• Title/Summary/Keyword: regeneration medium

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Effects of Plant Growth Regulators on Haploid Production of Wheat (밀 반수체 생산에서 식물생장조절제의 영향)

  • Kim, Young Jin;Lee, Kwang Won;Kim, Hag Sin;Cheong, Young Keun;Park, Kwang Geun;Kim, Kee Jong
    • Korean Journal of Breeding Science
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    • v.43 no.1
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    • pp.50-55
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    • 2011
  • The introduction of doubled haploid (DH) approach into breeding programs has reduced the times and population sizes required for the production of pure lines. We carried out the experiment for development on effective method of producing haploid in wheat. Emasculated spikelets of wheat were pollinated with maize pollen and cultured in the solution containing 40 g/L sucrose and 2,4-D, NAA, 2,4,5-T and dicamba 24 h after pollination, and then incubated until embryo rescue. Fourteen to sixteen days after pollination, the embryos are excised and cultured in half-strength MS basal medium supplemented with 20 g/L sucrose and 1 mg/L NAA. The type of plant growth regulators was found to be most significant in production of haploid plants. The application of synthetic auxins to pollinated florets, stimulates haploid embryo development to a stage where the embryos can be rescued onto nutrient media. The percentage of seeded florets was significantly affected by 100 mg/L 2,4-D, 150 mg/L 2,4,5-T and 50 mg/L dicamba. The percentage of embryos formed was significantly increased by treatment with 2,4-D and 2,4,5-T at 100 mg/L, and dicamba at 50 mg/L, but the treatments with 150 mg/L 2,4-D inhibited embryo development and plant regeneration. The optimum application time of plant growth regulators was 24 hrs after pollination.

Effect of Developmental Stage of Pollen Grain and Temperature Pretreatment on Anther Culturein Lilium Asiatic Hybrid 'Dreamland' (Lilium Asiatic hybrid 'Dreamland'의 약배양에 미치는 화분 발육단계와 온도 전처리의 영향)

  • Park, Young Ae;Hwang, Yoon Jung;Park, In Sook;Suh, Dong Hee;Jeon, Su Min;Yeo, Kum-Bok;Lee, Ga Young;Chung, Jae-Dong;Lim, Ki Byung
    • FLOWER RESEARCH JOURNAL
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    • v.16 no.1
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    • pp.12-16
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    • 2008
  • This study was carried out to produce haploid plants to verify a systematic breeding program and genetic analysis. Effect of developmental stage of pollen grains and pre-treatment temperature on production of haploid plants was investigated. Microscopic investigation of the explants (Lilium Asiatic hybrid 'reamland' revealed that the length of flower bud at 23.0-24.9, 25.0-26.9, and 27.0-28.9 mm long coincided with tetrad, uninucleate, and binucleate, respectively. When the efficiency of the anther culture from microgametogenetic stages was tested, late uninucleate to early binucleate stage, having the length of 23.0 to 28.0 mm long flower bud, was the best. The frequencies of the callus induction and plant regeneration from the stage mentioned above were 17.8 and 6.7%, respectively. When calli were cultured on the MS medium containing picloram and zeatin at $25^{\circ}C$, shoots were obtained. Roots of regenerated plantlets were confirmed as haploid through an microscopic observation.

Isolation and identification of goose skeletal muscle satellite cells and preliminary study on the function of C1q and tumor necrosis factor-related protein 3 gene

  • Wang, Han;He, Ke;Zeng, Xuehua;Zhou, Xiaolong;Yan, Feifei;Yang, Songbai;Zhao, Ayong
    • Animal Bioscience
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    • v.34 no.6
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    • pp.1078-1087
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    • 2021
  • Objective: Skeletal muscle satellite cells (SMSCs) are significant for the growth, regeneration, and maintenance of skeletal muscle after birth. However, currently, few studies have been performed on the isolation, culture and inducing differentiation of goose muscle satellite cells. Previous studies have shown that C1q and tumor necrosis factor-related protein 3 (CTRP3) participated in the process of muscle growth and development, but its role in the goose skeletal muscle development is not yet clear. This study aimed to isolate, culture, and identify the goose SMSCs in vitro. Additionally, to explore the function of CTRP3 in goose SMSCs. Methods: Goose SMSCs were isolated using 0.25% trypsin from leg muscle (LM) of 15 to 20 day fertilized goose eggs. Cell differentiation was induced by transferring the cells to differentiation medium with 2% horse serum and 1% penicillin streptomycin. Immunofluorescence staining of Desmin and Pax7 was used to identify goose SMSCs. Quantitative realtime polymerase chain reaction and western blot were applied to explore developmental expression profile of CTRP3 in LM and the regulation of CTRP3 on myosin heavy chains (MyHC), myogenin (MyoG) expression and Notch signaling pathway related genes expression. Results: The goose SMSCs were successfully isolated and cultured. The expression of Pax7 and Desmin were observed in the isolated cells. The expression of CTRP3 decreased significantly during leg muscle development. Overexpression of CTRP3 could enhance the expression of two myogenic differentiation marker genes, MyHC and MyoG. But knockdown of CTRP3 suppressed their expression. Furthermore, CTRP3 could repress the mRNA level of Notch signaling pathway-related genes, notch receptor 1, notch receptor 2 and hairy/enhancer-of-split related with YRPW motif 1, which previously showed a negative regulation in myoblast differentiation. Conclusion: These findings provide a useful cell model for the future research on goose muscle development and suggest that CTRP3 may play an essential role in skeletal muscle growth of goose.

A Comparative Case Study on Urban Strategy for Flexible Smart Shrinking City (유연한 스마트 축소도시를 위한 국내·외 도시전략 비교 연구)

  • Choi, Wan-Young;Lee, Heewon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.8
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    • pp.634-641
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    • 2019
  • The world's population reached 7.7 billion in 2019. Despite this trend, not all cities are growing. Most of the growing population is driven to large cities for convenience and jobs. This phenomenon has led to a decrease in the population of small and medium-sized cities, and in certain cities, a decrease in employment as well as recession have resulted. Korea is no exception. The nation's population is concentrated in the metropolitan cities. To solve these problems, researchers started to study how to deal with the contraction of a city, i.e., a shrinking city, rather than focusing on only the growth of a city. In this paper, demographically outstanding declined domestic cities were selected as shrinking cities and their characteristics were analyzed. The concept of smart shrinking cities was then defined for the selected cities, which were chosen as a good case in overseas countries to solve the shrinking city problem. Through this process, the strategic differences were compared between domestic and oversea cases. As a result, a modified strategy for the smart shrinking city concept, FSSC (flexible smart shrinking city), is proposed as an alternative strategy that can save resources and cost.

Somatic Embryogenesis and Plant Regeneration with Embryogenic Tissue Lines in Pinus densiflora (소나무 배발생조직 라인 별 체세포배 유도 및 식물체 재분화)

  • Kim, Yong Wook;Shin, Han Na;Moon, Heung Kyu
    • Journal of Korean Society of Forest Science
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    • v.100 no.3
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    • pp.449-454
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    • 2011
  • This experiment was conducted to evaluate growth rate among 15 embryogenic tissue lines (ETLs), comparison of maturation on somatic embryos (SEs) with 13 ETLs and efficiency with various concentrations of gelrite on SEs germination in Japanese red pine (Pinus densiflora). In comparison of ETLs growth rate (folds) with 15 lines, the 05-4 line (5.3 folds) showed the highest rate, on the other hand, the lowest one was recorded in the line of 05-37 (1.4 folds). The 13 ETLs were tested for the extent of SEs production. The best production was recorded in the line of 05-4 (39.8/90 mg F.W.). However, most of ETLs, except 5 lines (05-4, 12, 21, 29 and 37), did not produce SEs at all, therefore, big differences in the ability of SEs production existed among the ETLs tested. Effects of various gelrite concentrations for SEs germination with 3 ETLs were also compared. The highest result was obtained from 0.2% gelrite concentration with 05-4 line (47.3%), there was a inclination that the rate of germination was gradually declined over 0.2% gelrite concentration with the 05-4 and 29 lines. respectively. In contrast, in the line of 05-37, no SEs germination occurred on medium with 0.1 or 0.2% gelrite. In conclusion, the growth rate, SEs production and germination frequency were appeared to deeply depended on the ETLs.

A simple mid-term preservation method (SMPM) of plant callus under low temperature conditions (저온 보존을 이용한 간편 중기 식물캘러스 저장법)

  • Park, Sung-Chul;Park, Su Hyun;Kim, Soyoung;Jeong, Yu Jeong;Kim, Cha Young;Jeong, Jae Cheol
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.187-192
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    • 2022
  • The repeated monthly or weekly subculture of plant callus is labor intensive and increases the risk of somaclonal variation from the parental callus line. The most effective method for preserving plant callus is cryopreservation, which involves storage in liquid nitrogen. However, this method cannot be applied to the callus of different plant species in the same manner, so it is difficult to develop a standardized cryopreservation method. In addition, the survival rate of the frozen callus after thawing and the regeneration rate after survival are uncertain. Therefore, it is necessary to develop a method to extend the subculture interval of plant callus in an active state. In this study, active plant calli of various species without freezing was incubated at 15℃ for 4 to 12 weeks without subculture. After 12 weeks, 8 lines of plant callus grew less than 2-fold when cultured at 25℃, but at least 2 times as much when cultured at 15℃. Moreover, total antioxidant activity did not differ significantly between plant callus recovered at 25℃ after culturing at 15℃ or at 25℃. These results show that the subculture interval can be extended at a temperature of 15℃ without need for modified medium composition or additional processes. In addition, positive results in all calli of several plant species are expected to reduce labor as well as somaclonal variation by increasing the subculture.

Carbofuran Cytotoxicity on Rat Fibroblast Cells and Compensation Effects of Phenobarbital Sodium and 3-Methycholanthrene on Its Toxicity (살충제 Carbofuran이 쥐의 NIH3T3 섬유모세포에 끼치는 독성 및 Phenobarbital Sodium과 3-Methylcholanthrene에 의한 보상효과)

  • Han, Seong-Soo;Rim, Yo-Sup
    • Korean Journal of Environmental Agriculture
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    • v.16 no.2
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    • pp.149-155
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    • 1997
  • This study was carried out to investigate the effects of phenobarbital sodium(PB) and 3-methylcholanthrene(3-MC) on carbofuran cytotoxicity and to develop antitoxic agents based on the effectivness. Experimental groups for carbofuran cytotoxicity were divided into five groups ; medium alone and four treatments of carbofuran (1, 25, 50 and $100{\mu}M)$, and those for compensation effects were divided into six groups ; medium alone, $IC_{50}$ carbofuran and four combinations of carbofuran and PB or 3-MC($IC_{50}$ carbofuran plus 1, 25, 50, $100{\mu}M$ of PB and 3-MC, respectively). After incubation for 48 hrs under the same conditions, MTT(Tetrazolium MTT), NR(Neutral red) and SRB(Sulforhodamine B protein) assay were performed. Fifty percentage inhibition of MTT, NR, and SRB against carbofuran in rat fibroblast cell were 60.7, 82.5 and $87.0{\mu}M$, respectively. At the combination treatments of $IC_{50}$ of carbofuran and $100{\mu}M$ of PB, the significant compensation effects were observed from the results of MTT and NR but not from that of SRB absorbance. And at the combination treatments of $IC_{50}$ of carbofuran and 3-MC, the relatively significant compensation effects were found at $50{\mu}M$ 3-MC from the results of MTT and at $100{\mu}M$ 3-MC from that of NR and SRB absorbances, respectively. From the results of light microscopy, combination treatments of $carbofuran(IC_{50})$ and PB or 3-MC showed good regeneration in carbofuran toxicity of rat fibroblast cells. These results suggest that PB or 3-MC can compensate the cytotoxity of carbofuran insecticide in rat NIH3T3 fibroblast cells.

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A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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Identification of Matrix Mineralization-Related Genes in Human Periodontal Ligament Cells Using cDNA Microarray (cDNA microarray에 의한 치주인대세포의 광물화 결절형성에 관여하는 유전자들의 분석)

  • Shin, Jae-Hee;Park, Jin-Woo;Yeo, Shin-Il;Noh, Woo-Chang;Kim, Moon-Kyu;Kim, Jung-Chul;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.447-463
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    • 2007
  • Periodontal ligament (PDL) cells have been known as multipotential cells, and as playing an important rolesin periodontal regeneration. The PDL cells are composed of heterogeneous cell populations which have the capacity to differentiate into either cementoblasts or osteoblasts, depending on needs and conditions. Therefore, PDL cells have the capacity to produce mineralized nodules in vitro in mineralization medium which include ascorbic acid, ${\beta}$-glycerophosphate and dexamethasone. In spite of these well-known osteoblast like properties of PDL cells, very little is known about the molecules involved in the formation of the mineralized nodules in the PDL cells. In the present study, we analysed gene-expression profiles during the mineralization process of cultured PDL cells by means of a cDNA microarray consisting of 3063 genes. Nodules of mineralized matrix were strongly stained with alizarin red S on the PDL cells cultured in the media with mineralization supplements. Among 3,063 genes analyzed, 35 were up-regulated more than two-fold at one or more time points in cells that developed matrix mineralization nodules, and 38 were down-regulated to less than half their normal level of expression. In accord with the morphological change we observed, several genes related to calcium-related or mineral metabolism were induced in PDL cells during osteogenesis, such as IGF-II and IGFBP-2. Proteogycan 1, fibulin-5, keratin 5, ,${\beta}$-actin, ${\alpha}$-smooth muscle actin and capping protein, and cytoskeleton and extracellular matrix proteins were up-regulated during mineralization. Several genes encoding proteins related to apoptosis weredifferentially expressed in PDL cells cultured in the medium containing mineralization supplements. Dkk-I and Nip3, which are apoptosis-inducing agents, were up-regulated, and Btf and TAXlBP1, which have an anti-apoptosis activity, were down-regulated during mineralization. Also periostin and S100 calciumbinding protein A4 were down-regulated during mineralization.

In vitro Culture and Acclimatization of Regenerated Plants of Liliem cernum $K_{OMAROV}$ (솔나리 기내배양 및 재분화 식물체의 토양순화)

  • Kim, H.K.;Lim, Jung-Dae;Hyun, Tae-Kyung;Lee, Hyeon-Yong;Lee, Jin-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.4
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    • pp.310-317
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    • 2001
  • The regenerated-bulblets placed in liquid free media resulted in good formation of roots and bulblets. On 1/4 MS free medium, roots and bulblets were predominantly induced. The 1/4 MS liquid medium supplemented with plant growth regulators was the best suitable condition for elongation of leaves and roots. Somatic embryos were frequently developed from embryogenic callus in liquid media with 2,4-D 1mg/ l . On free liquid media, the viability of callus reduced. As the salt strength of MS media reduces, the viability of callus reduced significantly. However, Leaves were induced from several callus clumps. When leaves, roots and bulb-scale segments were placed on MS media containing NAA 1mg/ l or 2,4-D 1mg/ l and various sucrose concentration, the best result about the differentiation, growth of leaf and the differentiation of leaf was obtained on MS media added 1.5% sucrose and 2,4-D 1mg/ l, 3% sucrose and NAA 1mg/ l, and 1.5% sucrose and NAA 1mg/ l, respectively. Also the better result differentiation, growth of root and differentiation of bulb was obtained on MS media with 6% sucrose and NAA 1mg/ l. Spermidine promoted the growth of leaf and the differentiation of bulb. However, spermine promoted the differentiation of leaf, the differentiation and the growth of root in MS solid media. On the MS liquid media, both spermine and spermidine stimulated organogenesis from bulb-scale segments. Regenerated plantlets were acclimatizated and grown in greenhouse in vermiculite + perlite (1 : 1 by volume) well. The optimal soil condition of rooting for plantlets regenerated was in peat moss.

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