• Title/Summary/Keyword: 3R regeneration

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In vitro Regeneration and Genetic Stability Analysis of the Regenerated Green Plants in Japanese Blood Grass (Imperata cylindrica 'Rubra') (홍띠 기내 재생과 재생 녹색식물체의 유전적 안정성)

  • Kang, In-Jin;Lee, Ye-Jin;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.34 no.2
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    • pp.156-165
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    • 2021
  • The in vitro regeneration was established, and the genetic stability among the mother plants (control) and the micropropagated green plants was evaluated using ISSR markers in Imperata cylindrica 'Rubra', Poaceae which containing important bioenergy plants. Green shoots were multiply induced from growing point culture via callus on MS medium supplemented with 0.01 mg/L NAA and 2 mg/L BA, and the shoots were proliferated on the MS medium with rooting. Rooted plantlets were transplanted to the pot with 100% survival rate. Using ISSR markers, somaclonal variation was analyzed in eight mother plants (control), ten green-regenerant cultivated at culture room (ReR) and ten green-regenerant cultivated at field condition (ReF). All ISSRs produced a total of 97 bands, and the scorable bands varied from one to seven with an average of 4.4 bands per primer. The polymorphism rate of ReRs and ReFs was 4.1% and 3.1% respectively, showing higher rate than that of control (0%). The genetic similarity matrix (GSM) among all accessions ranged from 0.919 to 1.0 with a mean of 0.972. According to the clustering analysis, ReFs and mother plants were divided into two independent groups. The results indicate that no clear genetic diversity was detected among regenerated plants, and ISSR markers were useful tool for identification of somaclonal variation of regenerants.

Effects of Forest Restoration Methods and Stand Structure on Microclimate in Burned Forest Stand (산불 피해지 복원 방법이 임분 내 미세 기후에 미치는 영향)

  • Kim, Jeong Hwan;Lim, Joo-Hoon;Park, Chanwoo;Kwon, Jino;Choi, Hyung Tae
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.17 no.3
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    • pp.207-216
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    • 2015
  • The study was conducted to determine the effects of forest restoration methods and stand structure on solar radiation, air temperature, relative humidity, soil temperature, and soil water content, based on volume, in forest stand after forest fire. The changes of the micro-climate elements in naturally and artificially restored forest after forest fire were measured in Goseong and Samcheok, Gangwon province. Pinus spp. were commonly appeared in ridges, barren lands or planted areas of the study sites while the other areas were dominated by Quercus spp. In the early stage, trees in the naturally regenerated site grow better than the trees in artificially rehabilitated site. However, the growth ratio rapidly decreased by time passed in natural regeneration area. The environmental conditions (solar radiation, air temperature, relative humidity, soil temperature and soil water content) were significantly different by the regions and the methods (p<.05). However, the coefficients of variations of the environmental conditions were not significantly different at 95% confidence level. As the coverage and tree height in crown layer increased, the relative humidity and soil water content were increased while the temperature and solar radiation were decreased. Especially, the relative humidity, solar radiation, and soil water content were clearly affected by the tree height and coverage ratio ($R^2$ means from 0.628 to 0.924). Even though the data should have collected at least more than 5 years in meteorological analysis, the two year results show some clear relationship between forest structure and microclimate elements.

Herbicide Resistant Cabbage (Brassica oleracea ssp. capitata) Plants by Agrobacterium-mediated Transformation

  • Lee, Yeon-Hee;Lee, Seung-Bum;Suh, Suk-Chul;Byun, Myung-Ok;Kim, Ho-Il
    • Journal of Plant Biotechnology
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    • v.2 no.1
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    • pp.35-41
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    • 2000
  • Transgenic cabbage (Brassica oleracea ssp. capitata) plants resistant to the commercial herbicide Bast $a^{R}$ were obtained by Agrobacterium tumefaciens - mediated transformation. Hypocotyl segments of in vitro grown plants were infected with Agrobacterium tumefaciens LBA 4404 harboring plasmid pMOG6-Bar which contains hpt and bar genes. Explants were cultured on callus induction medium (MS basal medium + 1 mg/L NAA + 2 mg/L BA + 2 mg/L AgN $O_3$+ 100 mg/L carbenicillin + 250 mg/L cefotaxime) supplemented with 15 mg/L hygromycin. Hygromycin resistant calluses were transferred to shoot regeneration medium (MS basal medium + 0.1 mg/L NAA + 2 mg/L BA + 3% sucrose + 2 mg/L AgN $O_3$+ 15 mg/L hygromycin + 250 mg/L cefotaxime + 100 mg/L carbenicillin). In order to induce roots, elongated shoots were placed on the MS medium without plant growth regulators and hygromycin. Southern blot analysis of several putative transgenic plants indicated that one to five intact copies of Apt and bar genes were incorporated into the genome. Expression of bar gene was confirmed by Northern blot analysis and by herbicide resistant phenotype. Seed progeny from self-pollinated transformants expressed the herbicide resistance and showed Mendelian segregation of the introduced gene.e.

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Importance of Strain Improvement and Control of Fungal cells Morphology for Enhanced Production of Protein-bound Polysaccharides(β-D-glucan) in Suspended Cultures of Phellinus linteus Mycelia (Phellinus linteus의 균사체 액상배양에서 단백다당체(β-D-glucan)의 생산성 향상을 위한 균주 개량과 배양형태 조절의 중요성)

  • Shin, Woo-Shik;Kwon, Yong Jung;Jeong, Yong-Seob;Chun, Gie-Taek
    • Korean Chemical Engineering Research
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    • v.47 no.2
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    • pp.220-229
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    • 2009
  • Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.

Development of L-Lysine Producing Strains by Intergeneric Protoplast Fusion of Brevibacterium flavum and Corynebacterium glutamicum (Brevibacterium flavum과 Corynebacterium glutamicum의 이속간 원형질체 융합에 의한 L-라이신 생산균주 개발)

  • Kyung, Ki-Cheon;Lim, Bun-Sam;Lee, Se-Yong;Chun, Moon-Jin
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.279-283
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    • 1985
  • As a method of breeding L-lysine producing strains, the intergeneric protoplast fusion between Brevibacterium flavum and Corynebacterium glutamicum was performed. As a results, Brevibacterium flavum ATCC 21528 R showed 99% of protoplast formation and 10% of regeneration frequencies when treated with 400$\mu\textrm{g}$/$m\ell$ of lysozyme for 12hrs. In Corynebacterium glutamicum ATCC 21514 S, 99% and 12% were obtained by treatment of 300$\mu\textrm{g}$/$m\ell$ lysozyme for 12 hrs. In intergeneric protoplast fusion between Brevibacterium flavum ATCC 21528 R and Corynebacterium glutamicum ATCC 21831 S, 1.0$\times$10$^{-6}$ of recombinant frequency per regenerable cells was observed by use of PEG 6000, 30%(w/v). Among the strains obtained KR$_{43}$ strain showed 12% higher productivity of L-lysine than the parental cell. Then, the activity of aspartokinase of KR$_{43}$ was about 13% higher than the parental cell.

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Strategies for the Development of Tourism Resources in the Inland Waters for the Revitalization of Special Tourist Zone - Focus on value frame analysis - (관광특구 활성화를 위한 내수면 관광자원화 전략 -가치프레임 분석을 중심으로-)

  • Cho, Si-Young;Lee, Kwang-Kug;Jun, Jae-Kyoon;Yhang, Wii-Joo
    • The Journal of Fisheries Business Administration
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    • v.50 no.3
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    • pp.59-71
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    • 2019
  • In terms of coastal and marine tourism, Busan's Haeundae had the nation's representativeness, and is pushing for many kinds of related policies to revitalize the special tourism zone. Due to a drop in the number of beach users, it is inevitable for the Haeundae Special Tourist Zone to face active responses to new trends emerging in the global tourism market. Therefore, the purpose of this study is to present a tourism strategy for making the Suyeong River as a more competitive ourist resource along the zone. First, a language network analysis is conducted through interviews to understand the ideas of interest groups for the river cruise activation project. Second, the frame structure of stakeholders is used to analyze solutions by comparing the similarities and differences in recognition frames of interest groups. Third, we intend to analyze the detailed frame types of stakeholders and present new alternatives based on the structure of relationships between types.

A Study on the Anti-wrinkle Properties of Cosmetics Containing Essential Oil from Chrysanthemum boreale MAKINO (산국 에센셜오일을 함유한 화장품의 주름개선 임상효능에 관한 연구)

  • Choi, In Ho;Hwang, Dae Il;Kim, Do Yoon;Kim, Ha Bin;Lee, Hwan Myung
    • Journal of Life Science
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    • v.29 no.4
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    • pp.442-446
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    • 2019
  • We recently reported the characteristics of Chrysanthemum boreale MAKINO essential oil (CBMEO) at different harvesting stages and its in vitro antioxidant, whitening, skin regeneration, and antiwrinkle properties. In the present study, we investigated the effectiveness of cosmetic creams containing 0.1% of CBMEO in terms of wrinkle reduction and safety. The participants were 22 women aged 42 to 60 y with natural wrinkles. All the participants applied the CBMEO-containing cream to the eye area for 8 weeks. Visual observations of the skin by a dermatologist were used to assess the safety of the product, and improvements in skin wrinkles (roughness) were analyzed using a skin visiometer. Neither the CBMEO-containing cream nor the placebo caused abnormal reactions, such as erythema or allergies, during the test period. The CBMEO-containing cream-exposed test regions showed a greater reduction in the roughness index as compared with those of the placebo-exposed control regions. The R1, R2, and R3 indices showed a statistically significant decrease in the test regions as compared with the placebo control regions after using the cream for 2 weeks (p<0.05). Therefore, this study demonstrated that CBMEO-containing cosmetics are safe for human use and that they have skin wrinkle improvement properties. In the future, CBMEO may be a promising product for use in the development of functional anti-wrinkle cosmetics.

Healing of the bone around pure titanium implants without primary bone contact (초기 골 접촉이 없는 순수 티타늄 임프란트 주위 골의 치유반응)

  • Ahn, Jae-Hyun;Kim, Heung-Joong;Park, Joo-Cheol;Han, Kyung-Yoon;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.29 no.1
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    • pp.233-249
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    • 1999
  • Primary fixation is one of the most important factor in establishing adequate osseointegration between implant and bone. To evaluate the initial healing response of bone around implants without primary bone contact, this study was designed to create considerable space between implant and bone in 5 mongrel dogs, about 1-year old. After 3 holes of 6.0mm in diameter were prepared at the femur neck of the dogs, commercially pure titanium thread type implants(STERI-$OSS^{(R)}$), 8mm in length and 3.8mm, 5.0mm and 6.0mm in diameter, were inserted. Implants were supported by only nonresorbable membrane($Teflon^{(R)}$), and the penetration of upper soft tissue into the gap was inhibited by it. The each implant was positioned in the center of the drilled hole. 9 implants with different diameters were inserted in 3 dogs for histologic observation, and 12 were inserted in 2 dogs for mobility test and removal torque test.Fluorescent dyes were injected in order of Doxycycline, Alizarin Red S, and Calcein at intervals of 2 weeks. At 4-, 8-, and 12-week after placement, 3 dogs were sacrificed for histologic observation, and at 8- and 12-week after placement, 2 dogs were sacrificed for mobility test using $Periotest^{(R)}$ (Simens AG, Bensheim, Germany) and torque test using Autograph AGS-1000D $series^{(R)}$(Japan). The result were as follows: 1. The wider the gap between bone and implant was, the less bone maturity was, and the later osseointegration was occurred. Trabecular direction of new bone around implant was changed from parallel to perpendicular to the implant, and the gap was filled with new bone, over time. 2. There was a decreasing tendency over time in the mobility of all implants, but the wider gap between bone and implant was, the smaller decrease of the mobility was. 3. There was a increasing tendency over time in the removal torque gauge of all implants, and the wider gap was, the smaller increase of the removal torque gauge was. The results suggest that osseointegration in case of implant without primary bone contact may be obtained by guided bone regeneration technique with prolonged healing period, but the time of second surgery should be considered carefully.

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Synthesis of Optically Active Monoesters via Kinetic Resolution by Chiral Co (Salen) Complex Immobilized on Mesoporous BEA (메조세공 BEA에 고정화된 키랄 Co살렌 착체의 동적분할을 통한 고광학순도의 키랄 모노에스테르 합성)

  • Choi, Seong Dae;Park, Geun Woo;Lee, Gyung Chan;Kim, Geon-Joong
    • Applied Chemistry for Engineering
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    • v.26 no.2
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    • pp.132-137
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    • 2015
  • BEA-zeolite was modified by alkaline solution to introduce mesoporosity in the crystals and the homogeneous chiral Co(III) salen was immobilized in the mesopores. The dinuclear chiral Co(salen)-$GaCl_3$ catalyst immobilized on mesoporous BEA-zeolite showed high activity for the regioselective ring opening of terminal epoxides by carboxylic acids. Various chiral monoester derivatives could be synthesized with moderate enantioselectivity (47~69 ee%) from racemic epoxides through above reaction. When the chiral (S)-ECH was used as a reactant, it was efficiently resolved by carboxylic acid with a high enantioselectivity in the presence of heterogenized chiral salen catalyst, and the ring opened product afforded optically pure monoester epoxide (R)-GB (up to 98 ee%) through the ring closing in the basic solution by elimination of HCl. The heterogeneous catalyst could be fabricated easily, and the catalytic activity was retained for several times reuse without any further regeneration step.

Development of Fusant Degrading Aniline and 4-chlorobiphenyl by Spheroplast Fusion between Pseudomonas sp. and Flavimonas oryzihabitans (Flavimonas oryzihabitans와 Pseudomonas sp.간 원형질체 융합에 의한 Aniline과 4-chlorobiphenyl 분해균주 개발)

  • 박형수;박용근;김무훈;고범준;조미영;김치경
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.259-266
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    • 2000
  • Spheroplast cell fusions were performed with Flavimonas oryzihabitans degrading aniline and Pseudomonas sp. degrading 4-chlorobiphenyl to develope the new fusant degrading aniline and 4-CBP and its characters were investigated. F. oryzihabitans was induced to antibiotic marker ($Cm^r$ by NTG treatment for the fusants selection. The results of spheroplast formation and regeneration frequencies of the strains treated with lysozyme-EDTA were 99% and 5.0~6.6%, respectively. Fusion products were treated with 40% (v/v) PEG 6000 and fusion frequency was $3.16{\times}10^{-4} $. The DNA content of fusant, F22 was approximately 2-fold compared with parents. The fusant was stable, and showed the mixed biochemical characteristics of the parent strains. F22 was similar to parent for cell growth pattern and degrading capacity on 5 mM aniline but cell growth rate of F22 was 1.5-fold higher than that of the parent on 10mM aniline. However 4-CBP degrading ability of F22 was slightly lower than that of parental strain.

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