• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.193-196
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• 2009

Pinellia koreana K-H Tae & J-H Kim is a recently discovered Korea endemic medicinal plant species whose natural habitat is rapidly destroyed by industrial development. Described in this paper are culture conditions for high frequency plant regeneration via bulblet formation from leaf explant cultures of P. koreana. Leaf explants formed white nodular structures and off-white calluses at a frequency of 91.2% when cultured on MS medium supplemented with 2 mg/L BA and 0.5 mg/L NAA. However, the frequency of white nodular structures and off-white calluses formation was slightly decreased with an increasing concentration of NAA up to 4 mg/L, where the frequency reached 31.7%. Most petiole explants did not form white nodular structures and off-white calluses except the combination treatment of 2 mg/L BA and 2 mg/L NAA. Upon transfer onto MS basal medium, over 90% of nodular structures gave rise to numerous bulblets and developed into plantlets. Plantlets regenerated from bulblets were transplanted to potting soil and grown to maturity at a survival rate of over 95% in a growth chamber. Therefore, the in vitro plant regeneration system of P. koreana obtained in this study will be useful for mass propagation and long-term preservation of genetic resources of P. koreana.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.176-176
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• 2017

Hydropriming had positive effects on the time for germination to reach 50%, the germination index, the time to final germination percentage, and the number of uniform seedlings with enlarged cotyledons in in vitro germination of small watermelon. In addition, the highest shoot initiation was obtained from hydroprimed cotyledonary nodes ($95{\pm}6%$), followed by non-primed cotyledonary nodes ($78{\pm}6%$), hydroprimed cotyledons ($72{\pm}4%$), and non-primed cotyledons ($48{\pm}4%$). Meanwhile, no shoots were initiated from hypocotyls. The total number of shoots that initiated from cotyledonary nodes and cotyledon explants was insignificant, indicating that both cotyledons and cotyledonary node were good sources for the in vitro culture. Choosing explant sources that favor tetraploidy should be the key for producing higher polyploidy plants; a total of 10.5% of tetraploid regenerants were entirely identified from cotyledon explants. Cotyledons with highly differentiated cells might show higher variations than cotyledonary nodes with more preexisting meristematic cells. Cells of cotyledon tissue might undergo changes in ploidy level during differentiation of the culture, or it might be that some of the variations were already present in the tissues of the donor plant. Morphological changes in fruit length of tetraploid regenerants are genotype-dependent.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.131-131
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• 2017

Though the Platycodon grandiflorum, has a broad range of pharmacologic properties, but the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two-dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}2-fold$) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, the frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). Taken together, the protein profile may provide insight clues for better understanding the characteristics of proteins and its metabolic activities in various explants of this essential medicinal plant P. grandiflorum.

• Korean Journal of Plant Tissue Culture
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• v.22 no.3
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• pp.131-135
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• 1995

Physiologically modified stem nodes derived from ex vitro and in vivo explants of hybrid aspen (Populus alba L.X P.grandidentata Michx. 'Crandon') were tested for their multiple shoot regeneration capacity using a broad spectrum dosage of cytokinins. Ex vitro derived stem nodes with excised axillary buds at the time of culture produced 11 to 13 multiple shoots on 20 to 30 $\mu$M zeatin containing Woody Plant Medium (WPM) after 6 weeks. Excision of axillary bud sprouts after 2 weeks of culture and culture of the remaining stem nodes on WPM with 1.0 to 2.0 $\mu$M BA or 10 to 30 $\mu$M zeatin produced 13 to 15 and 7 to 8 shoots per explant, respectively, Multiple tiny shoots were produced when in vivo derived stem nodes (on which all leaves were removed) were cultured on WPM with 30 to 50 $\mu$M 2iP or 20 to 50 $\mu$M zeatin. The greatest number of multiple tiny shoot proliferation (32 to 50 shoots per explant) were obtained when the explants were cultured on media containing 20 $\mu$M zeatin. Successful transplanting of these multiple shoots into the greenhouse and/or nursery was achieved.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.435-439
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• 2000

Explants of lettuce (Lactuca sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring ${\gamma}$-tocopherol methyltransferase (${\gamma}$-TMT) gene from Arabidopsis thaliana. These explants were transferred to MS medium supplemented with 50 mg/L kanamycin, 500 mg/L carbenicillin, 0.1 mg/L NAA and 0.5 mg/L BA. After 4 weeks, kanamycin resistant shoots were obtained from the explants on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 50 mg/L kanamycin and 250 mg/L carbenicillin. Stable incorporation of the Arabidopsis ${\gamma}$-TMT cDNA into lettuce genomic DNA was confirmed by PCR and Southern analysis. HPLC analysis showed that $\alpha$- to ${\gamma}$-tocopherol ratio increased over four fold in a transgenic lettuce line indicating successful expression of the transgenic Arabidopsis ${\gamma}$-TMT in lettuce.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.469-474
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• 2000

Effect of the antibiotic carbenicillin on callus and shoot formation from the leaf disc culture of tobacco (Nicotiana tabacum L. cv. BY-4) was examined. The number of shoot induced from the leaf explants was decreased as the amount of carbenicillin increased from 250 mg/L to 2,000 mg/L on MS medium containing 0.5 mg/L of BAP or kinetin. In addition, calli formation from the leaf explants was increased by the treatment of 250 mg/L ∼ 500 mg/L carbenicillin with or without 0.5 mg/L of 2,4-D or NAA. However, the fresh weight of 4-week-cultured explants was decreased with increasing carbenicillin from 250 mg/L to 2,000 mg/L on MS medium containing 0.5 mg/L of 2,4-D or NAA. These results indicate that carbenicillin has an auxin-like effect, such as promoting callus formation and inhibiting shoot induction. It leads to the conclusion that the auxin-like property should be taken into account for the production of transgenic plants via Agrobacterium-mediated transformation.

• Journal of Plant Biotechnology
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• v.43 no.2
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• pp.240-247
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• 2016

In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having 'pBIN 19' plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with $500{\mu}l$ of bacterial solution after drying in sterile cabin for 35 min were used.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.60 no.1
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• pp.97-106
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• 2015

Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}$ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

• Korean Journal of Plant Tissue Culture
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• v.28 no.5
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• pp.277-281
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• 2001

Embryogenic callus induction and plant regeneration methods were developed for native Kentucky bluegrass (Poa pratenes L.) ecotypes. Mature caryopses and immature inflorescences (20 mm in length) of 4 native ecotypes and 5 foreign cultivars were plated on MS medium (30 g/L sucrose, 3 g/L Phytagel) supplemented with 1 mg/L 2,4-D, and cultured in the dark at 24$^{\circ}C$. Most explants formed calli, but more embryogenic calli were induced from the explants of immature inflorescences than caryopses which produced mostly non-embryogenic rooty calli. In P77 ecotypes, immature inflorescence explants formed embryogenic calli with the rate of 62~95%, and those of field-grown plants were more efficient than greenhouse-grown ones in embryogenic callus induction. Plantlets were regenerated from the embryogenic calli when they were transferred to hormone-free MS medium, and grew to maturity without morphological variations in greenhouse.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.335-342
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• 2017

Purple passion fruit (Passiflora edulis Sims) is one of the introduced tropical plants, an increasing interest has arisen due to its distinctive taste and attractive flavor. It is expected that passion fruit production and planted area will increase gradually in the years ahead because of high profitability and consumer's demands of healthful ingredients. So we tried to investigate the effect of plant growth regulators and antioxidants on in vitro plant regeneration and callus induction from leaf explants of passion fruit for an establishment of optimal mass propagation system. Young leaf explants of purple passion fruit were cultured in Murashige and Skoog (MS) medium containing different growth regulators and antioxidant additives to induce the shoot organogenesis. After 8 weeks, the highest embryogenic callus formation rate was obtained in MS medium supplemented with $1mg{\cdot}L^{-1}$ 6-benzylaminopurine (BAP) and $2mg{\cdot}L^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), furthermore, the shoot development via organogenesis was also observed. Silver nitrate ($AgNO_3$), which was added into the medium to minimize the adverse effects of leached phenolics, was effective for reduction of medium browning and sudden explant death. In the medium supplemented with $1mg{\cdot}L^{-1}$ BAP and $1mg{\cdot}L^{-1}$ gibberellic acid ($GA_3$), shoots were most vigorously regenerated and elongated. Most shoots rooted successfully in half strength medium with $1mg{\cdot}L^{-1}$ indol-3 acetic acid (IAA), and more than 90% of plantlets survived after 4-month acclimatization period.