• Title/Summary/Keyword: 2.5%25 sucrose

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A successful regeneration from shoot tips of Fragaria x ananassa DUCH following cryopreservation by droplet-vitrification (작은방울유리화법을 이용한 딸기 생장점 초저온동결보존)

  • Lee, Young-yi;Baek, Hyung-Jin;Yoon, Mun-Seop;Balaraju, Kotnala;Song, Jae-young
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.80-80
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    • 2019
  • This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of (Fragaria x ananassa DUCH. cvs. 'Derunoka' and 'Jumbo pure berry'. The shoot tips of strawberry were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7M). Precultured explants were treated with loading solution (LS, C4) containing glycerol 17.5% and sucrose 17.5% for 40 min and exposed to dehydration solution (B1) containing 50% of glycerol and 50% of sucrose for 60 min at $25^{\circ}C$, and then transferred onto droplets containing $2.5{\mu}l$ PVS3 on sterilized aluminum foils ($4cm{\times}0.5cm$) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regeneration rate (%) was obtained when shoot tips were precultured with treatment-2 (exposing of shoot tips to MS + 0.3M Sucrose for 30 h and then treated with MS+0.5 M sucrose for 16 h) at $25^{\circ}C$ in both the cultivars. The viability of cooled samples, followed by culturing on MS medium for 4 weeks was 77.8% and 60.0% for 'Derunoka' and 'Jumbo pure berry', respectively. This result shows droplet-vitrification would be a promising method for cryobanking strawberry germplasm.

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A Successful Regeneration from Shoot Tips of Chrysanthemum morifolium (Ramat.) following Cryopreservation by Droplet-vitrification

  • Yi, Jung-Yoon;Balaraju, Kotnala;Baek, Hyung-Jin;Yoon, Mun-Seop;Kim, Haeng-Hoon;Lee, Young-Yi
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.675-683
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    • 2018
  • This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of Chrysanthemum morifolium (Ramat.) cvs. 'Borami' and 'Yes morning'. The shoot tips of Chrysanthemum were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7 M). Precultured explants were treated with loading solution (LS, C6) containing glycerol 20% and sucrose 20% for 30 min and exposed to dehydration solution (B5) containing 40% of glycerol and 40% of sucrose for 60 min at $25^{\circ}C$, and then transferred onto droplets containing $2.5{\mu}l$ PVS3 on sterilized aluminum foils ($4cm{\times}0.5cm$) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regeneration rate (%) was obtained when shoot tips were precultured with treatment-2 (exposing of shoot tips to MS + 0.3M sucrose for 30 h and then treated with MS+0.5 M sucrose for 16 h) at $25^{\circ}C$ in both the cultivars. The viability of cooled samples, followed by culturing on $NH_4NO_3$-free MS medium for first 5 days was increased to two-fold (80.7%) regrowth rate over those cultured on normal MS medium or MS medium containing plant growth regulators. This result shows droplet-vitrification would be a promising method for cryobanking chrysanthemum germplasm.

Studies on the survival Rate after Slow and Ultrarapid Frozen-Thawing of Porcine Embryos (돼지 수정란의 완만 및 초급속 동결 융해후의 생존성에 관한 연구)

  • 이봉구;김상근;이규승
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.117-123
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    • 1992
  • This Study was carried out ot investigate the effects of concentration and equilibration time of cryoprotective aagents on survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and 0.25M sucrose were slowly freezed(from 2$0^{\circ}C$ to -7$^{\circ}C$/-1$^{\circ}C$/min., from -7$^{\circ}C$ to -35$^{\circ}C$/-0.2$^{\circ}C$/min., from -35$^{\circ}C$ to -38$^{\circ}C$/-0.3$^{\circ}C$/min.) by Cell Freezer and directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rate to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after slow frozen-thawing in the freezing medium of 0.25M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol+2.0M propanediol was 80.6, 84.7, 75.0 or 78.8%, respectively. 2. The survival rates of porcine embryos after slow frozen-thawing in the freezing medium of 0.50M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol+2.0M propanediol was 80.9, 82.4, 73.1 or 77.1%, respectively. 3. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol+2.0M propanediol was 65.3, 68.6, 63.2 or 59.9%, respectively. 4. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.50M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0 propanediol or 2.0M glycerol+2.0M propanediol was 67.5, 62.9, 56.9, or 62.8%, respectively. 5. The higher survival rate of porcine embryos was attained at the short period ofequilibration time(5min.) in the freezing medium added 0.25M sucrose and 3.0 DMSO compared to those of 10 or 20min. equilibration time in the same condition.

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Cryopreservation of in vitro Grown Shoot Tips of Strawberry (Fragaria × ananassa Duch.) Genetic Resources by Droplet-vitrification

  • Lee, Young-Yi;Balaraju, Kotnala;Song, Jae-Young;Yi, Jung-Yoon;Lee, Sun-Yi;Lee, Jung-Ro;Yoon, Munsup;Kim, Haeng-Hoon
    • Korean Journal of Plant Resources
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    • v.32 no.6
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    • pp.689-697
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    • 2019
  • This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of strawberry (Fragaria × ananassa Duch.) cvs. 'Wonkyo3114' and 'Gurumi40'. The shoot tips were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.5M). Precultured explants were osmoprotected with loading solution (LS, C4) containing 20% glycerol and 20% sucrose for 40 min and exposed to dehydration solution (B5) containing 40% glycerol and 40% sucrose for 40 min at 25℃, Subsequently, the explants were transferred onto droplets containing 2.5 μL PVS3 on sterilized aluminum foils (4 cm × 0.5 cm) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regrowth rate (%) in both the cultivars was obtained when the shoot tips were precultured with MS + 0.3M sucrose for 40 h at 25℃. The cryopreserved shoots tips exhibited 55% regrowth rate by culturing in NH4NO3-free MS medium supplemented with 3% sucrose, 1.0 g/L casein, 1.0 mg/L GA3, and 0.5 mg/L BA for 5 weeks and in MS medium supplemented with 0.5 mg/L GA3 for 8 weeks. This result shows that droplet-vitrification could be employed as a promising method for cryostorage of strawberry germplasm.

Effects of Membrane Filter and Sucrose Concentrations on the Growth of Balloon Flower (Platycodon grandiflorum A. DC.) Plantlets In Vitro (Membrane Filter와 Sucrose 농도가 도라지(Platycodon grandiflorum A. DC.) 기내 배양묘의 생장에 미치는 영향)

  • Choi, So-Ra;Kim, Myung-Jun;Eun, Jong-Seon;Ahn, Min-Sil;Lim, Hoi-Chun;Ryu, Jeong;You, Dong-Hyun
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.209-217
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    • 2004
  • The shoots of balloon flower (Platycodon grandiflorum A. DC.) in vitro germinated from seeds were cultured on MS basal medium containing 0.1 mg/L NAA under the various sucrose concentrations and with/without membrane filter (MF) on the lid of vessel. The growth responses were checked to obtain healthy plantlets. The $CO_2$ and $C_2$H$_4$ concentration in vessel without MF were higher than those with MF. The $CO_2$ concentration without MF was increased as days in culture went by whereas the $C_2$H$_4$ concentration was decreased. The plant growth with MF and high sucrose concentration was good. Fresh and dry weight of plantlets cultured in sucrose 4.5% with MF were higher than those in no sucrose without MF. Also the content of chlorophyll of plantlets cultured with MF was high and the content of sugar was shown a similar results and a remarkable difference between MF treatments, especially. Stomata cultured with MF was closer than that without MF and mesophyll of leaf were more developed with MF or in high sucrose concentration. When the plantlets were transplanted in the pot at 25$\pm$2$^{\circ}C$, 75% relative humidity and low PPFD (photosynthetic photon flux density), the percentage of survival after 13 days without MF was 0% but it was 100% with MF regardless of sucrose concentrations.

Comparison of the Pain-relieving Effects of Human Milk, Sucrose, and Distilled Water during Examinations for Retinopathy of Prematurity: A Randomized Controlled Trial

  • Jang, Eun Kyung;Lee, Hyejung;Jo, Keum Sik;Lee, Sung Mi;Seo, Hyun Jin;Huh, Eun Joo
    • Child Health Nursing Research
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    • v.25 no.3
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    • pp.255-261
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    • 2019
  • Purpose: This study compared the pain-relieving effects of human milk, sucrose, and distilled water during examinations for retinopathy of prematurity. Methods: Forty-five preterm infants were randomly assigned to receive a pacifier dipped in one of three solutions: human milk (n=14), 24% sucrose (n=15), or distilled water (n=16), 2 minutes before an eye examination. Their pain score, pulse rate, and oxygen saturation were measured at three time points: 5 minutes before the examination, 30 seconds after speculum introduction, and 2 minutes after the examination. Results: The infants' mean gestational age and weight at birth were $33.1{\pm}2.1$ weeks and $1,842{\pm}470g$, respectively. There were no between-group differences in pain relief during the eye examination. The pain score significantly increased both during (p<.001) and after the examinations (p=.003). Oxygen saturation decreased during the examinations (p<.001); however, the infants in the 24% sucrose group showed higher oxygen saturation (p=.047) during the examinations than the infants in the other groups. Conclusion: Sucking on a pacifier dipped in human milk or 24% sucrose did not reduce the pain associated with eye examinations in preterm infants. Pacifiers dipped in sucrose can be used to maintain better oxygen saturation during these examinations.

Effect of Exposure to Vitrification Solutions on Maturation and Cleavage Rates of Immature Porcine Oocytes in Vitro

  • Park, I. K.;H. B. Song
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.113-113
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    • 2003
  • This study was conducted to investigate the effect of vitrification solution(VS) on in vitro developmental competence of immature porcine oocytes. The immature porcine oocytes were exposed to the following vitrification solution, at RT. 1) EFS sol. : 20% ethylene glycol (EG) 3 min, 40% EG + 18%(w/v) Ficoll(MV70, 000) + 0.3 M sucrose 30 sec, 2) GE sol. : 10% glycerol 5 min, 10% G + 20% EG 5 min, 25% G +25% EG 30 sec, 3) EG sol : 1.5M EG 2.5 min, 5.5 M EG + 1.0 M sucrose 30 sec. Oocytes were immediately transferred into 1.0 M, 0.5 M, 0.25 M, 0125 M, 0 M sucrose solution for 2.5 min each at RT. After removal of VS, immature oocytes were matured in vitro and subsequently all oocytes were subjected to IVF followed in vitro culture for 7 days. Maturation rates of oocytes were 38.8%, 44.5%, 22.4% and 57.6%, in EFS, EG, GE and Control, respectively, maturation rates of oocytes in EG and Control was significantly higher than EFS and GE(P<0.01). Fertilization rates of oocytes in Control was significantly higher than other treated groups(P<0.05), but no difference were observed among treated groups. Polyspermic rates were no significant difference among four groups. Cleavage rates of oocytes were 21.9%, 47.1%, 19.0% and 65.9%, in EFS, EG, GE and Control, respectively, cleavage rates of oocytes in EG and Control was significantly higher than EFS and GE(P<0.05), but blastocyst formation rates were no significant difference among four groups. These results suggested that the use of EG solution could be a great challenge for reaching a successful vitrification of immature porcine oocytes.

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Isolation and Characterization of Oil Degrading Bacteria from Southern Sea of Korea (남해안 해수로부터 원유 분해 세균의 분리 및 특성)

  • 김학주;김봉조;공재열;구헌서
    • KSBB Journal
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    • v.15 no.1
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    • pp.27-34
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    • 2000
  • A marine bacterium having a high oil-degrading activity was isolated form the oil-polluted southern sea of Korea, and was identified as Pseudomonas aeruginosa and was named Pseudomonas aeruginosa BYK-2. The optimal tmeperatur, culture time, pH and NaCl concentration for biosurfactant production and cell growth showed $25^{\circ}C$, 48h, 7.0 and 0%(w/v), respectively. After cultivation at $25^{\circ}C$, 180 rpm in 250 mL erlenmeyer flask for 7days, 1%(w/v) arabian light crude oil and bunker C oil which are considered to be hardly degradable compounds were degraded 92.1%(w/w) and 76%(w/w) respectively. And then, cell adherence was measured on various carbon sources. The cell adherence indicated over 80% on hydrocarbons(arabian light crude oil, kuwait curde oil, bunker C oil, n-paraffine, n-hexadecane, n-tetradecane) as carbon sources. Lecithin among fatty acids(oleic acid, olive oil, lecithin) showed highest cell adherence of 91.5%. The cell adherence of sugars(arabinose, trehalose, dextrose, galactose, lactose, fructose, maltose, sorbitol, sucrose) observed to be less than 70% except for arabinose, galactose, sorbitol and sucrose.

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Effects of sucrose and MS salts in the addition of liquid medium on in vitro bulblet growth of Lilium oriental hybrid 'Casa Blanca' (액체배지 첨가시 sucrose 및 MS 염류가 Lilium Oriental Hybrid 'Casa Blanca' 의 기내 자구비대에 미치는 영향)

  • 한봉희;예병우;백기엽;염미란
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.239-242
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    • 2001
  • The effects of post-cultural addition of liquid medium addition to stimulate in vitro bulblet growth in Lilium oriental hybrid 'Casa Blanca' were investigated. The sections of bulblets with swollen basal plate (7 mm$\times$12 mm) were cultured on medium containing 60 g/L sucrose and 1 g/L activated charcoal for two months in dark, and then, liquid medium was added into the same vessels. The addition of liquid medium stimulated the growth of bulblets remarkably, compared to no addition of liquid medium. The liquid medium supplemented with 120 g/L sucrose and double strength of MS salts were the most effective on the growth of in vitro bulblets.

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In vitro shoot regeneration from leaf tissue of "Whangkeumbae" pear(Pyrus pyrifolia Nakai) (황금배(Pyrus pyrifolia Nakai) 잎 조직으로부터 기내 신초 재분화)

  • Chun, Jae An;Do, Kyung Ran;Kim, Se Hee;Cho, Kang-Hee;Kim, Hyun Ran;Hwang, Hae Sung;Shin, Il Sheob
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.288-294
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    • 2012
  • In order to establish an efficient adventitious shoot regeneration conditions from leaf explants for Asian pear 'Whangkeumbae', the effect of concentration and kinds of plant growth regulator and carbon source was investigated. Leaf explants of cultures grown on Murashige and Skoog (MS) medium containing 8 g/L plant agar were used. When the medium contained 0.25 mg/L thidiazuron (TDZ) and 0.3 mg/L indolebutyric acid (IBA), the adventitious shoot regeneration rate (ASRR) was greater as 61.1% than others treated and higher TDZ concentrations (2.5 and 5 mg/L) treatment significantly reduced the ASRR. As the effect of IBA and indoleacetic acid (IAA) concentration on the ASRR, 0.5 mg/L TDZ plus different concentration of IAA exhibited relatively high ASRR and 0.5 mg/L TDZ plus 0.3 mg/L IAA showed the highest ASRR of 76.7%. Also the effect of sucrose and sorbitol as carbon source on regeneration was examined. The highest ASRR and the most shoots per explants averaged 94.4% and 3.49 by treatment of 30 mg/L sorbitol, respectably. Sorbitol is considered better carbon source than sucrose for shoot regeneration of 'Whangkeumbae' pear.