• Journal of Plant Biotechnology
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• v.6 no.3
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• pp.171-179
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• 2004

Hormonal requirements inducing different regeneration pathways with particular emphasis on somatic embryo-genesis in Alhagi graecorum were studied. While combination of 0.5 $\mu{M}$ 2,4-dichlorophenoxyacetic acid (2,4-D), 2.5 $\mu{M}$ 6-benzylaminopurine (BAP) and 5 $\mu{M}$ 1-naphthaleneacetic acid (NAA) in MS medium induced callus formation and callus maintenance from internodal explants, each alone or in combination with other induced distinct regeneration pathway. Adventitious bud formation was induced on MS medium supplemented with 2.5 $\mu{M}$ BAP. It was improved when 2.5 $\mu{M}$ BAP was used in combination with 5 $\mu{M}$ NAA. MS medium containing 0.5 $\mu{M}$ 2,4-D or 5 $\mu{M}$ NAA induced the formation of abnormal direct somatic embryos. While increase of 2,4-D concentration (1.125-9) resulted in the formation of viable embryogenic mass, increase of NAA did not change its effect. NAA should be used in combination with 2,4-D even at low concentration (0.5 $\mu{M}$) to form embryogenic mass. In A. gaecorum, the role of 2,4-D as trigger of somatic embryogenesis and BAP as trigger of adventitious bud formation was deduced, but for maximum yield certain auxin-cytokinin ratio should be applied. Embryogenic masses characterized by high water content, low peroxidase activity, and low number of peroxidase and glutamate oxaloacetate transaminase bands in comparison with calli obtained under conditions stimulating adventitious bud formation. The resulted differential gene expression, which could be detected by native-PAGE patterns, could be used as marker for organogenic pathway in A. graecorum.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.37-48
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• 2010

[ ${\Delta}^1$ ]pyrroline-5-carboxylate synthetase (P5CS) is a proline biosynthetic pathway enzyme and is known for conferring enhanced salt and drought stress in transgenics carrying this gene in a variety of plant species; however, the wild-type P5CS is subjected to feedback control. Therefore, in the present study, we used a mutagenized version of this osmoregulatory gene-P5CSF129A, which is not subjected to feedback control, for producing transgenic indica rice plants of cultivar Karjat-3 via Agrobacterium tumefaciens. We have used two types of explants for this purpose, namely mature embryo-derived callus and shoot apices. Various parameters for transformation were optimized including antibiotic concentration for selection, duration of cocultivation, addition of phenolic compound, and bacterial culture density. The resultant primary transgenic plants showed more enhanced proline accumulation than their non-transformed counterparts. This proline level was particularly enhanced in the transgenic plants of next generation ($T_1$) under 150 mM NaCl stress. The higher proline level shown by transgenic plants was associated with better biomass production and growth performance under salt stress and lower extent of lipid peroxidation, indicating that overproduction of proline may have a role in counteracting the negative effect of salt stress and higher maintenance of cellular integrity and basic physiological processes under stress.

• The Korea Journal of Herbology
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• v.26 no.3
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• pp.83-90
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• 2011

Objectives : This study was performed to investigate the influence of black ginseng radix extracts (BG) and ginsenoside Rg3, Rg5 on basic fibroblast growth factor (bFGF) induced proliferation, migration and capillary tubule-like formation of human umbilical vein endothelial cells (HUVECs). Methods : HUVECs were cultured with BG and ginsenoside Rg3, Rg5 at different concentrations (60, 125, 250, 500, $1,000{\mu}g/m\ell$) for 2 day In the presence of bFGF, respectively. XTT was used to detect the proliferation. Migration and tube formations were examined to detect the antiangiogenesis. Also, the chick embryo chorioallantoic membrane (CAM) assay was performed to detect the antiangiogenesis. Results : BG and ginsenoside Rg3, Rg5 significantly inhibited bFGF-induced endothelial cell proliferation and migration in a dose-dependent manner. Tube formation in bFGF-induced HUVECs were suppressed by BG and ginsenoside Rg3, Rg5. Moreover, BG and ginsenoside Rg3, Rg5 (30-$50{\mu}g$/egg) inhibited new blood vessel formation on the growing CAM. Conclusions:Based on the present results, it can be suggested that BG has a potential chemopreventive agent via antiangiogenesis.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.4
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• pp.240-246
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• 2016

Objective: The study aimed to investigate the efficacy of intrauterine instillation of granulocyte colony-stimulating factor (G-CSF) on the day of ovulation triggering or oocyte retrieval in infertile women with a thin endometrium. Methods: Fifty women whose endometrial thickness (EMT) was ${\leq}8mm$ at the time of triggering during at least one previous in vitro fertilization (IVF) cycle and an index IVF cycle were selected. On the day of triggering (n = 12) or oocyte retrieval (n = 38), $300{\mu}g$ of G-CSF was instilled into the uterine cavity. Results: In the 50 index IVF cycles, the mean EMT was $7.2{\pm}0.6mm$ on the triggering day and increased to $8.5{\pm}1.5mm$ on the embryo transfer day (p< 0.001). The overall clinical pregnancy rate was 22.0%, the implantation rate was 15.9%, and the ongoing pregnancy rate was 20%. The clinical pregnancy rate (41.7% vs. 15.8%), the implantation rate (26.7% vs. 11.7%), and the ongoing pregnancy rate (41.7% vs. 13.2%) were higher when G-CSF was instilled on the triggering day than when it was instilled on the retrieval day, although this tendency was likewise not statistically significant. Aspects of the stimulation process and mean changes in EMT were similar in women who became pregnant and women who did not. Conclusion: Intrauterine instillation of G-CSF enhanced endometrial development and resulted in an acceptable pregnancy rate. Instillation of G-CSF on the triggering day showed better outcomes. G-CSF instillation should be considered as a strategy for inducing endometrial growth and good pregnancy results in infertile women with a thin endometrium.

• Korean Journal of Plant Resources
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• v.19 no.3
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• pp.427-431
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• 2006

In vitro high-frequency plant regeneration of Muscari comosum var. plumosum through somatic embryogenesis was obtained via two developmental pathways: direct embryos and multiple shoots regenerated from embryogenic callus. Flower bud with pedicel, receptacle, petal and ovary wall, floral stalk and leaf as explants were cultured in MS medium supplemented with various plant growth regulators. Embryos formed directly from pedicel, receptacle and floral stalk. Depending on explant sources, the optimal medium was MS medium supplemented with 0.2 mg/L IBA and 0.3 mg/L BA, 3.0 mg/L IBA and 3.0 mg/L BA, and MS-free medium for pedicel, receptacle, and floral stalk, respectively. Multiple shoots regenerated from embryogenic cal]i which was initiated from petal, ovary and leaf were observed in MS medium with different concentrations and combinations of hormone. The most suitable medium for each type of explant was 3.0 mg/L IBA and 3.0 mg/L BA(petal and ovary) and 5.0 mg/L IBA and 5.0 mg/L BA (leaf) Furthermore, the combination of 0.1 mg/L 2,4-D and 1.0 mg/L BA was also good for all sources of explants not only for direct embryo formation, but also, for embryogenic callus induction.

• Journal of Plant Biotechnology
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• v.33 no.2
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• pp.99-104
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• 2006

This study was performed to investigate regeneration of plants differentiated from hybrid embryos between L. longifilorum Georgia and L. elegans Kakutanohikari. In addition, proliferation of callus and process of differentiation were investigated by histological observation. The germination of hybrid embryos was observed in 86 individuals from 48 slice cultures. Plant regeneration was effective on a medium supplemented with 1 mg/L HPh, and only callus proliferation was the highest in combination of 0.1 mg/L HPh and 1 mg/L BA. Also, plant regeneration was the most effective on a medium supplemented with 50 mg/L pyridoxine. We concluded that somatic embryos were formed from procambium of callus and proliferation of embryonic or proembryonic cells were stimulated with NAA from procambial cells.

• Clinical and Experimental Reproductive Medicine
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• v.13 no.2
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• pp.101-112
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• 1986

It is now common practice to attempt ovarian hyperstimulation in vitro fertilization and embryo transfer (IVF-ET) to promote the development of multiple preovulatory follicles and to maximize the number of mature egg available. There are several drugs for hyperstimulation such as clomiphene citrate only, clomiphene citrate and human menopausal gonadotropin (HMG) and HMG only. Accumlated experience has shown that the hyperstimulation of the ovary in IVF-ET results in high pregnancy rate. But the hyperstimulation of the ovary in IVF-ET may cause the hyperandrogenism, so we must consider the adverse effect on pregnancy rate of the hyperandrogenism. Little is known about the functional significance of androgen for the follicular growth, however, the hyperandrogenism might interfere with oocyte maturation. The aim of the present investigation was to determine the serum profiles of estradiol, androstenedione and testosterone during the hyperstimulated menstrual cycles in IVF. The results were summarized as follows: 1. There was a gradual increase in the mean levels of serum estradiol, androstenedione, and testosterone approaching follicular maturation. 2. The mean serum estradiol levels in the hyperstimulated groups were significantly higher than that in the control group in late follicular phase and ovum retrieval (ovulation) day (p<0.01). 3. The mean serum androstenedione levels in the clomiphene citrate groups were significantly higher than that in the control group in late follicular phase (p<0.01). There was no statistically significant different in the mean serum androstenedione levels between the control group and the HMG group (p>0.05). 4. There was no statistically significant difference in the mean levels of testosterone among each group (p>0.05). 5. There was no statistically significant different in the mean levels of estradiol, androstenedione and testosterone between the fertilized patients and non-fertilized patients in clomiphene citrate and HMG group (p>0.05).

• Clinical and Experimental Reproductive Medicine
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• v.13 no.1
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• pp.67-75
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• 1986

Follicle monitoring in the normal and clomiphene·stimulated cycles were analyzed in the Seoul IVF and ET (In vitro fertilization and embryo transfer) program. Ovarian follicular diameters were measured by the real·time sector scanner and plasma estradiol levels were assayed by radioimmunoassay methods during periovulatory period. The maximum follicular sizes of the clomiphene-stimulated and normal cycles were 21.1+-3.4mm and 19.2+-0.8mm, respectively. The peak levels of plasma estradiol in the clomiphene-stimulated and normal cycles were 10538+-553.6ng/ml and 298.3+-39.6pg/ml, respectively. Daily growth rate of the follicular diameters of the clomiphene-stimulated and normal cycles were 2.1mm and 1.9mm, respectively. Mean follicular number of the clomiphene-simulated and normal cycles were 2.28+-1.12 and 1.12+-0.21, respectively. There was a good statistical correlation between the mean follicular diameters and the plasma estradiol levels in the normal ovulatory and c1omiphene-stimulated ovulatory menstrual cycles (p<0.05). Our data revealad that the mean follicular diameter and the plasma estradiol level prior to HCG administration in IVF and program should reach at the level of 17.8+-3.0mm and 949.4+-487.1 pg/ml, respectively.

• Proceedings of the Botanical Society of Korea Conference
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• 1987.07a
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• pp.213-237
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• 1987

In vitro flowering system may minimize the confounded influence of non-floral meristem parts of plants in studying the relationship of a given treatment and flowering responses. We have induced flower buds from plantlets regenerated from zygotic embryo-derived somatic embryos of ginseng, which circumvented the normal 2-year juvenile period before flowering. The result suggests that the adulthood of ginseng root explants in the experiment previously conducted by Chang and Hsing (1980; Nature 284: 341-342) is not prerequired to flowering of plantlets regenerated through somatic embryogenesis. We have also induced flower buds from elongated axillary brandches from cotyledonary nodes by culturing ginseng zygotic embryos, seedlings, and excised cotyledonary nodes. It was found that 6-benzyladenine (BA) supplemented to the medium was essential for flowering, whereas abscisic acid (ABA) was inhibitory. Gibberellic acid(GA3) was also required for flowering when ABA was present with BA in the medium. The results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respective-ly, in the induction of flowering of ginseng. Tran Thanh Van (1980; Int. Rev. Cytol., Suppl. IIA: 175-194) has developed the "thin cell layer system" in which the induction of shoots, roots, or flower buds from epidermal layer explants were controlled by culture conditions and exogenous growth regulators in the medium, Utilizing the thin cell layer system, Meeks-Wagner et al. (1989; The Plant Cell 1: 25-35) have cloned genes specifically expressed during floral evocation. However, the system is too tedious for obtaining a sufficient amount of plant materials for biochmical and molecular biological studies of flowering. We have developed a garlic callus culture system and one obvious advantaging over the thin cell layer system is that an abundant cells committed to develope into flower buds proliferate. When the above cells were compared by two-dimensional gel electrophoresis with those which have just lost the competence for developing into flower buds, a few putative proteins specific to floral evocation were detected. The garlic callus culture system can be further explored for elucidation of the molecular biological mechanism of floral evocation and morphogenesis.hogenesis.

• Korean Journal of Medicinal Crop Science
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• v.1 no.2
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• pp.184-190
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• 1993

This study was carried out to investigate the appropriate medium and constitutionof growth regulators for somatic embryogenesis for development of rapid mass propagation system via somatic embrygenesis in Rehmennia glutinosa. Embryogenic callus formation from leaf explant was more effective when 4mg / l BA with 0.5mg / l NAA than that of treated with only auxins or cytokinins. LS medium was suitable for embryogenic callus formation. LS medium with 4mg / l BA with 0.5mg / l NAA was effective for the maintenance and proliferation of embryogenic callus. In suspension culture, addition of 1mg / l BA to LS medium was proper for somatic embryogenesis. The highest rate of shoot developement form cotyledon stage embryo was obtained in 1/2 LS medium and plantlet survived by 75% after transplanted to the soil. after 4 weeks.