• Journal of Korean Society of Forest Science
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• v.100 no.2
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• pp.172-177
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• 2011

This study was conducted to establish the optimal condition for in vitro propagation of wild Cynanchum wilfordii. The highest in vitro seed germination rate of 91.6% was obtained from the seed treated with gibberellic acid ($GA_{3}$) (100 ppm) for 24 hours followed by cultured on Woody Plant Medium (WPM) media supplemented with 1.0 mg/L 6-benzyl adenine (BA). The best shoot height obtained (5.2 cm) in medium with 1.0 mg/L $GA_{3}$, but the plants was grown abnormally and eventually died. The highest number (2.4) was shown when shoot were in cultured on the media including 0.1 mg/L BA after 4 weeks. Root induction from shoot obtained in vitro culture was effective on ventilation and without plant growth regulators (PGRs) and root length was highly developed (2 cm) at 0.1 mg/L naphthalene acetic acid (NAA). The highest survival rate (70%) was when plantlet grew pre-ventilation of in vitro condition.

• Plant Biotechnology Reports
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• v.2 no.3
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• pp.179-189
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• 2008

We established an efficient plant regeneration system for Catharanthus roseus L. (G.) Don through somatic embryogenesis. Embryogenic callus was induced from hypocotyl of seed germinated in vitro. Somatic embryogenesis in Catharanthus has been categorized into three distinct stages: (1) initiation and proliferation of embryo; (2) maturation, and; (3) germination or plantlet conversion. Beside plant growth regulators, various stages of embryogenesis were screened for their response to a wide variety of factors (pH, gelrite, light, sugar alcohols, polyethyleneglycol and amino acids), which affect embryogenesis. All of the tested factors had a small to marked influence on embryogeny and eventual conversion to plantlets. The plantlets were acclimatized successfully in a greenhouse. To our knowledge, this is the first report describing a detailed study of various cultural factors which regulate embryogenesis in C. roseus. The results discussed in this paper may be used in mass propagation to produce medicinal raw material, and the embryo precursor cells could be used in genetic modification programmes that aim to improve the alkaloid yield as well.

• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.124-129
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• 2009

In general, the root color of Codonopsis lanceolata is white, but red or blue-colored root is found at a low frequency in nature. Red or blue-colored roots have scarcity value, thus farmers wish to produce colored roots. The factors for determining the color of roots are unclear whether the color is controlled by genetically or simply by environmentally such as soil environment. Using in vitro culture system which is advantageous for setting of the same culture condition, we analyzed the physiological and morphological characteristics and genetic differences among red-, blue- and white lines of C. lanceolata. In the red colored roots, stems of in vitro cultured plantlet were colored in dark red pigment. Histological analysis revealed that the red pigment was accumulated in the outer cortex layer of the stem and determined as anthocyanin. Chlorophyll contents in red root lines were higher than those in white- and blue root lines. Plantlets from red roots were smaller in both shoot length and total leaf area than those from white- and blue roots. Genetic differences among the three different colored C. lanceolata were determined by RAPD (Randomly Amplified Polymorphic DNA) analysis. Each line of colored roots had clear DNA polymorphism. These results indicate that the occurrence of red- and blue colored roots in nature was determined by genetic factors rather than soil enviromental conditions.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.245-250
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• 2003

An efficient procedure was developed for adventitious shoot bud induction and plantlet regeneration from various explants of the ten genotypes of Pepper (Capsicum annuum L.) using Thidiazuron (TDZ). Among various treatments at 1.0-3.0 mg/L TDZ Induced maximum number of adventitious shoots depending upon the explant type and genotype compared to other treatments. Among the explants tested, leaf induced maximum number of adventitious shoots than the cotyledons. TDZ-mediated organo-genesis was possible in 10 pepper cultivars, the extent of the response being genotype-dependent. Of the ten genotypes tested, C. annuum cvs CA960, $G_4$ and X-235 were produced maximum number of adventitious shoots and Sell was the least, and all other genotypes gave moderate response. Elongation of multiple shoots was observed on medium supplemented with SA (0.05 mg/L) in combination of IAA (0.05 mg/L). Differences in ability for in vitro shoot regeneration and elongation depend upon the variety and explant type. The elongated shoots were success. Fully rooted on MS medium containing at 1.0 mG/L IAA. Plantlets regenerated from different explants of ten genotypes were found to be diploid (2n=24) and were devoid of any chromosomal aberrations. Regenerated plants were successfully established in soil where 85-90% of them developed into morphologically normal and fertile plants.

• Journal of Korean Society of Forest Science
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• v.71 no.1
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• pp.45-49
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• 1985

Protoplast-source meterial and enzyme strength had a significant influence on protoplast yield from hybrid poplar, Populus alba ${\times}$ P. grandidentata. The yield of protoplasts from in vitro culture of 1 month-old plantlets was more than that from greenhouse grown 4 month-old stock plant. In vitro cultured plantlets regulary produced more viable protoplasts with E-I enzyme solution (0.5% cellulase and 0.1% macerase) than those with E-II enzyme solution (1.0% cellulase and 0.2% macerase) after overnight incubation. The mean yield of protoplasts from in vitro cultured plantlets was $4{\times}10^6$ with E-I enzyme solution. Cell division was observed in these protoplast cultures after 7-10 days. Protoplast-derived hybrid poplar cells survived over 3 weeks in culture and some continuous cell divisions were evident. Other aspects associated with protoplasts from in vitro cultured plantlet are also discussed.

• Journal of Plant Biotechnology
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• v.45 no.1
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• pp.55-62
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• 2018

In the present study in vitro mutagenesis was used to study the effect of gamma irradiation and EMS on callus induction, morphogenesis and production of multiple shoots from different explants of Citrullus colocynthis (L.) Schrad. Gamma radiations (5 kR to 20 kR) and certain chemicals have been effected on plant growth developments and changes of biochemical metabolisms in plants. Murashige and Skoog (MS) medium containing with auxins such as NAA, IAA, 2,4-D (0.5 ~ 2.0 mg/l), cytokinines BAP, kn TDZ, (0.5 ~ 2.5 mg/l), L-Glutamic acid (1 ~ 2 mg/l) and Coconut milk (10 ~ 20%). After 5 weeks on induction media, explants and callus (EC) were exposed to 5 kR, 10 kR, 15 kR and 20kR, of gamma radiation and treated with 1, 2, 3, 4 and 5 mM ethyl methane sulphonate (EMS) for 30 min. The highest percentage of callusing was observed (70%) stem irradiated with 5 kR and significantly decrease in fresh and dry weight of callus in the below 4 kR doses and above 20 kR doses, there was a progressive decrease in the fresh weight and dry weights when compared to control callus. Maximum percentage of plantlet regeneration (59%) was induced from callus exposed to 15 kR gamma irradiation on MS media fortified with 2.0 mg/l 2,4-D + 2.0 mg/l BAP + 2.0 mg/l L-glutamic acid. Increase in gamma irradiation dose above 15 kR and 5 mM EMS reduced regeneration capacity of callus. Doses higher than 20 kR and 7 mM EMS was lethal to micropropagated plants of Citurullus colocynthis.

• The Plant Pathology Journal
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• v.19 no.3
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• pp.159-161
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• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

• Journal of Biosystems Engineering
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• v.32 no.2 s.121
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• pp.109-114
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• 2007

In mass production of seed-potato plantlets, the processes for in vitro propagation and ex vitro acclimatization with a high cost should be improved by a culture system with environmental control using scaled-up culture vessels. The experiment was conducted to design a hydroponic culture system for enhancement of growth and development of seed-potato (Solanum tuberosum) plantlets cultured under photoautotrophic (without sugar in culture medium) conditions with controlled light intensity and ventilation rate. The culture system was consisted of scaled-up culture vessels, ventilation pipes, a multi-cell tray and an environmental control system (ECS) for optimum controlling in temperature, light intensity, ventilation rate, and culture-medium supply. Growth and development of the plantlets was significantly increased under the ECS compared with a conventional culture system (CCS) of photomixotrophic culture (with sugar in culture medium) using small scale vessels. For 21 days, leaf area of the plantlets was expanded more than 2 times, and number of internodes also approximately 4 times greate. under the ECS. In addition, the photoautotrophic growth in sweetpotato (Ipomoea batatas) and chrysanthemum (Chrysanthemum morifolium) plantlets was greater more than 2 times compared with the CCS.

• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.263-267
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• 2003

This study was conducted to evaluate the effect of thidazuron(TDZ) on shoot proliferation and growth from axillary buds of 20-years-old Corylopsis coreana. Shoots proliferation was effectively achieved on WPM(Woody Plant Medium) supplemented with 0.03∼0.1mg/L TDZ. The highest shoot number(6.5$\pm$0.7) was obtained on 0.1mg/L TDZ treatment. On the TDZ medium shoots formed as clusters less than 1cm in height and therefore needed to subculture on GA$_{3}$ containing medium to induce elongation. In consecutive cultures, phenolic compounds were excreted at the proximal part of the explants and inhibited growth of the explants. Growth inhibition by the compounds was overcome using liquid and paper bridge culture system. About 60％ of the elongated shoots rooted on half- strength MS medium containing IBA. Generally, IBA was mire effective on in vitro rooting than NAA with optimal range of 0.5mg/L to 1.0mg/L. Rooted plantlets were transferred in an artificial soil(vermculite) and acclimatized in high humidity greenhouse condition. Survival rate differed greatly depending on rooting types of the explants. Two types of rooting were observed. The first type was direct rooting from the explants. The second type was callus formation followed by rooting from the callus. The explants showing the 1st type rooting survived can be multiplicated in vitro by TDZ treatment followed by elongation with GA$_{3}$ and rooting with IBA.

• Journal of Plant Biotechnology
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• v.8 no.1
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• pp.27-35
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• 2006

Plantlet regeneration in Asteracantha longifolia(L.) Nees (Acanthaceae), a medicinal herb has been achieved from seedling explants on basal MS medium. Three different seedling explants including node, internode and leaf segments on used. Of these three explant, leaf explants gave better response for both callus mediated organogenesis and direct multiple shoot induction. Number of explants showing differentiation of shout buds was higher on MS media supplemented with BA compared to kinetin. MS medium fortified with BA ($2.0mgl^{-1}$) and NAA ($0.5mgl^{-1}$) was found to be most suitable for both callus mediated organogenesis and elongation of shouts. The elongated shoots were successfully routed on MS medium fortified with NAA or IBA. Among them $0.1mgl^{-1}$ NAA or $0.2mgl^{-1}$ IBA provides better response for rhizogenesis. Regenerated plantlets were successfully established in soil where 85.4% or them developed into morphologically normal and fertile plants. RAPD profiling using four decamer primers confirmed the genetic uniformity of the regenerated plantlets and substantiated the efficacy and suitability of this protocol for in vitro propagation of A. longifolia.