• Title/Summary/Keyword: in vitro propagation

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Effect of Medium Components and Culture Methods on Shoots Regeneration from Athyrium niponicum (개고사리의 기내 포자체 재생에 미치는 배지구성물질 및 배양방법의 영향)

  • Shin, So-Lim;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.24 no.2
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    • pp.113-120
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    • 2011
  • Present studies are carried out to find media components and culture methods for in vitro propagation of Athyrium niponicum and to establish the optimal economic masspropagation systems. Among pinnae, petiole and rhizome segments only rhizome segments produced young plants. Rhizome segments showed vigorous plant regeneration on 1/2MS medium and supplement to 1% sucrose and 50 $mg{\cdot}L^{-1}$ $NaH_2PO_4$ were promoted the plant regeneration from rhizome segments. Kinetin was better than BA for plant regeneration and combination with 2 ${\mu}M$ kinetin and 5 ${\mu}M$ IBA was most efficient for plant regeneration. Solid or liquid medium with or without 0.1% qactivated charcoal in modified 1/2MS medium (1% sucrose, 50 $mg{\cdot}L^{-1}$ $NaH_2PO_4$, 2 ${\mu}M$ kinetin, 5 ${\mu}M$ IBA, pH 5.8) were used to find the optimal culture methods. The plant regeneration from rhizome segments were most vigorous on solid medium without activated charcoal. The addition of activated charcoal were inhibited the plant regeneration from rhizome segments not only on solid medium but also liquid stationary or suspension culture.

In-Vitro Thrombosis Detection of Mechanical Valve using Artificial Neural Network (인공신경망을 이용한 기계식 판막의 생체외 모의 혈전현상 검출)

  • 이혁수;이상훈
    • Journal of Biomedical Engineering Research
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    • v.18 no.4
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    • pp.429-438
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    • 1997
  • Mechanical valve is one of the most widely used implantable artificial organs of which the reliability is so important that its failure means the death of patient. Therefore early noninvasive detection is essentially required, though mechanical valve failure with thrombosis is the most common. The objective of this paper is to detect the thrombosis formation by spectral analysis and neural network. Using microphone and amplifier, we measured the sound from the mechanical valve which is attached to the pneumatic ventricular assist device. The sound was sampled by A/D converter(DaqBook 100) and the periodogram is the main algorithm for obtaining spectrum. We made the thrombosis models using pellethane and silicon and they are thrombosis model on the valvular disk, around the sewing ring and fibrous tissue growth across the orifice of valve. The performance of the measurment system was tested firstly using 1 KHz sinusoidal wave. The measurement system detected well 1KHz spectrum as expected. The spectrum of normal and 5 kinds of thrombotic valve were obtained and primary and secondary peak appeared in each spectrum waveform. We find that the secondary peak changes according to the thrombosis model. So to distinguish the secondary peak of normal and thrombotic valve quantatively, 3 layer back propagation neural network, which contains 7, 000 input node, 20 hidden layer and 1 output was employed The trained neural network can distinguish normal and valve with more than 90% probability. As a conclusion, the noninvasive monitoring of implanted mechanical valve is possible by analysing the acoustical spectrum using neural network algorithm and this method will be applied to the performance evaluation of other implantable artificial organs.

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Establishment of optimal conditions for micropropagation by node culture and multiple shoots formation from sucker explants of thornless Blackberry (Rubus fruticosus L. cv. BB21) (가시없는 블랙베리(Rubus fruticosus L. cv. BB21)의 근맹아를 이용한 다경유도와 절간배양을 통한 식물체 증식조건의 확립)

  • Lee, Kang Seop;Kim, Hyo Jin;Park, Dae Hyun;Oh, Seung Cheol;Cho, Han Jig;Kim, Ee Youb
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.110-116
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    • 2018
  • This study was conducted to develop a simple, rapid, and reliable method for in vitro propagation of disease-free and true-to-type clones from sucker explants of thornless blackberry (Rubus fruticosus L. ${\times}$ R. parvifolius L.). To induce multiple shoots, the sucker explants were sterilized in 1% NaOCl solution, and then were aseptically cultured on the full and 1/2 MS solid medium supplemented with BAP (0.1, 0.5, 1.0, 2.0 mg/L). After six weeks of culture, the highest frequency (85.4%) of shoot formation from sucker explants was obtained on the full-strength MS medium with 1.0 mg/L BAP. Node explants obtained from multiple shoots were cultured on the various media of full- or half-strength of AD, B5, MS, SH, QL, WPM media, respectively. After 30 days of culture, plant growth was good on the half-AD, half-QL medium. After 90 days of culture, plant growth was good on the full MS and full SH medium. The survival rate of the plantlets after transfer to plastic pots containing soil mixture (sand: soil: vermiculite was 1:1:1, vol.) in the greenhouse was 98%. The results indicate that a multiple-shoot procedure can be applied for an efficient mass propagation of Rubus fruticosus L. ${\times}$ R. parvifolius L.

Plant Production from Desiccated Somatic Embryos of Acanthopanax chiisanensis (지리오가피 (Acanthopanax chiisanensis) 체세포배의 건조처리를 통한 식물체 증식)

  • Lee, Kang-Seop;Bang, Keuk-Soo;Choi, Yong-Eui;Ahn, Byung-Yong
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.381-385
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    • 2003
  • An efficient method of plant regeneration from Acanthopanax chiisanensis somatic embryos was developed. Cotyledonary somatic embryos were obtained in liquid Murashige and Skoog (MS) medium from embryogenic cell suspension cultures. They were desiccated for 0 to 72 hr and then cultured on MS medium containing NAA, BA, GA$_3$, (0-0.5mg/L). The highest multiple shoots formation (100%) was obtained from 72 hr desiccated somatic embryos on ifs medium with 0.5mg/L NAA+0.5mg/L BA or 0.5 mg/L NAA+0.5mg/L BA+0.5mg/L GA$_3$ after 6 weeks culture. Plant conversion from multiple shoots was not high. The highest plant conversion from multiple shoots was obtained on 1/3MS medium with 1.0mg/L GA$_3$. Converted plantlets were transferred to ex vitro condition and the highest survival rate (70%) of the plantlets was obtained on plastic pots containing vermiculite and sand. These results indicate that micropropagation procedure can be applied for an efficient mass propagation of Acanthopanax chiisanensis.

Mass Propagation of Hypolepis punctata (Thunb.) Mett. Using In Vitro Culture Techniques (조직배양을 이용한 점고사리의 대량증식 방법)

  • Park, Kyungtae;Jang, Bo Kook;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.57-57
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    • 2019
  • 본 연구는 관상 및 조경용으로 개발이 가능한 남방계 양치식물인 점고사리[Hypolepis punctata (Thunb.) Mett.]의 전엽체 증식 및 포자체 형성에 적합한 배양조건을 구명하고자 수행되었다. 무가온 온실에서 성숙한 포자엽을 채집한 후 포자를 기내에서 발아시켜 전엽체를 획득하였으며, 8주 간격으로 계대배양 하여 실험의 재료로 사용하였다. 배지종류에 따른 전엽체의 증식 및 형태형성을 확인하고자, 배양된 전엽체 0.3g을 메스로 잘게 다진 후, 농도를 1/4, 1/2, 1, 2배로 조절한 MS배지에 8주간 배양하였다. 이후 선발된 배지를 기준으로 sucrose, 활성탄, 질소급원의 농도를 조절하여 전엽체의 증식과 형태형성을 확인하였다. 그 결과, 1MS배지에서 전엽체의 생체중이 초기 접종량인 0.3g에 비해 10.7배 증가한 3.2g으로 가장 높은 증가율을 보였다. 형태관찰에서도 장정기의 형성이 관찰되었으며, 전엽체의 쿠션조직이 비교적 잘 발달하였다. 전엽체 증식에 가장 좋은 효과를 보인 1MS배지를 기준으로 sucrose의 농도를 0-4%로 달리하여 실험한 결과, 1%의 처리구에서 6.7g으로 가장 높은 생체중을 보였다. 활성탄의 농도를 0-0.8%로 첨가한 네 처리구 중에서는 0.8%의 처리구에서 14.2g으로 무처리구에 비해 생체중이 2배 이상 증가하였다. 전엽체의 형태 또한 정상적인 발달을 보였다. 질소급원의 비율을 30-120mM로 조절한 배지에서는 60mM의 처리구에서 4.9g으로 가장 높은 생체중을보였다. 이후 포자체 형성을 위한 최적의 토양조건을 구명하고자, 원예상토, 피트모스, 펄라이트 및 마사토의 비율을 달리하여 5종류의 배양토를 조성하였다. 혼합된 토양은 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진 하였으며, 배양된 전엽체 1g을 증류수와 함께 10초간 분쇄한 다음 준비된 토양표면에 분주하여 재배하였다. 12주간의 재배 결과, 원예상토를 단용한 토양에서 포자체의 수가 포트당 250.0개로 가장 많이 형성되었으며, 포자체의 생육 또한 다른 처리구에 비해 우수한 결과를 보였다. 한편 피트모스가 혼합된 토양에서는 포자체가 형성되지 않았다. 따라서 점고사리의 전엽체 대량증식에 적합한 배지는 sucrose 1%와 질소급원의 농도를 60mM, 활성탄을 0.8% 첨가한 1MS배지로 판단되며, 포자체 대량생산을 위해서는 원예상토를 단용한 토양이 적합하다고 판단된다.

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Stability of Human Centromeric Alphoid DNA Repeat during Propagation in Recombination-Deficient Yeast Strains (효모의 재조합 변이주를 이용한 인간 Centromeric Alphoid DNA Repeat의 안정성에 관한 연구)

  • Kim, Kwang-Sup;Shin, Young-Sun;Lee, Sang-Yeop;Ahn, Eun-Kyung;Do, Eun-Ju;Park, In-Ho;Leem, Sun-Hee;SunWoo, Yang-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.243-249
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    • 2007
  • The centromere is a highly differentiated structure of the chromosome that fulfills a multitude of essential mitotic and meiotic functions. Alphoid DNA (${\alpha}$-satellite) is the most abundant family of repeated DNA found at the centromere of all human chromosomes, and chromosomes of primates in general. The most important parts in the development of Human Artificial Chromosomes (HACs), are the isolation and maintenance of stability of centromeric region. For isolation of this region, we could use the targeting hook with alphoid DNA repeat and cloned by Transformation-Associated Recombination (TAR) cloning technique in yeast Saccharomyces cerevisiae. The method includes rolling-circle amplification (RCA) of repeats in vitro to 5 kb-length and elongation of the RCA products by homologous recombination in yeast. Four types of $35\;kb{\sim}50\;kb$ of centromeric DNA repeat arrays (2, 4, 5, 6 mer) are used to examine the stability of repeats in homologous recombination mutant strains (rad51, rad52, and rad54). Following the transformation into wild type, rad51 and rad54 mutant strains, there were frequent changes in inserted size. A rad52 mutant strain showed extremely low transformation frequency, but increased stability of centromeric DNA repeat arrays at least 3 times higher than other strains. Based on these results, the incidence of large mutations could be reduced using a rad52 mutant strain in maintenance of centromeric DNA repeat arrays. This genetic method may use more general application in the maintenance of tandem repeats in construction of HAC.

In Vitro Plant Regeneration for Mass Propagation of Epimedium koreanum Nakai (삼지구엽초의 다량번식을 위한 기내 식물체 분화)

  • Han, Young-Hee;Choi, Byoung-Ryourl;Soh, Ho-Seob;Lee, Seong-Jae;Choi, Young-Jin;Kim, Se-Young
    • Horticultural Science & Technology
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    • v.18 no.6
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    • pp.834-838
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    • 2000
  • As an endeavor to establish a micropropagation system for Epimedium koreanum Nakai., this study was carried out to define methods to disinfect its explants and media for callus induction, proliferation and plant regeneration. The lowest infection rates by fungi or bacteria on apical and axillary bud explants of rhizome were observed when they were immerged in 0.3% NaOCl solution for 20 min after soaked in 0.1% $AgNO_3$ solution for 30 min, but leaf explants were seldom infected with fungi or bacteria by this disinfectant method. The highest rate of plantlet formation was obtained from the explants disinfected in 0.3% NaOCl solution for 20 min after soaked in 0.1% $AgNO_3$ solution for 60 min for tip buds and in 0.1 % $AgNO_3$ solution for 30 min for axillary buds of rhizome. Induction rate of callus was the highest from the explants disinfectd in 0.3% NaOCl solution for 20 min after soaked in 0.2% $AgNO_3$ solution for 15 min. Callus growth was proper in a modified 1/2 MS medium including half strength of $NH_4NO_3$ with $0.02-0.2mg{\cdot}L^{-1}$ BA and $2.0mg{\cdot}L^{-1}$ NAA. Low rate of plantlet regeneration was obtained in 1/2 UM or 1/2 White medium with $2.0mg{\cdot}L^{-1}$ BA and $0.2mg{\cdot}L^{-1}$ AA.

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The effect of reduced thickness in different regions on the fracture resistance of monolithic zirconia crowns (다양한 부위에서의 감소된 두께가 지르코니아 크라운의 파절 저항에 미치는 영향)

  • Abukabbos, Layla;Park, Je Uk;Lee, Wonsup
    • The Journal of Korean Academy of Prosthodontics
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    • v.60 no.2
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    • pp.135-142
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    • 2022
  • Purpose. This study aims to evaluate the combined effect of reduced thickness in different regions on the fracture resistance of monolithic zirconia crowns. Materials and methods. Seven nickel-chromium dies were generated from a 3D model of mandibular first molar using the digital scanner with the following geometries: 1.5 mm occlusal reduction, 1.0 mm deep chamfer. Based on the abutment model, Zirconia blocks (Luxen Zirconia) were selected to fabricate Sixty-three zirconia crowns with occlusal thicknesses of 0.3 mm, 0.5 mm, and 1.5 mm, and different axial thicknesses of 0.3 mm, 0.5 mm, and 1.0 mm. All crowns were cemented by resin cement. Next, the crowns were subjected to load-to-fracture test until fracture using an electronic universal testing machine. In addition, fracture patterns were observed with a scanning electron microscope (SEM). Two-way ANOVA and the Tuckey HSD test for post hoc analysis were used for statistical analysis (P < .05). Results. The mean values of fracture resistancerecorded was higher than the average biting force in the posterior region. The two-way ANOVA showed that the occlusal and axial thickness affected the fracture resistance significantly (P < .05). However, the effect of axial thickness on fracture resistance did not show a statistical difference when thicker than 0.5 mm. The observed failure modes were partial or complete fracture depending on the severity of crack propagation. Conclusion. Within the limitations of the present study, the CAD-CAM monolithic zirconia crown with extremely reduced thickness showed adequate fracture resistance to withstand occlusal load in molar regions. In addition, both occlusal and axial thickness affected the fracture resistance of the zirconia crown and showed different results as combined.

High frequency somatic embryogenesis through leaf explant-derived callus culture in Muscari armeniacum cv. 'Early Giant' (무스카리 'Early Giant' 잎 절편 유래 캘러스 배양을 통한 고빈도 체세포배 발생)

  • Lee, Hyang-Bun;Jeon, Su-Min;Chung, Mi-Young;Han, Jeung-Sul;Kim, Chang-Kil;Lim, Ki-Byung;Chung, Jae-Dong
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.69-74
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    • 2012
  • Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.

Studies on the Mass-Propagation of Potato (Solanum tuberosum L.) by In-vitro Culture. I. Effects of Phytohormons on the Callus Induction and the Organ Differentiation from Potato Meristem tip (기내배양에 의한 감자(Solanum tuberosum L.)의 대량번식(大量繁殖)에 관한 연구(硏究) I. 몇가지 생장조절물질(生長調節物質)이 생장점(生長點)으로부터의 Callus 및 기관분화(器官分化)에 미치는 영향(影響))

  • Kim, Choong Soo;Jo, Jae Seong;Choi, Chang Yeol
    • Korean Journal of Agricultural Science
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    • v.7 no.2
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    • pp.59-64
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    • 1980
  • These experiments were carried out to define the effects of 2.4-D, NAA and Benzyladenine on the differentiation and growth of the organs and the induction of callus from the potato meristem. The results were summarized as follows ; 1. The differentiation and growth of the shoots from the potato meristem was promoted in increased concentration of Benzyladenine but the callus was not induced on the M.S. medium containing Benzyladenine. 2. On the M.S. medium containing NAA 0.5 mg/l or higher concentration of NAA, the shoots were not initiated but the callus were induced from potato meristem. The growth of callus was promoted in increased concentration of NAA. 3. The roots were initiated from 50% of potato meristems planted on the M.S. medium containing more than 0.1 mg/l of NAA but the roots were pot initiated on the medium containing 2.4-D. 4. The shoot growth was significantly increased by increasing of 2.4-D concentration up to 0.1 mg/l, but the shoots were not initiated on the medium containing 2.4-D more than 1.0 mg/l. 5. For the induction and growth of the callus from potato meristem, NAA was more effective than 2.4-D and the most effective medium was M.S. medium supplemented with 2.0 mg/l of NAA. 6. The M.S. mediums supplemented with BA 2.0 mg/l and NAA 0.1 mg/l or BA 1.0 mg/l and 2.4-D 0.1 mg/l showed good results for entire plant regeneration from potato meristem.

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