• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.379-387
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• 2017

Various sizes (0.2 ~ 1.2 mm) and developmental stages (referred to as Stage 1 ~ 3) of apical and lateral meristems were excised, together or separately, directly from dormant buds of apple 'Hongro'. They were mixed infected by Apple scar skin viroid (ASSVd), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV), which are major viruses attacking apples. A total of 31 callus lines (> 10 mm in diameter) were obtained by culturing the explants on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA), and they were subjected to RT-PCR analysis for virus detection. A high rate of virus elimination (expressed as the percentage of calli that did not amplify during RT-PCR, i.e., RT-PCR negative calli per total number of calli obtained) was achieved for ACLSV (100%), ASSVd (93.7%), and ASPV (93.7%), whereas it was only 25.8% for ASGV. ASPV was detected in the presence of 2 ~ 3 bracts. Simultaneous virus elimination of ASSVd, ASPV, ACLSV, and ASGV occurred during the meristem culture, in which the early stages of the dormant buds (Stage 1) were used, because ASGV was mostly eliminated during that stage. The results of the present study will be valuable for the production of virus-free apple trees.

• Horticultural Science & Technology
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• v.28 no.4
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• pp.678-684
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• 2010

In an attempt to improve biomass and bioactive compound production, we cultured adventitious roots of $Eleutherococcus$ $koreanum$ in 250 mL Erlenmeyer flasks using Murashige and Skoog (MS) medium with different concentrations of auxins (IBA, NAA, IAA) and cytokinins (BA, kinetin, TDZ). Root biomass (fresh and dry weight) was enhanced at $5mg{\cdot}L^{-1}$ indole-3-butyric acid (IBA) after 5 weeks of culture. The content of total phenolics and flavonoids was also increased with $5mg{\cdot}L^{-1}$ IBA compared to ${\alpha}$-naphtalene acetic acid (NAA) or indole-3-acetic acid (IAA) treatments. The combination of $5mg{\cdot}L^{-1}$ IBA with $0.1mg{\cdot}L^{-1}$ thidiazuron (TDZ; N-phenyl-N'-1,2,3,-thidiazol-5-ylurea) enhanced root fresh and dry weight (1.4- and 1.6-fold, respectively) as well as the content of total phenolics and flavonoids compared to the relative control (without cytokinin). On the contrary, $N_6$-benzyladenine (BA) and 6-furfurylaminopurine (kinetin) did not significantly affect root biomass and bioactive compound production in adventitious roots of $E.$ $koreanum$. These results suggested that $5mg{\cdot}L^{-1}$ IBA combination with $0.1mg{\cdot}L^{-1}$ TDZ supplementation was most suitable for both biomass and bioactive compound production from adventitious roots of $E.$ $koreanum$.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.3
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• pp.204-208
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• 2006

Utilization of synthetic benzyladenine (BA) for bean sprout production should be reduced or minimized because of elevating production cost and not ascertaining action mechanism to human body. The study was done to compare the effects of BA concentrations under different watering methods (overspraying or underwatering) on growth and morphological characteristics of mungbean sprouts. Seeds of cv. Zhong-Lu 1 were soaked for 5 hours in the solutions with different BA concentrations (0, 10, 20, 30, 40, 50 ppm) before 4 hour aeration, and then were cultured for 6 days by both watering systems. Their morphological characters, fresh and dry weights were measured. Regardless of watering methods, lateral roots were sharply dropped over 30 ppm BA concentrations, and hypocotyl, root and total lengths were shortened with increased BA concentrations although ratios of hypocotyl to root lengths and hypocotyl diameters were enlarged with their increment. Total fresh weights were increased up to 20 ppm in overspraying method but up to 30 ppm in underwatering method mainly due to increment of hypocotyl fresh weights. The sprouts were faster grown in overspraying method than in underwatering method because the former showed longer lengths of hypocotyl and root, and total fresh weights.

• Korean Journal of Food Science and Technology
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• v.14 no.2
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• pp.179-183
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• 1982

This experiment was carried out to clarify the optimal concentrations of growth regulators for callus induction and the condition of callus culture of leaf tissue taken from Stevia rebaudiana Bertoni. The content of stevioside, sweetening component, in leaf-derived callus of stevia was also investigated. It was shown that the optimal concentrations of benzyladenine (BA) and ${\alpha}-naphthalene$acetic acid (NAA) for callus induction were $10^{-6}M$ and $10^{-5}M$, respectively. Reculture of these calli in media (Linsmaier and Skoog) supplemented with BA $10^{-6}M$ and NAA $10^{-5}M$ resulted in profuse calli 15 to 20 days after incubation. When sweetening components produced by callus were extracted and identified by TLC, stevioside appeared to have Rf value 0.50 in TLC which was exactly same as standard stevioside. Stevioside content obtained by TLC-FID analyzer was 260mg per 100g on the basis of dry weight.

• Journal of The Korean Society of Grassland and Forage Science
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• v.26 no.2
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• pp.69-76
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• 2006

To optimize tissue culture responses for genetic transformation of Kentucky bluegrass, the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of a cultivar 'Newport' as explant tissues. The optimal concentration of 2,4-D (2.4-dichloro phenoxy acetic acid) for the induction of embryogenic callus from mature seed was 3 mg/L. Plant regeneration frequency was 54% when embryogenic callus was cultured on the regeneration medium supplemented with 1 mg/L 2,4-D and 3 mg/L of BA (6-benzyladenine). Addition of 1 g/L of casein hydrolysate and 500 mg/L of L-proline improved frequencies of embryogenic callus induction and plant regeneration up to 60.8% and 58.3%, respectively. Regenerated plants were grown normally when shoots transplanted to the soil. A rapid and efficient plant regeneration system established in this study. We suggest that the results may be useful for molecular breeding of Kentucky bluegrass through genetic transformation.

• Korean Journal of Medicinal Crop Science
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• v.3 no.2
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• pp.111-115
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• 1995

This study was carried out to investigate the effect of some plant growth regulators on the germination and seedling growth of Wasabia japonica Matsum seeds. The seeds were soaked in different concentration of indolacetic acid(LAA), indolbutyric acid(IBA), $GA_3\;GA_4$, Benzyladenine(BA) and kinetin, and the germination percent, the average germination day and the growth of seedlings were measured. The seeds were released from dormancy and the germination percent was considerably increased when the seedcoat was removed. $GA_3,\;GA_4,$ BA and kinetin were effective in breaking the dormancy, but LAA and IBA were not effective. $GA_4$, BA and kinetin were more effective in accelerating germination of dormancy seed than $GA_3$. The applications of BA combined with $GA_3$ l00ppm showed higher germination percent than that of BA alone within the range 50ppm and 100ppm. Root, hypocotyl and petiol of seedling overgrew by the application of $GA_3\;and\;GA_4$. But effects of BA and kinetin were good on seedling growth compared with those of $GA_3\;and\;GA_4$. Therefore, BA and kinetin could be use for dormancy breaking of seed instead of $GA_3$.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.239-244
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• 2015

Calanthe striata f. sieboldii Decne. ex Regel is a terrestrial orchid with beautiful flowers arranged in racemose inflorescences. This species is threatened due to over-collection and loss of suitable habitats. Asymbiotic germination is useful in the conservation efforts to re-establish plants in the wild, and for commercial propagation. In this study, we investigate the effects of sodium hypochlorite (NaOCl), three types of culture media (Phytomax Orchid Maintenance - POM, Seed Germination Maintenance - SGM, and Murashige and Skoog 1962 - MS), and plant growth regulators on embryo swelling, protocorm formation, and embryo diameter of C. striata f. sieboldii. Treatment with 1% NaOCl for 30 min greatly enhanced embryo swelling (28.3%), embryo diameter ($205.8{\mu}m$), and embryo protocorm formation (54.8%) compared to seeds without NaOCl treatment (embryo swelling 8.5%, embryo diameter $14.6{\mu}m$, and protocorm formation 13.4%) on POM medium. Protocorm formation on POM medium supplemented with 1.0 mg/L N6-benzyladenine (BA) (95.6%) was better than the control (54.5%). Additionally, the effects of activated charcoal (AC) and sucrose on seedling growth in in vitro culture were examined. The protocorm converted into healthy plants with well-developed shoot primordia on the POM medium with AC and sucrose. The most suitable conditions for seedling growth after 10 weeks of culture were the POM medium with AC or sucrose. These results show effective asymbiotic seed germination and in vitro seedling growth of C. striata f. sieboldii.

• Journal of Plant Biotechnology
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• v.34 no.2
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• pp.139-144
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• 2007

Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.

• Korean Journal of Medicinal Crop Science
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• v.14 no.4
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• pp.255-260
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• 2006

Culture conditions for high frequency plant regeneration via somatic embryogenesis from embryogenic cell suspension cultures of Hovenia dulcis are described. Germinated somatic embryos were selected for induction of secondary embryogenesis. Friable embryogenic cells were induced directly from somatic embryos when transfer to 1/3 MS solid or liquid medium lacking plant growth regulators. The temperature strongly effected on induction of secondary embryognesis than other conditions in culture. All somatic embryos produced friable embryogenic cell clumps within 10 days when germinated somatic embryos cultured in 1/3 MS medium at $30^{\circ}C$ in suspension culture. No somatic embryos formed from embryogenic cell suspension cultures at $18^{\circ}C$. Numerous somatic embryos were induced and subsequently developed uniformly into germination stage from suspended cell clumps after 4 weeks of culture on $18^{\circ}C$. Plantlets conversion were observed on $18^{\circ}C$ when germinated somatic embryos were transferred to 1/3 MS solid medium without plant growth regulators or supplemented with 0.1-0.5 mg/l benzyladenine.

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.187-193
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• 2005

In an effort to optimize tissue culture conditions for genetic transformation of tall fescue (Festuca arundinacea Schreb.), an efficient plant regeneration system from seed-derived calli was established. MS medium containing 6 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.1 mg/L benzyladenine (BA) were optimal for embryogenic callus formation from mature seed and had a strong effect on successive plant regeneration. The plant regeneration frequency above 50% was observed when embryogenic calli induced in this medium were transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Among several basic media, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. 'Kentucky-31' showed to have high frequencies of embryogenic callus induction and plant regeneration up to 58.3 and 50%, respectively. Addition of sucrose to the regeneration medium as a carbon source increased regeneration frequency up to 55%. A short tissue culture period and high-frequency regeneration system established in this study will be useful for molecular breeding of tall fescue through genetic transformation.