• Plant Biotechnology Reports
• /
• v.2 no.3
• /
• pp.171-177
• /
• 2008

This paper reports an improved protocol for isolation, culture and regeneration of Lotus corniculatus protoplasts. A range of parameters which influence the isolation of L. corniculatus protoplasts were investigated, i.e., enzyme combination, tissue type, incubation period and osmolarity level. Of three enzyme combinations tested, the highest yield of viable protoplasts was achieved with the combination of 2% Cellulase Onozuka RS, 1% Macerozyme R-10, 0.5% Driselase and 0.2% Pectolyase. The use of etiolated cotyledon tissue as a source for protoplast isolation proved vital in obtaining substantially higher protoplast yields than previously reported. Culture of the protoplasts on a nitrocellulose membrane with a Lolium perenne feeder-layer on the sequential series of PEL medium was highly successful in the formation of microcolonies with plating efficiencies 3-10 times greater than previous studies. Shoot regeneration and intact plants were achieved from 46% of protoplast-derived cell colonies.

• Journal of Korean Society of Forest Science
• /
• v.95 no.2
• /
• pp.155-159
• /
• 2006

An efficient method for in vitro propagation of the medicinal plant Hovenia duleis, was established. Plantlets for micropropagation of H. dulcis were obtained from in vitro germinated seeds. The effectiveness of various levels of cytokinins (BAP, Kinetin and TDZ) on multiple shoot formation from stem nodes was tested. BAP (1.0 mg/L) treatment induced highest number of multiple shoots. The growth pattern of plantlet on various culture media was undertaken. The shoot elongation was optimal on 2MS basal medium without growth regulators. The in vitro rooting ability of H. dulcis shoots was examined with two-auxins IAA and IBA. The IAA (1.0 mg/L) treatments induced earliest rooting with maximum number of roots and root growth. Rooted shoots were transferred directly to small pots with artificial soil and such established plant exhibited a normal growth pattern similar to wild plantlet.

• Journal of Plant Biotechnology
• /
• v.36 no.4
• /
• pp.403-406
• /
• 2009

This study was conducted to develop an effective production of callus induction and plant regeneration system for garlic transformation. The best callus production occurred on in vitro root segment initially cultured on MS medium with 1.0 mg/L 2,4-D and 0.2 mg/L IAA in both ‘Danyang' and ‘Euseong'. The frequency of callus formation were 81.2% ‘Danyang' and 76.1% ‘Euseong'. Eight weeks after callus induction, callus lines were transferred to regeneration medium during 7 weeks. The best shoot regeneration medium was MS supplemented with 5 mg/L Kinetin and 1 mg/L NAA for ‘Danyang' and MS supplemented with 10 mg/L BAP for ‘Euseong'. The frequency of shoot regeneration were 51.5% ‘Danyang' and 56.6% ‘Euseong' The plantlets were acclimatized and transferred to the greenhouse with almost survival. This in vitro regeneration system should be useful for garlic transformation.

• Horticultural Science & Technology
• /
• v.32 no.5
• /
• pp.694-701
• /
• 2014

Allium hookeri L. (Alliaceae family) is an important ethnomedicinal plant native to the Himalayan region of Asia. The aim of this research was to establish a high-frequency plant regeneration system for in vitro propagation of A. hookeri. Among the tissue types examined, receptacle explants derived from immature flower buds showed the highest regeneration rate of shoots ($93.33{\pm}4.63%$), roots ($76.67{\pm}7.85%$), and calli ($80.00{\pm}7.43%$) when cultured on Gamborg B5 (B5) medium containing $10{\mu}M$ 6-benzylaminopurine (BA) + $1{\mu}M$ naphthalene acetic acid (NAA), $0.5{\mu}M$ BA + $5{\mu}M$ NAA, and $1{\mu}M$ BA + $10{\mu}M$ NAA, respectively. Shoot multiplication was superior when cultured in liquid rather than on solid medium and relatively high concentrations of BA, ranging from 5 to $10{\mu}M$. Efficient bulblet formation following root induction from shoot clumps was achieved with culture in liquid B5 medium containing 7% (w/v) sucrose. Regenerated bulblets were successfully acclimatized to ex vitro conditions with a greater than 95% survival rate. By this method, a maximum of 62 plantlets per receptacle could be propagated within 9 weeks of initial culture. The in vitro propagation system established in this study will promote A. hookeri biotechnology, including large-scale production of healthy and aseptic clones, preserving parental genotypes with desirable traits, and genetic manipulation to enhance medicinal value.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.5
• /
• pp.299-302
• /
• 1994

Immature zygotic embryos (up to 4mm in length) were cultured on MS medium supplemented with 0.5 to 8mg/L 2,4-D. Up to 87% of them formed somatic embryos on the plumule without producing an intervening callus. The site of somatic embryo formation was confirmed by culturing plumule explants, which consisted of shoot apical meristem domes with 1 or 2 leaf primordia excised from 2-week-old seedlings. When the concentration of 2,4-D was increased over 4 mg/L, the plumule explants produced nonembryogenic calli only, whereas the distal end of the cotyledons directly formed numerous somatic embryos at frequencies of up to 60%. Upon transfer onto MS basal medium,2 out of 15 somatic embryos converted into plantlets. The plantlets were potted to a soil mixture and grown to maturity in a phytotron.

• Journal of Plant Biotechnology
• /
• v.37 no.4
• /
• pp.465-471
• /
• 2010

We assessed whether the cow1 mutant defects are associated with growth of Tos17 and T-DNA insertional rice in blue light (BL). Growth of oscow1 mutants which encoded a member of the YUCCA protein family was retarded in BL. Root to shoot ratios of the mutants were reduced about 2 times lower in the absence of NAA and about 2.5 times lower in the presence of NAA; the shoot growth was not significantly changed by NAA addition. Photosynthetic activity of the mutants was however inhibited in high light. Pigment analysis showed significant difference between wild-type (Chl a:b = 3.02) and mutants (3.84). Carotenoid contents of the mutants were also decreased considerably, implying the involvement of cow1 in pigment formation. These findings lead us to suggest that the growth retardation of oscow1 mutant plants by BL results from the difference of photosynthetic activity in part.

• Korean Journal of Plant Resources
• /
• v.21 no.5
• /
• pp.368-373
• /
• 2008

In order to investigate the effects of low temperature pretreatment of floral bud and plant growth regulators on anther-derived callus and shoot differentiation, anthers were cultured on 1/2 MS medium supplemented with 2,4-D, NAA, BA and TDZ. This plant depends on the plant growth regulators, for these anthers couldn't respond on 1/2 MS medium without plant growth regulators. 2,4-D was a prerequisite substance in this experiment, especially 52.6% of callus formation on MS medium with 2.0mg/L 2,4-D alone. However, the optimum medium was on 1/2 MS medium with 0.1 mg/L 2,4-D and 1.0mg/L BA for continuous growth and shoot differentiation from the anther. Calli derived from on MS medium with 2.0mg/L 2,4-D transferred to the 1/2MS medium with TDZ and BA. TDZ were less superior to BA, only one anther could produce shoot on MS media with 1.0mg/L TDZ. On the other hand, when the calli transferred to the medium with 3.0mg/L BA, adventitious shoots were proliferated, subsequently, regenerated shoots elongated from the embryogenic calli. After floral buds of one week before anthesis were incubated at $5^{\circ}C$ refrigerator for eight or fifteen days, anthers seperated from floral buds were cultured on 1/2MS medium supplemented with 0.1mg/L 2,4-D and 1.0mg/L BA. Callusing and shoot differentiation on anthers from treated at $5^{\circ}C$ for eight days were more effective than those of fifteen days or control.

• Korean Journal of Weed Science
• /
• v.6 no.1
• /
• pp.25-32
• /
• 1986

This experiment was conducted to evaluate effect of various hormones on callus induction, and on plantlet formation on various media, and to detect of Londax [Methyl 2-[[[[[(4,6-dimethoxy pyrimidin-2-yl) amino] carbonyl] amono] sulfonyl) methyl] benzoate] and Basta[Ammonium-(3-amino-3-carboxy-propyl)-methyl phosphinate] on callus growth and reaction of succinate dehydrogenase in callus against TTC, using various species such as Eleocharis kuroguwai, Cyperus serotinus, Oryza sativa (samgangbyeo) and Echinochloa crusgalli P. Beauv. var. caudata Kitagawa. The optimal levels of 2,4-D in MS medium seems to be different among species tested, 2.0 ppm for rice and E. crusgalli, 1.0 ppm for Eleocharis kuroguwai, and 4.0 ppm for C. serotinus derived callus from shoot-tip. In case of combination of 2,4-D with BA, 1.0 plus 0.3 ppm appeared the most appropriate level to induce callus from rice and E. kuroguwai, and I.0 plus 0.1 ppm for C. serotinus and E. crusgalli. When 2,4-D treated with TIBA, 1.0 plus 0.5 ppm appeared the most appropriate rates to induce callus derived from seeds of rice, E. crusgalli, seeds of C. serotinus and E. kuroguwai, 1.0 plus 0.3 ppm for shoot-tip of C. serotinus. Positive reaction of succinate dehydrogenase against TTC was observed regardless of calli and herbicides tested, indicating that they all are alive, and these herbicides were not able to kill the calli tested within the short period of time 20 hrs treatment. Regardless of plant species used, the rate of plantlet formation from callus was very low. However, some plantlet formed from E. crusgalli at 0.8 ppm of 2,4-D plus 8.0 ppm of kinetin, and from E. kuroguwai at 1.6 ppm of 2,4-D plus 16.0 ppm of kinetin, showing effectiveness of 2,4-D with kinetin mixture treatment. No callus was induced from C. serotinus treated with Basta from $10^{-6}M$ to $10^{-3}M$. In general, rice was the least susceptible to Basta among plant species tested, followed by E. crusgalli, and E. kuroguwai. In Londax treatment, rice showed the least inhibition rate in callus growth. Callus was induced from rice even at $10^{-3}M$ of Londax. However, $10^{-3}M$ of Londax completely inhibited callus induction from the test species. Rice showed most tolerant to both herbicides, indicating the existence of different responses among plant species.

• Korean Journal of Plant Taxonomy
• /
• v.49 no.1
• /
• pp.1-7
• /
• 2019

Early leaves within the Viola albida complex were investigated by scanning electron microscopy in order to determine the morphological segments during morphogenesis. The early leaf development of V. albida var. albida could be morphologically divided into the eight stages in the following order: I, the initiation of shoot germination; II, the conical growth directionally of the leaf; III, the adaxial and abaxial formation of the leaf; IV, the initiation of the stipule; V, the formation of a transitional zone between the leaf blade and petiole; VI, the expansion of the upper part of the leaf blade; VII, the formation of almost all parts of the early leaf; VIII, the early simple leaf. Viola albida var. takahashii differs from V. albida var. albida by additional stages, i.e., V-1, the initiation of the first lateral lobe at the both lateral parts of the leaf after the stage V and an early lobed leaf. Viola albida var. chaerophylloides is also distinguished from two taxa by two developmental features, V-2, the initiation of a second lateral lobe below of the first lateral lobe, and an early palmately compound leaf. These findings suggest that the Viola albida complex would be in the process of peramorphosis, showing developmental changes in a chain of events, leading to a different leaf shape. These data would also be useful for isolating genes that give rise to different leaf morphogenesis outcomes among the taxa in the Viola albida complex.

• Journal of Plant Biotechnology
• /
• v.44 no.4
• /
• pp.484-489
• /
• 2017

Lilies as cut flowers are one of the most popular ornamental plants in South Korea. It is necessary to develop lily cultivars with high qualities. Therefore, highly efficient propagation systems are needed following release of elite cultivars. In this study, we used taurine treatment to improve the growth conditions including shoot and bulb formation, fresh weight gain, and reduction of rooting and browning. We experimentally evaluated the effect of taurine as a growth stimulator, at concentrations of 0, 2.5, 5, 10, 15 and 20 mg/l. The results showed that 20 mg of taurine enhanced shoot formation by 85% and increased fresh weight 5.5-fold, which was higher than the approximately four-fold increase in the control. In addition, multiple bulb formation rate was increased by 80% and rooting by 82% following exposure to 20 mg/l of taurine. The efficiency of taurine treatment was higher than that of control with 50% multiple bulb formation rate and 60% rooting rate. The browning was 10.6% at 2.5 mg/l of taurine when compared with 0.8% at 20 mg/l. Taurine showed a positive effect on the overall growth of lily plants in terms of increased fresh weight, shoot formation rate, rooting, and formation of multiple bulbs, indicating that taurine can be used as an alternative to amino acids or as an antioxidant such as citrate and vitamin C in plant tissue culture.