of opine-positive clones which were considered to have been transformed. Mannopine and agropine were presented in hairy root clones while mannopine was presented in two hairy root clones. Eight hairy root clones were selected and cultured in MS, B5 and WP media. Each of hairy root clones was showed a difference in branch pattern and growth rate. The best culture medium and culture conditions of hairy roots were in $\frac{1}{2}$MS(3% sucrose, pH 5.7) liquid medium at 25$^\circ C$, 70 rpm under dark, the growth rate in $\frac{1}{2}$MS liquid medium was increased with 210-fold more than that of inoculated hairy roots and with 2-fold more than that in MS liquid medium. Also, the adequate condition for hairy root growth was such that concentration of KH$_2PO$_4 was 1.25mM and the ratio of NH${_4}{^+}$ : NO${_3}{^-}$ was 1 to 3 in MS medium. The presence of pyrrolizidine alkaloids, monocrotaline, in the hairy roots was detected by TLC.

• Journal of Plant Biology
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• 제39권3호
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• pp.161-166
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• 1996

The hairy root cultures of Centella asiatica were established by infection leaf explants with Agrobacterium rhizogenes A4, 15834 in 1/2 Murashing and skoog liquid medium supplemented with 50 $\mu$M acetosyringone. The induced hairy roots were subjected to paper electrophoresis for the detection of opine and opine-positive clones which were considered to have been transformed. Five hairy root clones were selected according to the different bacterial strains used, growth rate and pattern. Among media tested, MS basal medium substituted phosphate concentration by 2.5mM K2HPO4 showed the highest growth rate in the dark condition.

• KSBB Journal
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• 제9권2호
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• pp.157-164
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• 1994

인삼의 잎, 줄기, 뿌리 절편에 Agrobatriumrhizogenes(stain $A_4$를 접종하여 형질이 전환된 조직(모상근)을 유도, 배양한 후 saponin 고생산능 clones을 선발하였으며, 아울러 형질전환 여부를 dot blot hybridization 과 opine 분석으로 확인하였다. 여러 부위에서 유도된 모상근은 모두 암조건의 MS 배지에서 활발히 생장하였으며, 이들 중 HB3 clone을 이용하여 동질화딘 모상근 clon으로 배양한 후 saponin 고생산능 clon을 선발한 결과 HB2-10 clone이 최대의 생장을 보였으며 총 saponin함량은 0.55wt%로 나타났다. 그러나 총 saponin함량이 가장 높은 clone은 비교적 생장이 느린 HB3-2 clone이었으며 0.74wt%의 함량을 나타내었다. 아울러 dot blot hybridization 결과 Ri-T-DNA가 식물체의genome 내로 삽입되어 있었으며, opine확인결과 모든 모상근 clone으로부터 argropine과 mannopine이 검출되었다.

• 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)

• 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.

• 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.

• Journal of Plant Biology
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• 제38권4호
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• pp.337-341
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• 1995

Hairy roots of Korean ballon flower (Platycodon grandiflorum A. DC) were induced from the root tissues infected with Agrobacterium rhizogenes ATCC 15834. Growth and polyacetylene [lobetyol (1), lobetyolin (2) and lobetyolinin (3)] production fo ten hairy root clones cultured in 1/4 Gamborg B5 (B5) liquid medium were determined. One selected hairy root clone (D6) grew well in hormone free-B5 liquid medium and showed maximum content of polyacetylenes at week 6 for 1 (0.375% dry wt) and at week 7 for 2 and 3 (3.030% and 0.206% dry wt, respectively) whose levels were much higher than those of the intact plant root (1:0.019%, 2:0.077% dry wt, 3 was not detected).

• 한국약용작물학회지
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• 제12권2호
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• pp.154-162
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• 2004

Hairy roots were induced from Korean ginseng (Panax ginseng C. A. Meyer) root explants and studied for their gene expression. A total of 3,000 ESTs (expressed sequence tags) from ginseng hairy root were determined and about 2,700 ESTs have a length of readable sequence, which result in 1,352 unique ESTs sequences. The 879 ESTs showed significant similarities to known nucleotide or amino acid sequences in other plant species, which were divided into eleven categories depending upon gene function. The remaining 473 sequences showed no significant matches, which are likely to be transcripts or to be matched to other organisms. The results indicated that the analysis of the ginseng hairy root ESTs by partial sequencing of random cDNA clones may be an efficient approach to isolate genes that are functional in ginseng root in a large scale. Our extensive EST analysis of genes expressed in ginseng hairy root not only contributes to the understanding of the dynamics of genome expression patterns in root organ but also adds data to the repertoire of all genomic genes.

• 식물조직배양학회지
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• 제24권3호
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• pp.137-142
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• 1997

모상근의 성장을 위한 최적배지 및 tropane alkaloids함량을 위한 최적배지를 구명하기 위하여, 독말풀(Datura stramonium var. tatula Torr.)의 잎과 줄기에 Agrobacterium spp.spp.를 접종하였다. 접종된 균주 중 모상근의 유도에 효과적인 균주는 Agrobacterium tumefaciens $\textrm{A}_{4}$ T와 A rhizogenes ATCC 15834이었다. SH (Schenk and Hildebrandt, 1972) 액체배지에서 선발된 23개의 clone중 성장이 양호한 clone은 DTLA9 이었으며, tropane alkaloids의 함량이 양호한 clone은 DTLE6 이었다. 이들 두 종류의 세포주를 9종류의 서로 다른 액체배지에 각각 배양하여 성장 및 tropane alkaloids의 함량을 조사한 결과, 모상근의 성장은 두 세포주 모두 SH와 MS배지(SH > MS배지순)에서 다른 7개의 배지에서 보다 양호하였다. 한편, tropane alkaloids의 함량은 두 세포주 공히 GD와 B5 배지(GD > B5배지순)에서 다른 7개의 배지에서 보다 높게 나타났다. 따라서 SH배지에서 모상근을 성장시킨 후, GD배지에 옮겨 2단계 연속배양하면 tropane alkaloids의 함량을 높일 수 있을 것으로 확인되었다.