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Optimization of particle gun-mediated transformation system in Cymbidium (유전자총을 이용한 형질전환 심비디움 식물체 생산체계 최적화)

  • Noh, Hee-Sun;Kim, Mi-Seon;Lee, Yu-Mi;Lee, Yi-Rae;Lee, Sang-Il;Kim, Jong-Bo
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.293-300
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    • 2011
  • This study is conducted to develop an efficient transformation system via particle bombardment with PLBs (Protocorm-like bodies) in Cymbidium. For this, pCAMBIA3301 vector which carries a herbicide-resistant bar gene and gus gene as a reporter gene was used for transformation with Cymbidium cultivars 'Youngflower ${\times}$ masako' line. To select transformants, proper concentration of herbicide, PPT (phosphinotricin), should be determined. As a result, 5 mg/l of PPT was selected as a proper concentration. Further, proper conditions for particle bombardment were determined to obtain a high frequency of transformation. Results showed that 1.0 ${\mu}g$ of DNA concentration, 1,100 and 1,350 psi for helium gas pressure, 1.0 ${\mu}m$ of gold particle and 6 cm of target distance showed the best result for the particle bombardment experiment. Also, pre-treatment with combination 0.2 M sorbitol and 0.2 M mannitol for 4 hrs prior to genetic transformation increased the transformation efficiency up to 2.5 times. Using transformation system developed in this study, 3.2 ~ 4.0 transgenic cymbidium plants can be produced from 100 bombarded PLBs on average. Putative transgenic plants produced in this system confirmed the presence of the bar gene by PCR analysis. Also, leaves from randomely selected five transgenic lines were applied for Basta solution (0.5% v/v) to check the resistance to the PPT herbicide. As a result, three of them showed resistance and one of them showed the strongest resistance with the maintenance of green color as non-transformed plants showed. Using this established transformation system, more genes of interests can be introduced into Cymbidium plants by genetic transformation in the future.

Improvement of Green-up of Zoysiagrass and Cool-season Grass during Early Spring in Korea (한국잔디 및 한지형 잔디의 초봄 그린업 촉진)

  • Lee Jae-Pil;Kim Doo-Hwan
    • Asian Journal of Turfgrass Science
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    • v.19 no.2
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    • pp.103-113
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    • 2005
  • This study was conducted to improve the green up of zoysiagrass(2. japonica) and cool-season grass($80\%$ Kentucky bluegrass+$20\%$ Perennial ryegrass) during early spring in Korea. Treatments fur zoysiagrass were control, Polyethylene film, Black screen, Black screen+polyethylene film, Green screen+polyethylene film, Polyethylene film+Black screen, Polyethylene film+Green screen, low mowing height(1.5-2cm) and homing. For cool-season grass, non-punched Polyethylene film, punched Polyethylene film treatments were included. Application dates of covering with Polyethylene film were Feb. 22, Feb. 28, March 7, and March 14. Green up was evaluated by visual color rating. The results are as follows; 1. The best method for improving green up of zoysiagrass were Polyethylene film and optimal covering day for zoysiagrass was on Feb. 22. 2. Low mowing height(1.5-2cm) and burning of zoysiagrass showed the faster greening 1$\sim$weeks before than control. .3 Non-punched Polyethylene film covering was best to improve green up of cool-season grass. More time of covering time with cool-season grass induces rapid green up.

Morphological Study on the Epithelial Cells in the Gall Blader of Vertebrates (각급 척추동물의 담낭 상피세포에 대한 형태학적 연구)

  • 노용태
    • The Korean Journal of Zoology
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    • v.17 no.2
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    • pp.57-68
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    • 1974
  • These experiments were performed in order to study histologically and histochemically on the epithelial cells of gall bladder in Carassius carassius, Bufo bufo gargarizans, Natrix tigrina lateralis, Urloncha striata var. domesticus and Bos taurus var. domesticus. The results of the observation were as follows: 1. There were different cell types in the epithelium of gall bladder in each animal and it could not be supported histochemically that the epithelia cells of gall bladder were divided into two cell types of the rod-shaped and barrel-shaped ones. 2. The epithelium of gall bladder in Carassius carassius, Bufo bufo gargarizans, Natrix tigrina lateralis, Urloncha striata var. domesticus and Bos taurus var. domesticus was simple columnar epithelium. 3. The eosinophilities of cytoplasm in the epithelial cells of gall bladder were in uniform stronger in the upper portion of nucleus in Carassius carassius, Bufo bufo gargarizans and Natrix tigrina lateralis than its other portions, and in Urloncha striata var. domesticus and Lepus cuniculus var. domesticus existed uniformly in all portions, but there were many non-eosinophilic cells in Bufo bufo gargarizans and many cells that weakly eosinophilic around nucleus in Bos taurus var. domesticus. 4. The periodic acid Schiff's reactivities in the epithelial cells of gall bladder were different in each other and the epithelial cells in PAS reaction were divided into two cell types of the dark and light ones. There presented the light cells of 6.4%, 4.3% and 3.7% of epithelial cell of gall bladder in Carassius carassius, Bufo bufo gargarizans and Urloncha striata var. domesticus for each other, but were not presented in Natrix tigrina lateralis and Bos taurus var. domesticus. 5. The ninhydrin-Schiff-active proteins were much in the epithelial cells of gall bladder in Bos taurus var. domesticus, Carassius carassius, Urloncha striata var. domesticus and Natrix tigrina lateralis in order and were much in epithelial cells in the upper portion of mucosal folds in Carassius carassius, Urloncha striata var. domesticus and Natrix tigrina lateralis, and the ninhydrin-Schiff-active protein of the epithelium of gall bladder in Bufo bufo gargarizans was uniformly distributed. 6. The epithelial cells of gall bladder in Carassius carassius, Natrix tigrina lateralis, Urloncha striata var. domesticus and Bos taurus var. domesticus had no stain reactivity or weak stain reactivity to neutral fat and all epithelial cells in Bufo bufo gargarizans had strong stain reactivity, though they were different in quantity of epithelial cell portion. 7. The stain reactivities to RNA and DNA were stronger in the epithelial cells of the upper portion of mucosal fold than in those of other portions.

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Breeding status and nest site characteristics of Black-faced Spoonbills Platalea minor on Chilsando Islands, Korea (칠산도의 저어새 번식 현황과 둥지장소 특성)

  • Kwon, In-Ki;Kang, Jung-Hoon;Lee, Ki-Sup;Lee, Ji-Yeon;Kim, In-Kyu;Yoo, Jeong-Chil
    • Korean Journal of Environment and Ecology
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    • v.29 no.5
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    • pp.703-709
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    • 2015
  • A breeding pair of the Black-faced Spoonbill Platalea minor was firstly recorded on Chilsando Islands, Younggwang, Jeollanamdo Province in 1991. Since the mid 2000s, breeding population on the breeding sites has gradually increased. This study was conducted to identify breeding status and nest site characteristics of the species from May to August, 2013 on Chilsando Islands. We recorded number of nests, length and width of the nest base, slope around the nests, nest materials, distances from the nearest nest, presence of nest cover and nesting area. In 2013 breeding season, 25 of 49 nests produced at least one successful fledging. A total of 55 youngs were successfully fledged and number of fledging per nest was 2.20 individuals. Nesting area was $77.8m^2$ and $93.4m^2$ for Sansando and Yuksando Islet, respectively. Soil and soil mixed with tree root were preferred for substrate of nest base over rock and Brassica napus was dominantly selected as nest materials by Black-faced Spoonbills. Nest characteristics of 22 nests in Sasando and Yuksando Islet varied $49.59{\pm}6.53cm$(mean${\pm}$SD) for length of nest base, $41.00{\pm}5.82cm$ for width of nest base, $20.85{\pm}9.96^{\circ}$ for slope above the nest, $34.09{\pm}17.75^{\circ}$ for slope below the nest and $130.82{\pm}84.17cm$ for distances from the nearest nest. Fifteen pairs (68.2%) occupied where nest cover existed. Nest cover were located in front of the nest for 5 pairs, back of the nest for 9 pairs and both front and back of the nest for 1 pair.

Evaluation of Germplasm and Development of SSR Markers for Marker-assisted Backcross in Tomato (분자마커 이용 여교잡 육종을 위한 토마토 유전자원 평가 및 SSR 마커 개발)

  • Hwang, Ji-Hyun;Kim, Hyuk-Jun;Chae, Young;Choi, Hak-Soon;Kim, Myung-Kwon;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.30 no.5
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    • pp.557-567
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    • 2012
  • This study was conducted to achieve basal information for the development of tomato cultivars with disease resistances through marker-assisted backcross (MAB). Ten inbred lines with TYLCV, late blight, bacterial wilt, or powdery mildew resistance and four adapted inbred lines with superior horticultural traits were collected, which can be useful as the donor parents and recurrent parents in MAB, respectively. Inbred lines collected were evaluated by molecular markers and bioassay for confirming their disease resistances. To develop DNA markers for selecting recurrent parent genome (background selection) in MAB, a total of 108 simple sequence repeat (SSR) primer sets (nine per chromosome at average) were selected from the tomato reference genetic maps posted on SOL Genomics Network. Genetic similarity and relationships among the inbred lines were assessed using a total of 303 polymorphic SSR markers. Similarity coefficient ranged from 0.33 to 0.80; the highest similarity coefficient (0.80) was found between bacterial wilt-resistant donor lines '10BA333' and '10BA424', and the lowest (0.33) between a late blight resistant-wild species L3708 (S. pimpinelliforium L.) and '10BA424'. UPGMA analysis grouped the inbred lines into three clusters based on the similarity coefficient 0.58. Most of the donor lines of the same resistance were closely related, indicating the possibility that these lines were developed using a common resistance source. Parent combinations (donor parent ${\times}$ recurrent parent) showing appropriate levels of genetic distance and SSR marker polymorphism for MAB were selected based on the dendrogram. These combinations included 'TYR1' ${\times}$ 'RPL1' for TYLCV, '10BA333' or '10BA424' ${\times}$ 'RPL2' for bacterial wilt, and 'KNU12' ${\times}$ 'AV107-4' or 'RPL2' for powdery mildew. For late blight, the wild species resistant line 'L3708' was distantly related to all recurrent parental lines, and a suitable parent combination for MAB was 'L3708' ${\times}$ 'AV107-4', which showed a similarity coefficient of 0.41 and 45 polymorphic SSR markers.

Analysis of the Genetic Diversity of Radish Germplasm through SSR Markers Derived from Chinese Cabbage (배추 SSR 마커를 이용한 무의 육성 계통 및 수집종의 유전적 다양성 분석)

  • Park, Suhyoung;Choi, Su Ryun;Lee, Jung-Soo;Nguyen, Van Dan;Kim, Sunggil;Lim, Yong Pyo
    • Horticultural Science & Technology
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    • v.31 no.4
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    • pp.457-466
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    • 2013
  • Since the early 1980s, the National Institute of Horticultural & Herbal Sciences has been breeding and collecting diverse radish breeds to select those samples with better horticultural characteristics, to ultimately expand and develop as good radish produce. Genetic diversity is a crucial factor in crop improvement and therefore it is very important to obtain various variations through sample collection. The collected samples were compared with one another in order to assess the level of diversity among the collections, and this procedure allowed for increased application of the gathered resources and aided in determining the direction to secure further samples. Towards this end, this experiment was conducted in order to examine whether the SSR markers derived from Chinese cabbage samples could be transferred to the radish samples. Among the radish breeding lines and introduced resources, 44 lines were used as materials to analyze the genotype using 22 SSR markers selected. As a result, the analysis showed that among all the selected markers, 'cnu_m139' and 'cnu_m289' were the most useful markers for diversity evaluation. The genetic relationship of the radish genetic resources showed that the geographic origins affected the diversity. Furthermore, the different types of radish groups were also determined by the year they were bred. This result demonstrated that there are differences between the older radish breeds and the more recently developed radish breeds. Even though a relatively small number of markers were used in the analysis, it was possible to distinguish whether the radish was bred 30 years ago or in the 2000s, and that the similar physical shapes comprised a particular group, showed that the SSR markers can indeed be successfully applied to to study the diversity within radish breeding lines. Through the results of this study, it can be concluded that the SSR marker developed for the Chinese cabbage can be applied to examine the genetic diversity and analyze the relationship (genetic resource determination) of radish.

The Study on the Dilution Time of Radioactive Tracer in Estradiol Measurement (Estradiol 검사 시 방사성 추적자의 희석시간에 대한 고찰)

  • Lee, Hae Yeon;Seo, Han Kyung;Jang, Yi Sun;Kim, Hee Jeoung
    • The Korean Journal of Nuclear Medicine Technology
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    • v.21 no.2
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    • pp.44-48
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    • 2017
  • Purpose Estradiol (E2) is a steroid hormone mainly produced in women and is a useful indicator for diagnosis of gynecological diseases, menstrual cycle, menopause, and precocious puberty. E2 measurement is performed by diluting the $^{125}I$ radioactive tracer and tracer buffer in the kit. However, It was not precisely specified when the period of tracer is available after activating. The purpose of this study was to determine the appropriate dilution time based on the measurement value with dilution time. Materials and Methods From December 2016 to February 2017, 60 E2 samples with concentrations ranging from 8 to 4577 pg/mL were divided into low, medium, and high concentrations. Dilution of the $^{125}I$ tracer was performed on a 230 RPM agitator for 30 minutes, 1 hour 30 minutes, and 2 hours 30 minutes, respectively. 24 hour dilution was gently shaken and refrigerated. To verify the difference and significance of the results according to the dilution time, a test of normality was performed using SPSS 18.0 and analyzed by Kruskal-Wallis test. The measured value according to the dilution time was compared with the interquartile range of the absolute error. Results The results of Kruskal-Wallis test were not significant (P>0.05). Measurement results are showed as interquartile range of absolute error. At low concentration, it is 0.052 between 1 hour 30 minutes and 2 hours 30 minutes, and 0.105 between 30 minutes and 1 hour 30 minutes. At medium concentration, 0.062 between 30 minutes and 1 hour 30 minutes, and 0.038 between 1 hour 30 minutes and 2 hours 30 minutes. At high concentration, it is 0.029 between 1 hour 30 minutes and 2 hours 30 minutes, and 0.06 between 2 hours 30 minutes and 24 hours. Conclusion There were no statistically significant differences. However, the change in the measured value is the smallest between 1 hour and 30 minutes to 2 hours and 30 minutes. Therefore, we recommend diluting time between 1 hour 30 minutes and 2 hours 30 minutes.

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A Quality Assurance Study for the Application of Cook/Chill System in School Foodservice Operation (II) - Pork Bulgogi (Broiled Sliced Pork with Sauces) - (학교급식에 Cook/Chill System 적용을 위한 품질보증연구(II) - 돼지불고기 -)

  • Kwak, Tong-Kyung;Moon, Hye-Kyung;Park, Hye-Won;Hong, Wan-Soo;Ryu, Kyung;Chang, Hye-Ja;Kim, Sung-Hee;Choi, Eun-Jung
    • Journal of Food Hygiene and Safety
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    • v.13 no.4
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    • pp.319-331
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    • 1998
  • The purposes of this study were to develop Hazard Analysis Critical Control Point plan applicable to cook/chilled Pork Bulkogi (broiled sliced pork with sauces) in school foodservice operations and to establish reasonable shelf-life limits by assessing food quality during chilled storage period of 5 days. During the product flow, time-temperature profile was recorded and microbiological analyses including mesophilic and psychrotrophic total plate counts, coliform, and fecal coliform and qualitative analyses of Salmonella and Listeria monocytogenes were done. Chemical analyses (pH, acid value, total volatile basic nitrogen), sensory evaluation, and quantitative analysis of thiamin were conducted for 5 days of chilled storage. The number of mesophiles in raw pork ($4.26{\pm}0.11\;Log\;CFU/g$), seasoning mixture ($5.97{\pm}O.04\;Log\;CFU/g$) and marinated pork ($5.56{\pm}0.21\;Log\;CFU/g$) were below the microbial standards for "requires further cooking" food items. Listeria monocytogenes was detected in seasoning mixture. After heating, the number of mesophiles ($5.17{\pm}0.04\;Log\;CFU/g$) were slightly reduced but it did not meet the microbial guidelines of $5\;Log\;CFU/g$ for "ready-to-eat" foods. No other microbes including pathogens were detected. By reheating the menu item after chilled storage, the number of mesophiles were reduced in every phase of 1st day ($4.62{\pm}0.22\;Log\;CFU/g$), 3rd day ($4.55{\pm}0.20\;Log\;CFU/g$) and 5th day ($4.25{\pm}0.16\;Log\;CFU/g$) of chilled storage, and the number of microbes was below the standard limits for "ready-to-eat" foods. At the fifth day of chilled storage, pH (p<0.05), acid value (p<0.01) and TVBN (p<0.05) showed significant increases. Sensory evaluation results did not show any significant change for 5 days of chilled storage. Thiamin content showed a decrease for 5 days of chilled storage. Consequently, the ideal shelflife recommended for Pork Bulkogi was within 3 days of chilled storage. CCPs for Pork Bulkogi were purchasing and receiving of raw meat and some seasoning ingredients, heating, chilling, chilled storage, reheating, and distribution.

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Evaluation of Microbiological Safety of Food Service Environment in Child Care Centers (어린이집 급식환경의 미생물학적 안전성 평가)

  • Lee, Han-Cheol;Jun, Se-Young;Ha, Heon-Ho;Song, Ju-Seok;Lee, Young-Ju;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.35 no.2
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    • pp.146-151
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    • 2020
  • This study investigated microbiological safety of employees' hands, dining tables, and indoor air in cooking areas and lunchrooms in child care centers. Microbiological tests were performed according to the Korea Food Code. Total numbers of aerobic bacteria and coliform bacteria were measured as 5.8±1.9 log CFU/hand and 4.0±2.4 log CFU/hand on employees' hands, and 4.3±3.0 log CFU/100 ㎠ and 2.6±3.3 log CFU/100 ㎠ on dining tables. Bacillus cereus were detected in two cases each of employees' hands and dining tables, respectively. The analysis of microbiological contamination of indoor air in chid care centers showed that the total numbers of aerobic bacteria and coliform bacterial were 28±7.2 CFU/plate and 3.1±2.9 CFU/plate, respectively. Bacillus cereus and Staphylococcus aureus were counted as 1.7±0.2 CFU/plate and 1.6±0.5 CFU/plate from the indoor air in child cate centers. These results indicate that indoor-air in child care centers is considered more safe compared to previous reports. In conclusion, it is necessary to carry out hygienic management using alcohol-based disinfectants before meals to remove microorganism contamination on dining tables and hands. In order to reduce microbial contamination in indoor air, it is also deemed necessary to freshen the sanitary caps, masks, and clothing of the catering staff with periodic ventilation of indoor air.

Study on the Quality Properties of Hardtack Added with Acorn Jelly Powder and Acorn Ethanol Extract (도토리묵가루 및 추출물을 첨가한 건빵의 품질특성)

  • Lee, Won-Kyoung;Kim, Seung-Hee;Choi, Chang-Suk;Cho, Soo-Muk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.3
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    • pp.376-382
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    • 2012
  • This study examined the effects of acorn (Quercus. acutissima CARR.) jelly powder (0%, 25%, 50%, and 75%) and acorn extract (0%, 0.1%, 0.5%, and 1.0%) addition on the quality characteristics of hardtack. In the sensory test, acorn jelly powder added group scored 50% higher than the other added group. Regarding hardtack color, L (lightness) and b (yellowness) values decreased with increasing acorn powder and acorn extract addition. Hardness of hardtack increased with added acorn jelly powder, but no significant difference was observed with acorn extract. The taste and texture of the hardtack 0.1% acorn extract added group significantly increased. Overall, preferences decreased with increasing acorn extract but not significantly. In conclusion, the results of this study suggest that addition of 50% acorn jelly powder in combination with addition of less than 0.1% acorn extract to hardtack was the most desirable.