• Journal of Plant Biotechnology
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• 제35권2호
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• pp.163-167
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• 2008

Adventitious buds appeared within 2 weeks on the base of the petiole explants and increased for two months. A maximum of regeneration (15.6%) was obtained on the medium containing $1.5\;{\mu}M$ TDZ in combination with $1\;{\mu}M$ IBA. To know which explants are the best for the induction of regeneration, three explants such as leaf, petiole and leaf-petiole were used. Among the explant types, the leaf-petiole explant was significantly more effective than leaf and petiole for promoting adventitious shoots, with leaf-petiole inducing at the highest regeneration frequency (33.7%). The regeneration frequency of adventitious shoots in the leaf-petiole explants was significantly affected by leaf size and the position of explants. The leaf-petiole smaller than 5 mm leaf in width was induced at the highest regeneration frequency (68.9%). The smaller leaf sizes, the greater regeneration frequency. Also when the leaves are nearer to the shoot tip, the regeneration frequency is higher. When the rooted micro-shoots were transferred to the soil after growing for 6 weeks in the media, the survival rate was 90%.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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• pp.85-89
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• 1999

Somatic embryogendesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology. This paper describes the direct somatic embryogenesis from zygotic embryos of Panax ginseng is reversely related to normal axis growth of zygotic embryos by the experiment of various chemical treatments. Under the normal growth condition, the apical tips of embryo axis produced an agar-diffusible substance, which suppressed somatic embryo development from cotyledons. Although the cells of zygotic embryos were released from the restraint of embryo axis, various factors were still involved for somatic embryo development. Electron microscopic observation revealed that the ultrastructure of cells of cotyledon epidermis markedly changed before initiation of embryonic cell division, probably indicating reprogramming events into the cells embryogenically determined state. Polar accumulation of endogenous auxin or cell-cell isolation by plasmolysis pre-treatment is the strong inducer for the somatic embryo development. The cells for the process of somatic embryogenesis might be determined by the physiological conditions fo explants and medium compositions. Direct somatic embryos from cotyledons fo ginseng were originated eithrer from single or multiple cells. The different cellular origin of somatic embryos was originated either from single or multiple cell. The different cellular origin of somatic embryos was depended on various developmental stages of cotyledons. Immature meristematic cotyledons produced multiple cell-derived somatic embryos, which developed into multiple embryos. While fully mature cotyledons produced single cell-derived single embryos with independent state. Plasmolysis pretreatment of cotyledons strongly enhanced single cell-derived somatic embryogenesis. Single embryos were converted into normal plantlets with shoot and roots, while multiple embryos were converted into only multiple shoots. GA3 or a chilling treatment was prerequisite for germination and plant conversion. Low concentration of ammonium ion in medium was necessary for balanced growth of root and shoot of plantlets. Therefore, using above procedures, successful plant regeneration of ginseng was accomplished through direct single embryogenesis, which makes it possible to produce genetically transformed ginseng efficently.

• 한국자원식물학회지
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• 제19권3호
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• pp.432-435
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• 2006

A rapid and mass propagation method for multiple shoots and plant regeneration using bulb scales of Muscari comosum var. plumosum were developed. In vitro different parts of bulb scale as explants were cultured on 11 kinds of MS (1962) media supplemented with various plant growth regulators to induce shoot and callus. A combination of 2.0 mg/L 6-BA and 2.0 mg/L IBA on MS medium was the most favorable and induced the highest production (80%) of shoot formation after 30 days. We also found that the middle part of bulb scale was the best for mass propagation of Muscari comosum var. plumosum of which production could reach 64.4%.

• Journal of Plant Biotechnology
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• 제31권1호
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• pp.67-72
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• 2004

고구마의 정단배양에 의한 배발생 캘러스로부터 대량증식 체계가 개발되어져 왔다. 고구마 정단분열조직은 1mg/L 2,4-D가 첨가된 MS배지에서 배양 4주 경에 최적의 배발생 캘러스가 형성되었다. 또한 2,4-D가 첨가된 배지에 casein을 첨가함으로써 고구마 신천미 품종의 배발생 효율을 최고 90%이상으로 2.4-D단독처리보다 현저하게 증가시켰다. 배발생 캘러스로부터 체세포배의 유도는 식물생장조절제가 제거된 MS 기본배지에서 효과적으로 형성되었으며 300∼500mg/L casein을 첨가한 배지에서는 더 높은 형성 빈도와 녹색의 단단한 체세포배가 발달하였다. 한편, 2mm이하의 체세포배로부터 이차 배발생 캘러스 형성 및 체세포배의 발달이 100∼300mg/L casein의 첨가에 의해 증가하였다 배발생 캘러스에서 얻어진 체세포배는 직접 MS기본배지에서 쉽게 각 기관이 형성되었으며, 발근과 shoot를 발달시켜 정상적인 식물체로 하여 토양에 성공적으로 옮겨 심을 수 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.80-80
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• 2019

This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of (Fragaria x ananassa DUCH. cvs. 'Derunoka' and 'Jumbo pure berry'. The shoot tips of strawberry were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7M). Precultured explants were treated with loading solution (LS, C4) containing glycerol 17.5% and sucrose 17.5% for 40 min and exposed to dehydration solution (B1) containing 50% of glycerol and 50% of sucrose for 60 min at $25^{\circ}C$, and then transferred onto droplets containing $2.5{\mu}l$ PVS3 on sterilized aluminum foils ($4cm{\times}0.5cm$) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regeneration rate (%) was obtained when shoot tips were precultured with treatment-2 (exposing of shoot tips to MS + 0.3M Sucrose for 30 h and then treated with MS+0.5 M sucrose for 16 h) at $25^{\circ}C$ in both the cultivars. The viability of cooled samples, followed by culturing on MS medium for 4 weeks was 77.8% and 60.0% for 'Derunoka' and 'Jumbo pure berry', respectively. This result shows droplet-vitrification would be a promising method for cryobanking strawberry germplasm.

• 한국자원식물학회지
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• 제31권6호
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• pp.675-683
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• 2018

This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of Chrysanthemum morifolium (Ramat.) cvs. 'Borami' and 'Yes morning'. The shoot tips of Chrysanthemum were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7 M). Precultured explants were treated with loading solution (LS, C6) containing glycerol 20% and sucrose 20% for 30 min and exposed to dehydration solution (B5) containing 40% of glycerol and 40% of sucrose for 60 min at $25^{\circ}C$, and then transferred onto droplets containing $2.5{\mu}l$ PVS3 on sterilized aluminum foils ($4cm{\times}0.5cm$) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regeneration rate (%) was obtained when shoot tips were precultured with treatment-2 (exposing of shoot tips to MS + 0.3M sucrose for 30 h and then treated with MS+0.5 M sucrose for 16 h) at $25^{\circ}C$ in both the cultivars. The viability of cooled samples, followed by culturing on $NH_4NO_3$-free MS medium for first 5 days was increased to two-fold (80.7%) regrowth rate over those cultured on normal MS medium or MS medium containing plant growth regulators. This result shows droplet-vitrification would be a promising method for cryobanking chrysanthemum germplasm.

• 한국자원식물학회지
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• 제18권1호
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• pp.154-160
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• 2005

유색 칼라 세 품종(Sunlight, Chiante, Pink Persuation)의 기내 급속증식체계를 확립코자 정단분 열조직을 배양함에 있어 callus, 신초 및 뿌리 분화에 미치는 식물생장조절물질의 효과와 sucrose가 기내 괴경 형성 및 비대에 미치는 효과를 조사하였다. 식물체는 캘러스를 통하거나 또는 치상체에서 직접 분화되는 두 경로로 형성되었으며 2.0 mg/L BA 단용 처리에서 Sunlight품종은 $53.3\%$ 캘러스를, Chiante 품종은 $56.7\%$ 직접 신초를 형성하였다. 2.0-3.0 mg/L BA 단용 처리는 품종별로 식물체재 분화 빈도가 $20-70\%$로 차이가 있었으나 세 품종 모두 3.0 mg/L. BA 단용 처리에서 신초 분화가 양호하였다. 다아체 형성에 미치는 cytokinin의 효과는 Sunlight품종은 1.0 mg/L 2ip 처리로 16개체를, Chiante 품종은 5.0 mg/L BA를 처리로 14개체를, Pink Persuasion품종은 1.0 mg/L BA처리로 12개체를 형성하였다. NAA는 세품종 뿌리분화에 효과가 없었으며 Sunlight 와 Chiante 품종은 1.0 mg/L IAA에서, Pink Persuasion 품종은 2.0 mg/L IBA가 효과적이었다. 괴경형성에 미치는 sucrose는 Sunlight 품종은 90 g/L, Chiante 및 Pink Persuasion 품종은 70 g/L 가 괴경 형성 및 비대에 효과적이었다.

• 한국자원식물학회지
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• 제17권1호
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• pp.1-6
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• 2004

쪽파의 정단분열조직을 LS기본배지에 zeatin(0.1, 0.5, 1.0, 3.0mg/L)과 NAA(0.1, 0.5, 1.0, 2.0, 5.0mgA)를 단독 및 혼합 첨가시키어 식물체 생산과 인경 형성율을 조사하였다. 정 단분열조직으로부터 신초 분화는 zeatin 0.1, 0.5, 1.0 mg/L 단독처리구와에 zeatin 0.5 mg/L에 NAA 1.0mg/L 혼합첨가구에서 가장 양호한 반응을 보였다. 뿌리분화율은 zeatin 0.5 mg/L에 NAA 1.0mg/L 혼합첨가구에서 가장 좋았다. 분화된 식물체로부터 정상적인 인경형성은 NAA단독첨가보다는 zeatin과의 혼합첨가가 효과적이었다. 특히 NAA 3.0mg/L에 zeatin 1.0mg/L를 혼합시켰을 때 정상적인 인경 형성에 가장 좋은 반응을 보였다.

• Journal of Plant Biotechnology
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• 제8권1호
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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.

• 한국약용작물학회지
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• 제13권6호
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• pp.273-277
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• 2005

지황 (R. glutinosa)의 조직절편으로부터 고효율의 식물체 재분화 시스템을 확립하기 위하여 기내에서 발아된 유묘의 잎을 절취하여 auxins과 cytokinins을 농도별로 단독 또는 혼합하여 MS배지에 치상한 후 신초의 형성율을 조사하였다. 잎 절편으로부터 부정아의 형성율은 1 mg/{\ell}\;BA와\;2mg/{\ell}$ IAA가 첨가된 MS배지에서 93.4%로 가장 높았다. 한편, 엽령에 따른 부정아의 형성율을 조사한 결과 기내에서 발아 후 6주가 지난 유식물체의 정단부위로부터 3번째의 잎을 사용함으로써 신초의 형성율을 98.4%까지 높일 수 있었다. 형성된 신초는 $0.1mg/{\ell}$ IBA가 첨가된 1/2MS배지로 옮겼을 때 4주 후 최대 발근율을 보여주었다.