• Korean Journal of Medicinal Crop Science
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• v.14 no.1
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• pp.23-26
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• 2006

One of micropropagation methods was investigated by using a multiple-shoots protocol. Multiple shoot formation was obtained from excised axillary buds of Hypericum erectum on half-strength or basal MS medium supplemented with TDZ or BA. The optimal combination of shoot multiplication for the production of more shoots with a suitable size was MS medium supplemented with $0.005\;mg{\cdot}L^{-1}$ TDZ (6.5 adventitious shoots per node). In vitro rooting was carried on half-strength MS medium with $1\;mg{\cdot}L^{-1}\;GA_3\;and\;0.5\;mg{\cdot}L^{-1}$ IBA treatment. In addition, the rooted cuttings were showed a better root growth in the greenhouse and survived in more than 90%. The results show that the species can be micropropagated effectively by the application of axillary bud culture systems.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.301-306
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• 2005

Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.223-228
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• 2008

This study was conducted to investigate an optimal condition for shoot proliferation and regenerate shoots from in vitro leaflet and embryogenic calli from in vitro roots in rose. The effect of BAP on shoot proliferation was somewhat different depending upon genotypes or gelling agents. Leaflets with petiole cut from donor shoots which had been cultured in MS medium supplemented with 0.1 $mg{\cdot}L^{-1}$ NAA for six weeks was effective for regeneration of adventitious buds(ABs) as well as shoot elongation of Rosa hybrida cv. Sweet Pink. Culturing seven leaflet explants per petri plate($100mm{\times}15mm$) was effective for regeneration of ABs. Embryogenesis was shown in the calli induced from roots of Rosa hybrida cv. Sweet Pink cultured in the SH medium supplemented with 11 $mg{\cdot}L^{-1}$ 2, 4-D for four weeks. Color of calli induced from roots was yellow although their color was a little different as type of basal medium.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.04a
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• pp.131-132
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• 2003

Leaf discs and apical meristems were cultured in Murashige and Skoog (MS) medium supplemented with cytokinin and auxin at different concentrations. Callus production was observed in all tested media after six days of incubation. Callus produced in the presence of high concentration of NAA (2.0mg/1) was fragile in texture and yellow in colour. Highest callus formation was observed from leaf discs in the medium supplemented with 1.0mg/1 NAA and 0.5 mg/l BAP in dark at $25{\pm}1{\circ}C$. Percentage of callus formation was 95% and mean callus fresh weight was 654.88 43.53 mg. Shoots were induced from the callus after 4 weeks in 1/2MS medium supplemented with BAP and kinetin both at 0.5mg/1. When elongated shoots were separated and transferred into multiplication medium (MS+0.5mg/1 BAP+0.5mg/1 kinetin) multiplication rate was 6.4 after 6 weeks. Higher concentrations of BAP caused callus production at the base. Direct shoot induction was observed from apical meristems in MS medium in the presence of 0.175 mg/1 IAA + 2.25mg/1 BAP and 0.175 mg/1 IAA + 3.0 mg/1 BAP in 16 hour day at $25{\pm}1{\circ}C$. Explants (apical meristems) elongated to form a single shoot forming a callus at the base. Adventitious buds were sprouted out from the base. Percentage explants which producing shoots was 28.57 and 65.5 respectively. Multiple shoot induction was also observed in the same media. Highest multiple shoot production was observed in the presence of 0.175 mg/l IAA and 3.0mg/l BAP, Mean number of shoots per explant was 5.36 and the mean shoot length was $16.66{\pm}4.15$mm. Shoots (20 30m length) were tested for root induction. Excised shoots were transferred into rooting media, which contains different concentrations of NAA and IAA. Best rooting performance was observed in 1/2MS medium supplemented with 0.1mg/1 NAA after 10 days of incubation in 16 hr photoperiod at $25{\pm}1{\circ}C$. Mean number of roots per shoot was 6 and the mean root length was 252mm. Rooted plantlets were transferred into sterile coir dust:sand (1:4) mixture and maintained in a humid chamber for two weeks, They were gradually exposed to the natural environment. After three weeks they were transferred to pots containing coir dust:sand (1:2) mixture for further development where the 90% survival was observed.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.77-85
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• 2004

Development of efficient plant regeneration and genetic transformation protocols (using the Particle Inflow micro-projectile Gun and the shoot-tips as target tissue) of Sorghum bicolor (L.) Moench in terms of expression of the reporter gene, $\beta$-glucuronidase(uidA) is reported here. Two Indian cultivars of sorghum were used in the study, viz. M-35-1 and CSV-15. Plant regeneration was achieved from one-week-old seedling shoot-tip explants via multiple-shoot-clumps and also somatic embryos. The multiple-shoot-clumps were produced on MS medium containing BA (0.5, 1.0 or 2.0 mg/$L^{-1}$), with biweekly subculture. Somatic embryos were directly produced on the enlarged dome shaped expansive structures that developed from shoot-tip explants (without any callus formation) when cultured on MS medium supplemented both with BA (0.5, 1.0 or 2.0 mg/$L^{-1}$) and 2,4-D (0.5 mg/$L^{-1}$). Whereas each multiple-shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing 1.0mg/$L^{-1}$ IBA and successfully transplanted to the glasshouse and grown to maturity with a survival rate of 92%. The plant regeneration efficiency of both the genotypes were similar. After the micro-projectile bombardment, expression of uidA gene was determined by scoring blue transformed cell sectors in the bombarded tissue by an in situ enzyme assay. The optimal conditions comprising a helium pressure of 2200 K Pa, the target distance of 11 cm with helium inlet fully opened and the use of osmoticum have been defined to aid our future strategies of genetic engineering in sorghum with genes for tolerance to biotic and abiotic stresses.

• 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.

• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

• Korean Journal of Plant Resources
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• v.18 no.1
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• pp.154-160
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• 2005

To establish rapid micropropagation through organogenesis from apices-derived callus or direct adventitious shoot of three calla lily cultivars(Zantedeschia spp, cv. Sunlight, cv. Chiante, cv. Pink Persuation) were cultured on Murashige and Skoog medium supplemented with different plant growth regulators. The formation rate of callus, organogenesis and in viかo tuber production among the three cultivars were tested. Callus was obtained from cvs. Sunlight, Chiante and Pink Persuasion; the best cultivar was Sunlight. Sunlight induced $53.3\%$ callus and Chiante had the highest rate of $56.7\%$ direct shoot regeneration on medium with 2.0 mg/L BA. Regeneration frequencies ranged from 20 to $70\%$ on medium with 2.0-3.0 mg/L BA. The highest percentage of regeneration and the greatest number of shoots were obtained on medium containing 3.0 mg/L BA in three cultivars. Cytokinins induced multiple shoot formation; 1.0 mg/L of 2ip, 5.0 mg/L of BA, and 1.0 m/L of BA induced 16, 14 and 12 multiple shoots in cvs. Sunlight, Chiante and Pink Persuasion, respectivly. 1.0 mg/L of IAA enhanced root growth in cvs. Sunlight and Chiante while cv. Pink Persuasion exhibited enhanced root growth at 2.0 mg/L of IBA. NAA, however, induced no change in root growth. The addition of 90 g/L sucrose enhanced in vitro tuber formation and following tuber expansion in cv. Sunlight, while 70 g/L of sucrose was effective in cvs. Chiante and Pink Persuasion.

• Korean Journal of Plant Resources
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• v.22 no.5
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• pp.394-402
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• 2009

Adventitious shoots were induced from pinnae, petiole and rhizome in Pteris cretica 'Wilsonii' in order to develop the efficient mass propagation method, using in vitro culture. Only homogenized rhizome segments could regenerate young sporophytes. Efficient regeneration of multiple shoots was obtained on the one-eighth strength MS medium containing 1% sucrose, and $50mg{\cdot}L^{-1}$ $NaH_2PO_4$. To achieve higher rate of regeneration from rhizome segments, rhizome segments were exposed to growth regulators for 2 months and then subcultured on hormone-free medium. The greatest shoot regeneration was obtained by $1{\mu}M$ kinetin with $5{\mu}M$ NAA. BA was effective in formation of GGB (kind of meristems), but they showed low shoot regeneration rate. Plants obtained from present experiments were transplanted to examine good environmental conditions for acclimation. Juvenile plants obtained by the one-eighth strength MS medium showed highest survival rate and vigorous growth at the seedling stage.

• Korean Journal of Plant Resources
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• v.21 no.2
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• pp.111-116
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• 2008

This study was carried out to induce plant regeneration via multiple shoot formation from sucker explants of Rubus fruticosus L. ${\times}$ R. parvifolius L. To induce adventitious shoots, the sucker explants were sterilized in 1.2% sodium hypochlorite (NaOCl) solution, and then were cultured on the full and 1/2 MS solid medium supplemented with BA (0.1, 0.5, 1.0, $2.0mg{\cdot}L^{-1}$). After 4 weeks of culture, the highest frequency (83.3%) of shoot formation from sucker explants was obtained from the full MS medium with $1.0mg{\cdot}L^{-1}$ BA. The highest shoot number (3.7) per explant was obtained from the full MS medium with $0.5mg{\cdot}L^{-1}$ BA. After 12 weeks of culture, the number of shoots (15.4) per explant was increased. The highest frequency (95%) of root formation was obtained from the 1/2 MS medium, when the explant with shoot were cultured on the full, 1/2 and 1/4 MS medium. The survival rate of the plantlets after transfer to plastic pots containing sand, soil, and vermiculite (1.1:1, vol.) was 95%. The results indicate that multiple shoot procedure can be applied for an efficient mass propagation of Rubus fruticosus L. ${\times}$ R. parvifolius L.