• Korean Journal of Organic Agriculture
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• v.29 no.4
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• pp.561-574
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• 2021

This study was conducted to evaluate the control efficacy of the organic farming materials (OFMs) on tuber rot of Gastrodia elata caused by Fusarium spp. The antifungal activities in vitro as well as the suppressive effect of 15 OFMs on the spore germination and germ tube growth by inoculating spore suspension on immature tubers in vivo were investigated. 7 OFMs inhibited the mycelial growth of Fusarium spp. and 7 of them were microbial agents. In the screening using immature tubers, 3 OFMs were very effective with control efficacy value of 70%. Among them, sulfur provided suppressive effect on both mycelial growth and spore germination against tuber rot of G. elata. Finally, 3 OFMs were selected to test the protective and curative effects, and all chosen OFMs significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, sulfur and Bacillus subtilis gave excellent protective control efficacy with control values of 93.2% and 86.9%, respectively, whereas its curative control effect was relatively low (73.3%, 60.2%). On the other hand, the preventive and curative effects of citronella + paraffin oil + ethyl alcohol were 73.3% and 67.0%, respectively. This study suggests that tuber rot of G. elata can be controlled by some OFMs in the rain shelter greenhouse under continuous cultivation condition and protective treatment is more important and efficient.

• Horticultural Science & Technology
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• v.19 no.1
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• pp.17-21
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• 2001

To develop a selection system for regenerating plants from transformed tissues, effects of four antibiotics (kanamycin, hygromycin, carbenicillin, cefotaxime) and herbicide (phosphinotricin) on shoot regeneration from cotyledon and hypocotyl explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis) were studied. For cotyledon, shoot induction was not significantly affected by kanamycin at $1mg{\cdot}L^{-1}$, but the number of shoots formed was significantly reduced at $2mg{\cdot}L^{-1}$, and no shoots were regenerated from any explants at $6mg{\cdot}L^{-1}$ or higher. Hypocotyl explants showed similar result as cotyledon. Kanamycin at $7mg{\cdot}L^{-1}$ may be adequate for selecting Chinese cabbage transformants. Hygromycin at $4mg{\cdot}L^{-1}$ or higher completely inhibited the growth and shoot regeneration of Chinese cabbage explants. Therefore, resistance gene to hygromycin may also be used as a selective marker for Chinese cabbage transformation. Carbenicillin and cefotaxime, the cephalosporin type of antibiotics, had little effect on shoot regeneration of Chinese cabbage explants. Since carbenicillin and cefotaxime have low toxicity to Chinese cabbage, they are suitable for use in tissue culture to eliminate Agrobacterium in transformation experiments after co-cultivation. Shoot regeneration from cotyledon and hypocotyl explants was significantly reduced in presence of $1mg{\cdot}L^{-1}$ phosphinotricin (PPT) and completely inhibited by $2mg{\cdot}L^{-1}$ or higher. PPT, same as antibiotics, may also be used to select transformed cells. Since Chinese cabbage is known to be recalcitrant to in vitro shoot regeneration compared to other Brassica species, even though lower levels of selectable markers result in more transformants but simultaneously allow more untransformed escapes to develop, lower levels of antibiotics and herbicides could be successfully used as a selectable marker to reduce selection pressure.

• The Korean Journal of Mycology
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• v.46 no.4
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• pp.479-490
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• 2018

Red pepper is seriously damaged by thrips (Thrips palmi) and anthracnose caused by Colletotrichum acutatum throughout its development. Because of biotic constraints, producers often depend on chemicals that are expensive and have adverse effects on the environment, operator, and beneficial insects. In addition, resistance is developed because of the repeated use of chemicals. In recent decades, the use of microorganisms in crop protection has become a credible alternative because it is eco-friendly. In this study, we aimed to select isolates with insecticidal and fungicidal activities against the pathogens that cause anthracnose and thrips. We treated T. palmi adults and juveniles with 13 strains of entomopathogenic fungi (isolated from the soil by using the insect-bait method), and 6 strains showed excellent insecticidal activity (70-100%) 5 days after the treatment. The selected isolates were cultured with C. acutatum to screen for the strain with excellent anti-fungal activities, among which an isolate FT333 showed more than 95% control efficacy against C. acutatum in vitro. The isolate was identified as Isaria javanica through its morphological characteristics and phylogenetic analysis of the ITS and ${\beta}-tubulin$ nucleotide sequences. The Isaria javanica FT333 isolate could be used effectively for dual bio-control of thrips and anthracnose during red pepper cultivation.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.1
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• pp.39-46
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• 2011

This study was carried out to determine the forage yield, quality, and regrowth of Italian ryegrass (IRG) and barley sown on 2 March 2009 in Suwon. The five treatments were two IRG cultivars (Kowinearly with early maturity and Kowinmaster with medium maturity), one barley cultivar (Yuyeon), and two mixtures (Kowinearly + Yuyeon and Kowinmaster + Yuyeon). The first harvest date was 26 May, which was at late heading, heading and early dough stage of Kowinearly, Kowinmaster and barley, respectively. Regrowth yield was investigated on 29 June. The heading dates of Kowinearly and Kowinmaster were 16 May and 22 May, respectively, and that of barley was 13 May. The dry matter (DM) percentage were 13.0~18.4% at first harvest, and 22.5~24.8% at regrowth in all treatments. The forage yield of barley and Kowinmaster + Yuyeon mixture at first harvest was higher than that of IRG (p<0.05), but higher regrowth yield was observed in IRG, and then IRG + barley mixtures (p<0.05). The crude protein (CP) content and in vitro DM digestibility (IVDMD) of IRG at first harvest were 16.7~17.1% and 78.3~80.4%, respectively, which were higher than those of barley (CP 12.2% and IVDMD 72.6%) and IRG + barley mixtures. The total yields of DM, CP and digestible DM were high in Kowinmaster + Yuyeon mixture as 11,628 kg, 1,669 kg and 8,457 kg per ha, respectively. In conclusion, spring seeding of IRG + barley mixtures and/or barley were recommended when early harvest. Regrowth of IRG sown in early spring was vigorous. Mixture cultivation of IRG and barley was effective, because of forage yield and stable production, and harvest at June instead of May was desirable for forage productivity of spring sown IRG and barley.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.151-157
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• 2003

Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS tells is 2.↑ $\times$ 10$^3$cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA(0.02%) solution for 15 min at 37$^{\circ}C$, however, our modified method was able to obtain about 6.9 $\times$ 10$^3$cel1s/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 $\times$ 10$^{7}$ cells was obtained in a serum-free medium. while a modified E-medium with mitomycin C-treated feeder tells produced a total of 6.3 $\times$ 10$^{7}$ Cel1s over 17 days When Starting With 7.5 $\times$ 10$^4$cells. Finally, We Confirmed the effectiveness of our ORS tell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro

• The Korean Journal of Mycology
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• v.41 no.4
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• pp.248-254
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• 2013

A Gram-negative bacterium was isolated from mushroom media that markedly reduces the level of extracellular toxins (i.e., tolaasins) produced by Pseudomonas tolaasii, the most destructive pathogen of cultivated mushrooms. The HC1 strain was selected as detoxifying tolaasin by bioassay on potato and it was identified Pseudomonas sp. by the cultural, morphological and physiological characteristics, and analysis of the 16S rRNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for P. tolaasii cell, was sufficient for detoxification in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Pleurotus ostreatus, Flammunia velutipes and Agaricus bisporus. Control efficacy of brown blotch of strain HC1 treatment was 69, 68 and 55% on Agaricus bisporus, Flammulina velutipes and Pleurotus ostreatus, respectively. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by P. tolaasii.

• Korean Journal of Plant Resources
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• v.13 no.1
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• pp.41-47
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• 2000

This study was conducted to evaluate development of seed pellet technique such as pellet polymer search, the shape formation and hardness, the germination and emergence rate of the pellet seeds for labor-saving and reducing production cost in onion(Allium cepa L.) cultivation. The pellet seeds shape formation was good such as kaolin, clay, ash, and gypsum, and clay was good shape formation but surface of pellet seed was cracked during the drying. PG(pearlite + gypsum) as pellet material and PVA as binder were the best among the material in consideration with shape and hardness together. The hardness of the pellet seeds was affected by polymers, the kinds and concentration of binders, and that degree was large at polymer. The high hardness polymers were gypsum and coal ash, but burned lime was the lowest hardness among the pellet material. The germination(GP) and emergence percentage(EP) of pellet seed with PG in vitro were the highest among the material, and that was 93.6, 91.8%, respectively. The EP of pellet seed with PG at 20, $25^{\circ}C$ were 91.3, 92.0%, respectively, The EP of pellet seed were over the 91%, and those did not show difference with field moisture capacity , and that of 5 and 6mm size seed were the highest as 92%, respectively. and other size seeds showed over 90%, too. The EP of pellet seed with PG was decreasing as increasing the sowing depth, and that of at 10mm sowing depth was the highest as 92.7%. The time to 50% emergence of that under 70% field moisture capacity was 158h, and that was delayed at 20h compare to control seed.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.163-169
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• 1999

This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.3
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• pp.261-268
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• 2011

This study was carried out to determine the forage production and quality of Italian ryegrass (IRG) and forage barley developed by Korea in Suwon, 2009~'10. The nine treatments were two IRG varieties (Kowinearly with early maturity and Kowinmaster with medium maturity), five barley varieties (Youngyang, Wooho, Yuyeon, Dami and Youho), and two mixtures (Kowinearly + Yuyeon and Kowinmaster + Yuyeon). The heading dates of Kowinearly and Kowinmaster were 14 May and 18 May, respectively. The growth stage of barley investigated at 22 May were late milk in Youngyang and Wooho, early dough in Dami and early to medium dough in Yuyeon and Youho. Plant length of IRG in IRG + barley mixtures was 117~118 cm, which was longer than those of IRG monoculture of 98~101 cm, and no lodging was found in mixtures. The dry matter (DM) percentage at harvest was 20.7~25.4% in all treatments. The botanical composition of IRG in mixtures was 43.1%. The percentage of spike per barley plant was become high according to progressed maturity, as a 35.7%, 44.1%, 54.8% and 57.2% in late milk, dough, yellowish and full ripeness stage, respectively, and the spike percentages of Youngyang and Wooho were tends to high. The crude protein (CP) content of IRG as 9.0~10.0% was higher than that of barley (7.0~8.5%), and the contents of NDF and ADF of barley were lower than those of IRG, and in vitro DM digestibility were 64.4% in Kowinearly, 64.1% in Kowinmaster, 64.5% in mixture, and 60.2% (Youho) to 66.4% (Wooho) in barley. The yields of DM, CP and in vitro digestible DM were high in Kowinmaster+barley mixture as a 11,508 kg, 1,046 kg and 7,422 kg per ha, respectively (p<0.05). However, no significant differences in forage yield were observed among cultivar of IRG, and barley, although Wooho was tends to high in digestibility and forage yield among five barley varieties. In conclusion, the mixture cultivation of IRG Kowinmaster + forage barley was recommended, because of preventing of IRG lodging, higher plant length of IRG, increasing of forage yield, and stable production. Selection of suitable winter forage species and variety for district, climate environment, and utilization type of farm was also important.