• Title/Summary/Keyword: 균주선발

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Characterization of a Restriction Endonuclease AspJI from Alcaligenes sp. J-482 (Alcaligenes sp. J-482 로부터 분리한 제한효소 AspJI의 특성)

  • Lee, Jeong-Taek;;Lim, Jai-Yun
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.285-290
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    • 1994
  • About 500 bacterial and fungal strains from a wide variety of natural habitats were screened for a new type II restriction endonuclease. Among the 500 species, we selected one species that produced a new restriction endonuclease. This strain has an optimum temperature of $30^{circ}C$ for growth. Morphological, cultural, and physiological characteristics were examined for identification of the isolated strain J-482. This strain was found to belong to the genus Alcaligenes. The restriction endonuclease was named as AspJI and partially purified from Alcaligenes sp. J-482 by DEAE-Sephadex A-50 column chromatography and gel filtration. Most of other nucleases were removed by the purification steps. The AspJI has a substrate specificity to ${lambda}$ DNA, pBR322 and Adenovirus-2 DNA. For its maximal activity, the isolated enzyme requires $MgCl_2$, which should be at least 12.5 mM and it does not need any other cofactors. It is maximally active in the absence of NaCl and is completely inactivated at 100 mM NaCl. The pH and temperature optima for activity were pH 7.5 and $37^{circ}C$, respectively. The DNA fragments generated by digesting ${lambda}$ DNA, pBR322, and Adenovirus-2 DNA with AspJI were the same as that produced by AatII. This suggests that AspJI is an isoschizomer of AatII.

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Development of a New PCR Method for Detection of Pectobacterium carotovorum (Pectobacterium carotovorum의 검출을 위한 PCR 진단법의 개발)

  • No, Ji-Na;Yoo, Mi-Sun;Park, Dong-Suk;Kim, Jeong-Gu;Yoon, Byoung-Su
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.306-311
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    • 2009
  • A new PCR method was developed to detect Pectobacterium carotovorum which is the causative agent of soft rot in Brassica pekinensis. A specific detection primer set based on Lytic murein transglycolase gene was designed and evaluated. Using ERB3_F (5'-TGC GAC ACC TCC TCA TCA CG-3') and ERB3_R (5'-CTT ATC ACG CTG TAA CCA GC-3') primers, 437 nucleotides long fragment was specifically amplified. The amplified products were observed in 52 out of 55 strains of P. carotovorum or Pectobacterium carotovorum subsp. carotovorum. On the other hand, no amplification was observed in 8 organisms including Chinese cabbage and potato. The optimal PCR condition for the ERB3_F/ERB3_R primer set was $58^{\circ}C$ for annealing and 15 mM for $MgCl_2$. With serially diluted templates, the specific PCR sensitivitie limit was $2\times10^3$ copies. Also, this method can be applied not only to DNA but also to field samples. This PCR method may be expected to be useful for specific detection of P. carotovorum.

Selection and Antifungal Activity of Antagonistic Bacterium Bacillus subtilis KMU-13 against Cucumber scab, Cladosporium cucumerinum KACC 40576 (검은별무늬병균 Cladosporium cucumerinum KACC 40576에 대한 길항균주 Bacillus subtilis KMU-13의 선발 및 항진균 활성)

  • Park Sung-Min;Lee Jun-Seuk;Park Chi-Duck;Lee Jung-Hun;Jung Hyuck-Jun;Yu Tae-Shick
    • KSBB Journal
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    • v.21 no.1 s.96
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    • pp.42-48
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    • 2006
  • Bacillus subtilis KMU-13 was isolated from the Lillehammer forest soils at Norway and shown a strong antifungal activity on cucumber scab, Cladosporium cucumerinum KACC 40576. B. subtilis KMU-13 produced a maximum level of antifungal substance under incubation aerobically at $30^{\circ}C$, 180 rpm for 48 hours in LB broth containing 0.5% maltose and 0.5% bactopeptone and initial pH adjusted to 6.0. Butanol extract of cultured broth was confirmed inhibitory zone by plate assay and Rf 0.64 value substance by thin layer chromatography (TLC) represented high antifungal activity against C. cucumerinum KACC 40576 and also shown fungal growth inhibitory activity against Botytis cinerea KACC 40573, C. gloeosporioides KACC 40804, D. byoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. soiani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.

Screening of Edible Mushrooms for the Production of Lovastatin and its HMG-CoA Reductase Inhibitory Activity (Lovastatin을 생산하는 식용버섯 선발과 HMG-CoA reductase 저해 효과)

  • Lee Jae-Won;Lee Soo-Min;Gwak Ki-Seob;Lee Ji-Yoon;Choi In-Gyu
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.83-88
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    • 2006
  • This research was performed to determine the production of lovastatin and its HMG-CoA reductase activity produced by fruit bodies and mycelial liquid cultures of domestic edible mushrooms (8 fungal strains). By deter-mining TLC analysis for the confirmation of the presence of lovastatin, all the extracts from fruit bodies and mycelial liquid culture showed same Rf value (0.46), whick was identical to that of the standard lovastatin. In order to extract lovastatin from fruit body, the mixture of water/acetonitrile/methanol was chosen as the most effective solvent. Extracts from fruit body and mycelial liquid culture of pleurotus ostreatus produced the high-est lovastatin 0.98 mg/g based on dry biomass, and 21.90 mg/L, respectively. In the inhibition rate of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, the highest was obtained in P. ostreatus as 67.8% among fruit bodies, and the rates of mycelial liquid culture extracts from P. ostreatus and Laetiporus sulphureus were 37.2% and 29.1%, respectively. Unusually L. sulphureus showed high inhibition rate with low content of lovastatin due to the contribution of campesterol and gamma-sitosterol with hypocholesterolemic activity as metabolites.

Simple Mass-screening Methods for Resistance of Tomato to Fusarium oxysporum f. sp. lycopersici (토마토 시들음병에 대한 간편 대량 저항성 검정법)

  • Park, Myung Soo;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.110-116
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    • 2013
  • This study was carried out to establish the simple mass-screening methods for resistant tomato to Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL). Root dip inoculation method has been used in many studies on the resistance of tomato to disease. On the other hand, in mass-screening for resistant tomato to Fusarium wilt, the inoculation method is time-consuming and laborious procedure. Disease development of two FOL isolates on two cultivars of tomato according to inoculation method including root dip, tip and scalpel methods were investigated. In compatible interaction, tomato seedlings of each cultivar inoculated by tip method showed the lower and more variable disease severities than by root dip method. Whereas the seedlings by scalpel method represented clear resistant and susceptible responses to Fusarium wilt as root dip method. The resistance degree of each cultivar inoculated with FOL isolates by scalpel method was hardly affected by the tested incubation temperature and inoculum concentration. On the basis of the results, we suggest scalpel inoculation method as an efficient mass-screening method for resistant of tomato cultivars to Fusarium wilt. Roots of tomato seedlings at two-leaf stage grown in plastic cell tray were injured with scalpel and then spore suspension (more than $1{\times}10^7\;conidia{\cdot}mL^{-1}$) of FOL was poured directly on the roots. The infected plants were cultivated in a growth room at $25-30^{\circ}C$ for 4 weeks with 12-hours light a day.

Entomopathogenic Fungi-mediated Pest Management and R&D Strategy (곤충병원성 진균을 활용한 해충 관리와 개발 전략)

  • Lee, Se Jin;Shin, Tae Young;Kim, Jong-Cheol;Kim, Jae Su
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.197-210
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    • 2022
  • Entomopathogenic fungi can be used to control a variety of sucking and chewing insects, with little effect on beneficial insects and natural enemies. Approximately 170 entomopathogenic fungal insecticides have been registered and used worldwide, with the recent focus being on the mode of action and mechanism of insect-fungal interactions. During the initial period of research and development, the industrialization of entomopathogenic fungi focused on the selection of strains with high virulence. However, improvement in productivity, including securing resistance to environmental stressors, is a major issue that needs to be solved. Although conidia are the primary application propagules, efforts are being made to overcome the limitations of blastospores to improve the economic feasibility of the production procedure. Fungal transformation is also being conducted to enhance insecticidal activity, and molecular biology is being used to investigate functions of various genes. In the fungi-based pest management market, global companies are setting up cooperative platforms with specialized biological companies in the form of M&As or partnerships with the aim of implementing a tank-mix strategy by combining chemical pesticides and entomopathogenic fungi. In this regard, understanding insect ecology in the field helps in providing more effective fungal applications in pest management, which can be used complementary to chemicals. In the future, when fungal applications are combined with digital farming technology, above-ground applications to control leaf-dwelling pests will be more effective. Therefore, for practical industrialization, it is necessary to secure clear research data on intellectual property rights.

Analysis of the Ergothioneine Content in the Fruiting Bodies of Sawdust-cultivated Lentinula edodes Cultivars in Korea (한국의 표고 톱밥재배 품종의 에르고티오네인 함량 분석)

  • Min-Jun Kim;Yeun Sug Jeong;Eunjin Kim;Yeongseon Jang;Kang-Hyeon Ka
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.301-310
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    • 2022
  • Lentinula edodes is a type of mushroom widely consumed in East Asia, and many of the components contained in them are used as raw materials for pharmaceuticals or as health supplements. Ergothioneine, a sulfur-containing amino acid in L. edodes, is attracting attention due to its antioxidant and anti-inflammatory effects. L. edodes generally contain an ergothioneine content of about 200 to 800 mg per kg; there are some differences in this value depending on the cultivation conditions. In this study, 24 domestic sawdust-cultivated L. edodes cultivars were cultured under the same conditions, and the morphological characteristics and ergothioneine content of the fruiting bodies were investigated. The yield and morphological characteristics of the fruiting bodies were different in each cultivar. Taehyanggo and Sanjo 713ho had the largest and thickest pileus. The pileus of L. edodes had a higher ergothioneine content than the stipe. Sanjo 715ho had the highest ergothioneine content in the pileus and the stipe at 1,225 mg/kg and 753 mg/kg, respectively. An ergothioneine content of more than 1,000 mg/kg in the pileus was observed in Bambithyang, Sansanhyang, Sulbaekhyang, Taehyanggo, Sanjo 705ho, Sanjo 709ho, Sanjo 715ho, and Sanjo 716ho. These cultivars can be used as parental strains to develop new cultivars with enhanced ergothioneine functionalities.

Biotransformation of Diterpenoids From Aralia continentalis Roots by the Genus Fusarium (곰팡이 Fusarium 속을 이용한 독활 뿌리 추출물로부터 디테르페노이드의 생물전환)

  • Keumok Moon;Seola Lee;Eunhye Jo;Areum Lee;Jaeho Cha
    • Journal of Life Science
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    • v.34 no.4
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    • pp.215-226
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    • 2024
  • Aralia continentalis is widely distributed in Far East Asian countries such as Korea, China, and Japan. A. continentalis has traditionally been used as an herbal remedy for various conditions, including analgesia, headache, inflammation, lameness, lumbago, rheumatism, and dental diseases in Korea. Previously, epi-continentalic acid, continentalic acid, and kaurenoic acid as major active biological compounds belonging to the diterpenoid class were identified. To synthesize diterpenoid derivatives with enhanced bioavailability, Fusarium fujikuroi was employed to biotransform diterpenoids due to its known antibacterial activity. This yielded two derivatives of kaurenoic acid, namely 16α-hydroxyent-kauran-2-on-19-oic acid and 2β, 16α-dihydroxy-ent-kauran-19-oic acid, with their chemical structures elucidated via NMR analysis. These derivatives exhibited increased polarity compared to kaur- enoic acid, as evidenced by their retention time on preparative HPLC using the ODS-A column and structural modifications. Evaluation of their antidiabetic activity targeting PTP1B, a negative regulator of the insulin signaling pathway, revealed inhibitory activities of 30.8% and 27.6%, respectively, at a concentration of 4 ㎍/ml. Additionally, both derivatives demonstrated low cytotoxicity, with an IC50 value 18 times higher than kaurenoic acid. Therefore, the augmented water solubility and reduced toxicity of 16α-hydroxy-ent-kauran-2-on-19-oic acid and 2β, 16α-dihydroxy-ent-kauran-19-oic acid, resulting from biotransformation by F. fujikuroi, render them promising candidates for industrial applications.

Evaluation of Garlic Germplasm for Resistance to Leaf Blight Caused by Stemphylium vesicarium (마늘 유전자원의 Stemphylium vesicarium에 의한 잎마름병 저항성 평가)

  • Jin Ju Lee;JiWon Han;Hun Kim;Jin-Cheol Kim;Gyung Ja Choi
    • Research in Plant Disease
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    • v.30 no.2
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    • pp.124-130
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    • 2024
  • Leaf blight caused by Stemphylium vesicarium is one of the most important fungal diseases of garlic (Allium sativum L.) worldwide, which results in a reduction of quality and yield. The breeding of resistant cultivars is an efficient approach to decrease the use of chemical fungicides and minimize crop losses. In this study, to find the resistant garlic resources against S. vesicarium, we evaluated the resistance degree of 20 garlic germplasms. To do this, garlic seedlings at four-leaf stage were rubbed with nonabsorbent cotton and then inoculated with spore suspension (3.0×105 spores/ml of potato dextrose broth) of S. vesicarium by spray method. Three to seven days after inoculation, the infected leaf area (%) of garlic seedling was measured. 'Daeseo' and 'Namdo' were included as susceptible and resistant control cultivars, respectively. After 3 to 7 days of incubation, the infected leaf area (%) of garlic seedling was measured. Our results showed that IT245512, IT245528, and IT244068 lines exhibited the highest resistance against S. vesicarium, whereas IT257134 and IT253043 lines were more susceptible than the susceptible cultivar 'Daeseo'. Based on the results, the resistant genetic resources selected in this study can be used a basic material for resistant garlic breeding system against leaf blight.

Characterization of a new commercial strain 'Goni' by intra-specific hyphal anastomosis in Pleurotus ostreatus (계통간 교잡에 의한 백색느타리 품종 '고니'의 육성 및 그 특성)

  • Yoo, Young Bok;Lee, Sang Cheol;Kim, Eun Jung;Kong, Won Sik;Jang, Kab Yeul;Shin, Pyung Gyun
    • Journal of Mushroom
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    • v.7 no.3
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    • pp.130-134
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    • 2009
  • To develop new white variety of Oyster mushroom, all white varieties which have been collected and kept in the lab were revived and screened their cultural characteristics. 84 intra-specific Oyster mushroom hybrids between the white-colored mutants Suhan and Wonhyeong were developed using hyphal anastomosis technique in 2007. The Po2007-63 ($2842-7{\times}0205-7$) was shown the best cultural characteristics, selected to be a new variety and named as 'Goni'. The new commercial strain, 'Goni' has white pilei and grows well under spring and autumn conditions in Korea. The fruiting bodies of 'Goni' are of an excellent quality in that not only the stipe is thick and long but also the pileus is small and hard. The optimum temperatures for mycelial growth and fruiting body development were $25-30^{\circ}C$ and $10-16^{\circ}C$, respectively. Time period required for the initiation of the first fruiting body is about 3 to 5 days depending on the temperatures. The shape of fruiting body is thin funnel shape. Fruiting body production per bottle was about $91{\pm}13$ g which is almost 97% quantity compared to that of other variety 'Miso'. Relatively low temperature incubation ($11^{\circ}C$) resulted in the development of better quality of 'Goni' mushrooms. When two different media including potato dextrose medium and mushroom complete medium were compared, the growth of mushroom were much faster in mushroom complete medium at $20-25^{\circ}C$, but not at $25^{\circ}C$. Similar results were observed with other variety 'Miso'. Analysis of the genetic characteristics of the new commercial strain 'Goni' showed a major DNA profile as that of the parental Suhan when primer URP 1 was used, but different to 'Miso' that was used as a control. When screens were performed with primer URP 2, DNA patterns were similar both to that of the parents and 'Miso'. This new variety of the white Oyster mushroom has a clean and fresh image that corresponds well to "health food". We therefore expect that this new strain will satisfy the consumers demand for variety and excellent mushrooms.

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