• Journal of Microbiology and Biotechnology
• /
• v.24 no.6
• /
• pp.823-834
• /
• 2014

In order to solve the shortage of natural Tripterygium wilfordii Hook. f. plant resource for the production of the important secondary metabolites triptolide and wilforine, hairy roots were induced from its root calli by Agrobacterium rhizogenes. Induced hairy roots not only could be maintained and grown well in hormone-free half-strength Murashige and Skoog medium but also could produce sufficient amounts of both triptolide and wilforine. Although hairy roots produced approximately 15% less triptolide than adventitious roots and 10% less wilforine than naturally grown roots, they could grow fast and could be a suitable system for producing both secondary metabolites compared with other tissues. Addition of $50{\mu}M$ methyl jasmonate (MeJA) could slightly affect hairy root growth, but dramatically stimulated the production of both triptolide and wilforine, whereas $50{\mu}M$ salicylic acid had no apparent effect on hairy root growth with slightly stimulatory effects on the production of both secondary metabolites. Addition of precursor nicotinic acid, isoleucine, or aspartic acid at the concentration of $500{\mu}M$ had varying effects on hairy root growth, but none of them had stimulatory effects on triptolide production, and only the former two had slightly beneficial effects on wilforine production. The majority of triptolide produced was secreted into the medium, whereas most of the produced wilforine was retained inside of hairy roots. Our studies provide a promising way to produce triptolide and wilforine in T. wilfordii hairy root cultures combined with MeJA treatment.

• Journal of Plant Biotechnology
• /
• v.36 no.3
• /
• pp.294-300
• /
• 2009

Activation tagging that uses T-DNA vectors containing multimerized transcriptional enhancers from the cauliflower mosaic virus (CaMV) 35S gene is a powerful tool to determine gene function in plants. This approach has been successfully applied in screening various types of mutations and cloning the corresponding genes. We generated an activation tagged hairy root pool of ginseng (Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the biosynthetic pathway of ginsenoside (triterpene saponin), which is known as the major active ingredient of the root. Quantitative and qualitative variation of ginsenoside in activation tagged hairy root lines were profiled using LC/MS. Metabolic profiling data enabled selection of a specific hairy root line which accumulated ginsenoside at a higher level than other lines. The relative expression level of several genes of triterpene biosynthetic pathway in the selected hairy root line was determined by real time RT-PCR. Overall results suggest that the activation tagged ginseng hairy root system described in this study would be useful in isolating genes involved in a complex metabolic pathway from genetically intractable plant species by metabolic profiling.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.1
• /
• pp.91-98
• /
• 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.

• Journal of the Korean Applied Science and Technology
• /
• v.35 no.4
• /
• pp.995-1002
• /
• 2018

Hairy root culture of ginseng is industrially prospected because the cultivation period of ginseng is relatively long. In this study, the effect of medium concentration and sucrose concentration on hairy root culture of ginseng was evaluated. The optimization of ginseng hairy roots transformed by Agrobacterium rhizogene were performed liquid medium. The MS(Murashinge & Skoog basal medium) concentration was selected with 1/2 strength MS and the optimal sucrose concentration was determined at 2-3%(w/v). At the optimum culture condition, The yield (the ratio of weight of grown hairy root cultures to weight of fresh ginseng hairy roots) and production rate of ginseng root were 19.42 times and 5.73 g/l-day. The major ginsenosides were Rb group, Re and Rg1. The produced total ginsenoside content in the solid medium was 9.87 (mg/g) and increased 1.34 times in the liquid medium (13.23 mg/g). In solid culture, the contents of ginsenosides Rb, Re and Rg1 were 2.14, 3.65 and 1.87 mg/g, respectively. In liquid culture, the contents of ginsenosides Rb, Re and Rg1 were 3.54, 4.12 and 2.63 mg/g, respectively.

• Journal of Plant Biology
• /
• v.38 no.4
• /
• pp.337-341
• /
• 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).

• Korean Journal of Medicinal Crop Science
• /
• v.12 no.2
• /
• pp.118-122
• /
• 2004

We have investigated that a couple of soft ferrite ceramic powders having a spinal structure have shown the effect on growth and secondary metabolites production of some medicinal plants cultured in vitro. The addition of the ceramic powders as bare state to culture medium has stimulated the growth of Achyranthes japonica callus and plantlet, adventitious root of Hyoscyamus niger and Platycodon grandiflorum hairy root about 65, 75, 150 and 50%, respectively. Whereas Centella asiatica callus and plantlet, Scopolia parviflora hairy root, and Hyoscyamus albus adventitious root were not affected markedly. Moreover, the ceramic powder has enhanced the growth of H. niger adventitious roots even under conditions of irradiating alone without any direct contact between ceramic powder and media. Based on growth stimulation effect, the ceramic powders have enhanced the gross production of tropane alkaloid in H. niger adventitious root, and polyacetylene in P. grandiflorum hairy root about 35 and 30%, respectively.

• Journal of Ginseng Research
• /
• v.20 no.3
• /
• pp.318-324
• /
• 1996

The effects of light on the growth and ginsenosides production were examined in the hairy roots of Panax ginsen C.A. Meyer induced by Agrobacterium rhizogines A4. The 9rowth of ginseng hairy roots in 1/2MS liquid medium was significantly decreased with an increment of light intensity (1,000~7,000 lux). The growth of hairy roots under 7,000 lux condition was decreased at 17% compared to the dark condition. The production of 7 ginsenosides in hairy root was very high in 3,500 lux condition. The production of ginsenoside-Rg, and Rf increased 3.3 and, 2.4 times respectively as compared to dark condition. The growth of hairy roots was inhibited by blue light, while ginsenosides production was increased. The sucrose demands of hairy roots was examined in light condition(3,500 lux). The growth of hairy roots in 1/2MS liquid medium with various sucrose concentrations(1~4%) was high in IVp sucrose, while ginsenosides production was high in 3% sucrose condition. The growth and ginsenosides production were high when hairy roots were cultured in dark condition for 1 week and then transferred to light condition(3,500 lux) for 4 weeks. It is suggested that ginsenosides production could be accelerated by light intensity of specific wavelength in cultures of ginseng hairy roots.

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

• Journal of Plant Biology
• /
• v.39 no.3
• /
• pp.161-166
• /
• 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.

• Journal of Plant Biology
• /
• v.33 no.3
• /
• pp.183-188
• /
• 1990

Induction and culture of hairy roots by Agrobacterium rhizogenes A4 were carried out in Platycodon grandiflorum DC. After 2-4 weeks of inoculation with Agrobacterium rhizogenes hairy roots were formed at root segments in the balloon flower. Optimized growth of hairy roots was obtained in hormone-free MS medium, 6% sucrose and pH 5.8. The pattern of ginsenoside in the transformed roots was not different with that in the ordinary roots.