• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.24-24
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• 2018

본 연구는 응달고사리[Cornopteris crenulatoserrulata (Makino) Nakai]의 전엽체 대량증식 및 포자체 형성을 위한 배지구성물질 및 배양토의 최적조건을 구명하고자 수행되었다. 실험재료는 기내에서 포자발아를 통해 획득한 전엽체를 계대배양하여 실험에 사용하였다. 배지구성물질이 전엽체의 증식 및 형태 형성에 미치는 영향을 알아보고자, 배지구성물질의 농도를 1/4, 1/2, 1배로 조절한 MS배지와 Knop배지를 조성하였다. 기내 배양된 전엽체 300mg을 메스로 다진 후 상기 5종의 배지에 배양하였다. 실험의 결과, 전엽체의 생체중은 Knop배지에서 3.93g으로 가장 많이 증가되었으며, 현미경으로 관찰한 결과 기관형성도 정상적으로 이루어졌다. 선행연구에서 선발된 Knop배지를 기준으로 sucrose(0, 0.5, 1.0, 2.0 및 3.0%) 및 활성탄(0, 0.1, 0.2 및 0.4%)의 농도를 달리 첨가하여 전엽체에 미치는 영향을 확인하였다. 실험결과, 0.5%의 sucrose를 첨가한 배지에서 생체중이 5.90g으로 증가하였으며, 활성탄은 첨가하지 않는 것이 효과적이었다. 포자체 형성을 위한 최적의 토양조건을 구명하고자, 원예상토와 피트모스, 펄라이트, 마사토 등 인공토양의 비율을 달리하여 8종류의 토양을 사각분에 충진하였으며, 기내에서 배양된 전엽체를 10초간 분쇄한 후 사각분에 1g씩 균일하게 분주하여 16주간 배양하였다. 배양결과, 원예상토를 단용으로 처리한 실험구에서 포자체가 171.5개로 가장 많이 발생하였으며, 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 125.0개, 원예상토, 피트모스, 마사토를 1:1:1(v:v:v)로 혼합한 토양에서 113.3개 순으로 포자체가 발생하였다. 포자체의 생육은 포자체의 형성이 가장 많았던 원예상토를 단용한 토양에 비해 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 생체중, 엽장, 엽폭, 근장 등이 우수하였다. 한편 피트모스가 혼합된 토양에서는 전엽체와 포자체 형성이 억제되는 경향을 보였다. 위 결과로 보아 응달고사리의 전엽체 기내배양에는 sucrose 0.5%가 첨가된 Knop배지가 적합하며, 포자체 대량생산을 위해서는 원예상토를 단용한 토양을 사용하는 것이 좋다고 판단되었다.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.74-81
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• 2022

Brassica napus, an oil crop that produces rapeseed oil, is an allotetraploid (AACC, 2n = 38) produced by natural hybridization between B. rapa and B. oleracea. In this study, microspore was cultured using the F₁ developed from a cross between 'EMS26' line with high oleic acid content and 'J8634-B-30' lines. The flower bud size showing the nuclear development at the late uninucleate and binucleate stage with high embryogenesis rate was 2.6 ~ 3.5 mm. Microspores were cultured using only this size and after then most microspore embryo developed into secondary embryos and then regeneration plants obtained from the developed multilobe. The analysis of the ploidy of the plants revealed that 66.7% and 27.8% of the total lines were tetraploids and octoploids, respectively. The sizes of stomatal cells in tetraploids, octoploids, and diploids were 25.5, 35.6, and 19.9 ㎛, respectively, indicating that ploidy level was positively correlated with cell size. Furthermore, 62 tetraploid doubled haploid (DH) lines were selected. The average oleic acid (C18:1) and linolenic acid (C18:3) concentrations of DH were 72.3% and 6.2%, respectively. Oleic acid and linolenic acid concentrations exceeded the two parental values in 5 and 14 DH lines, respectively, suggesting that these two fatty acids had transgressive segregation. Therefore, the DH population can be utilized for the biosynthesis of unsaturated fatty acids in rapeseed and related genes. It can also be used as a breeding material for varieties with high oleic acid concentrations.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.257-261
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• 1998

The repeated subcultures of in vitro plant materials in wasabi became highly vitrified and the capacity for multiple shoot formation from the vitrified plant materials was very low. In order to improve the quality of in vitro propagated planting materials, the experiments were carried out using culture vessels capped with membrane filter(MF). When vitrified shoots were cultured on MS medium with 0.2mg/L BA in the vessels with MF or without MF for 60 days, the shoots in the vessels with MF did not vitrified. In contrast, the shoots grown in the vessels without MF vitrified at 65%. The stomates of vitrified leaves were circular and inflated, whereas those of normal leaves acclimatizated in the vessels with MF were ovate in shape. The hardened shoots were also cultured on MS media without sucrose containing 0.01mg/L IBA in vessels with(photoautotrophic culture) or without(control) MF. Sucrose was necessary for survival of the in vitro plantlets in the vessels without MF. After 20 days of culture, the shoots in the vessels without MF on the sucrose-free media turned yellow and died. But the shoots in the vessels with MF in the sucrose-free media produced a lot of roots. When shoots were cultured on MS medium with 2% sucrose containing 0.01mg/L IBA in the vessels with(photomixotrophic culture) or without(heterotrophic culture) MF, best growth occured in photomixotrophic culture.

• Horticultural Science & Technology
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• v.32 no.5
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• pp.684-693
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• 2014

Abiotic stress conditions such as cold, drought, and salinity trigger physiological and morphological changes and yield loss in plants. Hence, plants adapt to adverse environments by developing tolerance through complex regulation of genes related to various metabolic processes. This study was conducted to construct a coexpression network for multidirectional analysis of salt-stress response genes in Brassica rapa (Chinese cabbage). To construct the coexpression network, we collected KBGP-24K microarray data from the B. rapa EST and microarray database (BrEMD) and performed time-based expression analyses of B. rapa plants. The constructed coexpression network model showed 1,853 nodes, 5,740 edges, and 142 connected components (correlation coefficient > 0.85). On the basis of the significantly expressed genes in the network, we concluded that the development of salt tolerance is closely related to the activation of $Na^+$ transport by reactive oxygen species signaling and the accumulation of proline in Chinese cabbage.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.49-49
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• 2018

본 연구는 관상가치가 높아 조경 및 관상소재로 개발이 가능한 큰개관중의 전엽체 증식 및 포자체 형성에 적합한 배양조건을 구명하고자 수행되었다. 실험재료는 무가온 온실에서 채집한 포자를 기내에서 발아시켜 전엽체를 획득한 후 8주 간격으로 계대배양하여 실험에 사용하였다. 배지종류에 따른 전엽체의 기내 증식 및 형태형성의 영향을 알아보고자 배양된 전엽체 300mg을 메스로 다진 다음 농도를 1/4, 1/2, 1, 2배로 조절한 MS와 Knop배지에 8주간 배양하였다. 그 결과, 1MS배지에서 전엽체의 생체중이 5.5g으로 가장 많이 증가하였다. 1MS를 제외한 타 처리구는 생체중의 증가수준이 1.1-3.0g에 머물러 1MS배지보다 저조한 수준을 보였다. 현미경을 이용한 전엽체의 관찰결과, 1MS배지는 엽육의 색이 녹색으로 생육이 양호하였다. 전엽체의 증식이 가장 저조하였던, 2MS와 1/4MS는 생육의 저조뿐만 아니라 노화현상도 관찰되었다. 포자체 형성을 위한 최적의 토양조건을 알아보고자, 원예상토, 피트모스, 펄라이트 및 마사토의 비율을 5종류로 달리하여 배양토를 혼합하였다. 혼합된 토양은 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하여 기내배양된 전엽체 1g을 증류수와 함께 10초간 분쇄한 다음 토양표면에 분주 후 10주간 재배하였다. 그 결과, 원예상토가 높은 비율로 첨가된 혼합조건에서 포자체의 형성이 우수하였다. 그중 원예상토와 마사토를 2:1(v:v)로 혼합한 토양, 원예상토와 펄라이트를 2:1(v:v)로 혼합한 토양에서 포트 당 각 357.0, 339.8개의 포자체가 형성되었다. 또한 형성된 포자체의 생육을 조사한 결과 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 생체중, 엽수, 엽장, 엽폭, 근수, 근장 및 SPAD value 등의 생육수치가 우수하였다. 따라서 큰개관중의 전엽체 증식에 적합한 배지는 1MS로 판단되었으며, 포자체 대량생산을 위해서는 원예상토와 마사토를 2:1(v:v)로 혼합한 토양이 적합하다고 판단된다.

• Korean Journal of Plant Tissue Culture
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• v.25 no.2
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• pp.115-118
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• 1998

This experiments were carried out to determine the optimum concentrations of plant growth regulators for the direct embryogenesis from the cotyledon culture of Paeonia lactiflora Pall. Zygotic embryos rescued from true seeds were developed abnormally in the medium containing ABA. But somatic embryogenesis from cotyledons of abnormal seedlings was induced more effectively. Also the somatic embryogenesis from cotyledons was promoted in the medium containing ABA. The frequency of embryogenesis was maximum(59.9%) from the cotyledons cultured on MS medium containing 0.5 mg/L ABA on which the frequency of somatic embryos with two cotyledons was 22.6%.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.23-23
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• 2018

본 연구는 참우드풀과 우드풀의 중간형으로 알려진 좀우드풀(Woodsia intermedia Tagawa)의 전엽체 증식 및 포자체 형성을 위한 기내 외 번식조건을 구명하고자 수행되었다. 실험재료는 포자를 발아시켜 획득한 전엽체를 8주 간격으로 계대배양하면서 확보하였다. 전엽체 증식에 적합한 배양조건을 탐색하고자, 전엽체 300mg을 다진 후 배지종류(Knop, 1/4, 1/2 및 1MS)와 배지구성물질(sucrose와 활성탄)의 농도를 달리하여 8주간 배양하였다. 배양실은 온도 $25{\pm}1.0^{\circ}C$와 광도 $30{\pm}1.0{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$(16/8h)로 조절되었다. 연구결과, Knop배지에서 생체중이 2.4g으로 전엽체의 증식이 가장 왕성하였다. 한편 MS계열 배지는 농도에 관계없이 매우 저조한 증식을 보였다. Sucrose는 0.5%를 첨가한 배지에서 생체중의 증가량이 가장 컸으며, 활성탄은 첨가농도에 관계없이 유사한 증식수준을 나타냈다. 포자체 형성에 적합한 토양조건을 확인하고자, 인공토양(원예상토, 피트모스, 펄라이트 및 마사토)의 비율을 달리하여 5종류의 혼합토양을 조성하였다. 혼합토양을 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하고, 준비된 전엽체 1g을 10초간 분쇄한 다음 토양표면에 분주하여 11주간 재배하였다. 재배환경은 온도 $25{\pm}1.0^{\circ}C$, 광도 $43{\pm}2.0{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$(16/8h) 및 습도 $72{\pm}2.0%$로 조절되었다. 연구결과, 모든 처리구에서 포자체의 형성이 확인되었으며, 특히 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 포트당 421.0개의 많은 포자체가 형성되었다. 다음으로 원예상토와 펄라이트를 2:1(v:v)로 혼용한 토양, 원예상토, 피트모스, 마사토를 1:1:1(v:v:v)로 혼합한 토양, 원예상토 단용, 원예상토, 피트모스, 펄라이트를 1:1:1(v:v:v)로 혼합한 토양 순으로 각 228.0, 203.3, 126.8, 91.5개 형성되었다. 따라서 좀우드풀의 포자체를 대량으로 생산하기 위해서는 원예상토와 마사토를 2:1(v:v)로 혼용한 토양에 전엽체를 분주하여 재배하는 방법이 효과적일 것으로 판단된다.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.1-6
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• 2000

This study was carried out to find the effects of plant growth regulators on callus formation, rooting and shooting from cotyledon explant in oriental me]on. Various combinations of 0.1 mg/L auxins (IAA, NAA) and 0.5, 1.0. 1.5, 2.0 mg/L cytokinins (BA, kinetin, zeatin) were treated to the MS basal medium, respectively. Callus was induced mort effectively as 2,437.0 mg (FW)/explant in MS medium supplemented with 0.1 mg/L NAA and 2.0 mg/L BA, but that was non-embryogenic callus as colored yellow white and broke easily. Root was induced most effectively at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L NAA and 0-5 mg/L kinetin. Shoots formed on cut part of vein at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L IAA and 2.0 mg/L BA, that were multiple shoots. in case of its concentration, BA and lower concentration of IAA and NAA (0.01 and 0.05 mg/L). respectively. shooting ratio was not increased. The result of treatment with BA 0-5 mg/L and IAA 0.1 mg/L, callus induced at a week, and shoot start to form multiple shoots about 3 weeks after inoculation. After 2 times subculture as 2 weeks intervals, divided shoots rooted and developed into intact plantlets at 10 weeks and then that grown normally on pots after acclimatization.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.900-910
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• 2015

This study aimed to investigate the efficiency of colchicine and oryzalin in inducing polyploidy in two Cymbidium hybrids [Showgirl 'Silky' and Mystery Island 'Silk Road' (Silk Road-4)]. Colchicine was used at concentrations ranging from 50 to $500mg{\cdot}L^{-1}$, with treatments lasting 1 to 3 weeks. Oryzalin was used at concentrations ranging from 3 to $20mg{\cdot}L^{-1}$, with treatments lasting 3 to 6 days or 1 to 3 weeks. The survival rate of PLBs was better in colchicine than in oryzalin solutions. The ploidy levels were screened using flow cytometry. In C. Showgirl 'Silky', the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (60%) and the 2-week treatment in $5mg{\cdot}L^{-1}$ oryzalin (46.7%). In C. Mystery Island 'Silk Road' (Silk Road-4), the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (16.7%) and the 3-day treatment in $10mg{\cdot}L^{-1}$ oryzalin (6.7%). Colchicine was more efficient than oryzalin in terms of polyploidy induction. Furthermore, pre-treatment, which entailed poking 10 times with forceps, improved the efficiency of chromosome doubling.

• Korean Journal of Plant Resources
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• v.21 no.4
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• pp.341-345
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• 2008

Two orchid species of Calanthe discolor Lindl. and C. discolor for. sieboldii (Decne.) Ohwi, which have different form flower color and size. They were crossed in mid April by artificial pollination, and the F1 hybrid seeds were collected mid October. Germination of seeds was investigated on pre-treatment of seeds and under the various environmental conditions. Germination was promoted by moisture absorption and ultrasonic treatment of seeds. Dark culture of F1 hybrid seeds enhanced germination and protocorm formation, and development into seedlings compared with light culture. Although, plant growth regulators such as NAA and BA had a slightly promotive effect on seed germination and protocorm growth, regenerated seeding were showed abnormal growth patterns. Regenerated F1 hybrid plantlets were successfully transferred to pot.