• 제목/요약/키워드: In vitro propagation

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멸종위기종 두메닥나무(Daphne pseudomezereum var. koreana)의 줄기 기내배양을 통한 식물체 생산 (Propagation of Endangered Species, Daphne pseudomezereum var. koreana via in vitro Bud Culture)

  • 추예린;박상희;정은주
    • 한국산림과학회지
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    • 제109권2호
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    • pp.189-194
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    • 2020
  • 두메닥나무 (Daphne pseudomezereum var. koreana)는 강원도, 전라북도, 경상도의 고산지대에 분포하는 한국 자생종이다. 최근 자생지가 축소되어 개체 수가 급격히 줄어들고 있어 멸종위기종으로 지정되었다. 두메닥나무의 추출물은 화장품 등에 유용한 소재로 이용되나 식물체의 번식 효율은 매우 낮은 실정이다. 본 연구는 두메닥나무의 유전자원 보존과 생물소재로 활용을 위해 대량증식법을 확립하고자 하였다. 온실에서 자란 두메닥나무의 액아를 포함한 줄기절편을 생장이 우수했던 WPM 배지를 기본 조성으로 하여 BA와 GA3를 혼용 처리한 4가지 배지에서 기내줄기의 정단의 유무에 따른 줄기생장과 신초유도를 비교하였다. 두메닥나무는 정아가 포함된 절편체는 BA 8μM + GA38μM 조성에서 줄기생장이 가장 우수하였으며, 정아가 존재하지 않을 때 BA 8μM + GA38μM 조성에서 신초유도가 가장 우수하였다. 반면 발근유도는 NAA 1μM, NAA 5μM, NAA 10μM 조성 모두 발근이 발생하지 않은 대신 잎과 줄기가 생장하여 효과가 없는 것으로 나타났다.

배지내 천연유기물 첨가가 딸기 "설향" 배양묘 기내 생육에 미치는 영향 (Effect of Natural Additives on In Vitro Growth Medium of Strawberry 'Seolhyang')

  • 권영희;이정관;김희규;김경옥;박재성;허윤선
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2019년도 춘계학술대회
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    • pp.55-55
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    • 2019
  • Strawberry which is the genus Fragaria under family Rosaceae is one of the most important fruit plants for both fresh consumption and food processing in the temperate and subtropical countries. Propagation of strawberry is achieved either through runners or by in vitro micropropagation. Meristem tips, generally obtained from runners of virus-free plants, are commonly used to establish in vitro cultures, which are employed for mass propagation or as a source of plant material for regeneration and transformation experiments. This study was conducted to determine the optimal natural additives strength to improve sprouting shoot rate of apical meristem of strawberry 'Seolhyang'. Strawberry apical meristem at size (0.2 mm to 0.3 mm) with leaf primordials were cultured on the 1/3MS(Murashige & Skoog) medium supplemented with five natural additives such as coconut milk, maple sap, banana powder and peptone. The sprouting ratio and growth characteristics were evaluated after eight weeks after in vitro culture. Shoot ratio of 'Seolhyang' apical meristem was 72.9% in 1/3MS medium supplemented with maple sap. On the other hand, the low shoot ratio was observed 47.7% in 1/3MS medium supplemented with banana powder. Shoot length was different as natural additives but numbers of leaf was not significantiy different among the natural additives. As a result, the sprouting ratio and plant growth were enhanced effectively in 1/3MS medium with maple sap compared to the others.

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Gametophyte Propagation and Sporophyte Formation of Asian Chain Fern [Woodwardia japonica (L. f.) Sm.] Under Various Medium Conditions In Vitro and Ex Vitro

  • Cho, Ju Sung;Jang, Bo Kook;Park, Kyungtae;Lee, Ha Min;Lee, Cheol Hee
    • 한국자원식물학회지
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    • 제32권6호
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    • pp.735-742
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    • 2019
  • This study investigated a suitable method that could be applied for Asian chain fern [Woodwardia japonica (L. f.) Sm.] to propagate gametophytes and promote sporophyte formation. The gametophytes used in all experiments were obtained from germinated spores in vitro and were subcultured at 8-week intervals. The most appropriate media for gametophyte propagation was identified by culturing 300 mg of gametophyte in Murashige and Skoog (MS) basal medium (1/8, 1/4, 1/2, 1, 2), and Knop medium for 8 weeks. As a result, fresh weight of the gametophyte was increased by 56.7-fold on MS medium. Moreover, antheridium formation as well as gametophyte growth was improved on MS medium, especially. To improve the sporophyte formation ex vitro, 1.0 g of gametophyte was ground with distilled water and spread on eight combinations onto four different culture mediums, such as bed soil, peat moss, perlite and decomposed granite. Then generation and growth of sporophytes were investigated after cultivation for 10 weeks. As a result of this experiment, peat moss had a promotive effect of sporophyte formation at single-use and mixed culture soils. In particular, a mixture of bed soil, peat moss and perlite in a 1:1:1 ratio (v/v/v) led to the accelerated formation (782.5 ea/pot) and the frond growth of sporophytes. This included increases in length and width of fronds. However, promotive effect of gametophyte growth and sporophyte formation was not found at single-use and treatment with high ratio of bed soil.

Rapid micropropagation of wild garlic (Allium victorialis var. platyphyllum) by the scooping method

  • Jeong, Mi Jin;Yong, Seong Hyeon;Kim, Do Hyeon;Park, Kwan Been;Kim, Hak Gon;Choi, Pil Son;Choi, Myung Suk
    • Journal of Plant Biotechnology
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    • 제49권3호
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    • pp.213-221
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    • 2022
  • Wild garlic (Allium victorialis var. platyphyllum, AVVP) is a nontimber forest product used as an edible and medicinal vegetable. AVVP is usually propagated form offspring bulbs but it takes a long time to harvest. Using tissue culture technology could overcome this problem. This study investigated the optimal conditions for shoot multiplication, root growth, and plant growth by scooping AVVP bulbs. AVVP bulbs harvested from Ulleung Island, Korea, the main producer of AVVP, were surface-sterilized and used for in vitro propagation. Shoot multiplication was performed by the scooping method. More than five multiple shoots were induced from scooped tissue in Quoirin and Lepoivre (QL) medium containing plant growth regulators (PGRs); the maximum number of multiple shoots were induced from scooped tissue in QL medium containing 0.45 μM thidiazuron (TDZ) after 16 weeks of culture. Roots were induced directly at the base of the shoots in all treatments. In vitro rooting depended on the type of PGRs, and the best root-inducing treatment was QL medium containing 9.84 μM indole-3-butyric acid (IBA). Plants with in vitro roots were transferred to pots containing artificial soil and successfully acclimatized for 4 weeks. The acclimatized plants showed a survival rate of 80% after 20 weeks and gradually promoted growth depending on the acclimatization period. The results of this study will be of great help to AVVP dissemination through sustainable mass propagation.

카사바 잎 절편 유래 체세포배 배양시 배지조성이 기내 식물체 재분화에 미치는 영향 (Effect of Medium Composition on in Vitro Shoot Regeneration from Leaves of Cassava (Manihot esculenta Crantz) Through Somatic Embryogenesis and Callus Induction)

  • 권영희;이정관;김희규;김경옥;김주형
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2020년도 춘계학술대회
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    • pp.19-19
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    • 2020
  • The Cassava(Manihot esculenta Crantz) is a tropical root crop, originally from Amazonia, that provides the staple food of an estimated 800 million people worldwide. It belongs to the family Euphorbiaceae which also includes rubber (Hevea brasiliensis) and castor bean (Ricinus communis). Among tropical crops, rice, sugarcane, maize and cassava are the most important sources of calories for human consumption. Problems in the propagation of cassava are virus diseases and low rates of seed germination. So we tried to optimize protocols for mass production of somatic embryo amenable to large-scale vegetative propagation of Cassava. After in vitro eight-week culture of leaves of Cassava, the medium which contained the 2,4-D, BAP and IBA showed the highest callus induction rate, embryogenesis callus formation rate and somatic embryo formation in Cassava culture. In the medium with GA3 and myo-inositol, shoots were most vigorously regenerated from somatic embryos of Cassava. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

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족제비고사리 전엽체의 기내배양 및 기외 포자체 형성에 미치는 제요인 (Several Factors Affecting on In vitro Culture of Prothallus and Ex Vitro Sporophyte Formation from Prothallus of Dyropteris varia (L.) O. Kuntze)

  • 정진아;이철희
    • 한국자원식물학회지
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    • 제19권2호
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    • pp.252-258
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    • 2006
  • 본 연구는 족제비고사리의 대량번식법을 개발하기 위하여 전엽체의 기내배양 및 기외 포자체 형성의 적정 조건을 구하고자 실시되었다. 기내에서 전엽체 증식은 MS배지의 무기물 조건에서 가장 활발하였고, $NH_4^+:NO_3^-$의 농도비가 10:50mM이고, 2%의 sucrose가 첨가된 배지에서 생장이 향상되었다. 전엽체 생장에 적합한 배지의 pH는 5.8이었으며, 적정 agar 농도는 $0.6{\sim}0.8%$로 조사되었다. 배지에 첨가된 생장조절물질(NA, IAA, 2,4-D, BAP, kinetin 및 2ip)은 대부분 농도가 높아질수록 전엽체의 생장을 더욱 저해하는 것으로 나타났다. 기내에서 증식된 전엽체를 토양 별로 기외이식한 결과, 상토 단용구에서 가장 많은 포자체가 형성되었으며, $GA_3\;100{\mu}M$에서 3시간 동안 침지처리 한 전엽체로부터 포자체의 형성이 현저히 촉진되었다.

Isolation and In Vitro Culture of Vascular Endothelial Cells from Mice

  • Choi, Shinkyu;Kim, Ji Aee;Kim, Kwan Chang;Suh, Suk Hyo
    • The Korean Journal of Physiology and Pharmacology
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    • 제19권1호
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    • pp.35-42
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    • 2015
  • In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

Optimization of in vitro seed germination of Taraxacum platycarpum

  • Lee, Jung-Hwan;Kim, Young-Kwan;Oh, Eun-Yi;Jung, Kuk-Young;Ko, Ki-Sung
    • 한국환경농학회지
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    • 제28권4호
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    • pp.403-408
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    • 2009
  • Dandelion (Taraxacum platycarpum) has been widely utilized for medicinal purposes. However, the dandelion seeds are relatively difficult to germinate under cultivation conditions, which hampers seedling propagation of dandelion plants and reduces the opportunity of usage of such a useful medicinal plant. Thus, in this study, in vitro conditions for the dandelion seed germination were optimized to enhance the germination rate. In seed washing steps, the sequential treatments with 20% of ethanol, 20% of NaOCl, and distilled water avoided microbial contamination with the highest in vitro germination rate (67.5%) from seeds sown in germination media. The media supplemented with 1.4 g/L of MS salts and 1% of sucrose significantly enhanced the germination rate compared to the media with 4.4 g/L of MS and 3% of sucrose. Sowing the seeds vertically in the optimized media supplement conditions, 1.4 g/L of MS salts and 1% of sucrose, gave the maximum in vitro germination rate (61%), which was almost three times higher than sowing seeds on a soil pot (23%). Our results indicate that the seed washing and sowing methods including germination medium supplements can be optimized to enhance in vitro seed germination of dandelion.