• Title/Summary/Keyword: Optimal culture

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Regeneration and Acclimatization of Plants Derived from Anther Cultures in Carrot (Daucus carota L.) (당근 약배양에 의한 식물체 재분화 및 순화)

  • Cho, Moon-Soo;Juang, Ue-Dong;Park, Sang-Gyu;Park, Yong
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.47-52
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    • 2003
  • Anthers from several lines of carrot (Daucus carota L.) were plated on the semi-solid B$_{5}$, basal medium supplemented with 2,4-D and NAA at two concentrations, 1.0 and 2.0 mg/L plus 0.2 mg/L BAP (benzylaminop-urine). Anthers of the most lines on the B$_{5}$ basal medium with 2,4-D showed higher percentages of callus formation than those with NAA. Particularly, in line 45477, highest percentages of callus formation (50%) were observed on B$_{5}$ medium with 1.0 mg/L 2,4-D plus 0.2 mg/L BAP. With 1.0 mg/L 2,4-D, two months was sufficient for initiation of callus development. Calli were regenerated into plantlets through embryogenesis onto regeneration medium without any growth regulators. When callus showing yellowish and soft structure was cultured, it yielded green plants at high regeneration rates, The response of anthers in callus induction and plant regeneration was different among lines investigated. Optimal callus induction and plant regeneration could be obtained through manipulating the concentration of growth regulators. Plantlets after transfer to perlite were grown successfully in greenhouse conditions. Anther culture of carrot will be used as a useful breeding tool in future.

Plant Regeneration via Adventitious Shoot Formation in Platycodon grandiflorum (Jacq. A. DC.) (도라지 (Platycodon grandiflorum (Jacq.) A. DC.) 부정아 형성을 통한 식물체 재분화)

  • Kim, Ju Young;Na, Hyun Sun;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.330-334
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    • 2017
  • To investigate optimal conditions for plant regeneration in Platycodon grandiflorum (Jacq. A. DC.).Both leaf and hypocotyl explants were cultured on Murashige& Skoog's (MS) medium supplemented with combinations of 0.1, 0.5, 1.0, or 2.0 mg/L cytokinins (BA and kinetin) and 1.0 mg/L 2,4-D for 6 weeks, respectively. According to the type of explant, the total shoot organogenesis (56.38%) in leaf explants was higher than in hypocotyls (28.20%). In comparison with kinetin and BA for the plant regeneration, the frequency (70.38%) of leaf explants was higher in combination with kinetin and 2,4-D than of BA with 2,4-D (42.38%), whereas the frequency (35.56%) of hypocotyls explants was higher in BA combination than kinetin combination (20.83%). Thehighest frequency (94.20%) was observed from the cultures of leaf explants on the MS medium supplemented with 1.0 mg/L kinetin and 1.0 mg/L 2,4-D. Upon transfer onto 1/2 MS basal medium containing 3% sucrose, shoots developed into plantlets with roots, and were well grown in soil in the greenhouse. These results lead us to speculate that the optimization of culture conditions was responsible for the mass propagation from in vitro cultures of Platycodon grandiflorum (Jacq. A. DC.).

Factors Influencing Efficient Agrobacterium-mediated Transformation of Panicum spp. (Agrobacterium법에 의한 Panicum속 식물들의 효과적인 형질전환에 영향을 미치는 요인)

  • Seo, Mi-Suk;Takahara, Manabu;Takamizo, Tadashi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.1
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    • pp.1-8
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    • 2011
  • Molecular techniques such as genetic transformation are powerful tools that can be used for the genetic modification of warm-season grasses. The P. meyerianum with high regeneration ability was used for establishing an Agrobacterium-mediated transformation system. We investigated various factors affecting Agrobacterium infection by examining GUS gene expression of pCAMBIA1304 vector. Among various concentration of acetosyringone and betaine tested for inoculation and co-cultivation, 10 mg/L acetosyringone and 60 mg/L betaine resulted in the highest transformation frequency in terms of GUS expression. The calli of 4 species of Panicum spp. with excellent tissue culture response were inoculated with Agrobacterium under the optimal infection conditions. The high activity of GUS gene was observed in all species and hygromycin-resistant calli expressing GFP were obtained in P. meyerianum, P. longijubatum, P. stapfianum and guineagrass Noh-PL1. Co-cultivated calli were transferred onto the selection medium containing hygromycin, and the hygromycin resistant calli were selected after 3 months. Hygromycin-resistant plantlets were then successfully regenerated from the calli and grown in a greenhouse. We confirmed stable insertion of hpt gene among the hygromycin-resistant plantlets of P. meyerianum by PCR analysis.

The Quality Characteristics of Beef Sausage with the Addition of Gynura procumbens (명월초 가루를 첨가한 우육 소시지의 최적화 연구)

  • Park, Young Il;Jeong, Hee Sun;Joo, Nami
    • Korean journal of food and cookery science
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    • v.31 no.4
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    • pp.395-404
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    • 2015
  • The aims of this study were to determine the optimal mixing condition for two different amounts of Gynura procumbens powder and olive oil for the preparation of beef sausage. The experiments were designed according to the central composite design of response surface methodology, which showed 10 experimental points including two replicates of Gynura procumbens powder and olive oil. The physicochemical and mechanical characteristics as well as the sensory properties were measured, and these values were applied to the mathematical optimization models. The results of the physiochemical and mechanical analysis of each sample, including chewiness, cohesiveness, color a, color b, moisture content, salinity and heating loss showed significant differences. The sensory measurements were significantly different in color, flavor, tenderness, texture and overall quality. The optimum formulation, which was calculated using the numerical and graphical method, was determined to be 2.1 g Gynura procumbens powder and 7.06 g of olive oil.

Characteristics of the alkaline protease from the moderate halophile, Halomonas sp. ES 10 (Halomonas sp. ES 10이 생산하는 alkaline protease의 특성)

  • Kim, Chan-Jo;Oh, Man-Jin;Choi, Seong-Hyun
    • Applied Biological Chemistry
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    • v.35 no.4
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    • pp.237-241
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    • 1992
  • The protease from Halomonas sp. ES 10 was purified by methanol precipitation, gel filtration on Sephadex G-150 and G-200, and ion exchange chromatography on DEAE-Sephadex A-50. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis. The specific activity of purified enzyme was 1,014 units/mg protein, and the yield of the total activity from the culture filtrate was 7%. The optimal temperature and pH for the enzyme activity were $35^{\circ}C$, and pH 11.0, respectively. And the enzyme was stable in the range of $pH\;7.5{\sim}11.0$. The residual activity of the enzyme was 70%, when the enzyme was incubated at $50^{\circ}C$ for 40 min. The Km value of the enzyme was 7.4 mg/ml to milk casein. $Li^+$, $Ca^{2+}$, SDS and Tween 80 were appeared to activators, whereas $Hg^{2+}$ and EDTA to inhibitors. The addition of DFP and PMSF showed the relative enzyme activities of 63% and 107%, respectively, suggesting that the enzyme may not belong to serine type protease. When the alkaline protease was treated with 0.5 M and 1 M NaCl, the relative enzyme activities were 95% and 65%, respectively. This enzyme showed 20% and 15% higher enzyme activity than that of Aspergillus oryzae (Sigma Chemical Company product, P4755) in the presence of 0.5 M and 1 M NaCl.

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Identification of characterization and statistical optimization of medium constituent for Bacillus subtilis SCJ4 isolated from Korean traditional fermented food (전통 장류 유래 Bacillus subtilis SCJ4의 특성확인 및 통계학적 방법을 이용한 배양조건 최적화)

  • Jeong, Su-Ji;Yang, Hee-Jong;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.48-60
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    • 2015
  • 612 strains isolated from Korean traditional fermented food in Sunchang and their investigated biochemical characterization and ability of biogenic amines non-producing. We selected the SCJ4 having various activity by measurement of extracellular enzyme, antioxidant and antimicrobial activities. Selected strain SCJ4 by 16S rRNA sequencing and biochemical characterization was named Bacillus subtilis SCJ4. And then, we investigated cell growth of SCJ4, and optimized of culture medium constituents using response surface methodology as statistically method. Response surface methodology used Plackett-Burman experimental design for screening of medium constituent. Tryptone, peptone and $MgSO_4$ as medium constituent improving cell growth selected. In order to find out optimal concentration on each constituent, we carried out central composite design. Consequently, optimized concentrations of tryptone, peptone and $MgSO_4$ were predicted to be 15.35 g/L, 12.235 g/L, and 3.5 g/L respectively. Through the model verification, we confirmed about 1.28-fold improvement of the dried cell weight from 0.8767 g/L to 1.1222 g/L when compared to basal medium.

Carbon Dioxide Fixation using Spirulina Platensis NIES 39 in Polyethylene Bag (Spirulina Platensis NIES 39를 이용한 Polyethylene Bag 반응기에서의 이산화탄소 고정화)

  • Kim, Young-Min;Kim, Ji-Youn;Lee, Sung-Mok;Ha, Jong-Myung;Kwon, Tae-Ho;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.21 no.3
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    • pp.272-277
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    • 2010
  • To replace current expensive photobioreactor, this study was conducted to develop low-cost photobioreactor made of polyethylene bag. In previous study, optimal culture conditions of Spirulina platensis NIES 39 have been established, and based on these, the study of biological carbon dioxide fixation has been conducted. The maximum growth was the biomass 2.677 g/L at conditions of 10% $CO_2$, 0.1 vvm. It was shown that $F_{CO_2}$ was 4.056 g $CO_2$/L and $R_{CO_2}$ was 0.312 g $CO_2$/L/day. But, compared with the data at conditions of 5% $CO_2$, 0.1 vvm, $FE_{CO_2}$ was shown 52.372% which is half of it. Regarding the effect of $CO_2$ following illumination, the growth revealed that the input conditions, for 10 min per 3 h, were excellent in the light. $CO_2$ in absent light. $CO_2$ concentration and flow rate were 5% $CO_2$, 0.1 vvm, respectively. Finally, the addition of $CO_2$ was ineffective in the absence of light.

Isolation of Serratia marcescens CK-3 against phytopathogenic fungi and its enzymatic properties (식물(植物) 병원류(病源惟) 사상균(絲狀菌)에 길항력(拮抗力)을 갖는 Serratia marcescens CK-3의 분리(分離) 및 효소적(酵素的) 성질(性質))

  • Kim, Yeong-Yil;Rhee, Young-Hwan;Kim, Kwang-Sik;Park, Hwa-Sung;Chun, Woo-Bock;Lee, Jae-Wha;Kim, Jong-Hyun
    • Applied Biological Chemistry
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    • v.34 no.1
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    • pp.54-60
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    • 1991
  • Serratia marcescens CK-3, decomposing chitin which is a mar component of cell wall in phyitopathogenic fungi, was isolated from the continuous cropping rhizosphere of pepper and cucumber and its enzymatic property was examined. S. marcescens CK-3 was found tn have an tagonistic effects against, Fusarium axysporum and Rhizoctonia solani and to have complex enzyme system such as chitinase, laminarinase, and proteinase. The preferable composition of the medium for production of chitinase was fond and was as follows : colloidal chitin 1.5%, tryptone 0.5%, glucose 1.0%, peptone 0.2%, $MgSO_4{\cdot}7H_2O\;0.1%,\;K_2HPO_4\;0.1%,\;and\;NaCl\;0.1%$(w/v), pH 6.8. The maximum enzyme production was observed after culture of 72 hours at $30^{\circ}C$ using a medium containing the above chemical composition. The optimal pH and temperature for in vitro activity of chitinase from S. marcescens CK-3 were pH 7.5 and $50^{\circ}C$, respectively. The enzyme activity in-creased by metal ions such as$Ag^+$ and $Mn^{++}$.

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Effects of Medium Compositions and Plant Growth Regulators on in vitro Organogenesis in Cultured Explants of Platycodon grandiflorum Species

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Kim, Hye-Rim;Moon, Young-Ja;Yoon, Ki-Hong;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.3
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    • pp.259-274
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    • 2017
  • Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on 1/4 MS for P. grandiflorum with wild and green petals and on 1/8 MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on 1/4 MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, 1/4 MS supplemented with $1mg\;L^{-1}$ 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on $0.5mg{\cdot}L^{-1}$ thidiazuron (TDZ) from double petal explants grown on 1/8 MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.

Molecular Cloning and Expression of Heat-stable Enterotoxin Gene from Swine Enterotoxigenic Escherichia coli (돼지에서 유래한 병원성 대장균의 내열성 장독소 생산유전자의 Cloning 및 발현)

  • 김교창;도대흥
    • Journal of Food Hygiene and Safety
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    • v.6 no.3
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    • pp.147-155
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    • 1991
  • Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

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