• 식물조직배양학회지
• /
• 제22권2호
• /
• pp.71-76
• /
• 1995

자리공 모상근에서 광처리에 따른 항산화효소의 활성을 조사하였다. Catalasa superoxide dismutasae, ascorbate oxidase (AO)의 활성은 광도가 2,000 lx까지 증가할수록 감소하였으며, 특히 AO활성은 2,000 lx에서 암상태보다 92% 감소하였다. Glutathione reductase, glutathione peroxidase (GPO), ascorbate peroxidase 그리고 peroxidase의 활성도는 500 lx까지는 광도가 높아질수록 증가하였으나 그 이상의 높은 광도에서는 현저하게 감소하였다. GPO의 활성은 AO처럼 2,000 lx에서 암상태보다 85%감소하였다. 광파장에 따른 항산화효소의 활성은 청색광의 파장에서 가장 많이 억제되었으며, AO의 활성은 25%까지 감소하였다. 청색광 파장의 광도에 따른 항산화효소의 활성도는 30 lx까지는 증가하다가 200 lx에서는 암상태보다 21-71%까지 감소하는 경향을 나타내었다. AO의 활성은 청색 파장의 광도(300-200 lx)가 증가할수록 급격히 감소하여 200 lx에서는 70%까지 억제되었다. 자리공 모상근의 항산화효소 활성은 청색광 파장의 높은 광도에서 주로 생성된 유해산소 의하여 억제되고 있음을 확인하였다. 광상태하에서 모상근의 생장과 betalain 합성을 향상시키기 위해서는 모상근에서 생성되는 산화제의 효율적인 제거가 요구됨이 시사되었다.

• 식물조직배양학회지
• /
• 제24권5호
• /
• pp.273-277
• /
• 1997

Rhizoctonia solani 추출물(elicitor)을 Hyoscyamus muticus 모상근배양에 처리하였을 때 sesquiterpene cyclase (SC)가 유도됨과 동시에 sesquiterpene화합물이 합성되어 배양액속에 축적하는 것으로 나타났다. 24시간 배양기간 동안 배양액으로부터 sesquiterpene을 추출ㆍ분리하여 배양할 경우, 추출ㆍ분리하지 않은 모상근배양보다 거의 두배의 함량이 생성되었다. Cyclase monoclonal antibody을 이용하여 immunoblot을 시도한 결과, 추출한 모상근배양과 추출하지 많은 모상근배양에서의 SC 절대량은 동일한 것으로 나타났으며, 효소활성도 현저한 차이가 없는 것으로 측정되었다. 이러한 결과는 terpenoid pathway에서 sesquiterpene 생합성이 이용가능한 기질의 량에 의해 조절되는 것으로 사료되며, feedback 조절은 sesquiterpene cyclase 효소에 앞서 일어나는 것으로 추정된다.

• Journal of Ginseng Research
• /
• 제24권2호
• /
• pp.74-78
• /
• 2000

인삼모상근의 생장과 ginsenosides의 함량을 높이기 위하여 생장조절제가 첨가되지 않은 1/2 MS 배지에 jasmonic acid와 methyl jasmonate의 농도와 처리시기를 달리하여 인삼모상근 KGHR울 세포주를 배양하였다. 인삼모상근 생장은jasmonic acid와 methyl jasmonate 모두 1$\mu\textrm{m}$ 농도에서 가장 양호하였으며 30$\mu\textrm{m}$ 이상 농도가 증가할수록 모상근생장이 감소하였다. 그러나 생장이 낮았던 jasmonic acid 10$\mu\textrm{m}$ 처리구와 methyl jasmonate 50$\mu\textrm{m}$에서 ginsenosides 함량과 생산성이 더 높았다. 배양시기별로 jasmonic acid와 methyl jasmonate의 처리 효과는 jasmonic acid는 배양 후4주, methyl jasmonate는 3주에 처리하는 것이 ginsenosides의 함량과 생산성을 높이는데 효과적이었다.

• 식물조직배양학회지
• /
• 제27권5호
• /
• pp.379-385
• /
• 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.

• 한국연초학회지
• /
• 제20권1호
• /
• pp.40-49
• /
• 1998

Protoplasts were isolated from mesophyll of tobacco(Nicotiana glauca) transformed with kanamycin-resistant gene (NPT II gene) and potato hairy root callus containing Ri plasmid of Agrobacterium rhiEogenes, and protoplasm fusion was made between the isolated protoplasts. The transgenic tobacco leaf tissue could grow on the media containing high concentrations of kanamycin, but not on the phytohormone-free media. On the other hand, the potato hairy root calli could be cultured on the phytohormone-free media but not on media containing more than 40 ㎍/ml kanamycin. In these conditions, the viability of both protoplasts were above 90%, These selection markers were used for the selection of protoplasts fused between the two, i.e. protoplast fusion was detected using selection media containing 100㎍/ml kanamycin and with no phytohormone. The mixture of 1.0% cellulase, 0.3% macerozyme, and 0.7M mannitol was best for the maximum protoplast production for tobacco, and that of 2.0% cellulase, 2.0% macerozyme, 1.0% dricelase, and 0.5M mannitol for potato. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution on the selectable medium. Cell walls were regenerated after 5 days in this medium, and colonies were alive until 4 weeks after cultural, but died after 6 weeks.

• 식물조직배양학회지
• /
• 제26권3호
• /
• pp.171-175
• /
• 1999

시호 모상근 (BFHR2 클론) 배양에서 생장과 saikosaponin 생산에 미치는 기본배지, 배지의 다량무기염농도 변화 및 IAA와 IBA의 효과를 조사하였다. 조사한 배양액 중 RCM 배지에 다량무기염농도를 2배로 증가시킨 배지에서 생장이 가장 양호하였다. 또한 2RCM배지에서는 saikosaponin인 a, c와 d의 생성을 확인하였으며, 반면에 MS배지에서는 saikosaponin의 생성이 극히 저조하였다. IBA는 0.5mg/L에서 약 60%의 생장촉진효과를 보였으나, IAA와 IBA두 가지 모두 0.01~5mg/L 까지의 농도에서 뿌리형태의 외적인 변화 없이 saikosaponin 생성을 강하게 억제하는 작용을 나타내었다.

• 원예과학기술지
• /
• 제18권6호
• /
• pp.802-807
• /
• 2000

Agrobacterium rhizogenes의 Ri t-DNA로 형질전환된 당근의 모상근에 Glomus sp.를 감염시킨후 arbuscular 균근균 포자를 Modified Strullu & Romand 배지상에서 12주 동안 기내증식하였다. 기내증식을 위한 접종원으로는 포자보다는 균근감염 뿌리가 더 좋았다. 당근 모상근과 arbuscular 균근균의 대치배양으로 증식된 Glomus sp.의 포자는 직경 약 $50{\mu}m$정도의 원형 또는 타원형의 포자로서 균사의 중간부위에서 형성되었다. 12주 동안 기내에서 대치 배양한 결과 plate 당 약 1,200개 정도의 arbuscular 균근균 포자를 생산하였다.

• Natural Product Sciences
• /
• 제25권3호
• /
• pp.233-237
• /
• 2019

Three Diels-Alder type adducts, guangsangon E (1), chalcomoracin (2) and sorocein I (3) were isolated from hairy root cultures of Morus macroura. The structures of the isolated compounds (1-3) were determined by spectroscopic method (NMR and MS), and spectral comparison to literature. Cytotoxic activities of the isolated compounds (1 - 3) were investigated against P-388 murine leukemia cell line. Guangsangon E (1) showed the most potent cytotoxicity against P-388 murine leukemia cell line with $IC_{50}$ value of $2.75{\pm}0.32{\mu}g/mL$. To the best of our knowledge, guangsangon E (1) and sorocein I (3) were reported for the first time from the tissue cultures of M. macroura.

• Journal of Ginseng Research
• /
• 제44권4호
• /
• pp.563-569
• /
• 2020

Background: White ginseng consists of the roots and rhizomes of the Panax species, and red ginseng is made by steaming and drying white ginseng. While red ginseng has both polar and nonpolar ginsenosides, previous studies showed white ginseng to have only polar ginsenosides. Because nonpolar ginsenosides are formed through the manufacture of red ginseng from white ginseng, researchers have generally thought that nonpolar ginsenosides do not exist in white ginseng. Methods: We developed a simultaneous quantitative method for six nonpolar ginsenosides in white ginseng using reverse-phase high-performance liquid chromatography coupled with integrated pulsed amperometric detection. The nonpolar ginsenosides of white ginseng were extracted for 4 h under reflux with 50% methanol. Results: Using the gradient elution system, all target components were completely separated within 50 min. Nonpolar ginsenosides were determined in the rhizome head (RH), main root (MR), lateral root, and hairy root (HR) of 6-year-old white ginseng samples obtained from several regions (Geumsan, Punggi, and Kanghwa). The total content in the HR of white ginseng was 37.8-56.8% of that in the HR of red ginseng. The total content in the MR of white ginseng was 5.9-24.3% of that in the MR of red ginseng. In addition, the total content in the RH of white ginseng was 28.5-35.8% of that in the HR of red ginseng Conclusion: It was confirmed that nonpolar ginsenosides known to be specific components of red ginseng were present at substantial concentrations in the HR or RH of white ginseng.

• Molecules and Cells
• /
• 제27권2호
• /
• pp.217-223
• /
• 2009

Development of symbiotic root nodules in legumes involves the induction and repression of numerous genes in conjunction with changes in the level of phytohormones. We have isolated several genes that exhibit differential expression patterns during the development of soybean nodules. One of such genes, which were repressed in mature nodules, was identified as a putative aldo/keto reductase and thus named Glycine max aldo/keto reductase 1 (GmAKR1). GmAKR1 appears to be a close relative of a yeast aldo/keto reductase YakC whose in vivo substrate has not been identified yet. The expression of GmAKR1 in soybean showed a root-specific expression pattern and inducibility by a synthetic auxin analogue 2,4-D, which appeared to be corroborated by presence of the root-specific element and the stress-response element in the promoter region. In addition, constitutive overexpression of GmAKR1 in transgenic soybean hairy roots inhibited nodule development, which suggests that it plays a negative role in the regulation of nodule development. One of the Arabidopsis orthologues of GmAKR1 is the ARF-GAP domain 2 protein, which is a potential negative regulator of vesicle trafficking; therefore GmAKR1 may have a similar function in the roots and nodules of legume plants.