• Korean Journal of Microbiology
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• v.30 no.6
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• pp.546-552
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• 1992

Ethanol-tolerant strain, S. eerevisiae BUI a26 ($alc^r thr^-$) and gJucoamylase-producing strain, S diastatieus AI5a6 (STA+ hom-) were prepared by means of genetic manipulation, Protoplast fusion was carried out to introduce STA gene from AI5a6 strain to BUla26 strain, Protoplast formation was shown at 0,8 M sorbitol and 200 Jig/ml to 400 Jig/ml zymolyase treatment for 2 hours incubation, Fusion frequency was $ 3.25 {\times} 10^{-3}$ to the regenerated protoplast number using PEG 6000 for 90 min incubation. The excellent fusants with genotype of STA- $alc^r thr^-$ hom+/STA+ ($alc^s thr^+$ hom- (2n), F7 and FIO, were selected by ethanol-tolerant, ethanol fermentation, and glucoamylase production tests, Glucoamylase production of AI5a6 showed 2,7 units, but 4.2 or 8.4 units for F7 or FIO fusant at $30^{\circ}C$, Ethanol fermentation from 32% glucose by BUla26 was 14,0%(v/v) in fermentaion medium for 5 days incubation, but 14.5% or 15,0% for F7 or FIO strain, respectively. Ethanol fermentation from 5% starch was 2,0% by F7, or 1.8% by FIO strain in fermentation medium for 5 days fermentation.

• Korean Journal of Medicinal Crop Science
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• v.22 no.6
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• pp.435-441
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• 2014

This study was conducted to investigate the quality of seeds, the germination rates and the days required for germination, to examine the patterns of protein expressions during the germination and to improve the techniques of managing and storing seeds and viability of the seeds of Lithospermum erythrorhizon Sieb. et Zucc. After collecting and harvesting seeds, they were classified to white and brown colors of seed coat through testing their seed size, weight, and quality. The germination rates, the days required for germination, and the protein expressions were examined with different colors of seed coats, storing temperatures and durations by treating the different plant growth regulators and primings. One hundred seed weight of white color was heavier about 1.17 g than those of brown one about 0.81 g. The germination rates in white color of seed coat was higher, 3.05 ~ 5.75%, than those in brown one. Its rates were decreased with getting longer in storage durations. There was no big differences on germination rates between storage temperatures. The plant growth regulator of $GA_3$ and Kinetin was affected to improve the seed germination. $GA_3$ increased the seed germination clearly at 25 ppm level, while kinetin increased it gradually from 25 to 100 ppm levels. In germination by seed primings, PEG6000 made higher germination rate with increasing their levels, whereas $KNO_3$ increased the germination until 100 mM level and then decreased it with 200 mM unlike PEG6000. The protein expressed during the seed germination were appeared more and clearer bands in the seed after germination, especially 20 ~ 30 kDa, compared to those in the seed before germination. These results showing more and clearer bands were positively related to the germination rates which were different by seed colors, storage temperatures and durations, and plant growth regulators and primings.

• Korean Journal of Medicinal Crop Science
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• v.23 no.1
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• pp.13-19
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• 2015

This study was conducted to improve the postharvest storage techniques of managing and storing seeds, to test qualities and viabilities of the seeds and to examine the germination rate and the protein expression of Achyranthes japonica Nakai. The seeds collected from different areas of Je-Cheon and Gwang-Ju were stored with different temperatures and durations. Two plant growth regulators and two seed priming were treated to investigate their effect on the germination rates and the days required for germination. The weight of one hundred seed collected in Gwang-Ju was heavier than those in Je-Cheon. Seed length collected in Gwang-Ju was also longer about 5.12 mm than those in Je-Cheon about 4.90 mm and seed width was longer in Gwang-Ju than those in Je-Cheon. The rates of seed germination in two different collection areas were higher about 2.9 to 13.0% in Gwang-Ju compared to those in Je-Cheon. Comparing its rates with the storing temperatures and durations, they were not clearly different in between $4^{\circ}C$ and $25^{\circ}C$ and they also were gradually decreased with getting longer storing durations. The germination rates treated by plant growth regulators were higher with $GA_3$ than those with Kinetin. The highest seed germination rate was appeared at 50 ppm of $GA_3$. Comparing its rates with different seed priming, they were relatively higher with $KNO_3$ than those with PEG6000. In protein expression patterns between before the germinating and after the germinating of seeds, more and clear bands were appeared in the seed after the germination compared to those before the germination of seeds, especially 10 ~ 20 kDa. These results showing more and clear bands were more clearly appeared in Gwang-Ju compared to Je-Cheon. Comparing the protein expression with plant growth regulators and seed primings, $GA_3$ was better expression than those with Kinetin and $KNO_3$ was better than those with PEG6000. More and clear bands were closely related to the germination rates of seeds and more detailed studies would be required.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2020.08a
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• pp.69-69
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• 2020

식물체가 건조 스트레스를 받으면 각 기관 물질 생산의 변이, 분화 및 발달 억제를 통해서 식물의 생산활동을 현저히 저하시켜, 식물의 생장, 형태, 개체발생 및 대사생리에 영향을 미치는 것으로 연구되어 왔다. 최근 기후 온난화로 인해 온도가 점진적으로 상승하고 가뭄과 같은 이상기상이 빈번하게 발생함에 따라 많은 노지 작물의 농업 생산성이 약화되고 있다. 배추는 우리나라의 대표적인 식품 중 하나인 김치의 주재료로 연중 안정적인 생산과 공급이 필요시 되지만, 배추의 경우 건조 조건에서 엽육조직의 붕괴와 같은 생리장해가 발생되기 때문에 최근 발생되고 있는 이상 기후의 영향으로 안정적인 생산이 어려워지고 있는 실정이다. 하지만, 배추에서는 이상 기후에 대응이 가능한 내건성 품종의 육성을 위한 연구는 미흡한 실정이다. 따라서 본 연구에서는 내건성이 높은 배추 개발을 위한 기초소재를 찾는 것을 목적으로 농촌진흥청 유전자원센터에서 보존하고 있는 재래종 배추 30계통을 분양받아 강릉원주대학교 생명과학대학 내의 조직배양실을 이용하여 연구를 수행하였다. 배지는 Tissue Culture Square Dish(125×125×20mm)에 Agar를 녹여 40ml씩 분주하여 고체배지를 조성한 후, 건조 처리구 Polyrthylene glycol 6000(PEG) 0%(Control), 20%(Mild Stress), 30%(Severe Stress)를 설정하여 60ml씩 추가 분주하여 배양기 28℃에서 15시간 처리를 하였다. 분양받은 각 계통의 종자는 1% 차아염소산 나트륨으로 10분간 종자표면을 살균한 후, 5번 정도 멸균수로 헹군 후, 표면 살균한 재래종 종자를 고체배지 시험관에 6립씩 치상하였다. 식물체 생육은 각 처리구별 3반복으로 하였으며 주간 12시간 주기, 광도 2,400Lux, 온도 20℃의 조직배양실에서 치상 후 7일간 생육하였다. 치상 후 7일간 생육시킨 식물체를 채취하여 지상부생체중, 지하부 생체중, 뿌리 길이를 측정하였으며, 지상부와 지하부로 나누어 50℃에서 72시간 건조시킨 후, 건물 생산량을 조사하였다. 본 실험 결과 건조 처리(PEG-6000)는 배추의 생장을 저해하였지만, 생육 저해 정도는 계통간의 차이가 있는 것을 관찰할 수 있었으며 무처리구와 스트레스 처리구간의 생장량 변화 정도를 기반으로 군집분석을 수행한 결과 'IT110483'과 'IT104903' 계통이 실험에 공시된 계통들 중 상대적으로 강한 건조 내성을 가진 것을 확인할 수 있었다. 따라서, 재래종 배추 계통 중 일부는 건조내성이 강한 새로운 배추 품종을 육성하는데 있어 유용하게 이용될 수 있을 것으로 사료된다.

• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.175-181
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• 1988

To produce L-lysine from cellulosic substrate, the intergeneric protoplast fusion between Cellulomonas flavigena and Corynebacterium glutamicum, Cellulomonas flavigena and Brevibacterium flavum was performed. The fusion frequencies were 1.9$\times$10$^{-6}$ to 2.1$\times$10$^{-6}$ for the regenerated protoplasts when two parental strains were treated with 30% of polyethyleneglycol (M.W.6000) containing 5 mM EDTA at 3$0^{\circ}C$ for 30 min. Two fusants, FCB3 and FCC 19 were finally selected by comparision of their genetic stability and L-lysine productivity. The properties of fusants-DNA con-tent, G＋C content and L-lysine productivity-were investigated. The DNA content of fusants was greater than those of the parental strain and their G＋C contents are equal to half of total G＋C con-tent of two parental strains. The fusants showed high productivity of L-lysine from carboxy methyl cellulose as substrate.

• The Korean Journal of Mycology
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• v.14 no.2
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• pp.165-174
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• 1986

Conditions for production, fusion and reversion of protoplasts of Aspergillus niger were investigated, and an attempt was made to enhance fusion frequency. Auxotrophic mutants and morphological mutants were induced by U.V. irradiation $(9.9\;erg/mm^2,\;13min)$ on Aspergillus niger. Maximum yield of protoplasts was obtained from 21 hr cultured mycelia by using 1% driselase in 0.6 M KCl or 0.6 M $NH_4Cl$ as osmotic stabilizer. The optimal temperature for mycelium digestion was $30^{\circ}C$, and the optimal pH was 6.0. Protoplasts produced at different digestion period showed heterogeneity in size and vacuole content. Maximal frequency of protoplasts reversion was obtained on 0.6 M KCl stabilized agar medium at pH 5.0. Reversion frequencies of protoplasts produced for 3 hr and 1 hr mycelial digestion were 8.0% and 15.3%, respectively. The optimal concentration of PEG(m.w. 6000) for protoplast fusion was 30%, and that of $CaCl_2$ was $1{\sim}50\;mM$. The optimal pH and period for the reaction of PEG solution were 8.0 and 10 minutes, respectively. Fusion frequencies between auxotrophic protoplasts produced for 3 hr-mycelial digestion were $0.06{\sim}0.42%$, and those for 1 hr-mycelial digestion were $0.09{\sim}0.54%$.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.10
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• pp.1389-1394
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• 2012

Sheanut cake (SNC), expeller (SNE) and solvent extractions (SNSE) samples were evaluated to determine their suitability in animal feeding. The CP content was highest in SNSE (16.2%) followed by SNE (14.7%) and SNC (11.6%). However, metabolizable energy (ME, MJ/kg) was maximum in SNC (8.2) followed by SNE (7.9) and SNSE (7.0). The tannin phenol content was about 7.0 per cent and mostly in the form of hydrolyzable tannin (HT), whereas condensed tannin (CT) was less than one per cent. The in vitro gas production profiles indicated similar y max (maximum potential of gas production) among the 3 by-products. However, the rate of degradation (k) was maximum in SNC followed by SNE and SNSE. The $t^{1/2}$ (time taken for reaching half asymptote) was lowest in SNC (14.4 h) followed by SNE (18.7 h) and SNSE (21.9 h). The increment in the in vitro gas volume (ml/200 mg DM) with PEG (polyethylene glycol)-6000 (as a tannin binder) addition was 12.0 in SNC, 9.6 in SNE and 11.0 in SNSE, respectively. The highest ratio of $CH_4$ (ml) reduction per ml of the total gas, an indicator of the potential of tannin, was recorded in SNE (0.482) followed by SNC (0.301) and SNSE (0.261). There was significant (p<0.05) reduction in entodinia population and total protozoa population. Differential protozoa counts revealed that Entodinia populations increased to a greater extent than Holotricha when PEG was added. This is the first report on the antimethanogenic property of sheanut byproducts. It could be concluded that all the three forms of SN byproducts are medium source of protein and energy for ruminants. There is a great potential for SN by-products to be incorporated in ruminant feeding not only as a source of energy and protein, but also to protect the protein from rumen degradation and suppress enteric methanogenesis.

• Korean Journal of Microbiology
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• v.28 no.1
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• pp.47-54
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• 1990

In order to investigate physiological characteristics of fusants by interspecific protoplast fusion of the genus Cellulomonas, protoplasts of Cellulomonas flavigena NCIB 12901 and Cellulomonas bibula NCIB 8142 were fused and cell wall regenerated. To give gene maker, C. bibula was treated with 500 ug/ml NTG for 1 hr and arginine requiring auxotrophic mutants were isolated. Protoplasts of the genus Cellulomonas were obtained by treatment with $600{\mu}{\textrm{g}}$/ml lysozyme, and 0.5M sorbitol was optimal for osmotic stabilizer on protoplast fromation. Protoplast fusion was enhanced by 40% PEG)M.W.6,000) containing 25 mM $CaCl_{2}$ at $30^{\circ}C$ for 30 min and fusion frequency between C. bibula and C. flavigena was $5\times 10^{-4}$. Processes of protoplast formation, cell wall regeneration and protoplast fusion were obsdrved by scanning electron microscope. By comparing enzyme activities of cellulase, exocellobiohydrolase, .betha.-glucosidase of the parent strains of Cellulomonas with those of thier mutants and fusants, fusants with increased enzyme activity were obtained. By the studies on nutritional requirement, antibiotic resistance, cellulolytic enzyme activities, type of peptidoglycan and motility of two mutants and fusants, fusants were proved to be recombinant of both mutant strains.

• Korean Journal of Microbiology
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• v.30 no.3
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• pp.207-212
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• 1992

To develope the new strains of microorganisms having the degradative ability for various aromatic hydrocarbons. spheroplast cell fusions were performed with Arthrobacter spp. degrading phthalate ester and Pseudomonas putida degrading alkylbenzen sulfonate(ABS) and the characteristics of the fusants were investigated. The spheroplasts of P. putia KUD15 and Arthrobacter sp. were formed effectively by lysozyme-EDTA treatment and by Ampicillin-lysozyme-EDTA treatment. respectively. The Spheroplast formation frequency and the regeneration frequency of the strains were 98-99% and 5-8%, respectively. For cell fusion. 40% PEG6000 was used as a fusogenic agent and the formation frequencies of fusion product were $1.8{\times}10^{4}-$2.9{\times}10^{4}$ Most of the fusants, which were selected in complemented antibiotics media showed the degradative ability in minimal selective medium added phthalate ester or ABS as sole carbon source. ABS degradation by fusant strain was increased about 20% with compared with the parental strain, while the degradative ability of phthalate ester was simillilar to that of parental strain.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.6
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• pp.507-512
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• 1993

Pre-sowing treatments of tobacco seeds (cv. NC82) were evaluated as a means of improving the speed of emergence, early seedling growth and uniformity of the seedlings. Seeds were treated through the use of polyethylene glycol 6000(PEG), solid matrix priming(SMP), chilling and dark-preincubati(DPI). Under the laboratory condition of 15$^{\circ}C$, all the treated seeds germinated more rapid and the time to 50% germination(T50) reduced than control. Among the treatments, SMP was the best to reduce the T50 for germination. All the treatments did not affect the final germination percentage, while the chilling showed of little benefit for germination and T50. Under the greenhouse condition, seedling from SMP emerged earlier and more uniformity and also greater mean fresh and dry weight showed than other treatments. The advantage of T50 from SMP was affected more in temperature than in water content of Agro-Lig during the treatment.