• Journal of Plant Biology
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• v.29 no.1
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• pp.41-51
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• 1986

The plants of Nicotiana tabacum L. cv. NC2326 were germinated in 10 cm D$\times$20 cm H polyethylene pot, and sand-cultured with Hoagland solution near by the window of laboratory room(26$\pm$5$^{\circ}C$). The growing plants were sprayed with various concentrations of ABA around 9 : 00 a.m. once in every two days for 12 weeks in summer. As the results, frequency of stomatal number, stomatal opening, chlorophyll content, respiration rate, and protein content in the leaves were decreased with the increasing of concentrations of ABA, respectively. The plant growth was inhibited by exogenous ABA, but leaf abscission was not found during the experimental period. The ratio of three to one in chlorophyll a to b was not altered by exogenous ABA. All the stomata were closed within three minutes by 100 $\mu\textrm{g}$ ml-1 ABA and within seven minutes by 1-10 $\mu\textrm{g}$ ml-1 ABA after the spraying of ABA, and then reopended after a few hours in 1-10 $\mu\textrm{g}$ ml-1 ABA and after 24 hours in 100 $\mu\textrm{g}$ ml-1 ABA. The polar movement of chloroplast within the guard cells was found in the higher concentrations of 10 and 100 $\mu\textrm{g}$ ml-1 ABA, but not found in the lower concentrations than 1 $\mu\textrm{g}$ ml-1 ABA. During the night and weak light, it was fond that the inhibition of respiration rate by the higher concentration of ABA was owing to firstly the stomatal closure by the spraying of ABA and secondly the decrease of stomatal frequency by the inhibition of stomatal development with exogenous ABA for the long period of 12 weeks. In the band number of leaf protein by the electrophoresis, most of the protein bands were disappeared by the higher concentration of 100 $\mu\textrm{g}$ ml-1 ABA, but were not altered by the lower concentration of ABA in comparison with control.

• Korean Journal of Weed Science
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• v.15 no.2
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• pp.154-159
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• 1995

Water extracts from Polygonum aviculare and Salix koreansis markedly inhibited the germination of lettuce and rice seeds, indicating the presence of biologically active substances. The biochemical substances such as salicylic and+vanillic acid, tannic acid + gallic acid, p-coumaric acid, p-cressol, sinapic acid and catechol etc. belonging to phenolic compounds were detected in the cultured cells, suggesting that the secondary metabolites can be synthesized in plant cell and tissue culture. In addition, fatty acid like linolenic acid and organic acid such as oxalic acid were presented in the highest amount, 3.7 mg/g and 14.288 mg/g, respectively, which seem to be related to exhibiting phytotoxicity of P. aviculare. Petroleum ether extract exhibited another potential relating to inhibitory effect which needs further investigation. Calli from two plant sources were easily introduced by uses of 1.0 mg/l of 2.4-D and 0.1 to 0.2 mg/l of BAP in MS basal medium which can be implemented for a large scale production through cell culture.

• Korean Journal of Weed Science
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• v.11 no.1
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• pp.68-73
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• 1991

This study was conducted to determine the appropriate culture conditions for developing photoautotrophic cells from tobacco using plant growth regulators, sucrose and light condition in LS medium. The greatest callus induction was observed in the LS medium supplemented with $10^{-5}$ M NAA and $10^{-4}$ M BA, showing 3.08 g of callus fresh weight determined at 30 days after incubation. The highest chlorophyll content of callus was obtained in the LS medium supplemented with $10^{-5}$M NAA and $10^{-6}$ M BA, showing 28.42${\mu}g/g$. In both light and dark conditions, callus induction increased as the concentration of sucrose increased from 0.5%to 3.0%. In particular, in the light condition, the greatest callus induction was made in the LS medium supplemented with 2% sucrose. On the other hand, the heighest chlorophyll content was observed at 0.5% sucrose and chlorophyll wasn't induced in dark condition. The chlorophyll content of callus cultured in LS medium containing 0.75% sucrose was similar to 1% sucrose, as far as the chlorophyll content of callus was concerned.

• Biomedical Science Letters
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• v.2 no.1
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• pp.121-126
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• 1996

Specific monoclonal antibodies(mAbs) against bis-indole alkaloids related to vinblastine were established to develop a simple and specific immunoassay system for the quantitation of Vinca alkaloids. Vinca alkaloids were extracted from tissue cultured cells of periwinkle plant (Vinca rosea L.). Spleen cells from Balb/c mice immunized with vinblastine-bovine serum albumin(VBL-BSA) conjugate as immunogen were fused with myeloma cells(Sp2/0-Ag.14) in the presence of polyethylene glycol. In the preliminary experiments, 32 clones which highly reacted with VBL-BSA conjugate were selected by ELISA(Enzyme-linked immunosorbent assay).These clones were further analyzed by inhibition assay of ELISA. The results obtained with two typical monoclonal antibodies, KN-1 and KN-2, were described. KN-1 exhibited considerable reactivities with soluble dimeric bis-indole alkaloids, whereas no cross reacted with monomeric bis-indole alkaloids. However KN-2 showed cross reactivity with mono- and di-meric bis-indole alkaloids. Furthermore, KN-1 was applied to the immunoassay system for determining the VBL amounts of in vitro cultured cell extracts. This assay system could detect dimeric vinca alkaloid as low as 0.05 nM.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.127-132
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• 1999

Protoplasts were isolated from the leaf mesophyll tissue of in vitro 4-weeks-old Arabidopsis thaliana and cultured in MS liquid medium supplemented with 2.0 mg/L NAA, 0.5 mg/L BAP and 9% mannitol in the dark at $25^{\circ}C$. When protoplast-derived microcolonies were dehydrated, the frequency of callus induction enhanced approximately 7-fold higher compared with non-dehydrated microcolonies in CP medium. Fifty callus lines were selected from dehydrated microcolonies. Shoots were efficiently initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium supplemented with 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for 4 weeks. Shoot regeneration frequencies (calli regenerating at least one shoot) were 3.5%~56%. Histological observations of shoot forming callus revealed that tracheary elements initiated from inner compact cells, and that meristemoids developed to shoot primordia and shoots. Roots were induced from these regenerating shoots on MS medium without phytohormones. These regenerants were successfully transplanted into potting soil. Morphological characterization of 50 protoplast-derived plants showed that the frequency of normal type was 78%.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.219-223
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• 1998

Anthers contanining uninucleate microspores of eight cultivars of pepper were cultured on MS medium supplemented with 0.004mg/L 2,4-D and 0.1mg/L kinetin, 3% sucrose and 0.2% Gelrite, kept at $35^{\circ}C$ for 24 h, and then cultured at $25^{\circ}C$ with a photoperiod of 16 h daylight for 40 days. Frequency of embryoid and callus formation was varied with cultivars. Embryoid formation was found in Cheongyang and Fushimi Amanaga, while callus formation was in California Wonder, Fushimi Amanaga and Geoseong. In anther culture medium supplemented with 1% activated charcoal, embryoid formation was found with 0.5% of frequency only in Cheongyang, while no callus formation was found. In 1/2MS medium, frequency of embryoid formation in Shishitou, Yatsufusa and Taka no Tsume was 1.2%, 0.4% and 0.4%, respectively. On the other hand, in 1/2 B5 medium, frequency of callus formation in Yatsufusa and Taka no Tsume was 2.8% and 2.7%, respectively. Embryoids transferred to hormone-free MS medium were developed to plantlets and acclimatized. The number of chromosomes in the root tip cells of the haploid plant was 2n=$\times$=12 in cv. Cheongyang.

• Journal of Plant Biotechnology
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• v.33 no.3
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• pp.195-207
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• 2006

Stem cells are the primordial, initial cells which usually divide asymmetrically giving rise to on the one hand self-renewals and on the other hand progenitor cells with potential for differentiation. Zygote (fertilized egg), with totipotency, deserves the top-ranking stem cell - he totipotent stem cell (TSC). Both the ICM (inner cell mass) taken from the 6 days-old human blastocyst and ESC (embryonic stem cell) derived from the in vitro cultured ICM have slightly less potency for differentiation than the zygote, and are termed pluripotent stem cells. Stem cells in the tissues and organs of fetus, infant, and adult have highly reduced potency and committed to produce only progenitor cells for particular tissues. These tissue-specific stem cells are called multipotent stem cells. These tissue-specific/committed multipotent stem cells, when placed in altered environment other than their original niche, can yield cells characteristic of the altered environment. These findings are certainly of potential interest from the clinical, therapeutic perspective. The controversial terminology 'somatic stem cell plasticity' coined by the stem cell community seems to have been proved true. Followings are some of the recent knowledges related to the stem cell. Just as the tissues of our body have their own multipotent stem cells, cancerous tumor has undifferentiated cells known as cancer stem cell (CSC). Each time CSC cleaves, it makes two daughter cells with different fate. One is endowed with immortality, the remarkable ability to divide indefinitely, while the other progeny cell divides occasionally but lives forever. In the cancer tumor, CSC is minority being as few as 3-5% of the tumor mass but it is the culprit behind the tumor-malignancy, metastasis, and recurrence of cancer. CSC is like a master print. As long as the original exists, copies can be made and the disease can persist. If the CSC is destroyed, cancer tumor can't grow. In the decades-long cancer therapy, efforts were focused on the reducing of the bulk of cancerous growth. How cancer therapy is changing to destroy the origin of tumor, the CSC. The next generation of treatments should be to recognize and target the root cause of cancerous growth, the CSC, rather than the reducing of the bulk of tumor, Now the strategy is to find a way to identify and isolate the stem cells. The surfaces of normal as well as the cancer stem cells are studded with proteins. In leukaemia stem cell, for example, protein CD 34 is identified. In the new treatment of cancer disease it is needed to look for protein unique to the CSC. Blocking the stem cell's source of nutrients might be another effective strategy. The mystery of sternness of stem cells has begun to be deciphered. ESC can replicate indefinitely and yet retains the potential to turn into any kind of differentiated cells. Polycomb group protein such as Suz 12 repress most of the regulatory genes which, activated, are turned to be developmental genes. These protein molecules keep the ESC in an undifferentiated state. Many of the regulator genes silenced by polycomb proteins are also occupied by such ESC transcription factors as Oct 4, Sox 2, and Nanog. Both polycomb and transcription factor proteins seem to cooperate to keep the ESC in an undifferentiated state, pluripotent, and self-renewable. A normal prion protein (PrP) is found throughout the body from blood to the brain. Prion diseases such as mad cow disease (bovine spongiform encephalopathy) are caused when a normal prion protein misfolds to give rise to PrP$^{SC}$ and assault brain tissue. Why has human body kept such a deadly and enigmatic protein? Although our body has preserved the prion protein, prion diseases are of rare occurrence. Deadly prion diseases have been intensively studied, but normal prion problems are not. Very few facts on the benefit of prion proteins have been known so far. It was found that PrP was hugely expressed on the stem cell surface of bone marrow and on the cells of neural progenitor, PrP seems to have some function in cell maturation and facilitate the division of stem cells and their self-renewal. PrP also might help guide the decision of neural progenitor cell to become a neuron.

• Journal of Korean Society of Forest Science
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• v.71 no.1
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• pp.45-49
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• 1985

Protoplast-source meterial and enzyme strength had a significant influence on protoplast yield from hybrid poplar, Populus alba ${\times}$ P. grandidentata. The yield of protoplasts from in vitro culture of 1 month-old plantlets was more than that from greenhouse grown 4 month-old stock plant. In vitro cultured plantlets regulary produced more viable protoplasts with E-I enzyme solution (0.5% cellulase and 0.1% macerase) than those with E-II enzyme solution (1.0% cellulase and 0.2% macerase) after overnight incubation. The mean yield of protoplasts from in vitro cultured plantlets was $4{\times}10^6$ with E-I enzyme solution. Cell division was observed in these protoplast cultures after 7-10 days. Protoplast-derived hybrid poplar cells survived over 3 weeks in culture and some continuous cell divisions were evident. Other aspects associated with protoplasts from in vitro cultured plantlet are also discussed.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.2
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• pp.137-142
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• 2008

Echinacea purpurea, an indian traditional plant medicine has been widely used as herbal remedy for the treatment of disease such as colds or other infections. However, Echinacea purpurea extracts recently have been applied as a cosmetic ingredient for skin care. We artificially cultured Echinacea purpurea by using the bioreactor culture system for this study. We induced callus from Echinacea purpurea and separated adventitious roots, harvested and extracted after cultured in bioreactors. Previously, several studies have been reported on anti-oxidant and immuno-enhancing effects of Echinacea purpurea extract but other efficacies were not well known. In this study, we investigated the whitening, anti-wrinkle and anti-oxidant effects to know applicable value of tissue-cultured Echinacea purpurea adventitious roots extract(TCEPARE) as a cosmetic ingredient. TCEPARE did not show cytotoxicity until a concentration of 2% and showed the anti-oxidative effect in DPPH and NBT tests. Also, the extract decreased tyrosinase expression in a dose-dependent manner and inhibited melanin synthesis in B16 melanoma cells. TCEPARE reduced protein levels of MMP-1, 2 secreted in culture medium or in cell lysates. From these results we suggest that TCEPARE has potential benefits applicable as to cosmetic ingredient for skin care products.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.13-19
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• 1998

We have reported previously that intact embryogenic cells can be used instead of protoplasts for electroporation-mediated transformation of zoysiagrass and rice. In this study, conditions of the tissue electroporation were examined to optimize the procedures. Embryogenic cell suspensions were established in liquid MS medium containing 2 mg/L of 2,4-D with embryogenic calluses induced from mature embryos of Z. japonica. The suspension-cultured cell clumps were electroporated with 35S-gusA expression vector DNA, and degrees of DNA introduction into the cells were determined by histological expression rates of the gusA marker gene. DNA transfer into the cell clumps occurred in wide range of voltage (100-400 V) and capacitance (10-1980 $\mu\textrm{F}$), but more in the ranges of 200-300 V and 330-800 $\mu\textrm{F}$ DNA concentrations higher than 6 $\mu\textrm{g}$/mL were adequate for GUS expression of the electroporated cells. DNA transfers were confirmed in all three embryogenic cell lines but only in one out of eleven non-embryogenic lines. Positive GUS expressions occurred with DNAs added even 20-40 h after pulse treatments. As a promoter of gusA, Act1 and Ubi1 were effective 7 and 5 times than 35S respectively in number of GUS expression units on electroporated cell clumps. Embryogenic cell clumps survived and regenerated into plantlets after pulse treatments of wide range of conditions.