• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.295-300
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• 2004

This article was conducted to induce the transgenic hairy roots and determine the effect of culture conditions on optimum growth of hairy roots by Agrobacterium rhizogenes strain, 15834 in Lycium chinense Miller. Hairy roots of L. chinense Miller. were induced from leaf segments by co-cultivation with A. rhizogenes. When the hairy roots were cultured in various MS medium strength and sucrose concentrations, the highest growth of hairy roots was observed in half-strength MS media supplemented with 3% sucrose, respectively. In air lift bioreactor cultures, the liquid medium contained with 1/2 MS and 3% sucrose was also the best for optimum growth of hairy roots.

• Proceedings of the Plant Resources Society of Korea Conference
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• 1999.10a
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• pp.46-57
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• 1999

Ginseng(Panax ginseng C.A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark(22$^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

• Korean Journal of Plant Tissue Culture
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• v.27 no.5
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• pp.379-385
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• 2000

tRNA and 7SL RNA based antisense vehicles were prepared by inserting conserved anti-viral and anti-viroid domains. Anti-PVS coat protein leader sequence (ACPL) and antistructural antihairpin domain of PSTVd (AHII) were inserted in tRNA cassette; anti- zing finger domain of PVS, AHII and anti hop latent viroid ribozyme were inserted in 7SL RNA gene isolated from A. thaliana. These constructs were shown to be transcribed both, in in vitro and in in vivo conditions. However, it followed from our work that closely linked position of PoIII reference genes and PoIIII antisense genes within T-DNA lead to the impairment of RNA expression in transgenic plants. To assay in vivo transcription of antisense genes, hairy root potato cultures were established using h. tumefaciens A4-24 bearing both, Ri plasmid and PoIII-promoterless plant expression vectors with antisense RNA genes. Expression of antisense RNA in transgenic potato tissues was proven by specific RT-PCR reactions.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.171-175
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• 1999

We examined the effects of various basal media and exogenous auxins on the growth and saikosaponin production in hairy root cultures of Bupleurum falcatum. Of the various media tested, 2RCM medium was superior for growth and saikosaponin content, in which saikosaponin a, c, and d was produced, however MS medium was resulted in extremely low level in saikosaponin production. By the treatment of 0.5 mg/L IBA the growth rate was increased by about 60% in 2RCM medium, whereas both IAA and IBA showed a potent inhibition effect in saikosaponin biosynthesis by the treatment of 0.01 mg/L~5mg/L.

• Journal of Ginseng Research
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• v.21 no.1
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• pp.28-34
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• 1997

The effects of light on the pigment production and chloroplast development were examined on ginseng hairy roots cultured in 1/2MS liquid medium. The chlorophyll and carotenoid production were increased from 1,000 to 3,500 lux condition, but decreased drastically in 7,000 lux condition. The anthocyanin production was significantly increased with increment light intensity(1,000∼7,000 lux). The thylakoid membrane of chloroplast was proplastid in dark condition and it began to develop into thylakoid membrane in 1,000 lux condition and then intact thylakoid membrane was developed in 3,500 lux condition. However, the development of thylakoid membrane in 7,000 lux condition was inhibited comparing to 3,500 lux condition. The total chlorophyll production in blue light condition were high comparing to other wavelength and same as 40% of total chlorophyll on white light(3,500 lux) condition. The chlorophyll and carotenoid production by sucrose concentration were high in 3% sucrose condition and anthocyanin production was high in 4% condition. The production of chlorophyll and carotenoid by light periods was high when explants were cultured in dark condition for 1 week and then transferred to light condition for 4 weeks. Our results suggest that pigment production and chloroplast development could be accelerated by light Intensity of specific wavelength in cultures of ginseng hairy root.

• Proceedings of the Botanical Society of Korea Conference
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• 1995.06a
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• pp.40-56
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• 1995

During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

• Korean Journal of Medicinal Crop Science
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• v.11 no.5
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• pp.358-363
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• 2003

ScopoIamine and hyoscyamine which belong to tropane alkaloids are the pharmaceutically valuable anticholinergic drugs. In order to increase the productivities, the effects of elicitation were investigated during hairy root cultures of Scopolia. parviflora. Biotic elicitors originated from 3 fungi and 1 yeast were prepared as homogenate and supernatant and added to 3-week-old cultures. Both of homogenate and supernatant of Candida albicans elicitors increased the scopolamine production. The production of hyoscyamine was enhanced by homogenate of Fusarium solani and supernatant of C. albicans. Most of the other fungal elicitors were also improved on the tropane alkaloid production compared to non-treatment. Among the elicitors tested, C. albicans was proved the optimal biotic elicitor on tropane alkaloids production. These results will be served mass production of tropane alkaloids by large-scale production.

• KSBB Journal
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• v.9 no.1
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• pp.26-34
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• 1994

When effects of exogenous hormone on hairy root cultures of Raphanus sativs cv. Chungpihongsim examined, the highest anthocyanin synthesis and disorganization were observed when 2, 4-D was supplemented to the culture medium Cytokinins showed early weak induction after transfer and ABA showed inhibitory effect and GA3 showed no effects in anthocyanin synthesis. Hormones except for 2, 4-D in 1 mg/$\ell$ concentration did not induce disorganization of hairy root and retarded growth of hairy root. Time-course changes in anthocyanin synthesis, phenylalanine ammonia-lyase activity and chalcone synthase activity were examined in culture condition contalning 2, 4-D and kinetin. In a medium containing 2, 4-D, anthocyanin synthesis began to increase on the 9th day and reaching maxima on the 18th day after transfer. Maximum peak of PAL activity appeared on the 3-9th day and another minor peak appeared on the 18th day. CHS activity increased from 9th day, reaching maximum on the 18th day and remained at a relatively high level for culture period. In a medium containing kinetin, anthocyanin synthesis increased temporarily on the 6-9th days, early days after transfer and maintained at a low level for remaining culture period. Peak of PAL activity appeared on the 6th day and CHS activity increased from the 6th days, reaching maxima about 18th day and remained at a relatively high level. In particular, addition of kinetin after preculture in hormone free medium for 2 weeks which was thought of wound healing period showed no effects in anthocyanin synthesis. This results showed that stimulation of anthocyanin synthesis by 2, 4-D and kinetin was meaningfully connected with changes of PAL, CHS activity, and then suggested rate-limiting role of CHS on anthocyanin synthesis in that there is close correlation between anthocyanin synthesis and changes of CHS activity in time-course. Besides, it is considered that cytoklnins involving kinetin stimulated anthocyanin synthesis be due to "wound response" by cutting of young roots, and that difference in time-course peak and PAL, CHS activities expressed by 2, 4-D and kinetin result from occurrence of isozyme which have different regulatory mechanism.mechanism.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.483-486
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• 2001

Growth and tropane alkaloids(hyoscyamine and scopolamine) contents were studied in root cultures from Scopolia paruiflora. All excised roots grew well, even without an addition of a growth regulators. Scopolia roots could be divided scopolamine-rich species. Among the culture media, SH medium was the best for tropane alkaloids production from the hairy roots, whereas the growth increased in SH medium. In precusor experiments, feeding of hyoscyamine was promotive for scopolamine formation. Abiotic elicitors made a slightly production promotion effect.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.188-188
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• 2003