• The Plant Pathology Journal
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• v.28 no.1
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• pp.20-31
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• 2012

Endophytic bacterial communities of tidal flat plants antagonistic to oomycete plant pathogens were studied by the isolation of 256 root colonizing endophytic bacteria from surface-disinfected root tissues of six plants ($Rosa$ $rugosa$, $Suaeda$ $maritima$, $Vitex$ $rotundifolia$, $Carex$ $scabrifolia$, $Glehnia$ $littoralis$ and $Elymus$ $mollis$) growing in a tidal flat area of Namhae Island, Korea. To understand the antagonistic potential, an $in$ $vitro$ antagonistic assay was performed to characterize and identify strains that were antagonistic to the oomycete plant pathogens $Phytophthora$ $capsici$ and $Pythium$ $ultimum$ from the total population. Nine percent of the total number of isolated bacteria exhibited in vitro inhibitory activity against target plant pathogenic oomycetes. Taxonomic and phylogenetic placement of the antagonistic bacteria was investigated by analysis of the 16S rRNA gene sequences. The sequence analysis classified the antagonistic strains into four major classes of the domain bacteria ($Firmicutes$, ${\alpha}-Proteobacteria$, ${\gamma}-Proteobacteria$ and $Actinomycetes$) and 10 different genera. Further production of secondary metabolites, hydrolytic enzymes and plant growth promoting traits were determined for the putative new species of antagonistic endophytic bacteria. These new strains could not be identified as known species of ${\alpha}-Proteobacteria$, and so may represent novel bacterial taxa. The unexpected high antagonistic bacterial diversity associated with the tidal flat plants may be indicative of their importance in tidal flat plants as a promising source of novel antimicrobial compounds and biocontrol agents.

• The Plant Pathology Journal
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• v.29 no.2
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• pp.125-135
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• 2013

In agro-ecosystems worldwide, some of the most important and devastating diseases are caused by soil-borne necrotrophic fungal pathogens, against which crop plants generally lack genetic resistance. However, plants have evolved approaches to protect themselves against pathogens by stimulating and supporting specific groups of beneficial microorganisms that have the ability to protect either by direct inhibition of the pathogen or by inducing resistance mechanisms in the plant. One of the best examples of protection of plant roots by antagonistic microbes occurs in soils that are suppressive to take-all disease of wheat. Take-all, caused by Gaeumannomyces graminis var. tritici, is the most economically important root disease of wheat worldwide. Take-all decline (TAD) is the spontaneous decline in incidence and severity of disease after a severe outbreak of take-all during continuous wheat or barley monoculture. TAD occurs worldwide, and in the United States and The Netherlands it results from a build-up of populations of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing fluorescent Pseudomonas spp. during wheat monoculture. The antibiotic 2,4-DAPG has a broad spectrum of activity and is especially active against the take-all pathogen. Based on genotype analysis by repetitive sequence-based-PCR analysis and restriction fragment length polymorphism of phlD, a key 2,4-DAPG biosynthesis gene, at least 22 genotypes of 2,4-DAPG producing fluorescent Pseudomonas spp. have been described worldwide. In this review, we provide an overview of G. graminis var. tritici, the take-all disease, Pseudomonas biocontrol agents, and mechanism of disease suppression.

• Korean journal of food and cookery science
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• v.24 no.1
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• pp.11-15
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• 2008

In searching for yeast to be utilized as biocontrol agents, a single yeast strain was isolated from Camellia sinensis based on its morphological, cultural, physiological, and biochemical properties, as well as by molecular techniques. This single strain was pink to red in color and designated as strain JY-1. The effects of temperature, pH, NaCl concentration, and ethanol concentration on the growth of the JY-l strain were examined for the JY-1. Growth occurred at temperatures ranging from 20 to $35^{\circ}C$, and between pH 3.0 and 12.0, with optimal growth at $25-30^{\circ}C$ and pH 5.0. The yeast also grew in the presence of 0-2% (w/v) NaCl and 0-4% (v/v) EtOH. The isolate was further classified based on biochemical characteristics using the VITEK system. The biochemical data obtained using this system were similar to those of Rhodotorula glutinis/Rhodotorula mucilaginosa (exhibiting a 93% matching level). Molecular phylogenetic analysis based on l8S rDNA sequences indicated that the yeast represented a basidiomycetous species, and its highest degree of sequence similarity was with Rhodosporidium azoricum, strain JCM11251 (99%).

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1741-1746
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• 2008

This study was conducted to select endophytic actinomycetes as biocontrol agents against Chinese cabbage clubroot caused by Plasmodiophora brassicae. A total of 81 endophytic actinomycetes were isolated from surface-sterilized roots of Chinese cabbage that was grown on paddy field and upland soils collected from various locations in Korea. By using 16S ribosomal DNA (rDNA) gene sequencing, they were classified to 8 actinobacterial genera. The genus Microbispora (67%) was most frequently isolated, followed by Streptomyces (12%) and Micromonospora (11%). Three of the 81 isolates, when inoculated in germinated Chinese cabbage seeds and then transplanted to pots, effectively suppressed the occurrence of a post-inoculated strain of P. brassicae in the pots. They showed control values of 58% for strain A004, 33% for strain A011, and 42% for strain A018. Based on cell wall components, morphological characteristics, and phylogenetic analyses, the three antagonistic isolates were identified as Microbispora rosea subsp. rosea (A004 and A011) and Streptomyces olivochromogenes (A018). Further researches on the field efficacy and action modes of the three actinomycetes are in progress.

• Journal of Forest and Environmental Science
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• v.33 no.4
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• pp.342-347
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• 2017

Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.

• The Korean Journal of Mycology
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• v.40 no.1
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• pp.44-48
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• 2012

Ginseng (Panax ginseng) is one of the most widely cultivated medicinal herb in Korea. However, yield losses reached up to 30~60 % due to various diseases during 3 or 5 years of ginseng cultivation. Therefore, successful production of ginseng roots depends primarily on the control of diseases. The objective of this study is to select potential multifunctional biocontrol agents from actinomycetes for the control of multiple ginseng diseases as an alternative to fungicides. Ninety three Streptomyces strains were selected and their ability to produce antibiotics, siderophore and lytic enzymes such as protease and cellulose were investigated. Eight of the isolates, strains A75, A501, 515, 523, A704, A1444, A3265 and A3283 produced cellulase and protease. These strains also produced siderophore and showed potent antifungal activity against Botrytis cinerea, Cylindrocarpon destructans, Collectotricum gloeosporioides, Phytophthora capsici and Rhizoctonia solani causing ginseng root rot.

• Journal of Ecology and Environment
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• v.44 no.4
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• pp.222-228
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• 2020

Background: Parthenium hysterophorus is a noxious invasive weed in tropical and subtropical regions of the world, including Nepal. Among 11 species of biological control agents released to control P. hysterophorus in Ausrtalia, winter rust Puccina abrupta var. partheniicola arrived fortuitously and has established in Kathmandu Valley, Nepal, nearly a decade back. However, the prevalence and effectiveness of this rust as a biological control agent in Kathmandu remain unknown. To address this knowledge gap, a roadside survey was done at an interval of 2 ± 0.25 km in Kathmandu Valley to assess the P. abrupta var. partheniicola incidence and its impacts on P. hysterophorus. Infested individuals of P. hysterophorus were further divided into four severity classes (very low, low, medium, and high), and rust incidence was calculated. The impact of the winter rust on the growth of P. hysterophorus was assessed by comparing biomass and seed output of infested and non-infested individuals. Results: Among 81 locations where P. hysterophorus was present in the Kathmandu Valley, winter rust infestation was observed at 98% locations. At some locations within Kathmandu Valley such as Tinkune, Kirtipur, Chabahil, Buddha Chowk, and Dhobighat, the impacts of the rust on P. hysterophorus were medium to high. Aboveground biomass and seed output of P. hysterophorus were reduced by 47% and 73%, respectively, due to winter rust infestation. The study indicates that winter rust incidence is widespread in Kathmandu Valley with very low to high levels of damages to P. hysterophorus depending on the localities. Conclusion: The rust has, therefore, a potential to reduce the growth performance of P. hysterophorus, and it can be used as a component of integrated management of P. hysterophorus by introducing to other suitable areas in Nepal.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.202-210
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• 2022

Aflatoxin contamination in rice has been documented in a number of studies, and has a high incidence in Asian countries, and as such, there has been a growing interest in alternative biocontrol strategies to address this issue. In this study, 147 strains of yeasts and yeast-like fungi were screened for their potential to produce volatile organic compounds (VOCs) active against Aspergillus flavus strains that produce aflatoxin B₁ (AFB₁). Five strains within four different genera showed greater than 50% growth inhibition of some strains of A. flavus. These were Anthracocystis sp. DMKU-PAL124, Aureobasidium sp. DMKU-PAL120, Aureobasidium sp. DMKU-PAL144, Rhodotorula sp. DMKU-PAL99, and Solicococcus keelungensis DMKU-PAL84. VOCs produced by these microorganisms ranged from 4 to 14 compounds and included alcohols, alkenes, aromatics, esters and furans. The major VOCs produced by the closely related Aureobasidium strains were found to bedistinct. Moreover, 2-phenylethanol was the most abundant compound generated by Aureobasidium sp. DMKU-PAL120, while methyl benzeneacetate was the major compound emitted from Aureobasidium sp. DMKU-PAL144. On the other hand, 2-methyl-1-butanol and 3-methyl-1-butanol were significant compounds produced by the other three genera. These antagonists apparently inhibited A. flavus sporulation and mycelial development. Additionally, the reduction of the AFB1 in the fungal-contaminated rice grains was observed after co-incubation with these VOC-producing strains and ranged from 37.7 ± 8.3% to 60.3 ± 3.4%. Our findings suggest that these same microorganisms are promising biological control agents for use against aflatoxin-producing fungi in rice and other agricultural products.

• Research in Plant Disease
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• v.29 no.2
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• pp.174-183
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• 2023

The limited effectiveness of current plant disease management treatments necessitates the development of new methods for controlling diseases using beneficial microbes. Demanding sustainable agriculture is increasingly highlighted as a biocontrol approach, particularly Streptomyces species known to produce a variety of antibiotic compounds and secondary metabolites. The Streptomyces globisporus SP6C4 strain and Streptomyces sp. S8 have been reported as potent antifungal agents and are gaining attention for improving crop growth in sustainable agriculture. In this study, we investigated the use of Streptomyces species formulations to enhance bacterial growth with nitrogen sources. Specifically, the addition of L-glutamic acid and L-cysteine resulted in earlier sporulation and bacterial growth in Streptomyces strains, respectively. This approach could expand the range of fermentation techniques in agriculture and be useful for controlling plant growth-promoting bacteria.

• 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.