• Journal of Mushroom
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• v.9 no.3
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• pp.91-95
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• 2011

Pleurotus ostreatus, the oyster mushroom, is one of the most important edible mushrooms. It is especially susceptible to bacterial blotch disease, which is caused by Pseudomonas tolaasii. In order to develop bacterial blotch disease-resistant transgenic mushroom, NUECIN cDNA, a gene for an antibacterial peptide cloned from Bombyx mori, was overexpressed in Pleurotus ostreatus. NUECIN cDNA was fused to the ${\beta}$-TUBULIN promoter of oyster mushroom and co-transformed with the pTRura3-2 vector into the uracil auxotrophic mutant strain. Twelve transformants containing the NUECIN gene were identified by genomic PCR and Southern blot analysis. NUECIN gene expression was confirmed by Northern blot analysis. Three transformants showed the transcriptional expression of the gene. However, we could not detect expression of the protein in the transformants. This study showed the possibility of transgenic mushroom development for disease resistance.

• Journal of Practical Agriculture & Fisheries Research
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• v.21 no.1
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• pp.135-148
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• 2019

King oyster mushroom(Pleurotus eryngii) is one of the most commercially important mushrooms in Korea. Development of fruit body and disease occurrence are sensitive to environmental conditions such as temperature, humidity, carbon dioxide(CO₂) concentration. The purpose of this study was to investigate the changes in the growth environment of king oyster mushroom by installing Airocide, an air purifier for the purpose of improving mushroom cultivation environment. The results of the environment conditions, identification of pathogenic organisms and pathogenesis during the cultivation were as follows. Airocide operation increased the CO₂ concentration of the cultivation room by more than 400 ppm on average, but the increase of CO₂ concentration at this level had little effect on the quality and growth of fruit body. Operation of the Airocide tended to reduce the air humidity of the cultivation room and required more humidification. In humidifying conditions, the Airocide has the effect of lowering the species and density of bacteria and reducing bacterial symptoms and abnormal fruiting body of mushroom. Pseudomonas sp., the mushroom pathogen, was isolated from the cultivation room without Airocide, resulting in serious disease and loss of yields, so that only about 83% of substrate could harvest normal fruiting bodies. No disease symptom caused by bacteria and fungi in the cultivation room with Airocide. Trichoderma sp., Penicillium sp. and Cladosporium sp. were isolated from all experimental conditions, but did not inhibit fruit growth or caused diseased.

• Journal of Applied Biological Chemistry
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• v.58 no.4
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• pp.349-354
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• 2015

Brown blotch disease in cultivated mushrooms is caused by Pseudomonas tolaasii, which secretes a lipodepsipeptide, tolaasin. Tolaasin is a pore-forming toxin in the cell membranes, thus destroying the fruiting body structure of mushroom. In this study, we isolated pathogenic bacteria from mushrooms that had symptoms of brown blotch disease. In order to identify these bacteria, their 16S rRNA genes were sequenced and analyzed. Pathogenic bacteria identified as Pseudomonas species were thirty five and classified into five subgroups: P1 to P5. Each subgroup showed different metabolic profile measured by API 20NE kit. Fifty percent of the bacteria were identified as P. tolaasii (P1 subgroup). All five subgroups caused the formation of brown blotches on mushroom tissues and the optimum temperature was 25oC, indicating that they may be able to secrete causal factors, such as tolaasin and similar peptide toxins. These results show that there are at least five different pathogenic Pseudomonas species as blotch-causing bacteria and, therefore, strains from the P2 to P5 subgroups should be also considered and studied as pathogens in order to improve the quality and yield of mushroom production.

• The Korean Journal of Mycology
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• v.44 no.4
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• pp.323-329
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• 2016

Water extract from spent mushroom substrate (WESMS) of Pleurotus eryngii suppressed bacterial wilt disease of tomato caused by Ralstonia solanacearum by 70% without any direct antibacterial activity against the pathogen. WESMS-treated tomato had increased contents of free phenolic compounds (increased by 3%) and total salicylic acid (increased by 75%), and significantly enhanced plant height, leaf number, and fresh weight compared to those of a water-treated tomato sample. These results suggest that the treatment of tomato with WESMS can suppress bacterial wilt disease by enhancing plant defense factors and overall plant health.

• Mycobiology
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• v.43 no.3
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• pp.311-318
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• 2015

Culture filtrates of six different edible mushroom species were screened for antimicrobial activity against tomato wilt bacteria Ralstonia solanacearum B3. Hericium erinaceus, Lentinula edodes (Sanjo 701), Grifola frondosa, and Hypsizygus marmoreus showed antibacterial activity against the bacteria. Water, n-butanol, and ethyl acetate extracts of spent mushroom substrate (SMS) of H. erinaceus exhibited high antibacterial activity against different phytopathogenic bacteria: Pectobacterium carotovorum subsp. carotovorum, Agrobacterium tumefaciens, R. solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. axonopodis pv. citiri, and X. axonopodis pv. glycine. Quantitative real-time PCR revealed that water extracts of SMS (WESMS) of H. erinaceus induced expressions of plant defense genes encoding ${\beta}$-1,3-glucanase (GluA) and pathogenesis-related protein-1a (PR-1a), associated with systemic acquired resistance. Furthermore, WESMS also suppressed tomato wilt disease caused by R. solanacearum by 85% in seedlings and promoted growth (height, leaf number, and fresh weight of the root and shoot) of tomato plants. These findings suggest the WESMS of H. erinaceus has the potential to suppress bacterial wilt disease of tomato through multiple effects including antibacterial activity, plant growth promotion, and defense gene induction.

• Journal of Mushroom
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• v.13 no.1
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• pp.16-20
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• 2015

Water extracts from spent mushroom substrate (SMSE)of edible mushrooms, Pleurotus eryngii, Hericium erinaceus and Lentinula edodes promoted growth of pepper seedling. Mycellial growth rate of Phythopthora capsici and Fusarium oxysporum was dramatically inhibited by 100% and 70% on PDA added with SMSE of H. erinaceus. SMSEs from H. erinaceus, P. eryngii, and L. edodes effectively reduced the disease severity of Phytophthora blight of pepper caused by Phytophthora capsici to 75%, 10% and 35%, respectively. These results suggested that SMSE from the mushrooms have dual effects that suppress phythopthora blight disease and promote plant growth of pepper.

• The Plant Pathology Journal
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• v.38 no.5
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• pp.472-481
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• 2022

Brown blotch disease, caused by Pseudomonas tolaasii, is one of the most serious diseases in mushroom cultivation, and its control remains an important issue. This study isolated and evaluated pathogen-specific bacteriophages for the biological control of the disease. In previous studies, 23 varieties of P. tolaasii were isolated from infected mushrooms with disease symptoms and classified into three subtypes, Ptα, Ptβ, and Ptγ, based on their 16S rRNA gene sequences analysis and pathogenic characters. In this study, 42 virulent bacteriophages were isolated against these pathogens and tested for their host range. Some phages could lyse more than two pathogens only within the corresponding subtype, and no phage exhibited a wide host range across different pathogen subtypes. To eliminate all pathogens of the Ptα, Ptβ, and Ptγ subtype, corresponding phages of one, six, and one strains were required, respectively. These phages were able to suppress the disease completely, as confirmed by the field-scale on-farm cultivation experiments. These results suggested that a cocktail of these eight phages is sufficient to control the disease induced by all 23 P. tolaasii pathogens. Additionally, the antibacterial effect of this phage cocktail persisted in the second cycle of mushroom growth on the cultivation bed.

• Mycobiology
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• v.40 no.3
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• pp.189-194
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• 2012

Four Cladobotryum isolates were collected from four different commercially grown mushroom types infected with cobweb disease in Cheongdo-gun and Chilgok-gun of Gyeongbuk Province, Korea in 2010. The isolates were identified as C. mycophilum from Agaricus bisporus and Pleurotus eryngii, C. varium from Flammulina velutipes and Hypsizygus marmoreus. The cultural characteristics of the four isolates were investigated using potato dextrose agar (PDA) media under nine different temperatures ranging from $5{\sim}32^{\circ}C$. Rapid growth of the isolates to colony diameters of 47~82 mm was observed at conditions of $18{\sim}22^{\circ}C$. No growth was observed at $32^{\circ}C$. C. mycophilum produced a yellowish red pigment while C. varium produced a cream colored pigment after cultivation for 25 days on PDA. Phylogenetic analysis of the internal transcribed spacer region and partial 28S rDNA from the four isolates confirmed they were C. mycophilum and C. varium. Cross pathogenicity tests revealed that the two isolates of C. mycophilum were highly pathogenic toward three mushroom types, but not toward H. marmoreus. The two isolates of C. varium were less pathogenic than those of C. mycophilum, but were pathogenic toward all mushroom types evaluated.

• Korean Journal Plant Pathology
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• v.14 no.2
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• pp.177-183
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• 1998

A DNA fragment which is involved in tolassin production was cloned to obtain a molecular marker of Pseudomonas tolaasii, a casual agent of bacterial brown blotch disease of oyster mushroom (Pleurotus ostreatus). Tolaasin is a lipodepsipeptide toxin and known as a primary disease determinant of the P. tolaasii. It is responsible for formation of white line in agar when P. tolaasii were cultured against white line reacting organisms (WLROs). White line negative mutants (WL-) were generated by conjugation between rifampicin resistant strain of P. tolaasii and E. coli carrying suicidal plasmid pSUP2021 : : Tn5. The ability of tolaasin production of the WL- mutants was examined by hemolysis test, pathogenicity test, and high pressure liquid chromatography (HPLC) analysis of culture filtrate. All of the WL- mutants were lost the ability of tolaasin production (Tol-). Genomic library of the Tol- mutant was constructed in pLAFR3 and the cosmid clone containing Tn5 was selected. DNA fragment fro franking region of Tn5 was cloned from the plasmid and used as a probe in Southern blot. DNA-DNA hybridization with the probe to total DNA from group of bacteria ecologically similar to P. tolaasii including WLORs, fluorescent Pseudomonads isolated from oyster mushroom, P. agarici, P. gingeri, and some of other species of Psedomonas showed that some of the tested bacteria do not have any hybridized band and others have bands sowing RFLP. The cloned DNA fragment or its nucleotide sequence will be useful in detection and identification of the P. tolaasii.

• Journal of Mushroom
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• v.2 no.2
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• pp.69-75
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• 2004

This study was carried out to investigate effect of $CaCO_3$ treatment on cultivation of oyster mushroom for suppression of green mold disease and for promotion of mycelial growth to stabilize mushroom production in field and laboratory experiment. Treatment of $CaCO_3$ in PDA media promoted mycelial growth of mushroom and suppressed that of green mold. Addition of $CaCO_3$ in rice straw substrate increased mushroom mycelial growth compared with control. In that case, growth of green mold increased up to treated 0.6% $CaCO_3$ but decreased in treatment beyond 0.8% $CaCO_3$. There were some differences on effect of $CaCO_3$ treatment according to green mold species. Trichoderma longibrachiatum was effected but T. virens was not effected by treated $CaCO_3$. Differences among mushroom strains by treated $CaCO_3$ were not shown. It is confirmed that treatment of $CaCO_3$ can promote mushroom mycelial growth but it's not clear in the field. In the result of field test, treatment of $CaCO_3$ in rice straw substrates tended to increase yield and decrease incidence of disease compared with non-treatment. These results suggest that $CaCO_3$ treatment on cultivation of oyster mushroom can be applied to take preventive steps against of green mold disease.