• Title/Summary/Keyword: spore analysis

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Isolation and Characterization of Antifungal Metabolites from Pterocarpus santalinus against Fusarium graminearum Causing Fusarium Head Blight on Wheat (자단향으로부터 밀 붉은곰팡이병균 Fusarium graminearum에 대한 항진균활성 물질의 분리 및 특성 규명)

  • Kim, Ji-In;Ha, Areum;Park, Ae Ran;Kim, Jin-Cheol
    • Research in Plant Disease
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    • v.23 no.3
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    • pp.268-277
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    • 2017
  • Fusarium head bight (FHB) is a devastating disease on major cereal crops worldwide which causes primarily by Fusarium graminearum. Synthetic fungicides are generally used in conventional agriculture to control FHB. Their prolonged usage has led to environmental issues and human health problems. This has prompted interest in developing environmentally friendly biofungicides, including botanical fungicides. In this study, a total 100 plant extracts were tested for antifungal activity against F. graminearum. The crude extract of Pterocarpus santalinus heartwood showed the strongest antifungal activity and contained two antifungal metabolites which were identified as ${\alpha}$-cedrol and widdrol by GC-MS analysis. ${\alpha}$-Cedrol and widdrol isolated from P. santalinus heartwood extract had 31.25 mg/l and 125 mg/l of minimal inhibitory concentration against the spore germination of F. graminearum, and also showed broad spectrum antifungal activities against various plant pathogens. In addition, the wettable powder type formulation of heartwood extract of P. santalinus decreased FHB incidence in dose-dependent manner and suppressed the development of FHB with control values of 87.2% at 250-fold dilution, similar to that of chemical fungicide (92.6% at 2,000-fold dilution). This study suggests that the heartwood extract of P. santalinus could be used as an effective biofungicide for the control of FHB.

A Thermostable Protease Produced from Bacillus sp. DF 218 (Bacillus sp. DF218이 생산하는 내열성 단백질 분해효소)

  • Lee, Joung-Hee;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.105-110
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    • 2004
  • Microorganism (strain DF 218) producing thermostable pretense was isolated from Korean soil and compost. It was Gram-positive, rod-shaped, aerobic, and spore-forming with yellowish white colony color, Temperature range for growth at pH 6.5 was $30-65^{\circ}C$, with optimum growth at $60^{\circ}C$. pH range for growth at $60^{\circ}C$ was 5-7 with optimum of 6.5, which indicates strain DF 218 to be thermophilic. The 16S rDNA sequence of strain DF 218 had 95% sequence similarity with that of Bacillus flexus. Based on physiological properties and phylogenetic analysis, we proposed the isolated strain as Bacillus sp. DF 218. Pretense was produced aerobically at $60^{\circ}C$ for 32 hr in a medium (pH 6.5) containing 1% each trypton, glucose, and NaCl. Its molecular weight was estimated as 61 kDa, with optimum temperature and pH of $60^{\circ}C$ and 7.5, respectively.

Isolation of Siderophore-producing Pseudomonas fluorescens GL7 and Its Biocontrol Activity against Root-rot Disease (Siderophore 생산성 생물방제균 Pseudomonas fluorescens GL7의 선발 및 식물근부병의 방제)

  • 이정목;임호성;장태현;김상달
    • Microbiology and Biotechnology Letters
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    • v.27 no.6
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    • pp.427-432
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    • 1999
  • For the development of a multifunctional biocontrol agent, the siderophore-producing strain GL7 was isolated from a rhizosphere on chrome azurol S agar. The GL7 was identified as a strain of Pseudomonas fluorescents on the basis of their reactions to standard physicochemcial tests from Bergey's manual, API diagnostic test, and fatty acid analysis. P. fluorescents GL7 considerably inhibited spore germination and hyphal growth of phytopathogenic fungus Funsarium solani in a dual culture. In pot trials of bean with P. fluorescens GL7, the disease incidence was significantly reduced down to 5% from 70% of incidence in the untreated control. P. fluorescens GL7 also enhanced plant growth to nearly 1.5 times than that of the untreated control, promoting elongation and development of the roots. These results suggest that the plant growth-promoting P. fluorescens GL7 can play an important role in the biological control of soil-borne plant disease in a rhizosphere.

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Characteristics of Microorganisms Contaminating Seafood Cooking Drips Exposed to Gamma Irradiation (감마선 조사된 수산 자숙액의 오염 미생물군 특성)

  • Choi, Jong-Il;Kim, Yeon-Joo;Kim, Jae-Hun;Chun, Byung-Soo;Ahn, Dong-Hyun;Kwon, Joong-Ho;Hwang, Young-Jung;Byun, Myung-Woo;Lee, Ju-Woon
    • Food Science and Preservation
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    • v.16 no.2
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    • pp.286-291
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    • 2009
  • Microorganisms in seafood cooking drips were counted and identified. Total viable cell counts were 6.40 and 3.10 log CFU/g in cooking drips of Hizikia fusiformis and Thunnus thynnus, respectively. However, microbial populations fell with increased irradiation doses. In H. fusiformis cooking drips, a 5-log reduction in total aerobic bacteria was obtained by irradiation with 5 kGy. In T. thynnus cooking drips, however, contaminating microorganisms were more resistant to gamma irradiation and only a 1-log reduction was seen. DNA sequence analysis showed that the principal contaminating microorganisms in H. fusiformis and T. thynnus cooking drips were Lactobacillus and Bacillus species, respectively. Therefore, the high irradiation resistance of T. thynnus cooking drips microbes may result from spore formation by Bacillus species.

A Thermostable Protease Produced from Bacillus sp. JE 375 Isolated from Korean Soil (한국의 토양으로부터 내열성 단백질 분해효소를 생산하는 Bacillus sp. JE 375의 선별)

  • Kim, Ji-Eun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.419-426
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    • 2006
  • A thermophilic microorganism, strain JE 375, which produces a thermostable protease, was isolated from soil and compost in Korea. This gram-positive, rod-shaped, catalase positive, motility positive, and hemolysis ${\beta}$ containing organism was implicated in glucose fermentation, mannitol fermentation, xylose oxidation, aerobic activity and spore formation. The color of the colony was yellowish white. The temperature range for growth at pH 6.5 was between 55 and $70^{\circ}C$, with an optimum growth temperature of $65^{\circ}C$. This result confirmed the strain JE 375 as a thermophilic microorganism. The enzyme was produced aerobically at $65^{\circ}C$ during 20 hr in a medium (pH 6.5) containing 1% trypton. 1% maltose, 0.5% yeast extract and 1% NaCl. The 16S rDNA of strain JE 375 had 97.6% sequence similarity with the 16S rDNA of Bacillus caldoxyloyticus. On the basis of biochemical and physiological properties and phylogenetic analysis, we named the isolated strain as Bacillus sp. JE 375. The thermostable protease from Bacillus sp. JE 375 had been partially purified and characterized. The molecular weight of the enzyme was deduced from SDS-PAGE and gel chromatography as 55 kDa and its optimal temperature was $60^{\circ}C$. The enzyme showed its highest activity at pH 7.5 and was stable from pH 7.0 to 8.0.

Interspecific Hybridization between Pleurotus ostreatus and Pleurotus sajor-caju by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리와 여름느타리버섯의 종간(種間) 교잡(交雜))

  • Yoo, Young-Bok;Lee, Haing-Sook
    • The Korean Journal of Mycology
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    • v.22 no.4
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    • pp.378-385
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    • 1994
  • Interspecific somatic hybrids were obtained by protoplast fusion between Pleurotus ostreatus and Pleurotus sajor-caju. The fusion products between incompatible strains did not form clamp connections. Fruiting body of the clampless fusants was induced by light-dark cycle on saw-dust-rice bran substrate in glass bottles. Out of them, seven somatic hybrids produced fruiting bodies of intermediate morphology of the two species. Light and low temperature were the initiating factors for the development of clamped hyphae from the clampless mycelial colonies. All of these basidiocarps had clamp connections. Eight fusants from the six crosses were analysed with the segregation of genetic characters by random spore isolates. In the three combinations, unexpected alleles were shown. Somatic hybrid between P188 (P. ostreatus 2-1 + P. sajor-caju 2-53) and P. florida 2-3 by triple cross produced fruiting bodies similar to those of fusant between P. ostreatus and P. florida. All the genetic charaters from the three strains were shown to segregate and recombine.

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Correlation between Biosurfactants and Antifungal Activity of a Biocontrol Bacterium, Bacillus amyloliquefaciens LM11 (생물적 방제균 Bacillus amyloliquefaciens LM11의 유래 생물계면활성물질과 항균활성과의 상관관계)

  • Kang, Beom Ryong;Kim, Yong Hwan;Nam, Hyo Song;Kim, Young Cheol
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.177-185
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    • 2017
  • Bacillus amyloliquefaciens LM11 was isolated from the feces of larvae of the rhino beetle and showed strong antifungal activities against various phytopathogenic fungi by producing biosurfactants. In this study, our overall goal was to determine relationship between biosurfactants produced from the LM11 strain and its role in growth inhibition of phytopathogenic fungi. Production and expression levels of B. amyloliquefaciens LM11 biosurfactants were significantly differed depending on growth phases. Transcriptional and biochemical analysis indicated that the biosurfactants of the LM11 strain were greatly enhanced in late log-phase to stationary phase. Inhibitions of phytopathogenic mycelial growth and spore germination were directly correlated (P<0.001, R=0.761) with concentrations of the LM11 cell-free culture filtrates. The minimum inhibitory surface tension of the culture filtrate of the B. amyloliquefaciens LM11 grown in stationary phase to inhibit mycelial growth of the phytopathogenic fungi was 38.5 mN/m (P<0.001, R=0.951-0.977). Our results indicated that the biosurfactants of B. amyloliquefaciens LM11 act as key antifungal metabolites in biocontrol of plant diseases, and measuring surface tension of the cell-free culture fluids can be used as an easy indicator for optimal usage of the biocontrol agents.

Stain improvement in the white button mushroom 'Seolgang' and its varietal characteristics in Agaricus bisporus

  • Lee, Byung-Joo;Lee, Mi-Ae;Kim, Yong-Gyun;Lee, Kwang-Won;Lim, Yong-Pyo;Lee, Byung-Eui;Song, Ho-Yeon
    • Journal of Mushroom
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    • v.10 no.4
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    • pp.151-159
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    • 2012
  • The button mushroom (Agaricus bisporus) is one of the most widely cultivated important edible mushroom species. In the breeding of new button mushroom, 'Seolgang' was developed by crossing two monokaryons 'CM020913-27' and 'SSU423-31'. Because of the secondarily homothallism, only a small percentage of the basidia produce 3 or 4 spores, which are mostly haploid (n) and do not fruit. Single spore cultures derived from these types of spores produce a vegetative mycelium that also contain a variable number of genetically identical nuclei per cell called monokaryon. The lack of clamp connections between monokaryon and dikaryon required a series of mycelial culture and fruiting test. After crossing, hybrids were cultivated on a small scale and on a commercial scale at a farm. For this, the spawn was made by a commercial spawn producer and the spawned compost by a commercial compost producer. Mycelial growth of 'Seolgang' on CDA was better at $20^{\circ}C$ and $25^{\circ}C$ when it was compared with that of '505 Ho'. The mature cap shape of new strain 'Seolgang' is oblate spheroid and the immature cap shape is round to oblate spheroid. The cap diameter was 41.2 mm on average. In comparison with white strain '505 Ho', the strain had a yield that was 9% higher. It produced fruiting bodies which had a higher weight on average per fruiting body and were 19% firmer with a good shelf life. Days of fruiting body were 3-4 days later than those of '505 Ho'. The physical characteristics such as elasticity, chewiness, adhesiveness were better than that of '505 Ho'. Genetic analysis of the new strain 'Seolgang' showed different profiles compared to '505 Ho', CM02913-27, SSU413-31, when RAPD primers A02 and O04 were used.

Physiological Characteristics of Bacillus spp. Isolated from Rice Straw as Cheonggukjang Starter (볏짚에서 분리한 청국장 starter용 Bacillus spp.의 생리적 특성)

  • Lee, Shin-Ho;Baek, Lag-Min;Park, La-Young
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.562-567
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    • 2008
  • The primary objective of this study was to select a probiotic starter for cheonggukjang using 60 strains isolated from rice straw. Among isolated strains, only 8 strains including strain B-59 evidenced proteolytic, amylolytic and soybean activity. These 8 strains were all gram-positive, spore-forming rods. The B-59 strain evidenced antimicrobial activity against Staphylococcus aureus, Listeria monocytogenes, Pseudomonas fluorescens, and Vibrio parahaemolyticus. The antimicrobial activity of isolated B-59 was verified by its ability to inhibit the growth of S. aureus, L. monocytogenes, P. fluorescens, and V. parahaemolyticus. The selected B-59 strain was indentified as Bacillus licheniformis, as shown by a result of 99.0% homology upon API kit analysis. The selected B-59 strain also displayed viability in pH 2.5 artificial gastric juice, artificial bile acid, NaCl (2, 4, 8, 16, 32%), and ethanol (4, 8, 16, 32%). The antioxidative activity of the strain B-59 culture was assessed via a DPPH free radical scavenging activity assay. The activity increased with an increasing in the fermentation time of strain B-59 for 20 hours.

Selection and Antagonistic Mechanism of Bacillus thuringiensis BK4 against Fusarium Wilt Disease of Tomato (토마토시들음병의 생물학적 방제를 위한 토착길항세균 Bacillus thuringiensis BK4의 선발과 길항기작)

  • Jung Hee-Kyoung;Kim Jin-Rack;Kim Bo-Kum;Yu Tae-Shik;Kim Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.194-199
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    • 2005
  • In oder to select an antifungal substance-producing antagonistic bacterium against Fusarium oxysporum casuing fusarium wilt on tomato, strain BK4 was isolated from local soil of Gyeoungbuk and was identified as Bacillus thuringiensis by 16s rDNA analysis, biochemical test, and Mcirolog TM 3.0 System. The antibiotic of B. thuringiensis BK 4 was highly produced at $30^{\circ}C$ in nutrient broth (pH 9.0). The crude antibiotic was even stable at $121^{\circ}C$ and more stable at slight alkalic condition than acid condition. It was also remained $50{\%}$ activity at pH 3.0. B. thuringiensis BK4 showed the inhibition of spore germination and the biocontrol ability against F. oxysporum causing fusarium wilt of tomato in vivo test. According to these results, B. thuringiensis BK4 was enough to use with a microbial agent for biocontrol against fusarium wilt.