• Journal of Plant Biotechnology
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• v.39 no.1
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• pp.69-74
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• 2012

Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.

• Journal of Bio-Environment Control
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• v.13 no.1
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• pp.51-55
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• 2004

The most commonly used inorganic nutrient compositions such as Murashige & Skoog medium have been optimized for heterotrophic growth. Therefore, they may not be optimal for photomixotrophic and photoautotrophic growth of plantlets. In photomixotrophic micropropagation, emdium sugar level is often lowered, while light and $CO_2$ levels in vessel are raised, and chlorophyllous explants are used to facilitate photosynthetic carbon acquisition. In a factorial experiment effect of addition (+) and omission(_) of organic materials (OM, 0.5 g ${\cdot}$ $m^{-3}$ each of thiamine, nicotinic acid and pyridoxine and 100 ${\cdot}$ $m^{-3}$ myo-inositiol) combined with three sucrose levels (0, 15, and 30 kg ${\cdot}$ $m^{-3}$) was tested on the growth of potato plantlets. Each of nodal cuttings with a leaf was cultured on 0.1${\times}$$10^{-4}m^{-3}$) MS agar ( 8 kg ${\cdot}$ $m^{-3}$) medium (pH 5.80 before autoclave) in glass test tubes (100 mm${\times}$25mm) capped with a sheet of transparent film with a 6 mm diameter gas permeable filter (5.1 air exchanges ${\cdot}$$h^{-1}$). Cultures were maintained in a room for 27 days at $23^{\circ}C$, 50% RH, 350-450${\mu}mol\;{\codt}\;mol^{-1}CO_2$, 16 h ${\cdot}$ $d^{-1}$ photoperiod at 13${\mu}mol\;{\codt}\;m\;{\codt}\;s^{-1}$ PPFD provided by white cool fluorescent lamps. Growth of potato plantlet in the +OM and -OM treatments were similar, while medium pH was 0.2 scale lower in the latter. Dry weight, % dry matter, and stem diameter enhanced, while shoot to root dry weight ratio, leaf area, chlorophyll concentration per gram dry weight, and medium pH decreased with increasing initial sucrose level. Interaction between OM and sucrose levels was observed in shoot length and medium pH. Results indicate that OM can be omitted from the medium without detrimental effect while addition of sucrose was beneficial for the photomixotrophic growth of potato plantlets under raised light and $CO_2$.

• Korean Journal of Medicinal Crop Science
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• v.4 no.2
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• pp.101-108
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• 1996

This study was conducted to investigate the effect of growth regulators and $GeO_2$ on the induction and proliferation of callus and the effect of $GeO_2$ and citric acid on the Ge content of callus from explants and plant, Angelica koreana Max. The results obtained were summarized as follows. The callus induction was most effective on MS (Murashige and Skoog) medium containing 1. 0ppm 2, 4 - D with petiolule. The proliferation of callus was most effective at 2. 0ppm 2, 4 - D on the medium, at 2. 5ppm $GeO_2$ on the medium containing 2. 0ppm 2, 4 - D, and at $0.\;1{\sim}1mM$citric acid on the medium at pH6 containing 2. 0ppm 2, 4 - D and 2. 5ppm $GeO_2.$ The more $GeO_2$ in MS medium up to 20ppm, the more Ge content in callus. Ge content in callus was highest when the medium was supplemented with 0. 1mM citric acid and the pH of medium was low. The Ge content in plant was high in order of leaf > root > stem. Application $GeO_2$ to the soil increased Ge content in plant and application of 1mM citric acid with $GeO_2$ resulted in increasing Ge content highest in plant, but application more than l0mM citric acid resulted in Ge content decreased. Application of $GeO_2$ increased Ge content in callus and plant but had a tendency to decrease some mineral content, on the other hand application of $0.\;1{\sim}1mM$ citric acid with $GeO_2$ had a tendency to increase mineral content.

• Journal of Plant Biotechnology
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• v.40 no.4
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• pp.210-216
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• 2013

Efficient regeneration methods and transformation system are a priority for successful application of genetic engineering to vegetative propagated plants such as grape (Vitis labrusca L.). This research is to establish shoot regeneration system from plant explants for 'Campbell Early', 'Tamnara', 'Heukgoosul', 'Heukbosek' using two types of plant growth regulators supplemented to medium. The highest adventitious shoot regeneration rate of 5% was achieved on a medium containing of Murashige and Skoog (MS) inorganic salts and Linsmaier and Skoog (LS) vitamins, 2 mg/L of TDZ and 0.1 mg/L of IBA. Leaf tissue of 'Campbell Early', was co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, gus gene as reporter gene and resistance to kanamycin as selective agent, the other Agrobacterium strains, GV3101 containing the vector pB7 WG2D carrying with mPAP1-D gene. mPAP1-D is a regulatory genes of the anthocyanin biosynthetic pathway. 'Campbell Early' harboring mPAP1-D gene was readily able to be selected by red color due to anthocyanin accumulation in the transformed shoot. These results might be helpful for further studies to enhance the transformation efficiency in grape.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.13-18
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• 2003

This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

• 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.