• Korean Journal of Plant Resources
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• v.26 no.2
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• pp.284-288
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• 2013

This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. The optimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploid plant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, the higher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. While individuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and no tetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulum Thunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05% for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.53-53
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• 2019

Cypripediums, popularly called lady's slippers or moccasin flowers, are the showiest and most sought after hardy terrestrial orchids, collected and grown by orchid and alpine plant enthusiasts alike. In Korea, 4 species of cypripedium are reported as Cypripedium japonicum, C. macranthos, C. guttatum, and C. calceolus. We had already reported the feasibilities of C. macranthos and C. guttatum with in vitro germination methods from immature seeds. The seeds of white lady's slipper orchid (Cypripedium macranthos Sw. alba) were collected 65 days after pollination in 2018. The green pods were sterilized with flame and sowed immediately on the POM(Phytomax orchid maintenance media(R), Sigma) supplemented with BAP 0, 0.5, 1.0 mg/L and NAA 0, 1, 2mg/L. The germination of seed was observed 90 days after sowing, and the plantlets were subcultured to the same media according to the size of the protocorm with 1~2, 2~3, 3~4, 5~6, 7~8mm. The time of the subculture to the new media seems to be critical factors of forming rhizoids which is the hairy root of the cypripediums. As a results, the protorms of the white lady's slipper orchid was successfully germinated in the POM media supplemented BAP 0.5 and NAA 1.0 mg/L. The roots and rhizoids were formed in 5~6mm protocorms subculture over 95% survival ratio. We also tried to subculture to liquid medium without activated charcoal, however the browning or malformation of the roots was observed in the root. The formation of shoots from the protocorm was effectively enhanced in the POM media with non-additives of plant growth regulators. These results indicate the possibility of high and stable production and practical industrialization of endangered white lady's slipper orchids.

• Korean Journal of Plant Resources
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• v.18 no.1
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• pp.123-130
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• 2005

High salt concentrations in the ginseng nursery soil environment of Korea is one of important reducing factors for the stable production of quality ginseng. These studies were accomplished for check the response on germination of ginseng seed, somatic embryogenesis of zygotic embryo, and biosynthesis of ginsenoside from ginseng hairy root against NaCl. Ratio of germination was at the $3\%\;and\;84.5\%$ on the basic media with 0.1M and free of NaCl repectedly, but $0\%$ at the upper of 0.2M NaCl. Somatic embryogenesis from zygotic embryo were the highest when immatured embryo was cultured on free of NaCl concentration, and which was intend to decrease at treatment of NaCl. However, in case of using the matured embryo, treatment of 0.05M NaCl resulted in better embryogenesis than NaCl free media. Red pigment was synthesized from ginseng hairy root cultured on the medium with various NaCl concentration(from 0.04 to 0.08M) and its pigment was analyzed as spectrum of anthocyane by spectrophoto- meter scanning. This cell line biosynthesized lots of crude saponin and total ginsenoside than other cell lines, also had 2 times of panaxadiol than panaxatriol.

• Journal of Plant Biotechnology
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• v.47 no.1
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• pp.53-65
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• 2020

Here, we describe an efficient and rapid protocol for the micropropagation of Thymus pallidus Cosson ex Batt., a very rare medicinal and aromatic plant in Morocco. After seed germination, we tested the effect of different macronutrients, cytokinins alone or in combination with gibberellic acid (GA₃) or auxins, on T. pallidus plantlet growth. We found that Margara macronutrients (N₃₀K) had the best effect on the in vitro development of the plantlets. The addition of 0.93 μM/L 1,3-diphenylurea (DPU), 0.46 μM/L adenine (Ad), and 0.46 and 0.93 μM/L kinetin (Kin) resulted in the best shoot multiplication and elongation. In addition, the combination of 0.46 μM/L Kin, DPU, or Ad with gibberellic acid, in particular, 0.46 μM/L Ad + 0.58 μM/L GA₃ and 0.46 μM/L Kin + 1.15 μM/L GA₃, led to better bud and shoot multiplication. Moreover, the integration of the combinations of 0.46 μM/L Kin and auxins, namely 0.46 μM/L Kin + 2.85 μM/L indole-3-acetic acid (IAA), 0.46 μM/L Kin + 2.85 or 5.71 μM/L indole-3-butyric acid (IBA), and 0.46 μM/L Kin + 0.3 or 0.57 μM/L 1-naphthaleneacetic acid (NAA), in the culture medium led to better root development and optimized aerial growth. Finally, the in vitro plants from the medium containing N₃₀K + 0.46 μM/L Kin + 2.85 μM/L IAA were successfully acclimatized; these plants served as a source for repeating in vitro culture.

• Korean Journal of Organic Agriculture
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• v.25 no.2
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• pp.345-355
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• 2017

This study was conducted to evaluate rice seed disinfection efficacy of loess-sulfur for the suppression of Bakanae disease caused by Fusarium fujikuroi. Rice seeds were treated at different concentrations of loess-sulfur, soaking time and temperature, and combination of hot-water treatment. Rice cultivar, Shindongjin harvested from Bakanae disease-infested area in 2015, was used. Loess-sulfur was treated as follows; concentration of undiluted solution, 2%, 1% and 0.5%; soaking time of 24 and 48 hours; treatment temperature of $20^{\circ}C$ and $30^{\circ}C$; hot water treatment or not. Optimal conditions of rice seed disinfection were selected soaking time of 48 hours and the suspension of 0.5% and 1% loess-sulfur by investigating seed germination and isolation frequency of Fusarium spp. on Komada agar medium in vitro, and were established 3 disinfection conditions as hot water ($60^{\circ}C$, 10 min.) + 1% loess-sulfur ($20^{\circ}C$, 48 hours), 1% loess-sulfur only ($30^{\circ}C$, 48 hours) and 1% loess-sulfur only ($20^{\circ}C$, 48 hours) through additional test in greenhouse. Above 3 conditions were verified by rice seedling box and paddy field test in the way of investigating Bakanae diseased plants (%) and healthy plants (%). Consequently, most effective rice seed disinfection conditions on Bakanae disease were combination of hot water and 1% loess-sulfur and loess-sulfur only at $30^{\circ}C$. Furthermore, treatments with these conditions showed control value of 100% were maintained from seedling to the heading stage in the field. However, treatment of 1% loess-sulfur only at $20^{\circ}C$ showed low control value of 78.2% in paddy field. Hot water only treatment turned out to be an effective disinfection method when conducted thoroughly with $60^{\circ}C$, 10 min. However, it was thought additional soaking process with loess-sulfur after hot water treatment served more high control effect against Bakanae disease when rice seeds were disinfected on a large scale. This results expected rice seed disinfection with loess-sulfur were effectively and easily usable method if farmers had only one of either hot water-disinfector or seed-disinfector. In addition, loess-sulfur is well-known to farmers, simple to manufacture method and cheap.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.149-149
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• 2017

The RING (Really Interesting New Gene) finger proteins are known to play crucial roles in various abiotic stresses in plants. In this study, we report on RING finger E3 ligase, ${\underline{O}ryza}$ ${\underline{s}ativa}$ ${\underline{s}alt$-, ${\underline{A}BA}$- and ${\underline{d}rounght}$ stress-${\underline{i}nduced}$ RING finger ${\underline{p}}rotein{\underline{1}}$ gene (OsSAD1). In vitro ubiquitination assay demonstrated that unlike OsSAD1, a single amino acid substitution ($OsSAD1^{C168A}$) of the RING domain showed no E3 ligase activity, supporting the notion that the activity of most E3s is specified by a RING domain. Result of Yeast-Two hybridization, In vivo protein degradation assay supports that OsSAD1 interacting with 3 substrate, OsSNAC2, OsGRAS44 and OsPIRIN1, and mediates proteolysis of 3 substrates via the 26S proteasome pathway. Subcellular localizations of OsSAD1 while approximately 62% of transient signals were detected in cytosol, 38% of signals were showed nucleus. However, transiently expression of OsSAD1 was detected in cytosol 30% while as 70% of nucleus under 200 mM salt treated rice protoplasts. Results of bimolecular fluorescence complementation (BiFC) showed that two nucleus-localized proteins (OsSNAC2 and OsGRAS44) interacted with OsSAD1 in the both cytosol and nucleus. Heterogeneous overexpression of OsSAD1 Heterogeneous overexpresssion of OsSAD1 in Arabidopsis exhibited sensitive phenotypes with respect to Salt-, mannitol-responsive seed germination, seedling growth. In ABA conditions, OsSAD1 overexpression plants showed highly tolerance phenotypes, such as root length and stomatal closure. Our findings suggest that the OsSAD1 may play a negative regulator in salt stress response by modulating levels of its target proteins.

• Korean Journal of Plant Resources
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• v.35 no.4
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• pp.508-514
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• 2022

Bulbophyllum auricomum Lindl. is very popular among orchid growers due to the attractive fragrance of its flowers and has become an endangered orchid in Myanmar. In this study, we carried out an aseptic technique that can be used to conserve this endangered orchid species. The seeds of B. auricomum Lindl. were obtained from artificial pollination and cultured in MS basal medium for seed germination. The effect of coconut water and BAP in MS basal medium on callus induction was investigated. The highest callus induction was found at 2.0 mg/L BAP. The maximum growth of protocorm-like bodies (PLBs) was evaluated, and the best response was observed on MS medium supplemented with 150 mL/L coconut water at pH 5.6. MS basal medium supplemented with 150 mL/L coconut water along with 2.0 mg/L BAP and 1.0 mg/L NAA (MCBN) showed the highest number of plantlets at 15℃ at the second week of culture. At the second and third week of culture, MS medium supplemented with 2.0 mg/L BAP and 1.0 mg/L NAA (MBN) showed the best result in terms of the number of leaves and the longest leaves at 15℃ and 25℃, respectively. The present study showed evidence of successful in vitro propagation of B. auricomum Lindl.

• Plant Biotechnology Reports
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• v.5 no.2
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• pp.147-156
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• 2011

Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

• Journal of Korean Society of Forest Science
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• v.7 no.1
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• pp.19-27
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• 1968

As a preliminary investigation to obtain useful auto- and allopolyploids a ten year old tetraploid of Pinus densiflora induced from the colchicine treated seed was observed on the cytological, morphological and physioa logical characters in the present study and the results can be summarized as follows. 1. The number of chromosome at the base of needle was 48 so that it was recognized as a tetraploid. 2. The needles were thicker and the number of them on an individual was less than the 2n plant. The needle combined to a single was appeared 2%. 3. No difference was found in the number of stomata rows on the central part of needle between the tetraploid and 2n plant, however, guard cells of tetraploid increased 36% in the long diameter compared with 2n plant. 4. Microsporangiate storbile developed normally showing the same size with 2n plant while the size of pollen grain increased about 20% larger than 2n plant. 5. Germination percentage in vitro increased until 17 hours but decreased after 26 hours compared with 2n plant. The longest length of pollen tube during the germination period appeared in the tetraploid pollen.

• Journal of Bio-Environment Control
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• v.22 no.2
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• pp.116-122
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• 2013

We investigated characteristics and self-incompatibility genotypes of 11 crabapple cultivars to introduce a new pollinizer of 'Fuji' apple tree in Korea. Flowering dates of eleven crabapples were two to seven days earlier than that of 'Fuji'. The rate of pollen germination in vitro was ranged from 85.6% to 98.0% except 'Virginia'. Controlled pollination treatment with each crabapples to 'Fuji' increased fruit set rate about 20.4% to 34.4%, the number of seed per fruit about 13.8% to 42.3% and fruit weight about 7.4% to 16.7% compared to open pollination. Tested crabapples were resistant to peach fruit moth, brown leaf spot and sooty blotch in general. A PCR amplification method using S-RNase primers was carry out in eleven crabapples. S-alleles, $S_3$, $S_5$, $S_9$, $S_{10}$, $S_{20}$, $S_{26} from six crabapples were determinated. Through sequencing analysis, $S_5$ ('Manchurian', 'Virginia') and $S_9$ ('Yantaishagou') showed 100% homologous to previous result. Based on our results, it was recommended that 'Manchurian', 'Hopa A', 'Hanyaehanakaidou', 'Spectabilis' could be promising pollinzers for 'Fuji' apple cultivar.