• Journal of Plant Biotechnology
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• 제43권1호
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• pp.104-109
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• 2016

Rosa rugosa is a medicinal, ornamental, and edible plant native to Eastern Asian countries, including Korea, Japan, and China. The aim of this study was to establish a system for biomass production and secondary metabolite accumulation during in vitro culture and acclimatization of Rosa rugosa. The highest rate of multiple shoot proliferation was achieved with $8.8{\mu}M$ benzyladenine (BA) (83.3%). However, the number of shoots (14.4 per explant) at $4.4{\mu}M$ BA was higher than that at $8.8{\mu}M$ BA. Compared to BA, a combination of thidiazuron (TDZ) and indole butyric acid (IBA) exhibited significantly lower shoot induction, with only 50.0~79.2% and 4.2~16.7% relative shoot formation, respectively. During acclimatization, shoots were sampled every week and their total phenolic contents were analyzed. Among various growth factors, fresh weight showed the most dramatic increase from the 3rd week (88.0 mg/plant) to 4th week (132.7 mg/plant). Total phenolics and flavonoids contents were the highest at $1^{st}$ week of acclimatization. Depending on developmental stages, total phenolics and flavonoids contents were higher in 1-yr-old shoots grown ex vitro than in those of older field-grown or in vitro-grown plants. Amongst different ages of field grown plants, 6-year-old plants, the oldest in this study, showed the lowest content in total phenolics.

• Journal of Plant Biotechnology
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• 제34권4호
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

• Plant Biotechnology Reports
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• 제4권1호
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• pp.53-59
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• 2010

A specific deleted version of ARABIDOPSIS RESPONSE REGULATOR1 (ARR1) lacking the signal receiver domain (1.152 amino acids)-coding sequence, referred to as $ARR1{\Delta}DDK$, was amplified using Arabidopsis thaliana cDNA prepared from adult leaves and transferred into the genome of Nicotiana tabacum cv. Samsun under the transcriptional control of a ${\beta}$-estradiol-inducible expression system. The ectopic expression of $ARR1{\Delta}DDK$ affected the morphology of transgenic seedlings and their segments in vitro. In the presence of an inducer, ${\beta}$-estradiol, ectopic expression of $ARR1{\Delta}DDK$ induced only the formation of soft, pseudo-bulbous tissue in the root tip region of intact seedlings, which appeared similar to callus generated on a hypocotyl segment in the presence of 2,4-D and 6-benzyladenine (BA), both at $1\;{\mu}M$. Those callus tissues on the root tip region could not generate shoots unless $1\;{\mu}M$ BA was supplied. In segment culture, ectopic expression of $ARR1{\Delta}DDK$ induced calluslike tissue around the cut-end of cotyledon and hypocotyl segments with occasional shoot formation, suggesting that the expression of $ARR1{\Delta}DDK$ could substitute for the effects of cytokinin on these segments. Additionally, treatment with only ${\beta}$-estradiol induced NtWUS, a WUS ortholog in tobacco, which was detected during the process of callus tissue formation in the root tip region and also in cotyledon or hypocotyl segments. These findings suggest that the NtWUS might be associated in the transdifferentiation process caused by the functional regulation of $ARR1{\Delta}DDK$ in transgenic tobacco seedlings.

• Journal of Plant Biotechnology
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• 제39권3호
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• pp.182-188
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• 2012

Iris dichotoma Pall. is an important endangered plant belonging to the family Iridaceae. A method was developed for the rapid micropropagation of I. dichotoma through plant regeneration from leaf, rhizome, and root explant-derived calli. Leaf, rhizome, and root segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; $0-3.0mg{\cdot}L^{-1}$) for callus induction. Callus production was highest at $1.0mg{\cdot}L^{-1}$ 2,4-D, where 73.8% and 45.5% of cultured rhizome and root cuttings, respectively, produced calli. The viable calli were maintained at an induced concentration of 2,4-D ($3.0mg{\cdot}L^{-1}$). They were then transferred to MS medium supplemented with various concentrations of 2,4-D ($0-3.0mg{\cdot}L^{-1}$) in combination with 6-benzyladenine (BA: 0, 1.0 and $3.0mg{\cdot}L^{-1}$) for adventitious shoot regeneration. The addition of a low concentration of 2,4-D into BA-containing medium significantly increased the frequency of shoot regeneration in leaf, rhizome, and root-derived calli. The highest number of adventitious shoots (26.4 per callus) formed at $0.5mg{\cdot}L^{-1}$ 2,4-D and 1.0 mg/l BA. For rooting of the shoots, half- strength MS medium supplemented with different concentrations of indole 3-butyric acid (IBA) $0-3.0mg{\cdot}L^{-1}$ was tested. The optimal results were observed using half-strength MS medium supplemented with $1.0mg{\cdot}L^{-1}$ IBA, on which 98% of the regenerated shoots developed roots with an average of 3.5 roots per shoot within 45 days. The plantlets raised in vitro were acclimatized and transferred to soil with 95% success. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

• 원예과학기술지
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• 제17권6호
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• pp.755-757
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• 1999

본 연구는 동양란 '철골소심'의 개화증진을 위하여 BA처리후 적정한 관리온도를 알아보기 위해 수행되었다. 개화율은 주야간의 기온이 $30/25^{\circ}C$을 유지했을 때 90%, $25/20^{\circ}C$와 대조구에서 50%로 나타났으며 $20/15^{\circ}C$처리에서는 전혀 개화가 되지 않았다. 기형화율은 $25/20^{\circ}C$에서는 나타나지 않았고 $30/25^{\circ}C$에서는 4%, 대조구에서는 9%로 나타났다. $30/25^{\circ}C$에서 화경장, 화경수 등이 증가하였고 화아품질이 향상되었다. BA 살포 후 철골소심의 개화 유도를 위한 한계 기온은 $20^{\circ}C$ 이상인 것으로 나타났다.

• 한국약용작물학회지
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• 제23권5호
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• pp.370-378
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• 2015

Background : Aruncus dioicus var. kamtschaticus, functional wild vegetable, is perennial herb and young leaves with soft texture are generally used as edible food. So the demand for the vegetable has increased recently. This study was carried out to determine the effects of temperature and shading on germination characteristics of A. dioicus. Methods and Results : The experiment was performed by temperature and shading treatments. Seed pre-treatment before the germination experiment was carried out by the storage in low temperature ($4^{\circ}C$) under wet condition (LTW) for 0, 15, 30, 45 and 60 days and shading treatment were 35%, 50%, and 75% under control, BA (6-benzyladenine) and $GA_3$ (gibberellic acid) condition for 24 hours. Increasing the length of the storage periods led to increases seed germination percent in low temperature ($4^{\circ}C$) under wet condition (LTW), germination rate of A. dioicus seed was the highest at $15^{\circ}C$ with 60 days of seed pre-treatment. In the case of seeds pre-treatment with LTW, the more temperature went up, the more days to 50% of Germination of Final Germination Rate ($T_{50}$) went down. As a result of surveying shading treatment, germination rate was the highest in control of 35% shading and the next higher was in control of 50% shading. Conclusions : It is concluded that the temperature and shading are important factors to produce A. dioicus. Also, We suggest these results as basic data of A. dioicus for sexual propagation.

• 한국환경과학회지
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• 제8권6호
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• pp.669-676
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• 1999

The effects of wounding and jasmonic acid(JA) on polyphenol oxidase(PPO) in tomato(Lycopersicon esculentum Mill.) seedlings were investigated. PPO was strongly induced by wounding or JA, and the response was also shown to be systemically induced by wounding. Mechanical wounding in cotyledon or leaf produced a signal that caused the concentration of PPO to increase in the unwounded cotyledon, in the first leaves but not in the second leaves. Severity of wounding and light intensity also affected wound induced change in PPO activity, JA showed a stimulatory effect on the loss of chlorophyll and the rapid increase in PPO activity. The PPO was clearly more active in the wounded leaves than in controls. The potency and specificity of the JA indicate a close relationship between JA and wound-induced changes in PPO in tomato species. JA and abscisic acid(ABA) acted similarly on both unwounded and wounded leaves, but the amount of PPO in the wounded leaves was always more than the respective controls. The highest increase in PPO activity occurred in woundand JA-induced leaves of seedlings kept under bright lighting. Benzyladenine(BA) completely abolished JA- and ABA-induced PPO activity. The results suggest that JA-induced PPO activity is due to de novo PPO synthesis. Histochemical tests for PPO in stems of wound- and JA -treated tomato plants indicate that PPO was localized primarily, in the. outer .cortex . and xylem parenchyma. It is concluded that exogenously applied JA acts as stress agents and PPO may be a component of the inducible anti-hervivore defense response.

• Journal of Ecology and Environment
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• 제30권3호
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• pp.271-276
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• 2007

A simple and efficient protocol was developed for culturing Cu-tolerant and Cu-accumulating plants via pre-adaptation to Cu during plant tissue culture. We induced multiple shoots from begonia (Begonia evansiana Andr.) leaf explants on MS medium supplemented with naphtaieneacetic acid and benzyladenine. After 3 months, small plantlets were transferred to MS medium supplemented with $100{\mu}M\;CuCl_2$ for pre-adaptation to Cu and cultured for 5 months. Then, these plantlets were individually planted in pots containing artificial soil. An additional 500 mg of Cu dissolved in 1/4 strength MS solution was applied to each pot during irrigation over the course of 2 months. We planted pre-adapted and control begonias in soil from the II-Kwang Mine, an abandoned Cu mine in Pusan, Korea, to examine their ability to tolerate and accumulate Cu for phytore-mediation. Pre-adapted begonias accumulated $1,200{\mu}g$ Cu/g dry root tissue over the course of 45 days. On the other hand, non-Cu-adapted controls accumulated only $85{\mu}g$ Cu/g dry root tissue. To enhance Cu extraction, chelating agents, ethylenediamine tetraacetic acid (EDTA)-dipotassiun and pyridine-2,6-dicarboxylic acid (PDA), were applied. While the chelating agents did not enhance accumulation of Cu in the roots of control begonias, EDTA application increased the level of Cu in the roots of pre-adapted begonias twofold (to $2,500{\mu}g$ Cu/g dry root tissue). Because pre-adapted begonias accumulated a large amount of Cu, mainly in their roots, they could be used for phytostabilization of Cu-contaminated soils. In addition, as a flowering plant, begonias can be used to create aesthetically pleasing remediation sites.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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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.

• Advances in Traditional Medicine
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• 제9권2호
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• pp.174-181
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• 2009

Pterocarpus marsupium, Clitoria ternatea, and Sanseveiria cylindrica are some of the important and endangered medicinal plant species of India. Despite of medicinal properties, antibacterial potential of the plants have not yet been explored. The present study was designed to optimize the in vitro technique for micropropagation and to screen the extracts from leaves and in vitro raised calli for antibacterial properties. Excised leaf-explants from the parent plants were surface sterilized and cultivated on Murashige & Skoog's (MS) medium containing $N^6$-benzyladenine (BA) in concentrations of 1, 2, 5, and $10{\mu}M$. Optimal growth of calli was noticed at a concentration of $5{\mu}M$, therefore the extracts from calli grown at this concentration were further studied for antibacterial activity. Both alcoholic and aqueous extracts from leaves of respective plants, and their in vitro raised calli were tested for antibacterial activity by agar well diffusion method against a range of Gram-positive and Gram-negative bacteria. Aqueous extracts showed antibacterial activity against limited number of bacterial species; notably the extracts of C. ternatea which showed antibacterial activity against Streptococcus pyogenes, Bacillus subtilis and Bacillus cereus. Alcoholic extracts of all three plants showed antibacterial activity against a wider range of bacteria. Among the Gram-positive bacteria, extracts from C. ternatea showed strong antibacterial activity against Bacillus spp., whereas the extracts of S. cylindrica showed good antibacterial potential for Staphylococcus aureus, S. epidermidis and S. pyogenes. The extracts from all three plants showed antibacterial activity against Gram-negative bacteria, including, Salmonella spp. and Shigella dysenteriae; organisms causing enteric fever and dysentery. In most of the cases, the extracts from respective calli showed comparable, and in some cases better, result in comparison to the extracts from parent leaves. To the best of our knowledge this is the first preliminary report on antibacterial potential, especially through calli extracts, of these plants; and in vitro cultivation of the explants may be used to obtain phytotherapeutic compounds.