• Research in Plant Disease
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• v.9 no.2
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• pp.94-98
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• 2003

To produce antibodies against Pseudomonas tolaasii and P. agarici, lyophilized P. tolaasii and P. agarici ($5{\times}10^7$ cfu/ml) and Freund, s adjuvant were immunized into rabbits 4 times. The specificity and sensitivity of the antibodies were evaluated by immunodiffusion test and indirect enzyme-linked immunosorbent assay (id ELISA). The ${\alpha}$-P. tolaasii antibody was very specific only against P. tolaasii, while ${\alpha}$-P. agarici antibody was not specific and showed a high cross reactivity toward P. tolaasii with detection limit concentration of $2{\times}10^3$ cfu/ml. However, the cross reactivities of ${\alpha}$-P. agarici antibody toward the related species including P. reactans were very low. Our results showed that ${\alpha}$-P. tolaasii and ${\alpha}$-P. agarici antibodies against P. tolaasii and P. agarici, respectively, might be useful for rapid and simple detection of the causal agents of bacterial brown and yellow blotches in cultivated oyster mushrooms.

• The Korean Journal of Mycology
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• v.44 no.3
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• pp.171-175
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• 2016

A gram-negative bacterium was isolated from spent substrates of Agaricus bisporus and showed significant antagonistic activity against Pseudomonas agarici. The bacterium was identified as Alcaligenes sp. based on cultural, biochemical, physiological characteristics and a 16S rRNA sequence analysis. The isolate is saprophytic, but not parasitic or pathogenic on cultivated mushroom, whereas it showed strong inhibitory effects against P. agarici cells in vitro. The control efficacy of Alcaligenes sp. HC12 against brown blotch of P. agarici was up to 63% on Agaricus bisporus. The suppressive bacterium may be useful for the development of biocontrol systems.

• Journal of Mushroom
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• v.17 no.1
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• pp.19-23
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• 2019

A gram-positive bacterium was isolated from the spent substrate of Agaricus bisporus that showed a marked antagonistic activity against Pseudomonas agarici. It was identified as Bacillus safensis HC42 based on its cultural, biochemical, and physiological characteristics, and 16S rRNA sequence. B. safensis HC42 was saprophytic, but not parasitic or pathogenic, in cultivated mushrooms and showed strong inhibition of P. agarici in vitro. Moreover, it showed a control efficacy of 66 % against browning disease caused by P. agarici in Agaricus bisporus. Therefore, B. safensis HC42 may be useful in the future for the development of a biocontrol system.

• The Korean Journal of Mycology
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• v.26 no.1 s.84
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• pp.60-68
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• 1998

The effect of P. agarici and P. tolaasii causing the bacterial disease of mushrooms on the mycelial growth and fruitbody formation of F. velutipes was evaluated in laboratory. When the pathogenic bacteria was inoculated simultaneously with F. velutipes or 5 days after inoculation of F. velutipes, they significantly deterred both mycelial growth and fruitbody formation of F. velutipes in sawdust culture and showed strong inhibition under high population density. They appeared to be tender or milky in exhibiting symptom on F. velutipes by inoculating the concentration of $10^2{\sim}10^6$ of unit/g media, and their growth seemed to be stopped under $10^8\;cfu/g$ media. On $10^2\;cfu/g$ media of P. agarici and $10^4\;cfu/g$ media of P. tolaasii, there was no effect on the fruitbody yield of F. velutipes. P. tolaasii was more suppressive in the mycelial growth of F. velutipes than P. agarici, while on fruitbodies formation of F. velutipes, P. agarici showed slightly higher inhibition than that of P. tolaasii. When the bacteria was inoculated 10 days after inoculation of F. velutipes, both mycelial growth and fruit body formation were not affected nearly.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.197-210
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• 1994

This experiment was carried out to study the cause of degeneration and rot of cultivated mushroom. Among 597 bacterial isolates derived from the rots of Button mushroom (Agaricus bisporus), Oyster mushroom (Pleurotus ostreatus) and Oak mushroom (Lentinus edodes) collected from markets of 5 cities (Seoul, Suwon, Taegu, Pohang and Pusan) in Korea (1991~1993), 111 bacterial isolates (18.5%) were proved as pathogenic bacteria. These pathogenic bacteria causing bacterial rots of cultivated mushrooms were identified as Pseudomonas tolasii, P. agarici, and Eriwinia sp., and the main causal bacteria were P. tolaasii. P. fluorescens and Klebsiella plenticola were confirmed as saprophytic non-pathogenic bacteria. One hundred fifty nine isolates (Group No. 39) of the 486 saprophytic bacterial isolates were classified as P. fluorescens, and this species was most often found rot area of cultivated mushrooms. P. tolaasii, the causal organism of bacterial blotch, was classified into two groups; One group can be differentiated from the other by the formation of white precipitation band by white line reacting organisms of Pseudomonas Agar F media. P. tolaasii attacked the cultivated mushrooms relatively well at lower incubation temperature such as 5$^{\circ}C$, but P. agarici rarely attack at below 1$0^{\circ}C$. The temperature for the infection commercial cultivated mushrooms by P. agarici was higher than that of P. tolaasii. Optimum temperature for the infection of mushrooms by P. tolaasii and P. agarici were 2$0^{\circ}C$ and $25^{\circ}C$, respectively.

• Journal of Mushroom
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• v.14 no.4
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• pp.191-196
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• 2016

This study was conducted to investigate optimum conditions for mass production of ntagonistic microbes Alcaligenes sp. HC12. Alcaligenes sp. HC12 had a potent biological control agent to control browning disease caused by Pseudomonas agarici. Alcaligenes sp. HC12 markedly showed the antagonistic activity against Pseudomonas agarici, the most destructive pathogen of cultivated mushrooms. To define the optimum conditions for the mass production of the Alcaligenes sp. HC12, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 9.0 and $30^{\circ}$, respectively. The optimal concentration of medium elements for the growth of pathogen inhibitor bacterium(Alcaligenes sp. HC12) was determined as follows: 0.5% dextrine, 1.5% yest extract, 1.0% $NaNO_3$, 0.5% $KH_2PO_4$, and 1.5% asparagine.

• Korean Journal Plant Pathology
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• v.14 no.4
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• pp.350-357
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• 1998

A total of 12 strains of fluorescent Pseudomonas isolated from the cultivated mushrooms such as Agaricus bisporus and Pleurotus ostreatus were collected. They consisted of pathogenic Pseudomonas spp. and epiphytic Pseudomonas spp. of the cultivated mushroom. To analyze the phylogenetic relationship of these strains, ITS I region, the 16S-23S intergenic spacer region in the ribosomal RNA (rRNA) operon, was cloned and sequenced. The spacer regions of these strains were 495∼527 nucleotides in length and contained the genes encoding isoleucine-tRNA (tRNAIle) and alanine-tRNA (tRNAAla). The reciprocal homologies of each ITS I sequence among these strains were in the range of 84.2%∼98.8%. According to the analysis of ITS I sequences, the fluorescent Pseudomonas spp. were phylogenetically classified into three clusters. Cluster I consisted of Pseudomonas fluorescens, P. tolaasii, P. gingeri’, and P.‘reactans’(WLRO). Cluster II comprised Pseudomonas fluorescens biovar C and F. Cluster III composed P. agarici. Cluster I and II could be classified into P. fluorescens complex. P. agarici formed an independent taxon clearly separable from P. florescens complex.

• The Korean Journal of Mycology
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• v.45 no.4
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• pp.370-376
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• 2017

Pseudomonads cause bacterial brown blotch disease, which causes great damage to the common mushroom Agaricus bisporus. The tolerance of A. bisporus to pseudomonads was tested and found to not be correlated with mycelium growth ability. The offsprings of the tolerant strain (ASI1085) to pseudomonads were not as tolerant as their parents in the mycelium stage. But, tolerance decreased compared to mycelium in the fruiting body. The offsprings of the weakly tolerant strain (ASI1321) were even more weak in the mycelium stage. It is presumed that the tolerance of the parents is transferred to later generations. The tolerance in the mycelium was not correlated in the fruiting body. Therefore, the browning of the fruiting body is thought to be induced by other factors. Pseudomonas tolaasii caused higher browning than Pseudomonas agarici. Pseudomonas reactans did not have a significant effect on the mycelium, but affected the browning of the fruit bodies. P. agarici had higher ability to inhibit mycelium growth than fruiting body growth.

• The Korean Journal of Mycology
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• v.22 no.2
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• pp.190-195
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• 1994

Studies were conducted to determine the potential of sodium hypochlorite(SHC) on the control of bacterial yellow blotch in cultivated oyster mushroom, Pleurotus ostreatus. SHC at the concentration of 80 ppm was effective on the control of Pseudomonas agarici causing yellow blotch in oyster mushroom except number 916 isolate. In vitro the mycelial growth was slightly inhibited at the concentration higher than 100 ppm of sodium hypochlorite, but retardation of the mycelial growth was soon recovered. Spray of SHC solution at the concentration of 40-50 ppm per day significantly reduced the incidence of the yellow blotch without impairing the growth of oyster mushroom in field culture. However, the higher concentration of SHC(67 ppm) induced yellow brown or dark gray in color and deformed cap and elongated stripe in morphology of fruiting body. Results indicate that periodical spray of sodium hypochlorite seems to be the recommendable method for protection against bacterial yellow blotch disease in oyster mushroom without reducing food quality.

• Journal of Food Hygiene and Safety
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• v.16 no.3
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• pp.232-240
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• 2001

One hundred twenty five bacterial isolates were obtained from the brown blotch-diseased oyster mushrooms collected from markets. Among them, 45 were determined as pathogenic bacteria and white line forming organisms(WLFO) were 6 strains and white line reaction organisms (WLRO) were 6 strains. All of the white line forming isolates were identified as Pseudomonas tolaasii which is a known pathogen of brown blotch disease of oyster mushroom by GC-MIS(Gas chromatography-microbial identification system). Six of the white line reacting organisms were identified as P. chlomraphis, P. fluorescens biotype A and type C. The rest of them were P gingeri, P. agarici, P. fluorescens biotype B, P. chloroyaphis, non-pathogenic P. tolaasii, P. putida biotype A and B etc. For spectrum of activity of tolaasin, culture filtrates from pathogenic isolates were examined by browning of mushroom tissue and pitting of mushroom caps. The weak pathogenic bacteria didn't induce browning or pitting of mushroom tissue. On the other hand, strong pathogenic isolates showed browning and pitting reaction on mushroom. An extracellular toxin produced by P. tolaasii, was investigated. The hemolysis activity test of 6 strains identified as P. tolaasii were 0.8∼0.9 at 600 nm and 3 strains of WLRO were 0.9∼1.0 and Pseudomonas app. were 1.0∼1.2. Observation of fresh mushroom tissue using confocal laser scanning microscopy was carried out for images of optical sectioning and vertical sectioning. Also images of brown blotch diseased oyster mushroom tissue after contamination P. tolaasii was obtained by CLSM.