• Title/Summary/Keyword: Preculture

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Cryopreservation of chrysanthemum shoot tips using the droplet-vitrification technique (작은방울-유리화법에 의한 국화 신초의 초저온동결보존)

  • Lee, Yoon-Keol;Park, Sang-Un;Kim, Haeng-Hoon
    • Korean Journal of Agricultural Science
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    • v.38 no.2
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    • pp.227-233
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    • 2011
  • This study aimed at developing cryopreservation protocol for chrysanthemum (Dendranthema grandiflora Tzelevcv. peak) shoot apices based on droplet-vitrification procedure, which is a combination of droplet-freezing and solution based vitrification. Progressive preculture of shoot apices in liquid MS medium supplemented with 0.3 and 0.7 M sucrose for 31 and 17 hours, respectively, was found optimum among preculture treatments tested. The composition of both loading and vitrification solutions significantly affected recovery growth of shoot tips before and after cryopreservation. Balancing glycerol and sucrose concentrations in the solutions was beneficial for recovery growth. The highest recovery after cryopreservation was observed when apical shoot tips were extracted from 4-week-old in vitro plantlets, progressively precultured with 0.3-0.5-0.7 M sucrose for 32-16-6 hours, respectively, then treated with loading solution comprising of 1.9 M glycerol + 0.5 M sucrose (35% PVS3 solution). Apices were then dehydrated with the vitrification solution consisted of 50% glycerol + 50% sucrose for 90 minutes then directly immersed in liquid nitrogen.

Preculture Condition for Stable Recovery of Strawberry (Fragaria × ananassa Duch.) Shoot Tips after Cryopreservation Using Droplet Vitrification

  • Lee, Young-Yi;Lee, Sun-Yi;Song, Jae-Young;Yoon, Munsup;Yi, Jung-Yoon;Lee, Jung-Ro;Kim, Haeng-Hoon
    • Korean Journal of Plant Resources
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    • v.33 no.6
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    • pp.675-681
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    • 2020
  • This study describes an efficient and stable droplet vitrification following cryopreservation of strawberry shoot tip (Fragaria × ananassa Duch.) accessions 'Massey' and 'MDUS3816'. The shoot tips were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7M). Precultured explants were osmoprotected with loading solution (LS, C4) containing 17.5% glycerol and 17.5% sucrose for 40 min and exposed to dehydration solution (B1) containing 50% glycerol and 50% sucrose for 40 min at 25oC. 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 0.3M sucrose for 30 h + 0.5M sucrose for 16 h at 25oC. The cryopreserved shoots tips exhibited 57.8 % recovery rate by culturing in NH4NO3-free MS medium supplemented with 3% sucrose, 1.0 g/L casein, 1.0mg/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. Variation was not observed in both of ploidy analysis and morphological investigation on plantlets of two accessions cryopreserved under variable preculture conditions.

Effect of Medium Conditions on Adventious Bud Formation of Mulberry Winter Bud Leaflet (배지조건이 뽕나무 동아엽편의 불정아형성에 미치는 영향)

  • 성규병;정정박명;김호락;구태원
    • Journal of Sericultural and Entomological Science
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    • v.37 no.1
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    • pp.6-9
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    • 1995
  • To establish a regeneration system of mulberry through the adventitious bud formation of winter bud leaflet, medium conditions were examined. Adventitious bud formation rate of mulberry winter bud leaflet on the medium containing different concentrations of BA and TDZ as the growth regulators was high on 5mM BA and 0.5mM TDZ. The effect of preculture on the medium containing BA or TDZ was affected not only by the concentration of growth regulators but the duration of preculture. Fructose as carbon source was effective on adventitious bud formation.

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The Effect of agrobacterium Density on Transformation Efficiency in Apple (Agrobacterium 농도가 사과 형질전화 효율에 미치는 영향)

  • Seong, Eun-Soo;Cha, Ji-Eun;Kim, Jeong-Hee;Park, Seong-Whan;Yu, Chang-Yeon;Song, Kwan-Jeong
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.215-219
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    • 2003
  • This study was conducted to find optimum bacterial density for improving the efficiency of transformation mediated by Agrobacterium tumefaciens in apples. Regeneration(15%) and transformation frequency(10%) were increased in resuspension-culture density $A_{600}$ 1.3 from preculture density $A_{600}$ 0.7 of Agrobacterium tumefaciens in ′Fuji′. In ′Gala′, 20% regeneration and 16% transformation frequency were observed at optimum bacterial density $A_{600}$ 0.7 form preculture density $A_{600}$ 1.3. ′Mclntosh as well as "Gala" were 25%regeneration and 10% transformation frequency. Hence a frequency optimum condition of bacterial density for the efficient transformation of apple could be depend on apple genotypes.

Mechanisms of Lipopolysaccharide-induced Lipopolysaccharide Tolerance in the Expression of TNF-$\alpha$ and IL-8 in Peripheral Blood Monocytes (말초 혈액 단핵구의 TNF-$\alpha$와 IL-8 발현에서 내독소에 대한 내성 기전에 관한 연구)

  • Park, Gye-Young;Kim, Jae-Yeol;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.601-610
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    • 1997
  • Background : Monocytes/macrophages play a central role in determining the host response during Gram-negative infection through secretion of a variety of mediators after stimulation of LPS. Even though cytokine production has been shown to play an important role in host defense during sepsis, cytokine release may also lead to tissue injury. Thus, regulation of macrophage response to LPS is critical for host survival during Gram-negative sepsis. In animals exposed to nonlethal doses of endotoxin, a characteristic hyporesponsiveness to subsequent administration of endotoxin has been observed. This phenomenon was known as 'LPS tolerance'. However, little information is available regarding the underlying mechanism of LPS tolerance. Method : Peripheral blood monocyte(PBMC) was isolated from peripheral blood of normal volunteers by adhesion purification method. To evaluate the conditions to obtain LPS tolerance, preculture was carried out with LPS at 10ng/ml for 24 hours. For stimulation, culture plates were washed two times and were stimulated with LPS at $1{\mu}g/ml$ for 4, 6 and 26 hours. To assess the underlying mechanisms of LPS tolerance, autologous serum, PMA, anti-CD14 Ab, Indomethacin or $PGF_2$ were added to preculture solution respectively. Cytokine concentrations in culture supernatants were measured using ELISA for TNF-$\alpha$ and IL-8 and mRNA of TNF-$\alpha$ and IL-8 were determined by Northern blot analysis. Results : The exposure of PBMC to low dose of LPS suppressed the cytokine production and mRNA expression of TNF-$\alpha$, but not IL-8. Anti-CD14 Ab partially recovered production of TNF-$\alpha$ which was suppressed by preculture with low dose LPS. The preculture with PMA induces LPS tolerance, as preculture with low dose LPS. Conclusion : LPS tolerance to TNF-$\alpha$ is regulated pretranslationally and is influenced by protein kinase C pathway and CD14.

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Regeneration of Plants from EMS-treated Immature Embryo Cultures in Soybean [Glycine max(L.) Merr.]

  • Van, Kyu-Jung;Jang, Hyun-Ju;Jang, Young-Eun;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.119-126
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    • 2008
  • Since somatic embryogenesis combined with ethylmethane sulfonate(EMS) treatments is the most efficient technique for mutagenesis, the embryogenic capacity of four soybean cultivars was evaluated at different EMS concentrations, treatment times, and preculture durations. Two to 4 mm long immature cotyledons were placed in induction medium after EMS treatment, and the numbers of somatic embryos formed per explant were counted four weeks after culture initiation. We observed genotypic differences in the efficiency of somatic embryogenesis from immature embryos among four cultivars treated with different concentrations of EMS for six hours. Cultivars, Sinpaldalkong 2 and Jack, displayed highly efficient somatic embryogenesis regardless of EMS concentration, whereas very low efficiency or no survival was observed in Jinju 1 and Iksannamulkong cultivars. Preculture duration did not influence the efficiency of somatic embryogenesis. Because Sinpaldalkong 2 exhibited the best somatic embryogenesis, much higher concentrations of EMS were used to test somatic embryo formation under different periods of time in this cultivar. Three and six hour treatments with both 1 and 2 mM EMS yielded higher embryo formation than longer periods of time. Increasing the time with embryos in 2 mM EMS caused a reduction in somatic embryogenesis in Sinpaldalkong 2, but many chlorophyll-deficient soybean variants were identified in the $M_1R_0$ and $M_2R_1$ generations. In addition to Jack, Sinpaldalkong 2 is a good genotype for plant regeneration from EMS-treated immature embryo cultures.

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Cryopreservation of in Vitro Grown Shoot Tips of Sweet Potato (Ipomoea batatas L.) by the Encapsulation-Vitrification Method

  • Yi, JungYoon;Lee, GiAn;Lee, YoungYi;Gwag, JaeGyun;Son, EunHo;Park, HongJae
    • Korean Journal of Plant Resources
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    • v.29 no.6
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    • pp.635-641
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    • 2016
  • Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulation-vitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at $25^{\circ}C$. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.

Studies on Some Properties of Cellulase Isolated from Pirieularia oryzae (Piricularia oryzae로부터 추출한 cellulase의 몇가지 성질에 대한 연구)

  • 전상윤
    • Korean Journal of Microbiology
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    • v.17 no.2
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    • pp.58-64
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    • 1979
  • Studies on some properties of cellulase isolated from Piricularia oryzae. Crude cellulases were prepared from dried rice plant powder (Tong-il, Pal-dal) culture of P. oryzae(N-2, C-8, T-2). The best yield of enzyme was obtained from the medium using Tong-il rice plant powder for P. oryzae cav. N-2 and 2%-sucrose concentration in preculture media. Two units of the enzyme were incubated at $60^{\circ}C$ for 1 hour with 1.0ml, 0.6% Na-CMC. The optimum temperature for the enzyme activity was at $60^{\circ}C$ and the optimum pH was at pH4.0. When Na-CMC was used as substrate the $K_m$ values of crude enzyme were calculated to be $1.05{\times}10^{-4}\;mM\;and\;V_{max}$ was 2.8 mmole/hour. A 10-fold partial purification was achieved by $(NH_4)_2SO_4$ precipitation followed by column chromatography on DEAE Sephadex A-25.

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Solid-State Fermentation of Rice by Monascus Purpureus

  • Lucas, Juergen;Schumacher, Jens;Kunz, Benno
    • Proceedings of the Korean Society of Food and Cookery Science Conference
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    • 1993.05a
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    • pp.149-159
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    • 1993
  • The concept of Solid-State Fermentation is briefly explained in comparison to other fermentation principles, and several types of fermenters are presented. A recently developed "Swing Reactor" for SSF is shown. When inoculated on rice, the mould Monascus purpureus forms red pigments, Which can be used as food colors (Ang-kak, Red Rice). By Response Surface Methodology, serveral factors have been optimized for maximal red colour formation. Showing that presoaking time of rice, pH of soaking water, age of preculture and inoculum size were not of importance within the observed limits. For a fermentation time of 7 days, start humidity is optimal at 34% and temperature is optimal at 28.8 C. These results of small scale fermentation could be transferred to the Swing Reactor.

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Effect of Phosphatase Activity on Product of $\beta$-galactosidase by E. coli P90C (Phosphatase 의 활성이 E. coli P90C의 $\beta$-galactosidase생성에 미치는 영향)

  • 최동원;심창환
    • The Korean Journal of Food And Nutrition
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    • v.8 no.1
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    • pp.1-5
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    • 1995
  • ATPase was the most available phosphatase in culture broth of 5. coli P90C. To measure the stable phosphatase activity it was necessary to react nth reaction reagent over 30 min and then we can get stable optical density at 410 m. Transfer time from preculture to main culture for the production of $\beta$-glactosidase was good after 3 hrs cultivation. Phosphatase activity was highest at log phase in main culture and as the cell begins to make $\beta$-galactosidase phosphatase activity begins to decrease.

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