• Title/Summary/Keyword: Bacterial culture tests

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Isolation and Culture Characteristics of a Bacterial Symbiont from Entomopathogenic Nematode Steinernema galseri (Steinernema glaseri 곤충병원선충으로부터 공생박테리아의 분리 및 배양특성)

  • 박선호;유연수
    • KSBB Journal
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    • v.14 no.2
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    • pp.198-204
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    • 1999
  • Asymbiotic bacterium with highly effective toxins was isolated from entomopathogenic nematode Steinernema glaseri which has been widely used against various soil-inhabiting pests. The symbiont of S. glaseri was identified as Xenorhabdus nematophilus sp. by using several biochemical and physiological tests. When this strain was released into the hemolymph of insect larva, it produced highly toxic substances and killed the larva within 2 days. Two colony forms that differed n some biochemical characteristics were observed when cultures in vitro. Phase l colonies were mucid and difficult to be dispersed in liquid. Phase II was not mucoid and was easily dispersed in liquid. It did not adsorb neutral red or bromothymol blue. Rod-shaped cell size was highly variable between two phases, ranging 2-10 ${\mu}{\textrm}{m}$. It was also found that only infective-stage nematodes can carry only primary-phase Xenorhabdus in their intestine.

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Characterization of Two Urease-Producing and Calcifying Bacillus spp. Isolated from Cement

  • Achal, Varenyam;Mukherjee, Abhijit;Reddy, M. Sudhakara
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1571-1576
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    • 2010
  • Two bacterial strains designated as CT2 and CT5 were isolated from highly alkaline cement samples using the enrichment culture technique. On the basis of various physiological tests and 16S rRNA sequence analysis, the bacteria were identified as Bacillus species. The urease production was 575.87 U/ml and 670.71 U/ml for CT2 and CT5, respectively. Calcite constituted 27.6% and 31% of the total weight of sand samples plugged by CT2 and CT5, respectively. Scanning electron micrography analysis revealed the direct involvement of these isolates in calcite precipitation. This is the first report of the isolation and identification of Bacillus species from cement. Based on the ability of these bacteria to tolerate the extreme environment of cement, they have potential to be used in remediating the cracks and fissures in various building or concrete structures.

Subcorneal pustular dermatosis in a dog (개에서 발생한 각질하 농포성 피부병 1례)

  • Park, Seong-Jun
    • Korean Journal of Veterinary Research
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    • v.53 no.2
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    • pp.125-127
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    • 2013
  • Canine Subcorneal pustular dermatosis (CSPD) represents a sterile, superficial, pustular skin disease of unknown cause but may be a variant of pemphigus foliaceus. A 7-year-old, intact female, mixed dog presented with 3-month history of pruritic multiple pustules and crusts. Direct smears from intact pustules revealed numerous nondegenerate neutrophils, some acantholytic cells, and bacterial culture was negative. Histologic examination of lesional skin showed subcorneal pustules filled with neutrophils and acantholytic cells. The direct immunofluorescence tests stained with IgG, IgA, IgM, C3 were negative. Oral administration of dapsone (1 mg/kg/q8h) was initiated and it was reduced to 1 mg/kg/q12h with good control of the lesions.

Isolation and Utilization of Antagonistic Pseudomonas fluorescens from Soils for the Protection of Soybean Sprouts Rot (콩나물 부패병 방제를 위해 토양으로부터 분리한 길항균 Pseudomonas fluorescens의 이용)

  • Kim, Jin-Ho;Joo, Gil-Jae;Choi, Yong-Hwa
    • Korean Journal of Environmental Agriculture
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    • v.20 no.1
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    • pp.50-56
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    • 2001
  • Thirty-three bacterial and fungal strains were isolated from the rotten soybeans and soybean sprouts to isolate pathogenic microorganisms which cause soybean sprouts rot during soybean sprouts cultivation. In pathogenicity tests of the isolates on soybean sprouts, two isolates(K-17 and K-28) caused soybean sprouts rot and were identified as Erwinia carotovora and Fusarium sp., respectively. To isolate antagonists aganist K-17 and K-28 pathogens, bacteria were isolated from various soybean-cultivated soils and screened by the inhibition zone method. A bacterial isolate(J-232) which inhibited growth of both pathogens was identified as Pseudomonas fluorescens and further examined. The culture filtrate of P. fluorescens J-232 (dilution rate of 500 times) inhibited the growth of Erwinia carotovora K-17 and Fusarium sp. K-28 both on potato dextrose agar medium and on soybean sprouts cultivated in vessel. The development of soybean sprouts rots was observed during cultivation by inoculation of soybean seeds with culture filtrate of both pathogens. The combined inoculation of soybean seeds with culture filtrate of antagonistic bacterium and that of pathogens prevented soybean sprouts rot, and the growth of soybean sprouts was similar to that of control. The soybean sprouts inoculated with antagonists culture filtrate alone did not develop soybean sprouts rot, and the growth of the seedlings was shown to be slightly promoted as compared with that of control.

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Biological Control of Soil-borne Diseases with Antagonistic Bacteria

  • Kim, Byung-Ryun;Hahm, Soo-Sang;Han, Kwang-Seop;Kim, Jong-Tae;Park, In-Hee
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.25-25
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    • 2016
  • Biological control has many advantages as a disease control method, particularly when compared with pesticides. One of the most important benefits is that biological control is an environmental friendly method and does not introduce pollutants into the environment. Another great advantage of this method is its selectivity. Selectivity is the important factor regarding the balance of agricultural ecosystems because a great damage to non target species can lead to the restriction of natural enemies' populations. The objective of this research was to evaluate the effects of several different bacterial isolates on the efficacy of biological control of soil borne diseases. White rot caused by Sclerotium cepivorum was reported to be severe disease of garlic and chive. The antifungal bacteria Burkholderia pyrrocinia CAB08106-4 was tested in field bioassays for its ability to suppress white rot disease. In field tests, B. pyrrocinia CAB08106-4 isolates suppressed white rot in garlic and chive, with the average control efficacies of 69.6% and 58.9%, respectively. In addition, when a culture filtrate of B. pyrrocinia CAB08106-4 was sprayed onto wounded garlic bulbs after inoculation with a Penicillium hirstum spore suspension in a cold storage room ($-2^{\circ}C$), blue mold disease on garlic bulbs was suppressed, with a control efficacy of 79.2%. These results suggested that B. pyrrocinia CAB08106-4 isolates could be used as effective biological control agents against both soil-borne and post-harvest diseases of Liliaceae. Chinese cabbage clubroot caused by Plasmodiophora brassicae was found to be highly virulent in Chinese cabbage, turnips, and cabbage. In this study, the endophytic bacterium Flavobacterium hercynium EPB-C313, which was isolated from Chinese cabbage tissues, was investigated for its antimicrobial activity by inactivating resting spores and its control effects on clubroot disease using bioassays. The bacterial cells, culture solutions, and culture filtrates of F. hercynium EPB-C313 inactivated the resting spores of P. brassicae, with the control efficacies of 90.4%, 36.8%, and 26.0%, respectively. Complex treatments greatly enhanced the control efficacy by 63.7% in a field of 50% diseased plants by incorporating pellets containing organic matter and F. hercynium EPB-C313 in soil, drenching seedlings with a culture solution of F. hercynium EPB-C313, and drenching soil for 10 days after planting. Soft rot caused by Pectobacterium carotovorum subsp. carotovorum was reported to be severe disease to Chinese cabbage in spring seasons. The antifungal bacterium, Bacillus sp. CAB12243-2 suppresses the soft rot disease on Chinese cabbage with 73.0% control efficacy in greenhouse assay. This isolate will increase the utilization of rhizobacteria species as biocontrol agents against soft rot disease of vegetable crops. Sclerotinia rot caused by Sclerotinia sclerotiorum has been reported on lettuce during winter. An antifungal isolate of Pseudomonas corrugata CAB07024-3 was tested in field bioassays for its ability to suppress scleritinia rot. This antagonistic microorganism showed four-year average effects of 63.1% of the control in the same field. Furthermore, P. corrugata CAB07024-3 has a wide antifungal spectrum against plant pathogens, including Sclerotinia sclerotiorum, Sclerotium cepivorum, Botrytis cinerea, Colletotrichum gloeosporioides, Phytophotra capsici, and Pythium myriotylum.

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Bacterial Blight Resistant Mid-late Maturing Rice 'Manbaek' with High Grain Quality (벼흰잎마름병 저항성 고품질 중만생 벼 '만백')

  • Park, Hyun-Su;Baek, Man-Kee;Kim, Bo-Kyeong;Kim, Ki-Young;Shin, Woon-Chul;Ko, Jae-Kwon;Nam, Jeong-Kwon;Kim, Woo-Jae;Cho, Young-Chan;Ko, Jong-Cheol;Kim, Jeong-Ju;Kim, Hyun-Soon
    • Korean Journal of Breeding Science
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    • v.49 no.3
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    • pp.235-244
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    • 2017
  • 'Manbaek' is a bacterial blight resistant mid-late maturing rice cultivar with high grain quality. 'Manbeak' was derived from anther culture using the backcross combination, $Hopum^*2/SR30075$. 'Hopum' is a mid-late maturing rice cultivar with high grain quality and 'SR30075' is a pyramid line carrying three bacterial blight resistance genes. 'Manbaek' was selected through the pedigree method, yield trials, and local adaptability tests. 'Manbeak' carrying two bacterial blight resistance gene Xa3 and xa5 showed high-level and broad-spectrum resistance against bacterial blight. 'Manbaek' was resistant to K3a, mostly virulent race in Korea, and exhibited resistance reaction against 16 Korean bacterial blight isolates. 'Manbaek' was a mid-late maturing rice. The heading date of 'Manbaek' was August 19th, which was 5 days later than that of 'Nampyeong'. Manbaek' was a lodging-tolerant rice with short culm and dark green leaf. Due to the low viviparous germination, 'Manbaek' could be a useful material to prevent pre-harvest sprouting. 'Mabeak' was resistant to bacterial blight and rice stripe virus, but susceptible to other virus diseases and insect pests. The yield of 'Manbaek' was similar to 'Nampyeong'. 'Manbaek' showed excellent grain appearance and good tastes of cooked rice, so that it could contribute to improving the quality of bacterial blight resistant cultivars. 'Manbaek', bacterial blight resistant cultivar with high grain quality, is suitable for the cultivation at bacterial blight prone area and has been utilized in the breeding programs for enhancing the resistance against bacterial blight (Registration No. 6069).

Isolation of Marine Bacteria Killing Red Tide Microalgae II. Isolation and Algicidal Properties of Pseudomonas sp. LG-2 Possessing Killing Activity for Dinoflagellate, Prorocentrum micans (적조생물 살조세균 탐색 II. 적조생물 Prorocentrum micans 살조세균 Pseudomonas sp. LG-2의 분리와 살조특성)

  • LEE Won-Jae;PARK Young-Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.6
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    • pp.852-858
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    • 1998
  • We have isolated a bacterial strain that tends to kill P. micans from the mixed culture of p. minns plus seawater filtrate (poresize, 0.8 $\mu$m) collected at Masan bay in July 1996, in which the mixed culture grown in the f/2 medium. According to the experimental results of the isolated bacterium such as fatty acids analysis, morphological and biochemical characteristic tests, the strain was supposed to be a Pseudomonas and then it was named as Pseudomonas sp. LG-2. The killing effect of Pseudomonas sp. LG-2 against P. micans was proportionally increased with the concentrations of culture filtrate (pore size, 0.8 $\mu$m) is well as with the number of bacterium inoculated. In the mixed culture inoculated with $1.3\times10^6$ cells/ml of Pseudomonas sp. LG-2, the number of P. micans (2,000 cells/ml) was gradually decreased and then killed below 100 cells/ml within 7 days. In addition, the culture filtrate with $30\%$ of final concentration revealed a significant killing effect against P. micans around 3 days after culture. In the relationship between killing effects and growth stage of Pseudomonas sp. LG-2, the culture filtrate at lag phase has little effects on P. micans. In constant, the culture filtrate at mid-log phase showed the killing effect by decreasing P. micans to 112 in number within 5 days. In particular, the culture filtrate at stationary phase showed a significant killing effect against P. micans in which the majority of it was killed after 3 day culture. The species specificity of killing effects of Pseudomonas sp. LG-2 against 5 species of dinoflagellate was only found in P. micans and Scrippsiella trochoidea.

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Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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Antibacterial properties of composite resins incorporating silver and zinc oxide nanoparticles on Streptococcus mutans and Lactobacillus

  • Kasraei, Shahin;Sami, Lida;Hendi, Sareh;AliKhani, Mohammad-Yousef;Rezaei-Soufi, Loghman;Khamverdi, Zahra
    • Restorative Dentistry and Endodontics
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    • v.39 no.2
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    • pp.109-114
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    • 2014
  • Objectives: Recurrent caries was partly ascribed to lack of antibacterial properties in composite resin. Silver and zinc nanoparticles are considered to be broad-spectrum antibacterial agents. The aim of the present study was to evaluate the antibacterial properties of composite resins containing 1% silver and zinc-oxide nanoparticles on Streptococcus mutans and Lactobacillus. Materials and Methods: Ninety discoid tablets containing 0%, 1% nano-silver and 1% nano zinc-oxide particles were prepared from flowable composite resin (n = 30). The antibacterial properties of composite resin discs were evaluated by direct contact test. Diluted solutions of Streptococcus mutans (PTCC 1683) and Lactobacillus (PTCC 1643) were prepared. 0.01 mL of each bacterial species was separately placed on the discs. The discs were transferred to liquid culture media and were incubated at $37^{\circ}C$ for 8 hr. 0.01 mL of each solution was cultured on blood agar and the colonies were counted. Data was analyzed with Kruskall-Wallis and Mann-Whitney U tests. Results: Composites containing nano zinc-oxide particles or silver nanoparticles exhibited higher antibacterial activity against Streptococcus mutans and Lactobacillus compared to the control group (p < 0.05). The effect of zinc-oxide on Streptococcus mutans was significantly higher than that of silver (p < 0.05). There were no significant differences in the antibacterial activity against Lactobacillus between composites containing silver nanoparticles and those containing zinc-oxide nanoparticles. Conclusions: Composite resins containing silver or zinc-oxide nanoparticles exhibited antibacterial activity against Streptococcus mutans and Lactobacillus.

Studies on the Pathogenic Pseudomonas Causing Bacterial Diseases of Cultivated Mushrooms in Korea 2. Bacteriological Characteristics of tolaasii Causing Mushroom Brown Blotch and White Line Reacting Organisms (인공재배 버섯에 병을 일으키는 Pseudomonas속 병원세균에 관한 연구 2. 버섯 세균성 갈색점무늬병의 병원세균 Pseudomonas tolaasii와 White Line 형성균의 세균학적 특성)

  • 김종완;권순익;강희진
    • Korean Journal Plant Pathology
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    • v.11 no.4
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    • pp.353-360
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    • 1995
  • Tests were performed on 232 bacterial strains (71 strains of Pseudomonas tolaasii and 161 white line reacting organisms, WLRO) isolated from cultivated mushrooms. As results, P. tolaasii was divided into 5 groups on the basis of the phenotypical characteristics for the strains, and group 3 was the major one including 48 (62%) out of the total 71 strains. WLRO were classified into 23 groups, and group 10 was the major group (65 strains, 30% of the total WLRO tested). A white line was well formed at 22$^{\circ}C$ and at 4 mm distance between P. tolaasii and WLRO colonies in their dual culture on Pseudomonas agar F medium within 36-hr incubation, but not formed at $25^{\circ}C$ even for 72-hr incubation. The morphological, cultural and biological properties of P. tolaasii group 3, and the main group of WLRO, group 10, were different only in the light of pathogenicity. Also group 2 of P. tolaasii had the characteristics similar to group 24 of WLRO which was pathogenic to cultivated mushrooms, suggesting the P. tollaasiii and WLRO strains may be the same species although their white line forming ability and pathogenicity were more or less different from one another. Subculture of the strains in P. tolaasii group 1 induced the variation of their pathogenicity, white line forming ability and utilization of sodium benzoate and sodium tartrate, and thus it can be inferred that they were converted to strains having the characteristics of group 3 or WLRO groups.

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