• Title/Summary/Keyword: 2.5% sucrose

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Effects of Carbon, Nitrogen Sources and pH on Direct Somatic Embryogenesis in Liquid Culture of Rehmannia glutinosa Lib. (지황의 액체배양에서 탄소원.질소원 및 pH가 직접 체세포배 형성에 미치는 영향)

  • Chae, Young-Am;Park, Ju-Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.1
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    • pp.1-6
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    • 1999
  • Basic informations for direct somatic embryo formation in Rehmannja glutinosa Lib. were obtained in 500ml erlenmyer flask. The ratio of ammonium to nitrate nitrogen of 825(mg/l) : 1900(mg/l) was proper condition for somatic embryo formation from stem and petiole explants and 3% sucrose was the most effective carbon source. Full strength MS medium with 2mg/l BA was better than LS medium for somatic embryogenesis. The initial pH 5.7 of medium(full strength MS with 2.0mg/l BA and 0.1mg/l NAA) was good for embryo production. Potassium ion was taken up rapidly within 2 weeks. while $Ca^{++}$ and $Mg^{++}$ ion contents were almost constant during culture period. Sucrose hydrolysis occurred throughout the culture, while glucose and fructose were absorbed simultaneously from the third week of culture.

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Optimum Conditions for the Protoplast Formation of Lactobacillus plantarum and Leuconostoc mesenteroides (Lactobacillus plantarum과 Leuconostoc mesenteroides의 Protoplast 생성의 최적 조건)

  • 전억한;박범식조재선
    • KSBB Journal
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    • v.9 no.2
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    • pp.191-199
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    • 1994
  • Protoplasts of both strains were produced by lysozyme digestion at $30^{\circ}C$ for 180min. Both strains were treated with $40{\mu}g$/ml of lysozyme in 30mM Tris-HCl buffer(pH 7.5) containing 10% sucrose at the late logarithmic growth phase. It was found that the efficiency of protoplast formation was high at $30^{\circ}C$ and pH 7.5 by measuring the decrease in absorbance. Optimum concentrations of sucrose $Ca^{2+}, \;Mg^{2+}$ for protoplast formation were determined to be 15%, 20mM and 6mM, respectively. Hydrolysis of cell wall and protoplast formation efficiency for L. plantarum showed better results than those for Leu. mesenteroides. The resistances to antibiotics erythromycin and chloramphenicols were chosen as the selection marker for the fusant between L. plantarum and Leu. mesenteroides. Production phase of protoplast in Leu. mesenteroides was also compared with L. plantarum in this paper.

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A Study on the Pullulan Production Using Whey (Whey를 이용한 Pullulan의 발효생산에 관한 연구)

  • 정영일;김동운
    • KSBB Journal
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    • v.9 no.2
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    • pp.91-97
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    • 1994
  • In this study, Pullulan Production from whey medium by Aureohsidium pullulans was investigated. When the Pullulan production from whey medium was carried outs final cell concentration was similar to sucrose basal medium but Pullulan concentration was less than 5g/$\ell$. For pullulan production from whey medium, adaptation culture technique was tried on lactose and galactose base medium When the adaptation culture technique was not applied, the maximum concentration of pullulan was 3.4g/$\ell$ for lactose, 2.5g/$\ell$ for galactose. Adapted strains produced 10~12g/$\ell$ from lactose, 10~11g/$\ell$ from galactose. The pullulan production from lactose basal medium was 13.5g/$\ell$ for lactose adapted strains and 18.6g/$\ell$ for galactose adapted strains. When the adapted strains were inoculated on whey medium, maximum pullulan production was obtained at initial pH 3.0.

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Genetic Recombination by Protoplast Fusion of Cellulomonas sp CS 1-1 (원형질체 융합법에 의한 cellulomonas sp. CS 1-1의 유전자 재조합)

  • 김병홍;이학주
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.309-314
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    • 1985
  • Autoxotrophic mutants of a cellulolytic baterium Cellulomonas sp. CS 1-1 were grown at $30^{\circ}C$ for 6hr using a complete medium containing 0.5M sucrose and for another 90 min after addition of 0.3 U/ml penicillin G, and were protoplasted by 0.2mg/ml lysozyme for 2hr. Prototrophic recombinants were obtained at the rates of $10^{-3}$ to $10^{-5}$by fusing the protoplasts in the presence of 40% polyethyleneglycol3350. Nystatin could be used to eliminate fungal contamination during the regeneration of the plotaplasts.

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체외수정시 Sucrose 첨가에 의한 돼지 난자의 Polyspermy 제어에 관한 연구

  • 이창현;박진기;민관식;임기순;양병철;김광식;이향흔;장원경;이훈택
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.85-85
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    • 2001
  • 본 연구는 돼지 난자의 체외 수정시 sucrose 첨가가 돼지 난자의 다정자 침입율에 미치는 영향을 조사하기 위하여 실시하였다. 돼지 난포란은 0.1% PVA, 3.05 mM D-glucose, 0.91 mM sodium pyruvate, 0.57 mM cysteine, 0.5 $\mu\textrm{g}$/$m\ell$ FSH, 0.5 $\mu\textrm{g}$/$m\ell$ LH, 및 10 ng/$m\ell$ EGF가 첨가된 TCM199 배양액에서 42-44 시간 동안 배양하여 체외 성숙을 유도하였으며, 2 mM caffeine과 0.1% BSA가 첨가된 체외수정용 mTBM 배양액에 3% sucorse를 첨가하여 6시간동안 수정을 유도하였다. 수정된 난자의 다정자 침입율을 조사하기 위해 체외수정 후 12, 13 및 14 시간대에 각각 1% orcein으로 염색하여 전핵 형성율을 조사하였다. 돼지 난자의 체외수정시 3% sucorse를 첨가하였을 때, 전핵 형성율을 조사한 결과 대조구에서 미수정, 1, 2, 및 3 PN 이상의 전핵 형성율은 각각 57.6 7.6, 9.7, 및 25%로, 처리구 74.2, 7.6, 13.6, 및 4.5% 에 비교하여 다정자 침입율이 유의하게 높았다(p<0.05). 그리고 3% sucorse를 첨가한 체외수정용 배양액에서 수정을 유도한 후 12, 13, 및 14 시간대별로 나누어 전핵 형성율을 조사한 결과, 수정 후 시간이 경과되면서 다정자 침입율이 증가되는 것을 볼 수 있었다 그러나 대조구 60.7%에 비해서는 각각 32.5, 36.0, 및 33.3%로 현저히 낮은 것을 볼 수 있었다.(Table Omitted). 또한, 3% sucorse가 첨가된 배양액에서 수정된 수정란의 발달율을 조사한 결과, 배반포 생산율은 처리구 21.8%로 대조구 6.9%에 비해 유의하게 높은 것을 확인 하였다. 따라서, 돼지 난자의 체외수정시 3% sucorse첨가는 체외수정란 생산에서 문제가 되고 있는 다정자 침입율을 저하시키고 배발달율을 향상시킴으로서 수정란의 체외 대량생산 효율성은 물론 수정란 이식시 수태율을 증가시킬 수 있을 것으로 사료된다.

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Improvement of In Vitro Seed Germination in Chicory (Cichorium intybus L.) and Garland Chrysanthemum (Chrysanthemum coronarium L.) (치커리(Cichorium intybus L.)와 쑥갓(Chrysanthemum coronarium L.)의 기내 종자발아 개선)

  • Hwang, Hyeon-Jeong;Choi, Geun-Won
    • Journal of Bio-Environment Control
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    • v.17 no.4
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    • pp.297-305
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    • 2008
  • A series of experiments were conducted to suggest efficient in vitro germination conditions to improve germination rate and aseptic seedling production in chicory and garland chrysanthemum. For this purpose, various levels of NaOCl concentration and soaking treatment time combinations were tasted. Fifteen minutes of soaking treatment with 5% NaOCl solution significantly improved germination and seedling formation percentages and prevented contamination in both chicory 'Precole' and 'Chiavari' and garland chrysanthemum 'Okiku 3'. There was no significant difference in germination response between MS and 1/2 MS medium compositions, but germination and seedling formation were delayed as sucrose concentration increased. Although petri-dish among various culture containers gave rise to improved germination responses in chicory 'Precole', there was no significant difference in germination responses among culture container types in chicory 'Chiavari' and garland chrysanthemum 'Okiku 3'. The ultrasonic treatment stimulated germination and seedling formation in both chicory cultivars but there was no effect in garland chrysanthemum 'Okiku 3'. The results suggested 15 minutes with 5% NaOCl for seed sterilization, 1/2 MS medium with no sucrose and petri-dish as proper medium composition and culture container, and 120 minutes with 5% NaOCl solution of ultrasonic treatment as promoting method for in vitro germination in chicory and garland chrysanthemum.

Production and properties of exoinulase from Streptomyces sp. S34 (Streptomyces sp. S34의 exoinulase 생산 및 성질)

  • Ha, Young-Ju;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.35 no.5
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    • pp.375-381
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    • 1992
  • An exoinulase-producing bacterium was isolated from soil, and identified as Streptomyces sp. The maximum inulase production was achieved when inulin as carbon source and soybean meal as organic nitrogen source were included in the culture. The exoinulase was considered to be a constitutive enzyme produced not only by inulin but also by soluble starch or glucose. The purified enzyme on DEAE-cellulose and Sephadex G-200 column showed the maximal activity at $pH\;5.5{\sim}6.0$ and $50^{\circ}C$, but lost 65% inulase activity at $50^{\circ}C$ after 1 hour incubation. This exoinulase was activated by $Mn^{+2}$, wherease more that 80% inactivation was observed with $Ag^+$, $Hg^{+2}$ and $Fe^{+3}$. The enzyme was possibly a metalloenzyme in that EDTA and 8-hydroxyquinoline inhibited the enzyme. Km values for inulin (16.51 mM) and sucrose (14.62 mM) were in very close range suggesting mostly equal affinity toward the subatrates. However, the maximum velocity for inulin was 10 times greater than for sucrose.

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Effects of Cryoprotectant, Warming Solution and Removal of Lipid on Viability of Porcine Nuclear Transfer Embryos Vitrified by Open Pulled Straw Method

  • Cong, Pei-Qing;Song, Eun-Sook;Kim, Eui-Sook;Li, Zhao-Hua;Zhang, Yong-Hua;Lee, Jang-Mi;Yi, Young-Joo;Park, Chang-Sik
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.103-108
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    • 2007
  • This study was carried out to investigate the effects of cryoprotectants, warming solution and removal of lipid on open pulled straw vitrification (OPS) method of porcine embryos produced by nuclear transfer (NT) of fetal fibroblasts. All solutions used during vitrification were prepared with holding medium consisting of 25 mM Hepes buffered TCM199 medium containing 20% fetal bovine serum (FBS) at $38.5^{\circ}C$. The blastocysts derived from NT with or without lipid were vitrified in each medium of different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Also, blastocysts after cryopreservation were warmed into different concentrations of sucrose in warming solution. The optimal concentrations of cryoprotectants in vitrification solution were 10% DMSO + 10% EG in vitrification solution 1 (VS1) and 20% DMSO + 20% EG in vitrification solution 2 (VS2). The optimal concentrations of sucrose were 0.3 M sucrose in warming solution 1 (WS1) and 0.15 M sucrose in warming solution 2 (WS2). lipid removal from oocytes before NT enhanced the viability of NT embryos after vitrification. Our results show that use of the OPS method in conjunction with lipid removal provides effective cryopreservation of porcine nuclear transfer embryos.

Optimal Culture Conditions for the Shoot Growth of Freesia 'Shiny Gold' during Bioreactor Culture

  • Kang, Ji Su;Jeong, Kyeong Jin;Choi, Youn Jung;Yun, Jae Gill
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.699-706
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    • 2017
  • For rapid production of freesia 'Shiny Gold' shoots by using a bioreactor, several culture conditions were investigated. Young shoots (< 1 cm) obtained from freesia corm section in vitro were used as plant materials for this experiment. As a basic experimental environment, 20 young shoots were inoculated into a 5 L balloon type bubble reactor which contained 1 L 1/2 strength MS medium supplemented with 30 g sucrose (3%), and the aeration was 0.1 vvm (vessel volumes per minute). The bioreactors were placed in a growth room with $23^{\circ}C$ temperature, 60% relative humidity and $60{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ light condition (16 h/8 h, day/night). The concentrations of MS media were set with 1/4, 1/2, 1 strength, medium volume 10, 20, 40%, sucrose concentration 3, 6, 9%, and aeration 0.1, 0.2, 0.4 vvm. After 4 weeks of cultivation, the growth indexes including the fresh and dry weight, and plant height were evaluated. At the same time, the consumption, pH, and EC of medium were estimated 4 weeks after incubating. The best results were achieved when 40 young shoots were incubated in a bioreactor in which 1 L of 1/2 strength MS medium supplemented with 6% sucrose was used for the rapid production of freesia shoots. The shoots were 17 cm in plant height and 1.0 g in fresh weight only 4 weeks after incubation which could be a good plant material suitable for corm enlargement in vitro. No correlation was observed between the growth of freesia shoots and the consumption, pH or EC of medium.

Effect of Different Conditions on the Hypoosmotic Swelling Test to Evaluate Functional Integrity of Canine Spermatozoa

  • 오진영;장현용;서정수;정유성;김종택;정희태;박춘근;김정익;양부근
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.277-277
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    • 2004
  • The purpose of this study was to investigate the effect of different conditions (osmolity, solution, incubation times, comparison of fresh and frozen/thawed semen and storage times) on the swelling of canine spermatozoa. Employing the hypoosmotic swelling test (HOST), the membrane integrity of spermatozoa in different solutions (sucrose, fructose, latose, Na-citrate, Na-citrate plus sucrose, Na-citrate plus fructose and Na-citrate plus lactose were 61.4%, 66.2%, 62.5%, 68.1%, 62.0%, 68.5% and 60.2%, respectively. (omitted)

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