• Korean Journal of Plant Resources
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• v.36 no.6
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• pp.571-579
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• 2023

In this study, cryopreservation by droplet-vitrification was applied to pear (Pyrus spp.) germplasm. We focused on the development and assessment of various strategies for the selection of suitable tissue, osmoprotection, and dehydration. We also evaluated post-thaw recovery of cryopreserved explants by droplet-vitrification. Preferentially, we tested the effects of preculture and loading treatments to determine which tissues were more suitable, either the apical shoot tips or the axillary buds. Apical shoot tips showed the better regrowth rate than in vitro axillary buds. The most effective techniques for cryopreservation were as follows. Shoots from in vitro seedlings which had been cultured for about 5-6 weeks were cold-hardened at 4℃ for one week, excised shoot tips were precultured on liquid MS medium including 0.3 M sucrose for 31 hours and 0.7 M sucrose for 17 hours, osmoprotected in loading solution (LS) for 40 min, and then cryoprotected in dehydration solution (PVS3) for 90 min. In addition, we found that regrowth rates of explants on regrowth medium after exposure to liquid nitrogen (LN) were higher than those on MS medium. Results indicated that the highest regrowth percentage was 95.6% for 'Bartlett' cultivar and 68.9% for 'BaeYun No.3' cultivar. Consequently, apical shoot tips of two pear cultivars, 'Bartlett' (P. communis) and 'BaeYun No.3' (P. pyrifolia), were successfully cryopreserved by droplet-vitrification. Results of this study show that the enhanced droplet-vitrification method described in the present study could be used as an effective means for long-term storage of pear genetic resources.

• Journal of the Korea Organic Resources Recycling Association
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• v.9 no.1
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• pp.49-55
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• 2001

For the production of probiotic feed enriched with viable yeasts, aerobic liquid culture of Kluyveromyces marxianus was attempted in pulverized residual food wastes. After the preliminary shaking culture result, the liquid food wastes was added with urea($0.5g/{\ell}$), o-phosphate($0.4g/{\ell}$ ), molasses($4g/{\ell}$), and yeast extract($1g/{\ell}$), and the fermentation was carried out in 2-litre jar fermenter. In 12 hours of aerobic mixed culture with Aspersillus oryzae, viable cell count of the yeast reached to the number of $1.4{\times}10^{10}/{\ell}$ in the cultured medium.

• Korean Journal of Food Science and Technology
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• v.42 no.1
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• pp.82-89
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• 2010

To obtain functional materials from a submerged culture of Hericium erinaceum, a suitable basal medium for flask culture was screened and the optimal culture conditions in a jar fermenter were investigated with the addition of ginseng extracts (GE) to the basal liquid medium. Of all tested basal liquid media, the mushroom complete medium (MCM) supplemented with 0.5% of GE produced the highest mycelial dry weight (MDW) of 5.91 g/L in the flask, which reached a plateau at $25^{\circ}C$, pH 5.5 after 10 days. The submerged culture conditions for the mass production of mycelia in a 50 L jar fermenter were also optimal at $25^{\circ}C$, pH 5.5, 120 rpm agitation speed and 0.4 vvm aeration rate. Under these conditions, the maximum MDW was produced, which reached a value of 4.28 g/L within 5 days. When we investigated the effects of the amount of GE in the MCM on the production of MDW in the jar fermenter, the addition of 5% GE (HE-GE-5) under the optimal culture conditions produced the maximum MDW (4.93 g/L). In addition, the crude polysaccharide of HE-GE-5 contained mainly neutral sugars (63.2%) with considerable amounts of uronic acid (19.3%) and a small amount of proteins (8.8%) and it had potent immunostimulation properties.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1327-1332
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• 2004

The hemA gene encoding 5-aminolevulinic acid synthase (ALAS) was cloned from Rhodobacter capsulatus, and its nucleotide sequence was determined. DNA sequencing data revealed one open reading frame coding for a protein with 401 amino acids that displayed high similarity to the amino acid sequences of other known ALASs. The hemA gene was then cloned and expressed under the control of constitutive promotor in E. coli. The recombinant E. coli strain was able to accumulate 5-aminolevulinic acid to 21 mM in the liquid medium supplemented with 45 mM glycine and 120 mM succinate. In addition, a marked effect of the pH of the culture medium on ALA production was observed, and the optimum pH for culture medium was determined to be 5.8-6.3.

• Journal of the Korean Wood Science and Technology
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• v.35 no.5
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• pp.109-117
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• 2007

Methyl trans-cinnamate is a significant flavor compound of Tricholoma matsutake. Attempts were made to produce this compound by culturing the mycelium using submerged culture. No methyl trans-cinnamate could be detected when the mycelium was cultured in the basal liquid medium. However, the addition of Pinus desiflora extracts to the medium, methyl trans-cinnamate was largely produced. To find out compounds or fractions inducing methyl trans-cinnamate, dichloromethane fraction obtained from the wood extracts of P. densiflora was subjected to column chromatography. Three sub-fractions were obtained from the $CH_2CI_2$ fraction. Submerged cultured mycelium treated with sub-fraction 1 has the highest content of methyl trans-cinnamate. Maximum methyl trans-cinnamate $(470.2{\mu}g/g)$ was obtained when the first sub-fraction of dichloromethane fraction of the P. densiflora wood extracts was added to the medium. This indicates that wood extracts of P. densiflora contains inducer of the methyl trans-cinnamate production in the T. matsutake submerged culture.

• Fisheries and Aquatic Sciences
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• v.11 no.3
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• pp.149-158
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• 2008

Labyrinthulids are unicellular heterotrophic marine microalgae. Two labyrinthulid strains, L4 and L75, which produce generous amounts of polyunsaturated fatty acids(PUFAs), were isolated from leaves floating in the coastal areas of Quang Ninh province, Vietnam, and Fukuoka Prefecture, Japan, respectively. Both strains had spindle-shaped cells surrounded by ectoplasmic networks. Numerous oil bodies were observed in each cell, mostly in the vicinity of cell membranes. When soybean oil or soybean lecithin was used as a carbon source with peptone as a nitrogen source, the proportion of PUFAs reached 25-30% and 50-56% for L4 and L75, respectively. After 14 days of growth at $25^{\circ}C$, L4 produced 0.3 mg PUFAs/g-agar in PYA-SBO medium and 0.6 mg PUFAs/g-agar in PYA-SBL medium. In comparison, L75 produced 0.2 mg PUFAs/g-agar in both types of media. The differences between the two strains included changes in cellular morphology and the capacity for attaching tightly to fibers when cultured in liquid PYA medium containing 2% SBL. In addition, when the strains were grown under the same conditions, L4 had a higher growth rate and produced more PUFAs than L75.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.1
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• pp.51-56
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• 1998

A bacteium, KP-6408, capable of hydrolyzing fibrin was isolated from Chungkook-jang, which was possibly identified as a strain of Bacillus sp. The effects of culture condition and medium composition on the enzyme production were investigated. Among nitrogen sources tested, yeast extract was the most effective for the enzyme production, and the level of the concentration for the optimal enzyme production was 0.2%(w/v). For carbon sources, glucose was the best for the enzyme production with the level of 2.0%(w/v). The enzyme was maximally produced by cultivating the enzyme production with the level of 2.0%(w/v). The enzyme was maximally produced by cultivating the organism at the liquid medium of the initial pH 8.0 and temperature of 4$0^{\circ}C$. In Chungkook-jang fermentation, the enzyme was maximally produced when incubated at 35$^{\circ}C$ for 24 hrs using soybean as a solid medium. The addition of various rice starch to the soybean in Chungkook-jang fermentation lowered the enzyme production.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.175-182
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• 2009

Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

• Korean Journal of Animal Reproduction
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• v.13 no.3
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• pp.134-140
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• 1989

Studies were conducted to seek reasonable methods of rapid freezing of mouse embryos using liquid nitrogen. The effects of the cryoprotectants concentration and the substitution of raffinose to sucrose in freezing and dilution medium on mouse embryo survival rates were determined using the FDA-test. The summarized results are as follows : 1. When 0.3M of sucrose was added into the freezing and dilution medium, FDA scores of embryos were 1.48(1.5M), 3.81(3.0M) and 4.10(4.5M). Higher FDA scores of embryos were obtained in 3.0M and 4.5M glycerol concentrations (P<0.05). 2. With the addition of 0.3M raffinose to the freezing and dilution medium, FDA scores of embryos did not significantly differ between glycerol levels ; 3.97(1.5M), 4.11(3.0M) and 3.54(4.5M). Higher scores of embryos existed in 3.0M glycerol concentration. 3. Concentration of sucrose or raffinose in freezing and dilution medium affected FDA scores of embryos. When sucrose concerations of 0.3, 0.5 and 1.0M were added to the freezing medium, FDA scores of embryos were 3.12, 2.38 and 0, respectively. However, when the same concentrations of raffinose were added to freezing medium, the FDA scores were 4.21, 2.91 and 0. In both cases, better FDA scores of embryos were attained in 0.3M of sucrose or raffinose (P<0.01).

• Korean Journal of Optics and Photonics
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• v.19 no.2
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• pp.150-158
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• 2008

The exact transmission coefficients at the interface between a uniaxial anisotropic medium and an isotropic medium at? oblique incidence are derived by applying the extended Jones matrix method. When the birefringence of the uniaxial anisotropic medium is small ($|n_e-n_o|\;{\ll}\;n_o,\;n_e$), the exact transmission coefficients are compared with those by the conventional extended Jones matrix method by Yeh et al. They showed an excellent agreement with each other. In addition, using the exact transmission coefficients, we calculated the polarization characteristics of a light through a uniaxial medium to an incident light with arbitrary polarization state at? oblique incidence. We compared the transmittances of an unpolarized light through a pair of crossed o-type polarizers by two different methods and calculated the transmittance as the variation of the optical constants of the polarizers to evaluate of the extinction ratio. The polarization analysis method using the exact transmission coefficients can be applied to polarization characteristics of a light through a uniaxial medium with large birefringence as well as to liquid crystals and to optical anisotropic material.