• Proceedings of the Microbiological Society of Korea Conference
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• 1997.04a
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• pp.62.2-62.2
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• 1997

• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.7-14
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• 1999

In order to survey anthraquinone productivity from in vitro root culture, transformed hairy roots of Rumex crispus were induced from leaf segments by infection with Agrobacterium rhizogenes strain $A_4$ and compared with adventitious roots. The optimum condition of adventitious root formation from leaf segments was 5 $\mu$M NAA added to MS medium. Mannopine was detected in the extract of hairy roots by paper electrophoresis, but not in adventitious roots. Secondary root tips of both adventitious roots and hairy roots elongated without lateral root branching in hormone free MS medium, but primary root tips showed more rapid growth with extensive lateral root branching. MS basal medium was the best for growth of the adventious roots and hairy roots for anthraquinone content. Adventitious root tips and hairy root tips cultured in liquid MS medium supplemented with 0.05 $\mu$M NAA and 0.1 $\mu$M kinetin (contained 5% sucrose) showed the maximal growth and anthraquinone content. Anthraquinone content of hairy roots was increased by the culture periods, but was reduced after 25 days of culture.

• Korean Journal of Pharmacognosy
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• v.22 no.1
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• pp.26-32
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• 1991

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 rhizogenes strain 15834. Hairy roots were induced by plasmid. Agropine and mannopine were detected in the hairy roots. The organization of hairy roots on the transactional morphology was undifferentiated. Culture on the medium containing hormone(IBA 2, kinetin 0.1mg/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.

• Journal of Plant Biology
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• v.35 no.1
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• pp.37-43
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• 1992

The hairy root culture of Raphanus sativus cv. Chungpihongsim was established by transformation with Agrobacterium rhizogenes $A_4$. The transformed roots grew well in adjusted Murashige and Skoog medium to 1/2 basal salts, pH 5.2, 3% sucrose. Agropine and mannopine, opine synthesized in the transformed tissue were detected in the extract of hairy roots. When 2, 4-D and kinetin were added in culture medium of hairy roots, the synthesis of anthocyanin was induced with disorganization of hairy root. Especially, addition of $0.45\;\mu\textrm{M}$ 2, 4-D and $2.3\;\mu\textrm{M}$ kinetin showed the maximum synthesis of anthocyanin. Pattern of anthocyanin synthesized in transformed roots was somewhat different from that of ordinary roots. However, aglycone part of all anthocyanin was identified as pelargonidin. The content of total anthocyanin in this sample was tentatively calculated 0.49 mg/g fresh weight.weight.

• KSBB Journal
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• v.9 no.2
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• pp.157-164
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• 1994

Hairy root clones of Panax ginseng were established by selection of some hairy roots formed on the leaf, stem and root segments transformed with Agrobacterium rhizogenes strain $A_4$. The transformed roots grew well in MS medium under the dark condition. To confirm the transformation with Ri-T-DNA, dot blot hybridization and opine analysis were Performed. Among four hairy roots induced from different part of ginseng, the HB3 hairy roots were examined for selection of high-saponin-producing clones. Four clones isolated from HB3 hairy root cultures displayed various phenotypes characterized by growth and total saponin content. Maximum growth was obtained for cultures of HB3-10 clone and the content of total saponin was 0.55 wt%. However, higher amount of total saponin was obtained with HB3-2 clone cultures(0.74 wt%) in spite of lower growth. Dot blot hybridization confirmed the introduction of Ri-T-DNA in the plant genome. In the opine test, agropine and mannopine were detected from all hairy root clones.

• KSBB Journal
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• v.13 no.2
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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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• v.36 no.1
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• pp.43-49
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• 1993

In order to survey possibility to produce saikosaponin from in vitro hairy root culture, culture of callus, adventitous root, and hairy root of Bupleurum falcatum L. were estabilished, and quantitative and qualitative aspects in saikosaponin extracted from these were compared with those of cultivated root. Callus grew well in MS medium containing 0.9 $\mu$M 2, 4-D. In contrast, both of adventitous root and hairy root grew well in hormone-free MS medium. However, hairy root showed more rapid growth with extensive lateral root branches, characteristics of lower content of water and softer than in adventitous root. Among the selected lines of adventitous root and hairy root were observed difference in the growth rate. Mannopine, one of opine synthesized in the transformed tissue with Agrobacterium rhizogenes. A4 were detected in the extract of hairy root lines. Pattern and content of crude saponin from adventitous and hairy root showed no difference, but somewhat difference from those of cultivated root. However, in callus, distinct production-aspect of saponin was not observed.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.123-128
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• 1995

Several factors affecting on transformation efficiency were studied to establish a Agrobacterium rhizogenes mediated system for the transformation of Populus species, and We could obtaine tansgenic plantlets expressing the introduced gene. Leaf sections were more sensitive than stem sections to kanamycin and thought to be better material for transformant screening. The bacterial density did not affect on the efficiency of transformation over the range of $4{\times}10^5{\sim}7{\times}10^9\;cfu$. The optimum period for co-cultivation was one day or shorter. Both of the optimum concentrations of cefotaxime and ampicillin in the medium were $250\;{\mu}g/ml$ for elimination of bacteria from the inoculated leaf sections. The addition of acetosyringone in the bacterial culture medium increased transformation rate, and the highest rate was obtained at $50\;{\mu}M$ of acetosyringone. The transformed galls could be selectively induced and gown on the growth regulator-free medium or on the medium containing $100\;{\mu}g/ml$ or higher contrition of kanamycin. The roots were induced from the galls incited by A. rhizogenes within 3 weeks on the growth regulator-free medium as well as on the medium containing growth regulators. The plantlets were regenerated from the galls cultured for 6 weeks on the medium containing 0.05 mg/ml of NAA and 0.5 mg/ml of BA. The expressions of the introduced opine gene in the transformed galls and plantlets were confirmed by the analysis of agropine and mannopine.

• Journal of Plant Biology
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• v.34 no.4
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• pp.261-266
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• 1991

The hairy root was induced form potato tuber disc by infection of a. rhizogenes. The detection of the agropine and mannopine by paper electrophoresis confirmed that induced hairy root was transformed by A. rhizogenes. The activity of peroxidase was the highest at 5 weeks and isozyme pattern of peroxidase revealed 3 cathodic bands and 2 anodic bands and new C4 band(pI 4.6) was observed at 7 weeks after cultivation in hairy root was isoelectric focusing. To study the effect of Ca2+ on cell wall formation in hairy root, channel blocker of Ca2+ was treated. The activity of $\beta$-glucan synthetase II(GS II) related to cell wall synthesis was inhibited by about 50% in diltiazem and flunarizine treatment than that of control, but stimulated in CaCl2 treatment. Therefore these results showed that Ca2+ might be an effective factor in the cell wall formation. The activity of GS II by NaF treatment was increased by about 30%. This result suggested that the activity GS II is changed through phosphorylation process.

• KSBB Journal
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• v.6 no.1
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• pp.27-33
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• 1991

Hairy root induced by A. rhizogenes from potato tuber (Solanm tuberosum L.) synthesized the agropine and mannopine which were demonstrated with paper electrophoresis. And the starch contents in hairy root were increased gradually following the developmental stage. But protein contents were decreased. The activity of ${\beta}-glucan$ synthetase II(GSII) which is related to the cell wall biosynthesis was stimulated in hairy root on the developmental stage. And chloropromazine did not influence the activity of GS II while verapamil inhibited about 60% of the activity GS II. Therefore, these results showed $Ca^{2+}$ to be effective factor in the cell wall formation. Isozyme pattern of peroxidase was investigated in the callus and hairy root induced from potato tuber.