• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.528-534
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• 2005

Hairy root cultures have demonstrated great promise in terms of their biosynthetic capability toward the production of secondary metabolites, but continue to constitute a major challenge with regard to large-scale cultures. In order to assess the possibility of conducting mass production of biomass, and the extraction of useful metabolites from Panax ginseng. P. ginseng hairy roots, transformed by Rhizobium rhizogenes KCTC 2744, were used in bioreactors of different types and sizes. The most effective mass production of hairy roots was achieved in several differently Sized air bubble bioreactors compared to all other bioreactor types. Hairy root growth was enhanced by aeration, and the production increased with increasing aeration rate in a 1 L bioreactor culture. It was determined that the hairy root growth rate could be substantially enhanced by increases in the aeration rate upto 0.5vvm, but at aeration rates above 0.5vvm, only slight promotions in growth rates were observed. In 20 L air bubble bioreactors, with a variety of inoculum sizes, the hairy roots exhibited the most robust growth rates with an inoculum size of 0.1% (w/v), within the range 0.1 to 0.7% (w/v). The specific growth rates of the hairy root decreased with increases in the inoculum size.

• Journal of Ginseng Research
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• 제20권3호
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• pp.318-324
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• 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.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.88-94
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• 2005

Plant tissue culture studies have been done for the preservation of medicinal plant resources and efficient production of pharmaceutically important secondary metabolites. Micropropagation methods for Cephaelis ipecacuanha have been established and these methods enabled much more efficient propagation of the plants than the conventional methods using seedling or layering. The C. ipecacuanha plants derived from tissue culture grew uniformly in the field and they showed higher alkaloid contents compared to the plants grown from seedlings. Hairy root cultures of C. ipecacuanha and Panax ginseng have been established by infection with Agrobacterium rhizogenes, and the production of important pharmaceuticals by these cultures have been successfully demonstrated. In the case of C. ipecacuanha, the highest alkaloid yields from the hairy roots cultured for 8 weeks were 2.75-fold cephaeline (5.5 mg) and one third emetine (0.7 mg) compared with those from the roots of one-year old plant propagated through shoot-tip culture and cultivated in a greenhouse (2.0 mg cephaeline and 2.0 mg emetine). In the case of P. ginseng, ginsenoside contents in the hairy roots optimally cultured for 4 weeks were much higher than those in the roots of 4-year old field-grown plant. Thus our medicinal plant tissue cultures demonstrate desirable properties. However, they are always exposed to danger of microbial contamination or unexpected trouble of culture facilities. Cryopreservation of plant tissue cultures is a reliable method for long-term preservation. Cryopreservation studies on these cultures are also presented.

• 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.

• 식물조직배양학회지
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• 제22권2호
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• pp.65-70
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• 1995

자리공 모상근에서 betalain의 합성에는 광이 필수이지만 생장률은 암상태 보다 광상태(1500 1X)에서 5배 정도 억제된다. 따라서 광상태에서 모상근의 생장이 억제되는 원인을 규명하기 위해서 항산화제 처리에 따른 효과를 조사하였다. 항산화제(ascorbic acid, glutathione, $\alpha$-tocopherol, sodium pyrosulfate, propylgallic acid) 처리에 따른 모상근의 생장은 암상태에서 무처리보다 1.2-1.4배, 광상태에서 약 1.3-1.9배 생장이 증가되었으며, betalain 합성은 무처리보다 1.2-2.1배 향상되었다. 자리공 모상근의 성장 및 betalain 합성에서 항산화제 복합 처리에 따른 효과는 나타나지 않았다. Betalain 합성 과정에서 항산화제의 효과는 청색 파장하에서 가장 높게 나타났다. 따라서 광상태에서 자리공 모상근의 생장과 betalain 합성을 촉진시키기 위해서는 청색 파장에서 적절한 항산화제의 개발이 요구된다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.250-253
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• 2000

We researched effects of growth on initial inoculum size, liquid volume, and medium feeding rate etc. Cell suspension inoculated at low cell concentrations showed a typical growth reduction, whereas root cultures showed an improvement in growth. In this paper, Hairy roots showed high growth rate at 0.4 % inoculum size and 100 mL liquid volume in 250 mL flask cultures.

• KSBB Journal
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• 제5권3호
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• pp.207-214
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• 1990

Growth properties of carrot hairy roots transformed by Agrobacterium rhizogenes were compared in flask and bioreactor. Oxygen transfer coefficient KLa was measured during the cultivation in bioreactor. In flask cultures, initially sucrose 30g/l was nearly exhausted after 20days. pH was dropped from initially 5.8 to 4.79 after 4 days, but it is stable after that time. Finally, after 28 days, hairy roots were grown about twelve times. In view of the results studied optimum conditions, hairy roots were maintained high growth rates in sucrose 50g/l, pH 5.8, total nitrogen 60mM. Also in bioreactor cultures, fixed stainless sieve in bottom and aerated 0.31 vvm, the results of cultivation by the use of sucrose 50g/l had grown about twenty-eight times and pH variations were liked in flask. As a results, growth rate of 1.756g fresh weight/day/g inoculum in bioreactor were higher about three times than 0.57g fresh weight/day/g inoculum in flask culture. KLa values were showed a tendency to decrease from 0.209 min-1 to 0.068 min-1.

• Journal of Microbiology and Biotechnology
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• 제24권6호
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• pp.823-834
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• 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.

• 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)

• 식물조직배양학회지
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• 제26권1호
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• pp.7-14
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• 1999

형질전환된 소리쟁이 모상근의 기내 배양시 그 생장 특성과 anthraquinone생산성을 조사하기 위하여 소리쟁이의 모상근을 Agrobacterium rhirogenes strain $A_4$ 접종에 의하여 잎 절편으로부터 유기하여 부정근과 비교하였다. 잎 절편에서의 부정근 형성은 5 $\mu$M NAA를 첨가한 MS배지에서 가장 양호하였다. Paper electrophoresis를 통한 opine분석 결과 모상근에서만 mannopine이 검출되고 부정근에서는 검출되지 않았다. 부정근과 모상근 모두에서 2차근인 측근 근단은 호르몬을 첨가하지 않은 배지에서 신장만 할 뿐 분지하지 않았으나, 1차근인 주근의 근단은 많은 측지를 내며 빠르게 생장하였다. Anthraquinone 함량 증대에 따른 부정근과 모상근의 생장에는 MS기본 배지가 가장 양호하였다. 부정근과 모상근 배양시 생장량과 anthraquinone함량은 5% sucrose를 포함한 0.05 $\mu$M NAA와 0.1 $\mu$M kinetin으로 조성된 MS 액체 배지에서 가장 높은 것으로 나타났으며, 모상근의 anthraquinone함량은 배양 일수에 따라 증가하다가 배양 25일 이후에는 감소하였다.