• Title/Summary/Keyword: 균주선발

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Isolation and characterization of lactic acid bacteria for use as silage additives (사일리지 제조를 위한 유산균 탐색 및 특성연구)

  • Ro, Yu-Mi;Lee, Gwan-Hyeong;Park, InCheol;Kim, Wan-Gyu;Han, Byeong-Hak;You, Jaehong;Ahn, Jae-Hyung
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.444-454
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    • 2016
  • Sixteen lactic acid bacterial strains were isolated from silage and cow dung samples, and characterized to identify their potential as silage additives. They were identified as the members of the genera Lactobacillus, Enterococcus, and Weissella, and clustered into nine groups based on the sequences of the genes for 16S rRNA, RNA polymerase alpha subunit, 60-kDa heat shock protein, and phenylalanyl-tRNA synthase alpha subunit. Among them, the three strains which were genetically similar to L. plantarum showed the fastest growth and pH decrease in MRS and rye extract media, the highest numbers of available carbohydrates, and the widest ranges of pH, temperature, and salinity for growth. In addition, they showed no amplified DNA products in the PCR examination targeting the genes for the production of biogenic amines, and the MRS media where they had been cultured showed relatively high inhibition effect against the growth of silage-spoiling microorganisms, including fungi, yeast, and clostridia. The results suggest that these strains are good candidates for silage additives. However, the rye extract media where the lactic acid bacteria had been cultured had no effect on or stimulated the growth of the silage-spoiling microorganisms, and the causes must be established for the practical use of the lactic acid bacteria as silage additives.

Control Effects against Mosquitoes Larva of Bacillus thuringiensis subsp. israelensis CAB199 isolate according to Different Formulations (Bacillus thuringiensis subsp. israelensis CAB199균주의 제형에 따른 모기유충방제 효과)

  • Seo, Mi-Ja;Gil, Yeong-Jong;Kim, Tae-Hwan;Kim, Hyung-Joong;Youn, Young-Nam;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.49 no.2
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    • pp.151-158
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    • 2010
  • Among 18 Bacillus thuringiensis isolates with spherical parasporal inclusion from soils, B. thuringiensis subsp. israelensis CAB199 was selected. It was showing over 90% mortality against Aedes aegypti and Culex pipiens moletus. It was confirmed that this B. thuringiensis subsp. israelensis CAB199 isolate also had a insecticidal activity against Culex inatomii that was occurred in the marsh. Because most of mosquito larva were primarily situated or shifted from under- to surface water, we need to select long floating formulations on surface water for controlling mosquito larva. It was tested the pesticidal and control effects in the laboratory and wetland with two formulation types of B. thuringiensis subsp. israelensis, for example, wettable power (WP) and suspension concentrate type (SC). Laboratory test showed that SC formulation type was relatively faster and more effective against 3 tested mosquito species, C. pipiens, Aedes aegypti, and C. inatomii. Otherwise, the control efficacy of SC formulation type was more rapidly appeared against C. inatomii in the wetland.

Selection of parental monokaryons from Korean Hypsizigus marmoreus by protoplast regeneration (원형질체 재생을 통한 느티만가닥버섯 단핵균주 선발)

  • Oh, Youn-Lee;Kong, Won-Sik;Jang, Kab-Yeul;Shin, Pyung-Gyun;Kim, Eun-Sun;Oh, Min ji;Choi, In-Geol
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.270-273
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    • 2015
  • Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

Isolation and Characterization of a Bacteroides Strain Utilizing Inulin from Pig Feces (돼지분변으로부터 Inulin이용 Bacteroides속 균주의 분리 및 특성)

  • Kim, Chang-Gon;Kim, Su-Il;Shin, Hyun-Kyung
    • Korean Journal of Food Science and Technology
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    • v.25 no.6
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    • pp.780-786
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    • 1993
  • About 7.7% of the total anaerobic bacteria in pig feces grew with clear zone around the colonies on the agar medium containing inulin as a sole carbon source. Among these bacteria, a strain with the strongest inulin-utilizing activity was isolated and identified as Bacteroides sp. based on its morphological and taxonomical characteristics. The isolate grew well with inulin, fructooligosaccharides or glucose as a sole carbon source, while its growth dropped to 50% of that obtained with glucose when soluble starch or sucrose are used. Since the inulase activity was found only when fructooligosaccharides or inulin was added to the growth medium, but not when glucose, sucrose or soluble starch was applied, the inulase production was considered to be induced by fructooligosaccharides or inulin. The highest inulase activity, 0.42 U/ml was detected with the inulin medium and 0.25 U/ml with fructooligosaccharides medium. The cell growth of the isolated strain increased with the amounts of inulin up to 2%(w/v) and maximum production of inulase was found in the cells fed 1% inulin. The inulase of the isolated Bacteroides sp. showed its maximum activity at pH $7.0{\sim}7.5\;and\;50{\sim}50^{\circ}C$ and was found to be an exoinulase judging by its mode of action.

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Identification of the Oligotrophic Bacteria Strain 7F Biocontrolling Phytophthora Blight Disease of Red-pepper (고추 역병 방제를 위한 저영양 길항세균 7F 균주의 동정)

  • Kim, Dong-Gwan;Yeo, Yun-Soo;Kwon, Soon-Wo;Jang, Kil-Su;Lee, Chang-Muk;Lee, Mi-Hye;Kim, Soo-Jin;Koo, Bon-Sung;Yoon, Sang-Hong
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.41-47
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    • 2010
  • A total of 10,753 oligotrophic bacteria were isolated from the cultivated soils of red-pepper infected by Phytophthora blight disease in various regions of Korea (Chungju, Anmyon, Taean, Andong, Eumsung and Goesan). Seven bacteria isolates among these collected resources were selected by the first screening of in vitro antagonistic assay against major several plant pathogenic fungi including Phytophthora capsici. Finally, strain 7F was selected by pot assay for a possible biological control agent against Phytophthora blight disease of pepper seedling in the greenhouse. Strain 7F was identified as Bacillus subtilis on the basis of its 16S rDNA sequence analysis and as standardized biochemical characteristics assay kits such as API20 NE. In the experiment of P. capsici zoospore infected red-pepper on the pot test, infection rate of red-pepper with nonetreatment to Phytophthora blight disease was 87%, while the rate was only 6% in the pot treated with strain 7F. This result indicated that the Bacillus subtilis strain 7F will be useful as a potential biocontrol agent for Phytophthora blight disease of red-pepper.

Antifungal Activity of Bacillus vallismortis 1A against Phytopathogen (식물병원균에 대한 Bacillus vallismortis 1A 균주의 항진균 활성)

  • Lee, Mi-Hye;Kim, Soo-Jin;Lee, Chang-Muk;Jang, Jae-Seon;Chang, Hai-Joong;Park, Min-Seon;Koo, Bon-Sung;Yoon, Sang-Hong;Yeo, Yun-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.5
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    • pp.362-368
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    • 2008
  • In order to isolate novel oligotrophic bacteria exhibiting antifungal activities, soils were collected from pepper-cultivated fields of Yeongyang, Jecheon, Nonsan, Eumsong and Goesan area in Korea. From soils in pepper cultivated area, a total of 9,354 strains were isolated as oligotrophic bacteria by the R2A dilution method. Among 9,354 oligotrohic bacteria candidates, 1A strain was selected by screening against Phytophthora capsici causing phytophthora blight of hot pepper in the greenhouse and field. The strain was identified as Bacillus vallismortis based on its 16S rDNA sequence and key characteristics as compared with those of authentic cultures of B. vallismortis(KACC 12149) and B. mojavensis(KACC 12096). The strain showed broad spectrum of antibiotic activity in vitro test, as revealed in its strong inhibitory activity to the genera Phytophthora, Collectotrichum, Botrytis and Fusarium, but not to Rhizoctonia and Magnaporthe. In pot experiments, infection rate of hot pepper in the non-treated pots was about 89%, while it was only 29% in the pots treated with 1A strain. The result indicated B. vallismortis 1A is a potential biocontrol agent for phytophthora blight of hot pepper

Biological Control of Sclerotinia sclerotiorum in Lettuce Using Antagonistic Bacteria (길항세균을 이용한 상추 균핵병의 생물학적 방제)

  • Chon, Bong-Goan;Park, Suji;Kim, Jin-Won
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.12-20
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    • 2013
  • To isolate antagonistic bacteria against sclerotinia rot of lettuce, caused by Sclerotinia sclerotiorum, soil samples were collected from the diseased greenhouse field in Namyangju city, Gyeong-gi province from 2007 to 2008. A total of 196 bacterial isolates were isolated using serial dilution method. In dual culture assay in vitro, 26 isolates showed more than 80% of inhibition rates of mycelial growth of S. sclerotiorum. Based on 16S rDNA sequence analysis, the 26 isolates were identified as Bacillus megaterium, B. cereus, B. subtilis, Arthrobacter nicotianae, A. ramosus, Pseudomonas filiscindens, Stenotrophomonas maltophilia, Brevibacterium frigoritolerans and Sphingobacterium faecium. The 26 isolates inhibited the mycelial growth of S. sclerotiorum up to 80% and the sclerotial germination 0-100%. In the greenhouse pot test of ten isolates conducted in summer, 2 isolates B. megaterium (DK6) and B. cereus (C210) showed control efficacy on sclerotia viability of S. sclerotiorum, 20% and 35%, respectively. In the greenhouse pot test in winter, the disease incidence of the control group was 80%, whereas those of 9 isolates among 26 were approximately 20%. From the result, the 9 isolates are expected as potentially antagonistic bacteria for biological control of sclerotinia rot of lettuce caused by S. sclerotiorum.

Cultivation Conditions for Mass Production of an Antagonistic Bacterium Stenotrophomonas maltophilia BW-13 (길항세균 Stenotrophomonas maltophilia BW-13의 대량배양을 위한 최적 배양조건)

  • Park Jong-Young;Kim Han-Woo;Kim Hyun-Ju;Chun Ok-Ju;Jung Soon-Je;Choi Woobong;Lee Seon-Woo;Moon Byung-Ju
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.158-161
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    • 2005
  • Stenotrophomonas maltophilia BW-13 is a potent biocontrol agent to control crisphead lettuce bottom rot caused by Rhizoctonia solani. To define the optimum conditions for the mass production of the S. maltophilia BW-13, we have investigated optimum culture conditions and effects of various carbon sources on the bacterial growth. The optimum initial pH and temperature were determined as pH $6.0\~7.0 and $35^{\circ}C$, respectively. For the selection of effective carbon source for the mass production, we tested the low molecular carbon sources such as sucrose, glucose, lactose, maltose, manose and the high molecular carbon source such as dough conditioner, rice bran, corn starch, sweet potato starch. As the results, the addition of dough conditioner in a basal medium ($1.25\%\;K_{2}HPO_4,\;0.38\%\;KH_{2}PO_4,\;0.01\%\;MgSO_4{\cdot}7H_{2}O,\;0.5\%\;Yeast extract$) was able to achieve higher cell density and the antifungal activity than others. Therefore, the basal medium containing $3\%$ dough conditioner (named as dough conditioner medium) was finally selected the optimized media for the mass production of BW-13 strain.

Studies on the Development of Acid Tolerant and Superior Nitrogen Fixation Symbionts for Pasture on Hilly Land -I. The Degrees of Acid Tolerance of Rhizobia from the Pastures in Yeongnam Area (야산(野山) 목초지용(牧草地用) 내산성(耐酸性) 우수(優秀) 질소고정균주(窒素固定菌株) 개발(開發) -I. 영남지방(嶺南地方) 목야지(牧野地) 근류균(根瘤菌)의 내산성정도(耐酸性程度))

  • Kang, Ui-Gwm;Choi, Ju-Hyeon;Cho, Kang-Jin;Jung, Yeun-Tae;Lee, Jae-Saeng
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.4
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    • pp.369-374
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    • 1987
  • This study was conducted to evaluate the degree of acid tolerance of indigenous Rhizobium in natural soils, and to select the superior Rhizobia for legume forage on hilly land. 153 Rhizobial strains isolated from two host genera, Medicago and Trifolium in the pastures of Yeongnam area were screened on acidic agar plates. The results obtained are summarized as follows: 1. The degrees of acid tolerance of R. meliloti and R. trifolii were ranged from 4.3 to pH 5.5 and from 4.0 to pH 5.5 respectively. 2. Among acid tolerant Rhizobia isolates, two strains of R. meliloti at pH 4.3 and three strains of R. trifolii at pH 4.0 were shown to have the activities of nodulation and nitrogenase. 3. R. trifolii strains were shown to survive about 80 to 99% at the pH range 4.8 to 5.0, while R. meliloti were about 35 to 47% at that pH range. Therefore, more acid tolerant R. meliloti should be developed for legume forage in Korean hilly land.

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Isolation of Antifungal Substances by Bacillus amyloliquefaciens IUB158-03 and Antagonistic Activity against Pathogenic Fungi (Bacillus amyloliquefaciens IUB158-03이 생산하는 항진균물질의 분리와 항균활성)

  • Kim, Hye-Young;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.96-103
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    • 2009
  • For the control of pathogenic microorganisms, Bacillus spp. were isolated from diseased pepper fruits in Korea. Among them, Bacillus sp. IUB158-03 showed high inhibitory effect on mycelial growth and spore germination of C. gloeosporioides and Botrytis cinerea. The strain was identified as B. amyloliquefaciens IUB158-03 based on its physiological, biochemical characteristics and Microlog analysis. The highest level of antifungal substances by B. amyloliquefaciens IUB158-03 were obtained when the bacterium was cultured in medium containing 2% soluble starch, 3% yeast extract, 0.5% tryptone, 0.5% $NH_4H_2PO_4$, and 1% NaCl (pH 6.0) at $25^{\circ}C$ for 72 hrs. The antifungal substances were purified by butanol extraction, silica gel column chromatography, preparative thin layer chromatography, and high performance liquid chromatography. The purified antifungal substance was confirmed $R_f$ 0.27 by TLC. This substance exhibited antifungal activity against Fusarium solani, Rhizoctonia solani, Botrytis cineria, Alternata alternaria of plant pathogenic fungi and Trichophyton mentagrophytes, Epidermophyton floccosum, Cryptococcus neoformans of human pathogenic fungi.