• Title/Summary/Keyword: Culture efficiency

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Development of a Reliable Technique to Eliminate Sweet potato leaf curl virus through Meristem Tip Culture Combined with Therapy of Infected Ipomoea Species

  • Cheong, Eun-Ju;Hurtt, Suzanne;Salih, Sarbagh;Li, Ruhui
    • Korean Journal of Plant Resources
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    • v.23 no.3
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    • pp.233-241
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    • 2010
  • In vitro elimination of Sweet potato leaf curl virus (SPLCV) from infected sweet potato is difficult due to low number of virus-free plants obtained from meristem tip culture and long growth period required for the virus detection. In this study, efficient production of the SPLCV-free sweet potato by in vitro therapy coupled with a PCR assay for virus detection was investigated. Infected shoots cultured on Murashige and Skoog medium were treated at three different temperatures for 7 weeks followed by meristem tip culture on the medium with or without ribavirin at 50 mg/L. The regenerated plantlets were tested for virus infection by a PCR assay. The results showed that the both heat- and cold-treatments, and addition of the ribavirin did not have significant effect on efficiency of the virus elimination. The meristem size, however, greatly affected the survival rate. Meristems sized over 0.4 mm survived better than smaller ones (0.2-0.3 mm). The PCR assay was approved to be a rapid, sensitive and reliable for the SPLCV detection in regenerated plantlets. Therefore, combination of cultivating meristem tips sized 0.4-0.5 mm on the medium at $22^{\circ}C$ without ribavirin and detection of SPLCV in the regenerated plantlets by the PCR assay was an efficient system for the SPLCV elimination from infected sweet potato.

High efficiency of homemade culture medium supplemented with GDF9-β in human oocytes for rescue in vitro maturation

  • Mohsenzadeh, Mehdi;Khalili, Mohammad Ali;Anbari, Fatemeh;Vatanparast, Mahboubeh
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.2
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    • pp.149-158
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    • 2022
  • Objective: Optimizing culture media for the incubation of immature oocytes is a vital strategy to increase the oocyte maturation rate during in vitro maturation (IVM) programs. This study evaluated the IVM and fertilization rates of human germinal vesicle (GV) and metaphase I (MI) oocytes using two different maturation media (commercial and homemade) with or without growth differentiation factor 9-β (GDF9-β). supplementation. Methods: Immature oocytes from intracytoplasmic sperm injection (ICSI) cycles were collected and assigned to one of two IVM culture media (commercial or homemade; cleavage-stage base). After maturation, MII oocytes were examined under an inverted microscope for the presence of the polar body, zona pellucida (ZP) birefringence, and meiotic spindle (MS) visualization after maturation in four conditions (commercial or homemade medium, with or without GDF9-β. ICSI was done for matured oocytes, and fertilization was confirmed by the visualization of two distinct pronuclei and two polar bodies. Results: No significant differences were found between the two culture media in terms of the time and rate of oocyte maturation or the rate of fertilization (p>0.05). Growth factor supplementation increased the 24-hour maturation rate for both GV and MI oocytes only in homemade medium. The maturation rate after 24 hours was higher for MI oocytes (p<0.05). Similar results were observed for MS visualization and ZP structure in both types of media (p>0.05). Conclusion: Higher rates of oocyte maturation and fertilization were observed after application of homemade medium supplemented with GDF9-β. Therefore, this combination may be recommended as an alternative for clinical IVM programs.

Search for the Direction of the Digital Cultural Psychology through Environmental Changes in Plays (놀이환경 변화를 통해 본 디지털문화심리학의 방향 모색)

  • Jang Ju Lee
    • Korean Journal of Culture and Social Issue
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    • v.17 no.1
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    • pp.131-154
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    • 2011
  • Rapid social changes due to digitization have created new psychological challenges of adapting and coping. Plays are in the center of these changes. Plays used to be collateral activities of labor in the traditional society and industrial society, but they have become key activities embracing economy, society, culture in a digital society that has been maximized productivity and efficiency. Still, the theoretical approach of plays is based on the industrial societies. Analyzing these points, the psychological impacts of players using multimedia, social networking, and digital games utilizing both of them was dealt in this article. I dealt with multimedia, social networking, and digital games as key characteristic of digital culture. The difference between traditional psychology and Digital Cultural Psychology was dealt in the perspective of self-concept, information processing, and sex role. Then the conclusion on the future directions of Digital Cultural Psychology and its limitations will be followed.

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Stable Expression and Efficient Secretion of hSCF and hINFγ Protein using Binary Vectors in Chlorella vulgaris (클로렐라에서 바이너리 벡터를 이용한 hSCF와 hINFγ 단백질의 안정적인 발현과 효율적인 분비)

  • Yu Jeong Jeong;Hee Gyung Min;Won Young Lee;Sung Chun Kim
    • Journal of Marine Bioscience and Biotechnology
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    • v.16 no.1
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    • pp.45-54
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    • 2024
  • Microalgae have great potential in the biomedical and pharmaceutical industries as a new type of bioreactor that can produce proteins for specific purposes, including recombinant proteins, pharmaceuticals, and industrial enzymes. Despite the production advantages and importance of microalgae-based expression systems, studies on secretion efficiency are limited. In this study, for stable expression and efficient secretion of the heterologous protein (human SCF and human INFγ) in Chlorella vulgaris, we constructed SP:hSCF:His and SP:hINFγ:His plant binary vectors using the signal peptide (SP) of Chlamydomonas reinhardtii, and we obtained stable transformants through the effective agrobacterium-mediated transformation of these vectors. Transformants with accurately inserted hSCF and hINFγ demonstrated stably increased mRNA and protein expression using RT-PCR and western blotting under the same culture conditions. Following the analysis of the proteins secreted into the culture medium using ELISA, it was confirmed that hINFγ was effectively produced in the transformed C. vulgaris culture medium. The overall findings indicate that the combination of heterologous protein and SP may be crucial for ensuring the expression and secretion of recombinant proteins in Chlorella culture systems.

Several Factors Affecting Transformation Efficiency of tall Fescue (톨페스큐의 효율적인 형질전환을 위한 몇 가지 요인의 영향)

  • 김진수;이상훈;이병현
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.3
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    • pp.237-242
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    • 2004
  • A system for the production of transgenic plants has been developed for tall fescue (Festuca arundinacea Schreb.) via Agrobacterium-mediated transformation of mature seed-derived embryogenic callus. Seed-derived calli were infected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding the hygromycin phosphotransferase (HPT), neomycin phosphotransferase II (NPTII) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. Inclusion of $200\mu\textrm{M}$ acetosyringone (AS) in inoculation and co-culture media lead to a increase in stable transformation efficiency. Transformation efficiency was increased when embryogenic calli were co-cultured for 5 days on the co-culture medium. The highest transformation efficiency was obtained when embryogenic calli were inoculated with Agyobacterium in the presence of 0.1% Tween20 and $200\mu\textrm{M}$ AS. Hygromycin resistant calli were developed into complete plants via somatic embryogenesis. GUS histochemical assay and Southern blot analysis of transgenic plants demonstrated that transgenes were successfully integrated into the genome of tall fescue.

Factors Influencing Agrobacterium-Mediated Transformation Efficiency in Perennial Ryegrass (Agrobacterium 매개에 의한 페레니얼 라이그라스의 형질전환에 영향을 미치는 요인)

  • Lee, Ki-Won;Kim, Ki-Yong;Lee, Joung-Kyong;Park, Hyung-Soo;Kim, Kyung-Hee;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.3
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    • pp.165-170
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    • 2009
  • A system for the production of transgenic plants has been developed for perennial ryegrass (Lolium perenne L.) via Agrobacterium-mediated transformation. Included in this study were two factors which may affect the gene transfer efficiency: concentrations of acetosyringone (AS, 0 to 300 ${\mu}M$), and co-culture period (1 to 7 days). Both factors were very important to achieve high efficiency gene transformation in the perennial ryegrass. The highest transformation efficiency was obtained when embryogenic calli were inoculated with Agrobacterium in the presence of 100 ${\mu}M$ AS with the culture medium for 5 days. Phosphinothricin resistant calli were developed with into complete plants. GUS histochemical assay, polymerase chain reaction (PCR) and Northern blot analysis of transgenic plants demonstrated that transgenes were integrated into the genome of perennial ryegrass. Using this protocol, it was possible to obtain transformants efficiently for further study.

Isolation and Characterization of Bacteria Capable of Degrading Bisphenol A (Bisphenol A 분해세균의 분리 및 특성)

  • 김희식;이영기;이완석;박찬선;윤병대;오희목
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.189-196
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    • 2001
  • Eighty-seven microbial strains capable of growing on bisphenol A (BPA) as a sole carbon source were isolated from soils, waste waters and sludges. Among them, three bacterial strains were finally selected as potential decomposers through measuring BPA-degradation efficiency by HPLC analysis. Two of these bacterial strains were identified as Serratia marcescens 1901 and S. marcescens 1902, and another was Pseudomonas putida 1401 by 16S rDNA partial sequences and based on morphological and physiological properties. They showed higher cell growth and BPA degradation in PAV (PAS medium containing vitamin mixtures) than in PAS medium. The degradation efficiencies of these bacterial strains were within a range of 20-40% in the PAV containing 500 mg/1 or 100 mg/l of BPA fur 3 days. S. marcescens 1901 showed higher degradation efficiency at 100 mg/1 of BPA than those of other selected strains, while S. marcescens 1902 and P. putida 1401 degraded a high concentration of BPA (500 mg/l) with a degradation efficiency of 40% for 3 days. The BPA degradation using a mixed culture of three selected strains showed the similar level of dog-radation efficiency with that using a pure culture.

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Analysis of a Sulfur-oxidizing Perchlorate-degrading Microbial Community (황 산화를 통해 퍼클로레이트를 분해하는 미생물 군집 분석)

  • Kim, Young-Hwa;Han, Kyoung-Rim;Hwang, Heejae;Kwon, Hyukjun;Kim, Yerim;Kim, Kwonwoo;Kim, Heejoo;Son, Myunghwa;Choi, Young-Ik;Sung, Nak-Chang;Ahn, Yeonghee
    • Journal of Life Science
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    • v.26 no.1
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    • pp.68-74
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    • 2016
  • Perchlorate (ClO4) is an emerging pollutant detected in surface water, soil, and groundwater. Previous studies provided experimental evidence of autotrophic ClO4 removal with elemental sulfur (S0) particles and activated sludge, which are inexpensive and easily available, respectively. In addition, ClO4 removal efficiency was shown to increase when an enrichment culture was used as an inoculum instead of activated sludge. PCR-DGGE was employed in the present study to investigate the microbial community in the enrichment culture that removed ClO4 autotrophically. Microorganisms in the enrichment culture showed 99.71% or more ClO4 removal efficiency after a 7-day incubation when the initial concentration was approximately 120 mg ClO4/l. Genomic DNA was isolated from the enriched culture and its inoculum (activated sludge), and used for PCR-DGGE analysis of 16S rRNA genes. Microbial compositions of the enrichment culture and the activated sludge were different, as determined by their different DGGE profiles. The difference in DGGE banding patterns suggests that environmental conditions of the enrichment culture caused a change in the microbial community composition of the inoculated activated sludge. Dominant DGGE bands in the enrichment culture sample were affiliated with the classes β-Proteobacteria, Bacteroidetes, and Spirochaetes. Further investigation is warranted to reveal the metabolic roles of the dominant populations in the ClO4 degradation process, along with their isolation.

Effect of Morphological Factors, Antibiotics and Agrobacterium Co-cultivation in the Efficiency of Somatic Embryogenesis of Eggplant (Solanum melongena L.)

  • Claudia Magioli;Erika Tarre;Krul, William-Robert;Elisabeth Mansur
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.19-25
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    • 2001
  • Induction of somatic embryogenesis from Brazilian eggplant variety F-100 was studied in response to four auxin types. NAA, at the optimal concentration of 54 $\mu\textrm{m}$, was the only one that resulted in the induction of somatic embryos in either leaf and cotyledon explant and, at murk lower intensity and frequency, in hypocotyl and epicotyl explants. The optimal temperatures for embryo induction were 28 and 35$^{\circ}C$ for cotyledon and leaf explants. Incubation at 22$^{\circ}C$ caused a significant reduction both in the frequency and intensity of induction. This system was used to study the effects of position and orientation of the tissue on the culture medium as well as of antibiotics and explant co-cultivation with Agrobacterium on the efficiency of somatic embryo induction. The intensity of embryo induction was greater in the midsections of cotyledons relative to apical and basal regions, when the abaxial surface was in contact with the culture medium. The presence of antibiotics resulted in approximately 40-60% reduction of embryo induction relative to control explants, which originated 335$\pm$26.6 embryos. Co-cultivation with Agrobacterium before treatment with antibiotics caused a more drastic reduction (80-99%). Ampicilin treatment after cocultivalion with Agrobacterium caused the least inhibitory effect, allowing the production of 60 embryos/explant.

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Effects of Liquid Culture of Agaricus blazei Murill on Protein, Mineral Levels and Enzyme Activities in Rats

  • Koh, Jin-Bog;Kim, Jai-Young
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.171-177
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    • 2004
  • This study was performed to investigate the effects of liquid culture of Agaricus blazei Murill on the weight gains, food efficiency ratios, serum protein and mineral levels, and serum enzyme activities in growing male rats. Sprague-Dawley rats (7 weeks old) were given four different types of diets for a succeeding period of 6 weeks, respectively: a normal diet group, a control diet group (normal diet + 15% lard + 0.5% cholesterol), a 30% or 40% A. blazei diet groups (control diet + 30% or 40% A. blazei in water) according to the levels of A. blazei supplementation. The body weight gains, food efficiency ratios, and the liver of the rats fed control diet, 30% or 40% A. blazei diets were significantly increased compared to rats fed the normal diet, but those of rats fed the 30% and 40% A. blazei diets were similar to those of rats fed the control diet. The concentrations of total protein, albumin, glucose, and hematocrit value in serum of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. The concentrations of urea and creatinine in serum of rats fed the 30% or 40% A. blazei diets were similar to those of rats fed the control diet. but the urea of rats fed the 30% and 40% A. blazei diets were significantly decreased compared to rats fed the normal diet. The concentrations of Ca, P, Fe and Mg, and $Ca^{++}$, $K^+$, $Na^+$ and $Cl^-$ of rats fed the control diet, 30% or 40% A. blazei diets were similar to those of rats fed the normal diet. There were no differences in the activities of aspartate aminotransferase, alanine aminotransferase, $\gamma$-glutamyltranspeptidase and alkaline phosphatase in the serum among the experimental groups. In conclusion, the rats fed the A. blazei maintained normal protein and mineral levels, and enzyme activities of serum. But the A. blazei feeding could not decrease the body and liver weights in the rats fed high cholesterol diets.

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