• Journal of Korean Society of Forest Science
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• v.89 no.4
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• pp.522-526
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• 2000

We tried to mass-propagate Prunus yedoensis from Jeju through tissue culture. We investigated the effect of bud collection time, the concentration of $NH_4NO_3$ in media and plant growth regulators(BAP, $GA_3$ and IBA) on the differentiation of winter buds. Buds, taken in February, flushed well regardless of various in vitro conditions. Bud flushing rate was significantly different depending on the collection time. BAP appeared to be effective on bud flushing. Sixty percent of buds taken in October flushed on the media containing $3.0mg/{\ell}$ BAR. No buds flushed on the medium supplemented with IBA. Buds, after flushing in BAP media, grew as foliated shoots and showed a rosette of leaves. When $GA_3$ supplemented to the BAP-containing media as a higher concentration than that of BAP, shoots elongated and developed into normal shoots. The combination of BAP $1.0mg/{\ell}$ and $GA_3$ $2.0mg/{\ell}$ is most recommendable for shoot elongation.

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.269-274
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• 2008

The purpose of this study was to determine the effects of Taxol pre-treatment to in vitro matured bovine oocytes, and sucrose and trehalose added to vitrification solution on spindle morphology and embryonic development following cryopreservation. Bovine oocytes were collected from ovaries and matured in tissue culture medium 199 (TCM 199) supplemented with 10% Fetal Bovine Serum (FBS), 0.05ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone and $1{\mu}g/ml$ estradiol for 22h in $39^{\circ}C$, 5% $CO_2$, TCM 199-HEPES containing 20% FBS was used as basic medium (BM) to prepare vitrification solution. Oocytes were pre-treated with $1\;{\mu}M$ Taxol in maturation medium for 15 min prior to vitrification. Oocytes were exposed to 1.6 M ethylene glycol (EG) and 1.3M dimethyl sulfoxide (DMSO) in BM and then were exposed to 3.2 M EG, 2.6 M DMSO and 0.5 M sucrose in BM or 3.2 M EG, 2.6 M DMSO and 0.5 M trehalose in BM. Oocytes with cumulus cells and oocytes without cumulus cells were considered as control 1 and control 2, respectively and held in TCM 199-HEPES at $39^{\circ}C$. Oocytes were frozen using modified solid surface vitrification and were stored in cryotubes in liquid nitrogen for more than 1 week. Frozen oocytes were thawed in TCM 199-HEPES containing 0.5 M, 0.25 M and 0.1 M sucrose in BM for 2 min, respectively or 0.5 M, 0.25 M and 0.1 M trehalose in BM for 2 min, respectively. Immunoflurorescence staining of oocytes was performed to assess spindle morphology and chromosome configuration of oocytes. The rates of cleavage and blastocyst were examined following in vitro fertilization. Normal spindle morphology rate of oocytes pre-treated with Taxol prior to vitrification was not higher than that of other vitrified groups. Taxol pre-treatment did not increase cleavage and blastocyst formation rates, although control groups showed significantly higher rates (p<0.05). Percentages of normal spindle and embryonic development were not significantly different among vitrified groups regardless of type of sugar. In conclusion, Taxol pre-treatment of oocytes before cryopreservation did not reduce the damage induced by vitrification and subsequently did not improve embryonic development following vitrification. Trehalose may be used as an alternative non-permeating cryoprotectant in vitrification solution.

• Journal of Embryo Transfer
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• v.23 no.4
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• pp.263-267
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• 2008

The purpose of this study was to determine toxic effect of sucrose and trehalose prior to cryopreservation on nuclear maturation and embryonic development in immature bovine oocytes. All cryoprotectant was prepared in tissue culture medium 199-HEPES (TCM 199-HEPES) with 10% fetal bovine serum (FBS). Immature oocytes were exposed to 1.2M ethylene glycol (EG) and 0.1M sucrose or 1.2M EG and 0.1M trehalose for 3 min and then were exposed to 3.2 M EG and 0.25 M sucrose or 3.2 M EG and 0.25 M trehalose for 1 min. Oocytes treated with cryoprotectants were exposed to 0.25 M sucrose or 0.25 M trehalose for 5 min and then 0.1 M sucrose or 0.1 M trehalose for 5 min. Depending on type of sugar added to cryopreservation solution, oocytes were allocated to sucrose group and trehalose group, respectively. Oocytes exposed to TCM 199-HEPES with 10% FBS were considered as control. Oocytes were cultured in TCM 199 supplemented with 10% FBS, 5 ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone, and $1\;{\mu}g/ml$ estradiol for 24 h in $39^{\circ}C$, 5% $CO_2$. Nuclear maturation was assessed by staining oocytes with 1% aceto-orcein. Oocytes were fertilized in vitro and were cultured in TCM 199 supplemented with 10% FBS, 5 mM sodium pyruvate, and antibiotics in $39^{\circ}C$, 5% $CO_2$. The rates of cleavage and blastocyst, and cell number in blastocyst were assessed. Metaphase II rates were not different among experimental groups regardless of type of sugar. The cleavage rate of trehalose group (73.3%) was significantly higher (p<0.05) than those of sucrose group (62.8%) and control group (60.8%). The blastocyst rate was significantly higher in trehalose group (p<0.05). Mean cell number in blastocyst were not different among experimental groups, although cell number of blastocyst in trehalose group was significantly higher on day 7 (p<0.05). In conclusion, sucrose and trehalose were not toxic to immature bovine oocytes prior to cryopreservation. In particular, trehalose was more effective on embryonic development.

• Research in Plant Disease
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• v.25 no.4
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• pp.220-225
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• 2019

During the year 2018, the symptoms of bunch rot on Shine Muscat (Vitis vinifera L.) were observed in Kimcheon-si, Gyeongbuk province in Korea. The disease appears on the Shine Muscat as a black rot due to prolific fungal sporulation after it has invaded into the Shine Muscat which look completely empty and dryness. Colonies of these fungi are present on the Shine Muscat skin from fruit setting and increase in amount from early season to harvest, while become peak at ripening stage. To isolate the causal agent, small fragments (2 to 3 mm) of decayed tissue from the lesion margin were placed onto potato dextrose agar (PDA) plates. Fungal colonies on PDA produced dense white aerial mycelium and then covered with dark black conidial heads. These heads were large and radiate, and vesicles were globose (2.12-32.0×2.0-3.1 ㎛). Based on morphological and cultural characteristics, this fungus was identified as Aspergillus tubingensis. To confirm its identity, the internal transcribed spacer, β-tubulin, and RNA polymerase II was sequenced for molecular identification. BLAST search indicated 99% identity with A. tubingensis. The pathogenicity test on healthy grape of Shine Muscat produced bunch rot, as the original symptoms. To select effective fungicides for the control of brunch rot, an in vitro antifungal activity of seven fungicides were evaluated against the growth of A. tubingensis. Five fungicides (dipenoconazole, tebuconazole, metconazole, iminoctadine, and captan) exhibited significantly strong suppression of the mycelial growth of A. tubingensis.

• Archives of Plastic Surgery
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• v.37 no.3
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• pp.227-232
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• 2010

Purpose: Continuous irrigation method is an important step in managing wound infection. V.A.C. devices have been used in intractable wounds for reducing discharge, improving local blood flow, and promoting healthy granulation tissue. We expect synergistic effects of reduced infection and more satisfactory, accelerated wound healing when using both methods simultaneously. This study evaluated continuous irrigation combined with V.A.C. appliance for treatment of infected chronic wounds. Methods: We reviewed data from 17 patients with infected intractable chronic wounds. V.A.C. device (Group A) was used in 9 patients, and V.A.C. with antibiotics irrigation (Group B) was used in 8 patients. We placed Mepitel$^{(R)}$ on the surface of wound and placed an irrigation and aspiration tube on each side. A sponge was placed on the Mepitel$^{(R)}$ and covered with film dressing. The wound was irrigated continuously with mixed antibiotics solution at the speed of 200 cc/hr and aspirated through the wall suction at the pressure of -125 mmHg. V.A.C. applied time, wound culture and wound size were compared between the two groups. Results: No complication were seen in two groups. Compared with Group A, in the Group B, V.A.C. applied time was shortened from 32.7 days to 25.6 days and showed efficacy in the reduction rate of wound size. No statistical differences were shown in bacterial reversion. Conclusion: V.A.C. appliance with continuous irrigation is an effective new method of managing infected chronic wounds and useful to reduce treatment duration and decrease wound size. Moreover it could be applied more widely to infected wound.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.1-5
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• 1998

This study describes the effect of the growth regulators such as 2,L-D and BA, on the structural abnormalities of somatic embryos derived from leaf explants of Angelica gigas Nakai, Also, the relationship between the cotyledon number of a somatic embryo and its germinability is explored. Embryogenic calli were selected from calli formed on explants cultured on MS solid basal medium supplemented with 0.5mg/L 2,4-D, 1mg/L 2,4-D, 1mg/L plus 0.1mg/L BA, and 1 mg/L 2,4-D plus 0.5mg/L BA. Cotyledonary abnormalities were observed in somatic embryos which were developed from embryogenic calli cultured on MS medium containing 1mg/L 2,4-D for 8 weeks and then subcultured on 2,4-D free MS medium for 3 weeks. The frequency of abnormalities was as follows: 22.8% one cotyledon, 42.5% two cotyledons, 16.8% three cotyledons, 7.8% four cotyledons, 1.8% five cotyledons, and 8.2% jar shaped cotyledon. In addition, ABA treatment indicated an improvement of the somatic embryo with normal cotyledon (65.3%). ABA was important role to the high production of normal somatic embryos. Two cotyledon embryos showed germinability 77.8%. However the germinability of somatic embryos with anomalous cotyledons was prominently low: One cotyledon, 62.5%; three cotyledons, 43.3%; four cotyledons, 60%; five cotyledons, 50% and jar shaped cotyledon, zero%. Thus, germinability was essentially, inversely proportional to cotyledon number.

• Korean Journal of Plant Tissue Culture
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• v.21 no.4
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• pp.207-213
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• 1994

In order to study the effect of cyclohexylamine (CHA) and polyamines during adventitious root formation, the correlation of adventitious root formation with endogenous polyamine content was investigated in inoculated soybean cotyledons at the rooting medium supplemented with CHA, spermidine and spermine. Adventitious root formation was inhibited in medium containing 10$^{-3}$ -10$^{-2}$ M CHA. Adventitious root formation was not formed in treatment with 10$^{-2}$ M CHA plus polyamine. It was inhibited in 2.5 $\times$ 10$^{-3}$ M CHA alone and reverved in 2.5 $\times$ 10$^{-3}$ M CHA plus 10$^{-4}$ M spermine. Putrescine content was lower in 10$^{-2}$ M CHA and 10$^{-2}$ M CHAplus 10$^{-4}$ M spermine treatment than in control after 3 days of incubation. However, it was higher in 2.5 $\times$ 10$^{-3}$ M CHA and 2.5 $\times$ 10$^{-3}$ M CHA plus 10$^{-4}$ M spermine treatment than in control. Spermine content was higher in all CHA treatments than in control, while spermidine content was lower than in control.

• Korean Journal of Plant Tissue Culture
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• v.22 no.1
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• pp.47-51
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• 1995

This study was conducted to investigate the possibility of obtaining platelets via somatic embryogenesis as a means of in vivo mass propagation in Paeonia lactiflora Pall.. When cultured on MS medium with 0.5 mg/L 2, 4-D, filament explants formed calli. UPon transfer to basal medium the calli gave rise to somatic embryos at a frequency of 38%. Additional of 3g/L activated charcoal enhanced the frequency to 60%. Mature embryos preheated at 5$^{\circ}C$ for 2 weeks had 37% germination on medium with 0.3 mg/L GA$_3$. The germinated embryos developed to complete plantlet when cultured on medium with 2mg/L BA.

• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.267-271
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• 1995

To improve regeneration efficiency of Brassica campestris ssp. pekinensis (chinese cabbage) in vitro, the effect of ehtylene inhibitors [AgNO$_3$ and silver thiosulfate (STS)] and optimal age of explantse were investigated. On the effect of ethylene inhibitors either 100 $\mu$M of AgNO$_3$ or 5 $\mu$M of STS enhanced shoot regeneration from cotyledons when it was added in basal shoot induction media(MS salts, B5 vitamine, sucrose 2%, BA 2.0mg/L, NAA 1.0mg/L). But at higher concentrations, AgNO$_3$ induced abnormal shoots, and STS greatly reduced regeneration frequency. On the other hand, the maximum regeneration rate was obtained from the cotyledons taken from 3-day old seedlings. However there was no distinctive effect among the containers used for cultivation. The most optimal condition of root induction was a minimal Murashige and Skoog media containing 0.1 mg/L NAA. In order to induce bolting and flowering from in vitro regenerated chinese cabbage, the plant were healed at 4$^{\circ}C$ for weeks in a cold chamber. When they were planted in pots, the plane produced phenotipically normal flowers and seeds. The overall results suggest that ethylene inhibitors promote regeneration of shoot from cotyledons of chinese cabbage without alleviating fertility.

• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.291-298
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• 1995

This study describes plant regeneration from leaf explant of Bupleurum falcatum through somatic embryogenesis, and the effect of 2,4-dichlorophenoxyacetic acid on somatic embryo abnormalities. The relationship between the cotyledon number of somatic embryo and its germinability is also described. Embryogenic calli were selected from calli formed on explants cultured on MS solid basal medium supplemented with 1 mg/L 2,4-D. Cotyledonary abnormalities were observed in somatic embryos which were developed from calli cultured on MS medium with 1 mg/L 2,4-D for 6-week and then subcultured on 2,4-D free MS medium for 3 weeks. The frequency of abnormalities was as follows: 7% of somatic embryos had one cotyledon, 65% of them had two cotyledons, 25% three cotyledons, 5% four cotyledons, 2% five cotyledons, and 3% trumpet-like cotyledons. The two cotyledon somatic embryos were germinated at a frequency of 80%. However the germination frequency of one cotyledon embryo and multicotyledonary embryo was lower than that of the two cotyledon somatic embryo. All of trumper-like somatic embryos did not germinate. Histological observations of multicotyledon embryo showed circular procambium in the root but pocambial strands in the cotyledonary node or upper hypocotyl. The number of the strands was equal to the cotyledon number.