• 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.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.137-142
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• 1997

In order to elucidate the optimum medium on the growth and tropane alkaloid production of hairy root, hairy root were induced by inoculating leaf and stem of Datura stramonium var. tatula Torr. with Agrobacterium spp. Both Agrobacterium tumefaciens $\textrm{A}_{4}$ T and A, rhizogenes ATCC 15834 among tested strains were effective on hairy root formation. Among 23 clones selected in SH (Schenk and Hildebrandt, 1972) liquid medium, DTLA9 clone was shown fast growth of hairy root and DTLE6 clone was shown high level production of tropane alkaloids. When both DTLA9 and DTLE6 clones were cultured in the GD (Gresshoff and Doy, 1972) medium, alkaloid production was higher than in 8 tested media. It was elucidated that optimum medium for root proliferation and for tropane alkaloid production is SH, GD medium, respectively.

• Korean Journal of Medicinal Crop Science
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• v.15 no.1
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• pp.46-50
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• 2007

Transformed hairy roots were induced from in vitro grown plantlets of Trichosanthes kirilowii by infection with Agrobacterium rhizogenes strain ATCC15834. Transformed hairy roots exhibited active growth with high branching of roots on plant growth regulators-free medium. Cloned line (TR-03) of hairy root was tested for its growth and extracellular protein accumulation in medium under various culture conditions. Among the culture media tested, a full-strength MS medium had a pronounced effect on root biomass and extracelluar protein accumulation in medium. The maximum root biomass (2.4 g DRW/flask) and extracellular total protein contents $(28.3ug/m\ell)$ in medium was obtained at inoculum size of 2 g (FRW) and in MS medium supplemented with 4% sucrose. In addition, the optimal shaking speed for root growth and extracellular protein accumulation in medium were 100 rpm. The total extracellualr protein concentration reached a maximum of $28.3ug/m\ell$ at 4 weeks and decreased thereafter. Protein translation inhibitory activity was observed in culture broths and reached levels of 21.3 unit. These studies demonstrate that the transformed hairy roots can be utilized for the in vitro production of ribosome-inactivating proteins.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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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.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.77-83
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• 1999

We have investigated the effect of culture conditions on tropane alkaloids (scopolamine, hyoscyamine) production in hairy root cultures of Hyoscyamus niger L. induced by Agrobacterium tumefaciens $A_4$T. SH medium was the best for tropane alkaloids production from the hairy root clones, HN18 and HN57. The optimum culture peroid was 5 weeks for HN18 clone and 6 weeks for HN57 clone, respectively. The optimum sucrose and dextrose concentrations in tropane alkaloids productivity were 3% and 2%, respectively. The growth of both HN18 and HN57 clones increased with as sucrose concentration increase up to 7% sucrose, but tropane alkaloid contents was significantly decreased. In the HN18 clone, the optimum concentration of sucrose for alkaloids productivity was 5% and those of dextrose was 2%. The productivity of tropane alkaloids for HN57 clone under dextrose treatments was quite a low level compared to sucrose treatments.

• KSBB Journal
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• v.21 no.2
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• pp.123-128
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• 2006

A transgenic hairy root clone AG-04 of Astragalus membranaceus was obtained following co-cultivation of leaf explants with Agrobacterium rhizogenes ATCC15834. This clone was examined for its growth and production of cyclolanostane-type saponins, astragalosides I, II, and III, under various culture conditions. Among the five basal media tested, Shenk and Hildebrandt(SH)(18) medium was best for roots growth and astragalosides production. The maximum root biomass was obtained at inoculum size of 500 mg FRW per flask, initial sucrose concentration of 3%, and shaking speeds of 90 rpm. The astagalosides production was promoted when the hairy root clone AG-04 was cultured at shaking speeds of 120 rpm and light irradiation of 18 h. Astragaloside contents was also stimulated with high initial sucrose concentration, and the maximum astargalosides contents of 6.21 %/g DRW was obtained at initial sucrose concentration of 6%. The addition of chitosan(100 mg/L) to the culture medium was significantly increased astragalosides production. This was 2.1 times higher than that obtained in a control culture without chitosan.

• Journal of Plant Biology
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• v.38 no.4
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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).

• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11b
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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)

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.143-148
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• 1997

The optimum concentrations of pH, sucrose and vitamins for the growth and tropane alkaloid production of hairy root clone DTLA9 (best growth line) were investigated. The optimum pH in growth and tropane alkaliod production of DTLA9 clone in SH (Schenk and Hildebrandt, 1972) basal medium without growth regulator were pH 6.3 and 6.5, respectively. Also, the optimum sucrose concentration in growth and tropane alkaliod production in the same medium were 3.0 and 2.8%, respectively. The optimum concentrations of ascorbic acid, D-pantothenate, nicotinic acid, pyridoxine, riboflavin, and thiamine on the growth of DTLA9 clone in SH basal medium without vitamins were 0.1 mM, 0.003 mM, 0.07 mM, 0.002 mM, 0.025 mM, and 0.01 mM, respectively. In particular, supplement of 0.1 mM ascorbic acid to SH basal medium without vitamins stimulated the tropane alkaloid production.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.65-71
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• 2003

This research aims the production of anti-tumor substances through in vitro culture of hairy roots transformed by Agrobacterium rhizogenes in Artemisia sylvatica MAX and the effect of culture conditions on optimum growth of hairy roots. We investigated the optimum medium, pH, carbon source, sucrose, light, Fe and polyamine conditions of various lines of hairy roots (NK3, NK4, YX. NK3-10) induced from Artemisia sylvatica to increase the optimum growth of hairy roots. MS medium was the best for optimum growth of hairy root clone, NK3-S10. The optimum culture period was 4 weeks for NK3-S10. The optimum sucrose concentration was 3.5%. The optimum concentration of FeSO$_4$, spermine and spermidine was 0.1 mM, 10 mM and 100 mM, respectively.