• Journal of Plant Biotechnology
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• v.7 no.1
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• pp.67-74
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• 2005

To overcome various disadvantages of conventional cul-ture vessels for micropropagation, a novel disposable vessel, the 'Vitron', made of a multi-layered $OTP^{(R)}$ film and supported by a polypropylene frame, was developed. The film possesses superior properties such as: high light transmittance, low water vapor transmittance and thermal stability and in particular, high gas-permeability. Single nodal explants, which were excised from the multiple shoots derived from shoot-tip culture, were cultured in Vitron and polycarbonate vessels on $3\%$ sugar-containing agar on MS medium and placed at 3000 ppm $CO_2$-enrichment at a low photosynthetic photon flux density (PPFD) ($45{\mu}mol\;m^{-2}\;s^{-1}$). The in vitro and ex vitro growth, and the net photosynthetic rate of in vitro and ex vitro plantlets were significantly enhanced in the Vitron compared to those cultured in a polycarbonate vessel. Explants that were cultured on the same MS medium under low PPFD at various $CO_2$ concentrations were also cultured at 3000 ppm $CO_2$- enrichment at various PPFD: 30, 45, 60, 75 and $90{\mu}mol\;m^{-2}\;s^{-1}$. The best in vitro and ex vitro growth obtained for 3000 ppm $CO_2$-enrichment at $75{\mu}mol\;m^{-2}\;s^{-1}$ PPFD. The novel Vitron vessel, when placed under the two conditions, may replace conventional culture vessels for the successful micropropagation of sweetpotato.

• Journal of Bio-Environment Control
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• v.14 no.4
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• pp.298-301
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• 2005

Potato (Solamum tuberosum 'Dejima') plantlets were investigated on culture type and initial quantity of inoculation in bioreactor and survival rate by hydroponics for mass production. rode stems (1 to 1.5cm in length) of potato plantlets multiplied in vitro were grown for 3 weeks in liquid Murashige and Skoog (MS) medium with sucrose $30 g\; L^{-1}$. When plantlets (80-node inoculation) were raised in 10L balloon type bubble (BB) bioreactor, the healthiest growth of plantlets was obtained from explants cultured in ebb & flow culture with medium supplied periodically 12 times per day. The suitable inoculation quantity of 20L BB bioreactor was 120 pieces of stem segments (mean 2.2g fresh weight) in ebb & flow culture. Number of nodal shoot was eight on the average. In controlled culture room, survival rate of plantlets at 7 days after stem cutting was above $70\%$ when they were acclimatized by hydroponics grown in deep flow and solid medium culture. The highest survival rate of the stem cutting plantlets was in nutrient solution adjusted to EC $1.4dS{\cdot}m^{-1}$. Stem cutting plantlets through one culture could be obtained $670\~900$, when plantlets were grown in ebb & flow culture during 3 weeks using a 20L bioreactor with initial 120 pieces of nodal segments. 11 is possible In do mass production of seedlings cultured in bioreactor and hydroponics.

• Korean Journal of Medicinal Crop Science
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• v.14 no.5
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• pp.278-281
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• 2006

The effect of plant growth regulators was investigated on in vitro shoot proliferation from axillary bud explants of Hypericum erectum. To determine the optimal cytokinin for proliferation of axillay buds, we carried out screening four cytokinins (BA, kinetin, 2iP, TDZ). When nodal segments were cultured on MS medium supplemented with $4.5\;{\mu}M$ TDZ (thidiazuron), a number of shoots were induced. Our results indicated that the addition of TDZ to culture medium resulted in the induction of significantly more axillary buds than in the addition of other cytokinins. The optimal concentration of TDZ for proliferation of axillary buds was $10\;{\mu}M$. 92% of shoots spontaneously rooted without any plant growth regulator (PGR) and formed whole plantlets within one month. More than 95% of these regenerants survived and they did not show any detectable variation in morphology or growth characteristics compared to their donor plants.

• Advances in Traditional Medicine
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• v.7 no.2
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• pp.197-204
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• 2007

The present study focused on the establishment of micropropagation protocol for the high value Pluchea (P.) indica (L.) Less., genotype, an important medicinal plant and evaluation of the diuretic activity of the leaf extract of the tissue cultured plant. Leaf explants, nodal segments and shoot tips were cultured in MS medium supplemented with auxin and cytokinin and their combinations. With the objective of inducing callus giving rise to new adult plants, naphthalene acetic acid was found to be most effective for (80%) for callus induction. The methanolic extract of leaves of the micropropagated P. indica was investigated for its diuretic activity in Wistar albino rats. Urinary excretion parameters were studied for evaluation of diuretic activity using Frusemide (20 mg/kg, p.o.) as standard. The extract showed significant diuretic activity at the doses of 100, 200 and 300 mg/kg. p.o. An oral acute toxicity study for the extract was carried out and the $LD_{50}$ value was found to be 2,825 mg/kg body weight.

• Journal of Forest and Environmental Science
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• v.29 no.4
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• pp.275-281
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• 2013

To establish in vitro nodal culture conditions of Echinosophora koreensis Nakai, one of rare and endangered species famous for beautiful flowers in the Korean Peninsula, the influence of plant growth regulators (PGRs) on shooting and rooting from in vitro shoots was investigated. In shoot multiplication, addition of 6-benzylaminopurine (BA) to the half-strength Driver and Kuniyuki's media in the range of 2.22 to 8.88 ${\mu}M $induced 2.5 to 2.7 shoots per axillary bud; and addition of 2.27 ${\mu}M $ thidiazuron (TDZ) produced 3.2 shoots, during 4 weeks of culture, while zeatin and isopentenyl adenine (2ip) were not effective on shoot multiplication as observed from several combination treatments of BA with other PGRs. Shoots established were smaller than 2 cm in length, in most of the treatments. while in BA 8.88 ${\mu}M $ treatment more than 30% of shoots were longer than 2 cm and shorter than 4 cm. In rooting, naphthalene acetic acid (NAA) from 5.37 to 21.48 ${\mu}M $ showed the rooting rate from 40.0 to 62.5%. Indole butyric acid (IBA) addition had little effect on rooting (<10%), although some roots in IBA-containing media were longer than those in NAA. Micropropagation from axillary buds of nodular explants was applicable and promising to multiplication and conservation of Echinosophora koreensis Nakai.

• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.149-153
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• 2005

In the present study, in vitro multiple shoot induction was achieved from cotyledonary node and stem nodal explants of cv. NARI-6 of safflower (Carthamus tinctorius L.). Among various growth regulators tested, MS salts and B5 vitamins supplemented with BA (6-Benzy-laminopurine) $17.76\;{\mu}M$ and KN (Kinetin) $6.96\;{\mu}M$ phytohormonal combination was found to be the most effective in initiating numerous shoot buds after 30 days of culture than BA ($4.44-44.39\;{\mu}M$) or KN ($2.32-46.40\;{\mu}M$) alone in the medium. In addition, 0.8% (w/v) agar (Hi-media) and 3.0% sucrose (w/v) was the optimum level for the formation of adventitious shoots. Further results showed the maximum shoot elongation occurred on MS medium with BA ($8.88\;{\mu}M$) and $GA_3$($11.56\;{\mu}M$) combinations. Efficient rooting occurred on quarter strength MS medium with NAA $10.74\;{\mu}M$. The regenerated plantlets were acclimatized and successfully transferred to the field.

• Horticultural Science & Technology
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• v.19 no.4
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• pp.459-464
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• 2001

Leaf disks from cultivar 'Kennebec' and one selection line (ND 860-2) were cultured on Murashige-Skoog medium with various combinations of indole acetic acid (IAA) and zeatin riboside. Shoots, roots and callus were induced at various combinations of plant growth regulator levels. The medium containing $3.5mg{\cdot}L^{-1}$ IAA and $4.0mg{\cdot}L^{-1}$ zeatin riboside produced the most plantlets. Rooted regenerants were grown in the greenhouse. The growth of regenerated plants obtained from the MS medium supplemented with $7.0mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside was significantly greater than those grown from nodal expalnts. In ND 860-2, a leaf chimera with chlorophyll deficient (light yellow) sectors was found in plants regenerated fiom leaf disks (grown on MS medium supplemented with $3.5mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside) but not in plants grown from nodal explants. The phenotypic variability was also observed in the tuber number, size and weight.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.175-182
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• 2009

Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.10a
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• pp.18-18
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• 2010

Molecular insights on the role of plant growth promoting rhizobacteria (PGPR) in potato tuberization are reported in the present study. The PGPRwere isolated from the soil collected from potato fields of Highland Agricultural Research Centre, Pyeongchang, Korea and they were identified to the genus level based on the 16S rRNA sequence analysis. These PGPR were heat-killed, filtered and the filtrates were addedindividually at a concentration of $10^7\;cfu\;mL^{-1}$ in MS (Murashige and Skoog's) medium supplemented with 7% (w/v) sucrose to study their influence on in vitro potato tuberization. Tuber initiation occurred early in untreated control, while tuber growth was pronounced in case of PGPR treatments. The control explants showed tuber formation as a result of sub-apical swelling of stolons while several sessile tubers formed directly in the axils of nodal cuttings in case of PGPR treatments, which is an indication of strong induction for tuberization. Theexplants cultured on MS medium supplemented with bacterial isolate 6 (Bacillus firmus strain 40) showed highest average tuber yield (Ca. 12.56 g per treatment) after 30 days of culture, which was 3 folds increase over the untreated control. A significant increase in lipoxygenase (LOX1) mRNA expression and activity of LOX enzyme were also detected in the tubers induced on PGPR treatments as compared to untreated control. This LOX expression level correlated with increased tuber growth and tuber yield. Further studies focused on the role of bacteria cell wall components, growth regulators and signal molecules released by PGPR are under investigation to elicit clues for PGPR-mediated signal pathway controlling potato tuberization.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.259-266
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• 2009

A plantlet regeneration protocol was developed on pot-grown mature plants of Elaeocarpus robustus Roxb. cv. Dwarf from nodal and leaf explants. The best yield of adventitious shoots was achieved from the leaf-derived calli in a modified MS ($MMS_1$, half strength of major salts, full strength of minor salts, and vitamins) medium containing $4.0{\mu}M$ BA + $4.0{\mu}M$ Kn + $0.5{\mu}M$ NAA + 15% coconut water (CW). The shoot multiplication rate was amplified about twofold per culture after the addition of 15% CW to the medium. The rate of shoot multiplication reached maximum at the 5th subculture, and it maintained this rate throughout the 3 subsequent subcultures. The best rooting in vitro was investigated by subculturing the microcuttings in an $MMS_2$ (half strength of both major salts and minor salts and full strength of vitamins) medium containing $1.0{\mu}M$ IBA in the dark for one initial week at $30^{\circ}C$, followed by subculturing them in a plant-growth regulator (PGR)-free medium in the light. The plantlets raised in vitro were successfully established under ex vitro conditions.