• Journal of Plant Biotechnology
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• 제36권2호
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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.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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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.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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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.

• 식물조직배양학회지
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• 제22권3호
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• pp.131-135
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• 1995

잡종 사기나무의 기내 대량증식을 목적으로 줄기절간 유래의 기내 및 기외시료를 이용하여 다경줄기 유도에 관여하는 식물생장 조절물질의 효과에 대하여 조사하였다. 기외시료의 측아를 제거한 다음 20에서 30$\mu$M 의 zeatin이 함유된 WPM 배지에 6주간 배양하였을때 각각 11개와 13개의 다경줄기가 유도되었다. 그러나 기외시료의 측아가 붙어있는 상태에서 기내배양하여 2주후에 측아를 제거하고 동일 배지에 배양하였을 경우에는 BA가 1.0에서 2.0 $\mu$M 함유된 배지에서도 13개와 15개의 다경줄기를 유도할 수 있었으며, 특히 이때 생산된 줄기의 상태는 아주 건전한 것으로 나타났다. 기내줄기를 사용하였을때는 20 $\mu$M의 zeatin을 처리하였을때 가장 많은 줄기를 생산할 수 있었으며, 생산된 조직배양묘는 일정기간 순화를 시킨 후 온실이나 포지에 식재가 가능하였다.

• 식물조직배양학회지
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• 제27권6호
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• pp.435-439
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• 2000

겨울 상추 품종인 청치마 상추의 자엽조직을 ${\gamma}$-TMT유전자가 도입된 A. tumefaciens LBA 4404와 공동배양한 후, 50 mg/L kanamycin, 500 mg/L carbenicillin, 0.1 mg/L NAA, 0.5mg/L BA가 첨가된 MS재분화 배지에 옮겨 약4주 배양하여 재분화된 신초를 얻었다. 재분화된 신초를 50 mg/L kanamycin, 250mg/L carbenicillin이 포함된 MS 기본 배지에 옮겨 발근된 소식물체를 얻었다. 선발된 형질전환 식물체의 genomic DNA에 대한 PCR분석과 Southern분석을 수행하여 애기장대 ${\gamma}$-TMT유전자 특이적 DNA단편이 상추의 genomic DNA내에 삽입되었음을 확인하였다. 선발된 상추 형질전환체 잎의 $\alpha$-tocopherol/${\gamma}$-tocopherol 함량 비율이 대조 식물체에 비하여 약 4배 증가하여 도입된 ${\gamma}$-TMT유전자가 안정적으로 발현함을 나타내었다.

• 식물조직배양학회지
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• 제27권6호
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• pp.469-474
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• 2000

Agrobacterium tumefaciens를 매개로 한 형질전환 과정에서 제균제로 사용되고 있는 항생제인 carbenicillin이 담배(Nicotiana tabacum L. cv. BY-4) 잎절편 배양시 기관분화에 미치는 영향을 검토하였다. BAP와 kinetin을 각각 0.5 mg/L 첨가한 MS 배지에 250 mg/L∼2000 mg/L의 carbenicillin을 처리한 배지에서 신초 수는 크게 감소하였다. 또한 carbenicillin을 단독첨가하거나 0.5 mg/L의 2,4-D 또는 NAA와 혼합첨가한 MS 배지에서 250 mg/L∼500mg/L농도의 carbenlcillin은 캘러스 형성을 증가시켰다. 반면 carbenicillin을 0.5 mg/L의 2,4-B, NAA와 각각 혼합첨가한 배지에서 배양 4주째 치상조직의 생중량은 carbenicillin의 농도가 250mg/L에서 2000mg/L로 높아질수록 감소하였다. 이러한 결과는 carbenicillin이 캘러스 형성을 촉진하고 신초형성을 억제하는 auxin과 유사한 효과를 가지고 있다는 것을 시사해 준다. 따라서 담배 잎절편에 Agrobacterium tumefaciens를 매개로 형질전환하고자 할 때 최적 재분화용 식물생장조절제의 농도는 carbenicillin이 제균제뿐만 아니라 auxin 유사효과를 가지는 물질임을 고려하여 결정하여야 한다.

• Journal of Plant Biotechnology
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• 제43권2호
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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.

• 한국작물학회지
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• 제60권1호
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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.

• 식물조직배양학회지
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• 제28권5호
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• pp.277-281
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• 2001

자생 왕포아풀 (Poa pratensis L.)의 배발생 캘러스 유도 및 식물체 재분화 조건을 규명하였다. 도입된 다섯 품종과 국내에서 수집한 네 가지 생태형의 미성숙 화서와 완숙종자를 sucrose 30 g/L, Phytagel 3 g/L와 2,4-D 1 mg/L가 함유된 MS배지에 치상하여 24$^{\circ}C$로 조절된 항온실에서 배양하면서 캘러스를 유도하였다. 대부분의 배양에서 캘러스가 형성되었으나 배발생 캘러스는 완숙종자를 절편체로 썼을 때보다 미숙화서에서, 온실에서 재배되어 형성된 화서보다는 자연 재배조건에서 형성된 화서에서 배발생 캘러스의 유도율이 높았다. 배발생 캘러스는 85~98%의 효율로 높게 유도되었으며 계대배양 및 액체 진탕배양을 통해 6개월 이상 장기 증식되었으며, 생장조절제가 함유되지 않은 MS 재분화 배지에 이식하면 다수의 식물체로 재분화되었다. 재분화 식물들은 온실재배에서도 정상적으로 생장을 하였다.

• Journal of Plant Biotechnology
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• 제44권3호
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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.