• 한국자원식물학회지
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• 제26권3호
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• pp.383-388
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• 2013

The research concerned of the regeneration of plants from embryos obtained from anther cultures of ginseng (Panax ginseng C. A. Meyer). The aim was to determine the influence of the regeneration medium on the efficiency of the regeneration process. We conducted to determine the optimum conditions such as cold pretreatment, plant growth regulators and carbon sources on anther culture of P. ginseng. Highest callus formation rate was obtained when flower buds pretreated at $4^{\circ}C$ for 1 day. Among the treated growth regulators with various degrees of concentration in Murashige and Skoog's (MS) medium, 4.53 ${\mu}M$ of 2.4-dichlorophenoxyacetic acid and 4.44 ${\mu}M$ of 6-benzylaminopurine gives the most responsive callus with the frequency of 73.89% and 129.53 g of fresh weight. When we used 3-9% of sucrose and maltose among the different kinds and various concentrations of carbohydrates, callus was formed highest 67.29% in the medium with 3% of sucrose. Shoots induced from callus supplemented with 28.9 ${\mu}M$ of gibberellic acid and rooted in Gamborg's B5 medium supplemented with 14.7 ${\mu}M$ of indole-3-butyric acid.

• 원예과학기술지
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• 제29권5호
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• pp.488-493
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• 2011

Callus culture initiation of strawberry (Fragaria${\times}$ananassa Duch.) was investigated at different Murashige and Skoog (MS) medium strengths, types and concentrations of plant growth regulators, and incorporating a cold pretreatment period to determine the optimal nutritional and environmental conditions. No high quality callus was induced on MS media without auxin regardless of medium strength. When 6-benzylaminopurine (BA) was combined with indole acetic acid (IAA), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D), high quality callus were highly induced compared to medium supplemented with auxin alone. When $0.5mg{\cdot}L^{-1}$ BA was combined with IAA, NAA, and 2,4-D, high quality callus induction was more effective than the medium supplemented with the other BA concentrations. The best combination of auxin and cytokinin for high quality callus induction was $1.0mg{\cdot}L^{-1}$ NAA and $0.5mg{\cdot}L^{-1}$ BA. Although the differences in callus induction were not significant, high quality callus induction at half strength MS medium was more effective than at full strength medium. When $30g{\cdot}L^{-1}$ sucrose was added to the half strength MS medium, the rate of high quality callus induction increased. The optimum cold pretreatment temperature and period for high quality callus induction were $4^{\circ}C$ and 72 h, respectively. Regeneration rate of high quality callus increased in MS medium supplemented with thidiazuron.

• Journal of Plant Biotechnology
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• 제34권3호
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• pp.189-195
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• 2007

Helicteres isora is medicinally important plant effective against asthma, diabetes, hypolipidemia, HIV, besides a good source of diosgenin. Seed dormancy and low rate of natural fruit production make this plant a perfect candidate for developing an in vitro method useful for its clonal propagation and further biotechnological developments. This is the first report on in vitro production of this plant. Nodal explants obtained from aseptically germinated seedlings were cultured on MS medium (Murashige and Skoog 1962) fortified with indole-3-acetic acid (IAA) ($0.57-22.83\;{\mu}M$), indole-3-butyric acid (IBA) ($0.41-16.58\;{\mu}M$), 6-benzylaminopurine (BA) ($0.44-17.75\;{\mu}M$) and kinetin (Kin) ($0.46-13.94\;{\mu}M$) either singly or in combinations of IAA + BA, IAA + Kin and BA + Kin. Combinations of cytokinins (BA and Kin) were most suitable for multiple shoot induction and $13.94\;{\mu}M\;Kin\;+\;13.31\;{\mu}M\;BA$ was optimum (79% frequency) associated with high number of microshoots (7.1 shoots per explant) after 20 days of culture. Maximum shoot elongation and proliferation (10 shoots per explant with 4.8 cm average height) was achieved on MS media containing $2.32\;{\mu}M\;Kin\;+\;2.22\;{\mu}M\;BA\;+\;2.85\;{\mu}M\;IAA$. High rooting frequency (70%) was achieved on MS medium (1/2 basal strength) fortified with $4.14\;{\mu}M$ IBA, while activated charcoal showed inhibitory effects on rooting. Hardening was done with 76% survival rate and these plants were growing without any visual defects and morphologically mimicking the naturally growing plants.

• Current Research on Agriculture and Life Sciences
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• 제10권
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• pp.157-164
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• 1992

본 연구는 Petunia hybrida $F_1$ hybrid를 약배양(葯培養)하여 반수체(半數體)를 얻을 목적으로 약(葯)의 치상시기(置床時期)와 배지(培地)의 종류(種類), 생장조절물질(生長調節物質)의 종류와 농도, 약(葯)의 전처리(前處理)와 품종(品種)에 따른 캘러스 형성률을 조사하였던 바 다음과 같은 결과를 얻었다. 약배양하기 전 0.8-1.2cm의 길이의 화뢰(花雷)가 달린 모본(母本)의 줄기를 $4^{\circ}C$에서 15일간 저온처리(低溫處理)하였다. 약을 채취하여 1/2 MS배지에 NAA 5.0mg/${\ell}$, BAP 0.5mg/${\ell}$를 첨가한 배지(sucrose 30g/${\ell}$, 2g/${\ell}$, pH 5.8)에서 배양 4주후 연녹색의 캘러스가 형성되었으며, 이들 캘러스로부터 배양 3주후 2ip 2.0mg/${\ell}$가 함유된 MS배지에서 식물체가 분화되었다.

• BMB Reports
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• 제32권1호
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• pp.51-59
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• 1999

The Korean radish cationic peroxidase (KRCP) promoter, comprising nucleotides -471 to +704 relative to the transcriptional initiation site, was fused to the GUS gene and transformed to tobacco BY-2 cells. We examined how auxin (2,4-dichlorophenoxyacetic acid, 2,4-D), cytokinin (6-benzylaminopurine, BAP), gibberellic acid ($GA_3$), abscisic acid (ABA), methyl jasmonate (MeJA), and phosphatidic acid (PA) affect the GUS expression in the presence or absence of 2,4-D in a modified LS medium. Exogenous 2,4-D or BAP greatly decreased the GUS expression regulated by the KRCP promoter in a modified LS medium containing 0.2 mg/l 2,4-D. $GA_3$ increased the GUS expression and ABA completely reduced the inductive effect of $GA_3$. The GUS expression was also increased dose-dependently by plant defense regulators, MeJA and PA. In contrast to the above results, auxin deprivation from the modified LS medium increased the GUS expression after treatment with exogenous 2,4-D whereas BAP still greatly decreased the GUS expression dose-dependently. $GA_3$ or MeJA slightly decreased the GUS expression. The data suggest that auxin deprivation changes the sensitivity of the suspension cells to exogenous chemicals and that the regulation of the KRCP promoter by 2,4-D, $GA_3$, and MeJA is dependent on auxin, whereas the regulation by BAP is not. This study will be valuable for understanding the function and expression mode of the Korean radish cationic peroxidase in Korean radish.

• Journal of Plant Biotechnology
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• 제36권2호
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• pp.174-178
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• 2009

Presently, we report a simple, reproducible and high frequency plant regeneration in Hibiscus syriacus L. using axillary buds. H. syriacus was regenerated from axillary buds directly or through a callus phase. Regenerated shoots were directly induced from young and fresh axillary buds cultured on Murashige and Skoog medium (MS) supplemented with 0.01 mg/L of the growth regulator thidiazuron (TDZ) after 2 weeks of culture. Directly induced shoots were transferred to hormone-free MS medium and root development was observed after 6 weeks. On the other hand, old and stale axillary buds were regenerated to shoots via callus induction on MS medium containing 0.01–2 mg/L TDZ after 4 weeks. A TDZ concentration of 0.01 mg/L was most effective in callus formation. Green callus was transferred to MS medium containing 0.01 mg/L α-naphthalene acetic acid (NAA) and 0.5 mg/L benzylaminopurine (BA). After 4 weeks, callus had developed into multiple shoots. Plantlets were formed from 10 week cultures of single shoots on hormone-free MS medium. Regenerated plantlets were cultured on MS medium for one month and then transferred to pots containing garden soil. Potted plants were acclimatized for one month and grown to maturity under greenhouse conditions. The present study has shown that various concentrations of plant growth regulator can be effective for in vitro plant regeneration of H. syriacus. The direct and indirect regeneration protocol presented here will be useful for understanding the manipulation and propagation of H. syriacus.

• Journal of Plant Biotechnology
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• 제47권3호
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• pp.235-241
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• 2020

This is the first study to establish a complete protocol for micropropagation of Rehmannia glutinosa from root segments. The study involved investigating the effect of plant growth regulators on in vitro shoot regeneration and rooting and identifying substrates supporting survival and growth performance of ex vitro seedlings. A Murashige and Skoog (MS) medium containing 30 g/L sucrose for shoot induction and 0.2 mg/L indole-3-acetic acid (IAA), 1 mg/L 6-benzylaminopurine (BAP), and 1 g/L polyvinylpyrrolidone (PVP) for shoot multiplication resulted in the highest number of shoots per explant and shoot height. Applying a medium containing 0.5 mg/L IAA and 1 g/L PVP yielded optimal rooting of the shoots grown in vitro. Compost enriched with microbial inoculants and perlite enhanced seedling growth better than that with organic biofertilizer-free substrates (soil and sand). We recommend the continuous production of micropropagated R. glutinosa seedlings from root segments under the aforementioned conditions as a possible propagation technique for crops of this species.

• Journal of Plant Biotechnology
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• 제44권3호
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• pp.335-342
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• 2017

Purple passion fruit (Passiflora edulis Sims) is one of the introduced tropical plants, an increasing interest has arisen due to its distinctive taste and attractive flavor. It is expected that passion fruit production and planted area will increase gradually in the years ahead because of high profitability and consumer's demands of healthful ingredients. So we tried to investigate the effect of plant growth regulators and antioxidants on in vitro plant regeneration and callus induction from leaf explants of passion fruit for an establishment of optimal mass propagation system. Young leaf explants of purple passion fruit were cultured in Murashige and Skoog (MS) medium containing different growth regulators and antioxidant additives to induce the shoot organogenesis. After 8 weeks, the highest embryogenic callus formation rate was obtained in MS medium supplemented with $1mg{\cdot}L^{-1}$ 6-benzylaminopurine (BAP) and $2mg{\cdot}L^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), furthermore, the shoot development via organogenesis was also observed. Silver nitrate ($AgNO_3$), which was added into the medium to minimize the adverse effects of leached phenolics, was effective for reduction of medium browning and sudden explant death. In the medium supplemented with $1mg{\cdot}L^{-1}$ BAP and $1mg{\cdot}L^{-1}$ gibberellic acid ($GA_3$), shoots were most vigorously regenerated and elongated. Most shoots rooted successfully in half strength medium with $1mg{\cdot}L^{-1}$ indol-3 acetic acid (IAA), and more than 90% of plantlets survived after 4-month acclimatization period.

• 한국작물학회지
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• 제48권1호
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• pp.38-43
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• 2003

Immature and mature embryos of 18 Korean wheat genotypes were cultured in vitro to develop an efficient method of callus formation and plant regeneration, and to compare the responses of both embryo cultures. Immature and mature embryos were placed on a solid agar medium containing the MS salts and vitamins, 30g/l maltose, 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), and amino acids. The developed calli were maintained on regeneration medium containing MS salts and B5 vitamins, 20 g/l sucrose, and the combination of two plant growth regulators, 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). Immature embryos in most genotypes showed high efficiency of callus induction except three genotypes; Eunpamil, Chunggemil, and Namhaemil, and significant differences among the genotypes. Plant regeneration of calli induced from immature embryos showed high efficiency in Geurumil (56.5%), Tapdongmil (50.5%), Gobunmil (45.5%), and Urimil(42.2%). The analysis of variance showed significant differences for regeneration frequency among the genotypes. Mature embryos showed low callus induction frequency compared with that in immature embryos, and significant differences among the genotypes. Plant regeneration of calli induced from mature embryos showed high efficiency in Keumkangmil (33.33%), Tapdongmil(28.13%), and Geurumil (27.78%). The analysis of variance showed significant differences for plant regeneration frequency among the genotypes.

• 통합자연과학논문집
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• 제4권2호
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• pp.113-120
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• 2011

Brassinosteroids (BRs) are plant steroid hormones that play essential roles in growth and development. Mutations in BR-signaling pathways cause defective in growth and development like dwarfism, male sterility, abnormal vascular development and photomorphogenesis. Transition from vegetative to reproductive growth is a critical phase change in the development of a flowering plant. In a screen of activation-tagged Arabidopsis, we identified a mutant named abz126 that displayed longer hypocotyls when grown in the dark on MS media containing brassinazole (Brz), an inhibitor of BRs biosynthesis. We have cloned the mutant locus using adapter ligation PCR walking and identified that a single T-DNA had been integrated into the ninth exon of the GIGANTEA (GI) gene, involved in controling flowering time. This insertion resulted in loss-of-function of the GI gene and caused the following phenotypes: long petioles, tall plant height, many rosette leaves and late flowering. RT-PCR assays on abz126 mutant showed that the T-DNA insertion in GIGANTEA led to the loss of mRNA expression of the GI gene. In the hormone dose response assay, abz126 mutant showed: 1) an insensitivity to paclobutrazole (PAC), 2) an altered response with 6-benzylaminopurine (BAP) and 3) insensitive to Brassinolide (BL). Based on these results, we propose that the late flowering and tall phenotypes displayed by the abz126 mutant are caused by a loss-of-function of the GI gene associated with brassinosteroid hormone signaling.