• Journal of Plant Biology
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• v.32 no.4
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• pp.313-322
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• 1989

A system was established for induction of multi-shoots and plant regeneration from mesophyll protoplasts of alfalfa, Medicago sativa L. cv. Vernal. Different hormonal effects were tested at each step of protoplast culture, i.e. cell division in modified Kao's liquid medium (K566-7). calli formation on SH semi solid medium, and multi-shoot regeneration from calli on SHa and SHb solid media. Frequency of multi-shoots and plant regeneration was affected by various combinations of phytohormones in final step. The evaluation of multi-shoots induction systems via protoplast culture was discused.

• Journal of Plant Biotechnology
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• v.50
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• pp.82-88
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• 2023

Shoot-tip culture was used to produce clonal plants of radish stock seeds. Using 6-benzyladenine (BA), the largest number of RA2 line multi-shoots were formed with an average of 14.67 shoots on 1.33 µM BA in early seedlings and 11.33 shoots on 1.78 µM BA in juvenile seedlings. The largest number of RA4 line multi-shoots were formed with an average of 11.67 shoots on 2.22 µM BA in early seedlings and 13.67 shoots on 1.33 µM BA in juvenile seedlings. There was little difference in the significance level by BA concentration in both lines. Using Thidiazuron (TDZ), the number of RA2 line multi-shoots increased with increasing TDZ concentration, forming the largest number of multi-shoots in 0.45 µM TDZ (7.0 and 3.0 multi-shoots for early and juvenile seedlings, respectively), but few multi-shoots were formed from TDZ 2.25 and 4.5 µM. RA4 line produced almost no multi-shoots in early seedlings, and 3.7 multi-shoots were produced in 0.23 and 0.45 µM TDZ in juvenile seedlings, but not at higher concentrations. Analysis of the tissue culture seedlings grown by cultivating the generated multi-shoots with Radish Foundation seeds using SSR marker revealed a weak pattern of mutation in the generated tissue culture seedlings, but there was no mutant. In addition, in terms of root roots, both RA2 and RA4 lines generally had the best rooting, number of roots, and degree of root development in 4.9 µM indol-3- butyric acid (IBA).

• Korean Journal of Medicinal Crop Science
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• v.7 no.4
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• pp.296-302
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• 1999

This experiment was conducted to study the effect of $CO_2$(0, 2, 500, 5, 000, 10, 000ppm) enrichment by enabling ventilation on micropropagation of multi-shoot and on the saponin contents in vitro in Korean ginseng (Panax ginseng C. A. Meyer). Embryo was cultured in Murashige and Skoog medium added 3mg/ l of Indolbutyric acid, Benzyladenin and Gibberellic acid $(GA_3)$, respectively. $CO_2$, enrichment had little effects on the number of adventitious buds and shoots originated from adventitious buds. The ratio of differentiated shoots to adventitious buds were about 50% in $CO_2$, enrichment treatment. The shoots originated from adventitious bud showed more rapid growth and had larger leaf area than the shoots originated from the leaf primordia did. The number of shoot primordia was the highest in 2, 500ppm of $CO_2$ enrichment treatment. On the contrary, 10,000ppm of $CO_2$, enrichment made smaller the number of shoot primordia and ratio of shoots to shoot primordia. The range of shoots differentiated was from shoot primordia were 15. 4 to 23. 9. The rate of dry weight of cultured shoots showed lowest (7. 5%) in control and highest (8. 59%) in 2, 500ppm of $CO_2$, enrichment. Rate of in vitro flower in control was 7.6% and that in 2500ppm of $CO_2$ was about twice (15.7-16.3%) as much as in control. Flower number per a embryo cultured was about 1.2-1.3. In the multi-shoots with callus enriched by 2, 500ppm of $CO_2$, the contents of crude saponin and ginsenosides in multi-shoots alone were higher than in multi-shoots with callus. The characteristics of ginsenosides in multi-shoots were especially the higher content of ginsenoside Rd, Re, and $Rg_1$.

• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.511-516
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• 2010

The process to acquire intron-GUS gene-expressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. 'Sweet Yellow' using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D = 0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with $250\;mg{\cdot}L^{-1}$ cefotaxim at $4^{\circ}C$ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$ to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim $250\;mg{\cdot}L^{-1}$. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.203-208
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• 2008

In order to micropropagate uniform plantlets of Philodendron cannifolium in vitro, the shoot tips were cultured on MS media supplemented with $0.5{\sim}10.0$ mg/L BA or $0.05{\sim}0.1$ mg/L thidiazuron(TDZ). The adventitious multi-bud clusters from basal part of shoots were formed on MS media containing $2.0{\sim}5.0$ mg/L BA or $0.05{\sim}0.1$ mg/L TDZ. But the shoots grown on MS media with TDZ showed necrosis by the lack of chlorophyll. The adventitious multi-bus clusters were cut into $5{\sim}7$ mm sections and cultured on MS media containing BA and TDZ for shoot proliferation. Shoots were proliferated vigorously on MS medium supplemented with $1.0{\sim}3.0$ mg/L BA with up to 30 shoots. But abnormally swollen hard calli were formed from basal parts of shoots on MS media with TDZ and high concentration of BA(10.0 mg/L). The proliferated shoots on same media also showed necrosis by the lack of chlorophyll. The shoot growth and rooting were favorable on MS media containing $0.5{\sim}2.0$ mg/L IBA. The rooted plantlets were acclimatizated effectively in soil mixed with perlite 1:vermiculite 1 or vermiculite alone. Fifteen mL of liquid medium containing 10 g/L activated charcoal and 30 g/L sucrose were added in same vessels after small shoots were proliferated to stimulate shoot growth and rooting. After 8 weeks in culture, the shoots were dipped into high concentration of IBA solution. and planted in soil mexed with perlite 1:vermiculite 1. The shoot growth and rooting were favorable in dipping treatments of $500{\sim}2,000$ ppm IBA solutions for 10 sec.

• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.25-32
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• 2001

Shoot tips of grape were cultured in uitro and tried to identify optimal culture conditions for regeneration, multiple shoot formation from meristemoid tissue and those subsequent elongation of multi-shoots. Healthy growing shoots were taken in early May, rinsed with running tap water, soaked in a neutral detergent and washed with soft brushing, and washed out with tap water, then sterilized with 10g Ca(ClO)$_2$/140 mL distilled water (Wilson's solution) for 5 min. Survival percentage of the cultures which were sterilized as above procedures was highly increased, compared with the other sterilized method. Propagation of multi-shoots from meristemoid showed a good response in 3/4 strength MS medium enriched with 0.1 mg/L NAA and 3.0 mg/L BA. Shoot elongation from multi-shooting clump well occurred in 3/4 strength MS medium supplemented with 80 mg/L adenine sulfate, 0.1 mg/L NAA and 1.0~2.0 mg/L BA.

• Journal of Plant Biotechnology
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• v.46 no.4
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• pp.303-309
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• 2019

A protocol for the in vitro propagation of Chamaecyparis obtusa was established in the present study. Multi-shoots were initiated from apical shoot explants from germinants after 10 weeks of culture on Litvay medium (LM) supplemented with different concentrations of cytokinin. The effects of pre-treatment with high concentrations of cytokinin and varying concentrations (0.2 to 5.0 mg/L) of zeatin on in vitro shoot elongation and shoot multiplication were investigated. Optimal shoot growth was achieved on LM medium, with over 10-mm shoots after 10 weeks of culture. In the anti-browning tests, ethanesulfonic acid triggered the least browning in the shoot tips. The highest multi-shoot induction was observed in the 0.5-mg/L zeatin treatments, which yielded 80% induction of shoots after 10 weeks of culture, and maximum shoot elongation was observed in the LM basal medium without the hormone. The highest rooting rates were 65% under 0.2 mg/L indole-3-butyric acid.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.10a
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• pp.162-162
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• 2003

• Journal of Plant Biotechnology
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• v.31 no.2
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• pp.115-119
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• 2004

In order to micropropagate uniform plantlets of Philodendron wend-imbe, the shoot tips were cultured on media supplemented with 0.5-10.0mg/L BA or 0.01-1.0 mg/L thidiazuron (TDZ). The multi-bud clusters from basal part of shoots formed vigorously on media containing 5.0-10.0 mg/L BA or 0.05-0.1 mg/L TDZ. Shoot formation from the bud cluster sections (5-7mm) was achieved favorably on medium with 5.0mg/L BA and 20 g/L sucrose. Lowering of sucrose in medium to 20 g/L was effective for the inhibition of callus growth from basal part of shoots. Growth of shoots and their rooting were favorable on media containing 1.0-2.0 mg/L IBA or 0.1mg/L NAA. The rooted plantlets were acclimatizated effectively in soil mixed with perlite 1: peat moss 1 or peat moss alone.

• Korean Journal of Plant Resources
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• v.18 no.3
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• pp.403-409
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• 2005

Mature nodal segments of 2-year-old greenhouse stock plant were cultured on Murashige and Skoog basal medium supplemented with the different kinds and various concentrations of cytokinins to produce multiple shoots in in vitro condition. The most adventitious shoots were produced from excised ends of nodal segments. The highest average number $(24.6\;{\pm}\;4.6)$ of shoots was produced with the combination of BA 1.0mg/L and TDZ 0.1mg/L in MS medium. In addition, several shoots were formed from lenticels of bark cambium with the same treatment. These concentrations promoted high shooting capability upto $94.6\%$ and NAA was the best cytokinin among five different PGR sources.