The evergreen oak tree, Quercus myrsinaefolia Blume, is not only economically important for wood, medicine, landscape trees, etc., but also becoming more important in terms of ecology due to climate change. However, asexual reproduction was difficult, so this study was conducted to establish the optimum conditions for micropropagation by shoot multiplication. The surface sterilized seeds of Q. myrsinaefolia were successfully germinated in WPM basal medium. BAP (1.0 mg/L) treatment was most effective for inducing multiple shoots. The highest induction rates of adventitious roots from the multiple shoots was shown in the treatment of 1.0 mg/L NAA. Both MS and WPM medium were most effective for growth of multiplied plantlets. For ex vitro acclimatization, the survival rates of multiplied plantlets were 100% in vermiculite and commercial soil. The results of this study can be used for proliferation and supply, and establishment of ex situ conservation of Q. myrsinaefolia elite trees.

This study was carried out to determine the basic mycelial culture conditions for Poria cocos growth. According to colony diameter and mycelial density, suitable media for mycelial growth were Malt yeast extract, Potato dextrose agar, Yeast extract agar, and Yeast malt agar. The optimum temperature for mycelial growth was between 25 and 35oC, and the optimum pH value was between 4 and 7. Carbon and nitrogen sources were fructose and yeast extract. The optimum C/N ratio was about 10 to 1 with 2% glucose. Other minor components for optimal growth were thiamine-HCl and nicotinamide as vitamins, acetic and lactic acid as organic acids, and MgSO4·7H2O and FeSO4·7H2O as mineral salts.

The in vitro culture of Toxoplasma gondii was evaluated using a JNUCK cell strain that was independently developed by our research team. The sensitivity of the JNUCK strain was compared with those of MDCK and vero cell strains, which were previously shown to be sensitive to T. gondii. Morphological observation and 3H-uracil absorption tests showed that JNUCK cell strain has the highest sensitivity, while the MDCK and vero cell strains showed lower sensitivities. The optimal culture conditions using the JNUCK cell strain were determined by inoculating T. gondii at multiplicity of infection (MOI) 1, MOI 5, and MOI 25 and the numbers of T. gondii were measured 24, 48, and 72 hours after inoculation. The optimal medium composition was determined by adding different concentrations of FBS to DMEM medium for inoculation at MOI 5, and then calculating the numbers of tachyzoites inside the cells and in the medium. The numbers of tachyzoites inside the cell were highest for 10% FBS, while the numbers of tachyzoites in the medium peaked after addition of 30% FBS. These studies confirmed that the optimal culture condition of T. gondii using the JNUCK cell strain was achieved by inoculation at MOI 5, using medium containing 20% FBS and culturing for 72 hours.

In vitro culture of murine embryos is an important step for in vitro production systems including in vitro fertilization and generations of genetically engineered mice. M16 is widely used commercialized culture media for the murine embryos. Compared to other media such as potassium simplex optimization medium, commercial M16 (Sigma) media lacks of amino acid, glutamine and antibiotics. In the present study, we optimized M16 based embryo culture system using commercialized antibiotics-glutamine or amino acids supplements. In vivo derived murine zygote were M16 media were supplemented with commercial Penicillin-Streptomycin-Glutamine solution (PSG; Gibco) or MEM Non- Essential Amino Acids solution (NEAA; Gibco) as experimental design. Addition of PSG did not improved cleavage and blastocyst rates. On the other hand, cleavage rate is not different between control and NEAA treated group, however, blastocyst formation is significantly (P<0.05) improved in NEAA treated group. Developmental competence between PSG and NEAA treated groups were also compared. Between two groups, cleavage rate was similar. However, blastocyst formation rate is significantly improved in NEAA treated group. Taken together, beneficial effect of NEAA on murine embryos development was confirmed. Effect of antibiotics and glutamine addition to M16 media is still not clear in the study.

본 연구에서는 식물성 프로바이오틱스에 해조류인 미역을 첨가하여 프로바이오틱스의 체내 안정성 및 면역원성을 향상 시킬 수 있는 고농도 배양법을 개발하고자 하였다. 미역 첨가 배지를 생산 수준으로 유산균을 배양하였을 때 유산균수는 18시간째 1022, 24시간째 109으로 고농도 배양이 가능하였다. 미역첨가배지로 배양된 유산균의열 안정성, 위산 및 담즙 안정성의 효과도 기본배지와 비교 하여 유의적인 증가를 확인할 수 있었다. 미역첨가배지로 배양된 프로바이오틱스는 생균 및 사균 모두 면역증강효과를 나타내는 것을 알 수 있었다. 배양만으로 기능성이 향상된 프리바이오틱스를 개발함에 따라, 다양한 프로바이오틱스의 기능성 향상의 기반 기술로서 활용이 가능할 것으로 사료 된다.

This study was conducted to reduce contamination ratio of oyster mushroom bottle cultivation. The optimalconditions of substrate sterilization for reducing of contamination ratio were at 121oC for 90min. In addition, UV-C irradiation isgood for lower contamination ratio to continue over 6 hours at cooling and inoculation room after sterilization. Thecontamination ratio and density of microorganisms of substrate were showed 0% after sterilization at 121oC for 90min.Trichoderma sp., main pathogen of mushrooms, was detected from substrate after sterilized during 2 or 4 hours at 101oC and 105oC,respectively. The amount of electricity used was the lowest at 121oC for 90min than that of other sterilization conditions. The UV-Cirradiation treatment was used UV-C lamp(40 watts) in the inoculation room(56m3). The density of bacteria did not detected after UV-C irradiation for 6 hours. And the death ratio of bacteria and Trichoderma sp. was 99.9% after UV-C irradiation for 6 hours. However,in the same UV-C irradiation time, the death ration of Cladosporium sp. was 90.9%. Therefore, the death ratio of fungi was lower thanthat of bacteria at the same UV-C irradiation treatment.

Sweetpotato whitefly (Bemisia tabaci), especially Q biotype, has been recognized one of the most destructive insect pests worldwide because of increased resistance to some insecticide groups requiring alternative strategies for its control. We studied the conidia production of entomopathogenic fungus Isaria javanica Pf04, which had been reported high virulence isolate against Q biotype of B. tabaci, using grain. Brown rice was most suitable for conidia mass production of the isolate of I. javanica. Conidia was produced high at 25 ~ 27.5℃. The isolate produced more spores when conidia suspension directly inoculated onto media than two-phase fermentation. When concentration of inoculum was high spore production was high, but increasing rate of conidia production was highest at low inoculum concentration (1×105 conidia/ml) as 6,700 times increase compared with 20 times increase at high inoculum concentration (1×108 conidia/ml). These results indicated that the isolate can produce more conidia with cheap agricultural product and can develop as a microbial pesticide to control sweetpotato whitefly.

Cellular microenvironment is an essential issue for regulating epithelial characteristics through the alteration of intricate signaling pathways and intercellular communications in different cell types. Thus, microenvironment influences tumor initiation, progression, and metastasis. This study aimed to investigate the relationship between microenvironment and epithelial property in HPV16 E6/E7-immortalized human oral keratinocytes (IHOKs). To investigate characteristics of IHOK cultured in different media, two media were used, which included keratinocyte growth media (KGM), F-medium composed of 3:1 ratio of DMEM and F-12 (P media) supplemented with 10% FBS and 1% penicillin/streptomycin. Proliferative property and invasive and migratory activity were observed. As results, proliferating activities of IHOK in different culture condition were changed. Likewise, migratory and invasive activities were also different depending on media types. These results suggest that cellular microenvironment can affect modification of biological properties of epithelial cells.

본 연구는 버들송이버섯의 균사 생육 최적조건을 구명하여 액체종균을 제조하고 또한 톱밥종균의 영양원을 선발하기 위하여 시험을 수행하였고, 선발된 종균에 대한 종균 접종량을 설정하여 병재배에 적합한 조건을 구명하고자 하였다. 버들송이버섯의 톱밥종균은 미송톱밥(70%)+밀기울(30%)의 비율, 포플러톱밥(80%)+옥수수가루(20%) 조건으로 배합할 때 균사 생육이 양호하였다. 액체종균에 대한 영양원 선발 및 첨가량은 백설탕 1.0~1.5%, yeast extract 0.7g/ℓ, 콩가루0.1.7g/ℓ, 무기물은 MgSO4·7H2O 0.3g/ℓ, KH2PO4 0.5g/ℓ, K2HPO 1.2g/ℓ가 최적 조건이었다. 이때 선발된 배지의 접종량은 액체 및 톱밥종균을 25㎖/850㎖과 20~25g/850㎖로 접종 할 때 자실체 발생이 양호하였다.

We investigated the cleavage rate and blastocyst yield for each culture condition to enhance tolerance of cryo-preservation of bovine IVF embryo with relatively lower cryo-tolerance compared to in vivo embryo. The cleavage rate and blastocysts yield for CR1aa, IVMD, IVD, CR1aa+10% FBS were 73.2, 69.3, 72.8, 68.5% and 44.1, 30.8, 33.3, 48.0%, respectively. The values did not differ among each treatments without serum. For embryo vitrification, In vivo and In vitro blastocysts were exposed to VS1(10% glycerin, 0.1 M glucose, 0.1 M sucrose, PEG 1%) for 5 min, and VS2 (10% glycerin, 10% EG, 0.2 M glucose, 0.2 M sucrose, PEG 2%) for 5 min and then VS3 (10% glycerin, 30% EG, 0.3 M glucose, 0.3 M sucrose, PEG 3%) for 1 min. The exposed embryos were then loaded into the 0.25 ml plastic straws and then plunged into liquid nitrogen. The straws were held for period of 1 to 2 weeks before thawing. In embryo viability, no differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos. whereas expansion-BL rates was significantly higher for in vivo-derived embryos (72.7%) when compared to in vitro-derived embryos (51.4%), respectively (P<0.05). In conclusion, our results indicate that combined use of CRIaa culture medium with vitrification might enhance tolerance of cryopreservation for bovine IVF embryo production.

현재까지 연구로는 흰목이 균사체에서 EPS 생산과 균사생장에 대한 적정 정치배양 조건이 연구되었다. 본 연구로부터 탄소원과 질소원의 처음 농도, 균사 형태와 발효조의 타입의 선택은 흰목이 균사체 EPS 생산에 가장 영향을 미친다는 것을 알게 되었다. 이들 결과는 공기주입식 반응기에서 EPS 생산성은 진탕탱크 반응기 보다 더 높았다는 점을 증명하였다. 또한 흰목이 균사의 정치배양의 생리적 생장에 대한 지식은 아직도 제한적이다.

항산화제는 산소의 저장고로서 무혈청 배양액에서 주요한 작용을 하며, 복합배지에서 유용한 첨가제로 알려져 있다. 따라서 본 연구는 한우 체외 수정란의 배양에 있어서 항산화제인 L-cysteine의 작용과 수정란의 발달 단계별 염색체의 분석을 통하여 체외 수정란의 배양 체계를 수립하고자 실시하였다. 한우 난포란의 체외 성숙은 0.1% PVA, 0.1 mM L-cysteine 첨가 시 체외 성숙율은 73.4%, 94.6%으로 각각 나타냈으며, 처리간에 유의적

본 연구는 mSOF(modified synthetic oviduct fluid medium) 배양액을 이용하여 와 배양 소적에서 일본 흑우의 수정란 생산 효율을 개선하기 위하여 수행하였다. 난구세포가 부착된 미성숙 난자는 각각 단독 배양조건( 소적) 및 그룹 배양 조건 ( 소적)에서 실시하였고 배양액은 TCM-199의 기본 배지에 10% FCS, 0.02IU/ml FSH와 를 첨가하여 사용하였다. 배반포 단계로 발육한 수정란은 1.5M ethylene