• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.266-272
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• 2018

A Polygonatum stenophyllum Maxim. is an important endangered plant belonging to the family Liliaceae. A method was developed for the rapid micropropagation of P. stenophyllum through plant regeneration from rhizome (1-year, 3-years, and 5-years) explant-derived calli. The rhizome segments were cultured in Murashige and Skoog (MS) medium supplemented with varying concentrations of 2,4-D (0, 0.5, 1.0, $1.5mg{\cdot}L^{-1}$) for callus induction. In media supplemented with $0.5mg{\cdot}L^{-1}$ of 2,4-D, 87% of 3-years rhizome produced callus. Subsequently, the callus was transferred to 1/2MS medium supplemented with various concentrations of IAA, IBA, NAA, and 2,4-D (0, 0.1, 0.5 and $1.0mg{\cdot}L^{-1}$) for adventitious shoot formation. The highest percentage of adventitious shoot induction (57%) was observed in 1/2MS medium containing $0.5mg{\cdot}L^{-1}$ of NAA. Elongation of the adventitious shoot was achieved in 1/2MS medium supplemented with $0.1mg{\cdot}L^{-1}$ of BA. Rooting was achieved in 1/2MS medium without any hormones. It is hypothesized that the stated in vitro propagation protocol will be useful for conservation and mass propagation of the endangered Polygonatum stenophyllum Maxim. for bioresources.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.320-324
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• 2017

Development of modern agricultural machinery and accompanying agricultural development cause soil compaction and reduce growth by stressing roots. Kalanchoe pinnata was used to investigate the impact of stress on rooting and changes in plant growth and reproduction. K. pinnata forms somatic embryos capable of asexual reproduction at the edge of leaves. Impact of root pressurization of K. pinnata on somatic embryogenesis and organ differentiation according to external stress factors was investigated by using a high concentration of agar and this phenomenon was studied histologically. Agar concentration in culture media ranged from 0.5%-1.5% to induce a compression effect on roots. The stem and leaf of K. pinnata were subjected to a microtechnique process to study changes in tissue. In vivo, K. pinnata produced 2nd and 3rd plantlets at edges of leaves from lack of water and excessive lighting conditions. In in vitro culture studies, the lower the concentration of agar, the higher the population and the higher the biomass, but plantlet did not occur in leaf bends. Conversely, as concentration of agar increased, increase in the number of individuals was low. Plantlet development occurred only in agar 1.5% medium. The difference in agar concentration was a stressor in the root of K. pinnata, and thus the pattern of asexual reproduction changed from the division method in root to a plantlet generation in leaf. This suggests root pressurization may act as stress and change in the plant reproduction pattern.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.62 no.3
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• pp.259-274
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• 2017

Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on 1/4 MS for P. grandiflorum with wild and green petals and on 1/8 MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on 1/4 MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, 1/4 MS supplemented with $1mg\;L^{-1}$ 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on $0.5mg{\cdot}L^{-1}$ thidiazuron (TDZ) from double petal explants grown on 1/8 MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.

• Korean Journal of Plant Resources
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• v.25 no.2
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• pp.277-284
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• 2012

In the present study, the effects of plant growth regulators on adventitious shoot and root formation of various explants of $in$ $vitro$ seedlings of $Solanum$ $nigrum$ L. were investigated to determine the optimum conditions for the high-efficiency plant regeneration of this species. The formation of adventitious shoots was higher in leaf explants than in cotyledon, hypocotyl, or epicotyl explants at low concentrations (0.5~2.0 mg $L^{-1}$ ) of 6-benzylaminopurine (BAP). The number of adventitious shoots and the shoot length were also higher in both leaf and cotyledon explants. In particular, 2.0 mg $L^{-1}$ BAP was most effective for stimulating the induction and multiplication of adventitious shoots. In terms of root formation and root development from shoots that were separated from multiple shoots, indole butyric acid (IBA) and indole acetic acid (IAA) were more effective than ${\alpha}$-naphthalene acetic acid (NAA). The percentage of rooting as well as the number of roots per shoot (4.0), root length (7.82 cm), and shoot length (8.76 cm) was highest on MS media supplemented with 0.05 mg $L^{-1}$ IAA. Furthermore, 100% of the regenerated plantlets survived when transplanted to compost soil. These results suggest that leaf explants are the best source for the high-efficiency regeneration of $S.$ $nigrum$ L., and that 2.0 mg $L^{-1}$ BAP and 0.05 mg $L^{-1}$ IAA are the best conditions for shoot and root induction, respectively.

• Journal of Plant Biotechnology
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• v.29 no.1
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• pp.45-49
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• 2002

The leaf and petiole segments of Farfugium japonica were cultured to investigate the influence of growth regulators on their callus induction and plant regeneration. The callus induction and growth showed a good response both leaf and petiole on MS media supplemented with 1∼2 mg/L 2,4-D and 1∼2 mg/L BA. Callus induction and growth were more effective in petiole segments than leaf one. The highest percentage of plant regeneration was obtained from 60-day-old calli on MS medium supplemented with 1 mg/L NAA and 2 mg/L BA. When subcultured to the same medium for about 60 days, multiple shoots were developed from regenerating callus. The shoots produced roots after transferring to rooting medium containing 0.5 mg/L IAA. The plantlets over 50 mm in height were successfully acclimatized in vermiculite, and the survival rate was over 95%.

• Journal of Plant Biotechnology
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• v.50
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• pp.176-182
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• 2023

Plant regeneration protocols for adventitious shoot organogenesis from apple (Malus domestica 'Fuji') leaf explants were developed in the present study. The effects of dark incubation periods in the early stages of culture, pre-treatment methods, the number of explants per culture container, the type of culture containers, and the orientation of the explants on culture media were evaluated to determine the optimal shoot regeneration conditions for 'Fuji' apple leaf explants. Light incubation of explants produced minimal response. However, dark incubation of explants for 4 weeks during the initial culture period enhanced shoot regeneration frequency. Comparing the number of explants per container, a higher percentage of shoot regeneration was obtained with nine explants per container compared with four explants per container. Pre-treatment, before culture, by dipping explants in a liquid regeneration medium containing 40 g/L of sorbitol for 2 hours produced the highest shoot formation rate, and the time of shoot formation was accelerated. The percentage of shoot regeneration and number of shoots per regenerating explant reached a maximum of 87.5% and 4.7, respectively. The regenerated shoots were elongated and rooted on a rooting medium of 1/4 MS with 0.2 mg/L IBA. The plantlets were successfully acclimatized, and the regenerated plants produced normal phenotypes.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• Journal of Bio-Environment Control
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• v.12 no.3
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• pp.152-159
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• 2003

This study was conducted to analyze the water consumption in closed transplants production system (CTPS) for the production of quality transplants and to investigate the effect of relative humidity on the water balance in CTPS. Potato (Solanum tuberosum L. cv. Dejima) plug seedlings were grown for 15 days at air temperature of 20$^{\circ}C$, relative humidity of 70%, photoperiod of 16/8 h, and photosynthetic photon flux (PPF) of 200 ${\mu}mol{\cdot}m^{-2}{\cdot}s^{-l}$ following rooting for 5 days in CTPS. Amount of humidified, dehumidified, irrigated and evapotranspirated water were 67.9 kg${\cdot}m^{-2},\;196.9{\cdot}m^{-2},\;44.3\;kg{\cdot}m^{-2},\;33.5\;kg{\cdot}m^{-2}$, respectively. Water content of media and plants were 1.2 kg${\cdot}m^{-2},\;6.9\;kg{\cdot}m^{-2}$, respectively. Three relative humidity levels of 60, 70, and 80% were provided to analyze the effect of humidity on the water balance in CTPS. Amount of humidified, dehumidified, irrigated, evapotranspiratad water and water contents of media and plants increased with increasing relative humidity. Since the water consumption required to produce plug seedlings in CTPS dec1eased with decreasing relative humidity, the available water utilization efficiency of CTPS increased with decreasing relative humidity. CTPS showed high available water utilization efficiency of 0.92 - 0.97 if dehumidified water in CTPS was recycled. The development of CTPS with recycling system of dehumidified water will not only reduce the water consuming for the production of transplants but contribute to the establishment of plant production economizing in water consumption.

• Journal of Plant Biotechnology
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• v.40 no.2
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• pp.79-87
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• 2013

The aim of this study was to establish plant regeneration from leaf explants of Sedum tosaense Makino, which is globally rare and endangered species. The leaf explants of S. tosaense were cultured on the MS medium supplemented with different concentration of BA and NAA for callus induction. Callus induction was showed the highest (100%) on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA. The highest number of shoots were regenerated when callus were cultured on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA for 5 weeks. The axillary bud were cultured on the MS media supplemented with combination of BA and NAA for in vitro propagation. The highest number of adventitious shoot (7.9 per explants) formed at $1.0mg{\cdot}L^{-1}$ NAA and $2.0mg{\cdot}L^{-1}$ BA. For rooting, MS medium supplemented with or without $2.0g{\cdot}L^{-1}$ activated charcoal was tested. The optimal results were observed using MS medium supplemented with $2.0g{\cdot}L^{-1}$ activated charcoal, on which 85.7 (No. of root), 4.6 cm (length of root). 1,200 ppm $CO_2$ and 350 ppm $CO_2$ were supplied for make certain the effects of $CO_2$ on pre-acclimatization by photoautotrophic culture. 1,200 ppm $CO_2$ treatment was established higher than 350 ppm $CO_2$ treatment. Soil acclimatization of in vitro plantlets was the best in mixture soil consisted of peat moss and perlite with 100% survival rate and they showed the maximum growth.