• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.968-975
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• 2018

In the course of screening for microbes with nematicidal activity, we found that Enterobacter asburiae HK169 displayed promising nematicidal activity against the root-knot nematode Meloidogyne incognita, along with plant growth-promoting properties. Soil drenching of a culture of HK169 reduced gall formation by 66% while also increasing root and shoot weights by 251% and 160%, respectively, compared with an untreated control. The cell-free culture filtrate of the HK169 culture killed all juveniles of M. incognita within 48 h. In addition, the nematicidal activity of the culture filtrate was dramatically reduced by a protease inhibitor, suggesting that proteolytic enzymes contribute to the nematicidal activity of HK169. In order to obtain genomic information about the HK169 isolate related to its nematicidal and plant growth-promoting activities, we sequenced and analyzed the whole genome of the HK169 isolate, and the resulting information provided evidence that the HK169 isolate has nematicidal and plant growth-promoting activities. Taken together, these observations enable the future application of E. asburiae HK169 as a biocontrol agent for nematode control and promote our understanding of the beneficial interactions between E. asburiae HK169 and plants.

• The Korean Journal of Mycology
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• v.41 no.4
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• pp.268-273
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• 2013

Bacillus amyloliquefaciens M27 was isolated from the cotton-waste compost for cultivation of oyster mushroom (Pleurotus ostreatus). B. amyloliquefaciens M27 is a biocontrol agent with antagonistic activities against a wide range of fungal pathogens. The aim of this work was to evaluate the possibility of exploiting antagonistic bacteria, B. amyloliquefaciens M27, in the biological control of the cucumber powdery mildew fungus, Podosphaera fusca. In greenhouse tests, the isolate was found to be very effective to control powdery mildew on cucumber leaves showing 4.0% diseased area, whereas diseased area in the control was 80.5%. The filtrate of the isolate cultured on MH and LB media were more effective for control of the disease than those cultured on TSB, NB, and KB media. When two, five, ten, 20, 50 and 100-fold diluted culture broth of isolate on LB media were treated, disease areas were 0%, 0%, 0%, 1.3%, 3.1% and 5.0%, respectively, whereas diseased area in the control was 60.0%. The filtrate of the isolate cultured on LB media was treated to cucumber plants on July, October and December just before the outbreak of the powdery mildew occurred. When 10-fold diluted filtrate of the isolate was treated, control efficacy was 88.9~98.9% in the treated seasons. The results showed that the culture filtrate of B. amyloliquefaciens M27 was very effective to control powdery mildew of cucumber.

• Journal of Environmental Science International
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• v.17 no.4
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• pp.439-449
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• 2008

Algicidal bacterium was isolated from sea water during the declining period of Cochlodinium polykrikoides blooms and this bacterium had a significant algicidal activity against C. polykrikoides. In this study, algicidal bacterium was identified on the basis of biochemical and chemotaxonomic characteristics, and analysis of 16S rDNA sequences. The algicidal bacterium showed 98.6% homology with Micrococcus luteus ATCC $381^T$. Therefore, this bacterium was designated Micrococcus luteus SY-13. The optimal culture conditions of the algicidal bacterium was $25^{\circ}C$, initial pH 8.0, and 3.0% NaCl concentration. M. luteus SY-13 is assumed to produce secondary metabolites which have algicidal activity. When 10% culture filtrate of this strain was applied to C. polykrikoides ($1.0\;{\times}\;10^4\;cells/ml$) cultures, over 98% of C, polykrikoides cells were destroyed within 6 hours. The culture filtrate of M. luteus SY-13 exhibited similar algicidal activity after heat-treatment at $121^{\circ}C$ for 15 min. While algicidal activity remained in filtrates with pH adjusted to 8.0, loss of algicidal activity occurred when the pHs of filtrates were adjusted to over 9.0 or heat-treated at $121{\times}180^{\circ}C$ for 1 hour. M. luteus SY-13 showed significant algicidal activities against C. polykrikoides (98.9%) and a wide algicidal range against various harmful algal bloom (HAB) species. However, there was no algicidal effect on diatom and marine livefood organisms except Isocrysis galbana. These results suggest that M. luteus SY-13 could be a candidate for use in the control of HABs.

• Mycobiology
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• v.43 no.2
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• pp.137-149
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• 2015

In the search for novel potent fungi-derived bioactive compounds for bioinsecticide applications, crude ethyl acetate culture filtrate extracts from 110 mangrove fungal endophytes were screened for their toxicity. Toxicity tests of all extracts against brine shrimp (Artemia salina) larvae were performed. The extracts with the highest toxicity were further examined for insecticidal activity against Spodoptera litura larvae and acetylcholinesterase (AChE) inhibition activity. The results showed that the extracts of five isolates exhibited the highest toxicity to brine shrimp at 50% lethal concentration ($LC_{50}$) values of 7.45 to 10.24 ppm. These five fungal isolates that obtained from Rhizophora mucronata were identified based on sequence data analysis of the internal transcribed spacer region of rDNA as Aspergillus oryzae (strain BPPTCC 6036), Emericella nidulans (strains BPPTCC 6035 and BPPTCC 6038), A. tamarii (strain BPPTCC 6037), and A. versicolor (strain BPPTCC 6039). The mean percentage of S. litura larval mortality following topical application of the five extracts ranged from 16.7% to 43.3%. In the AChE inhibition assay, the inhibition rates of the five extracts ranged from 40.7% to 48.9%, while eserine (positive control) had an inhibition rate of 96.8%, at a concentration of 100 ppm. The extracts used were crude extracts, so their potential as sources of AChE inhibition compounds makes them likely candidates as neurotoxins. The high-performance liquid chromatography profiles of the five extracts differed, indicating variations in their chemical constituents. This study highlights the potential of culture filtrate ethyl acetate extracts of mangrove fungal endophytes as a source of new potential bioactive compounds for bioinsecticide applications.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.324-332
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• 2016

A bacterial strain capable of hydrolyzing xylan was isolated from fermented soybean paste obtained from a domestic Buddhist temple, using enrichment culture with rice straw as a carbon source. The isolate, named YB-1301, was identified as Bacillus safensis on the basis of its DNA gyrase subunit B gene (gyrB) sequence. The xylanase productivity of strain YB-1301 was drastically increased when it was grown in the presence of wheat bran or various xylans. In particular, the maximum xylanase productivity reached above 340 U/ml in the culture filtrate from LB broth supplemented with only birchwood xylan at shake-flask level. The xylanase production was significantly induced by xylans at the stationary growth phase in LB medium containing xylan, whereas only a small amount of xylanase was constitutively produced from cells grown in LB medium with no addition of xylan. Furthermore, xylanase biosynthesis was induced more rapidly by the enzymatically hydrolyzed products of xylan than by the non-hydrolyzed xylan. In addition, the xylanase in the culture filtrate of B. safensis YB-1301 was found to have optimal activity at 55℃ and pH 6.5–7.0.

• The Plant Pathology Journal
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• v.32 no.3
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• pp.228-241
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• 2016

In our previous study, we reported that a novel endophytic bacterium Bacillus oryzicola YC7007 has suppressed bacterial diseases of rice via induced systemic resistance and antibiotic production. This endophytic strain, B. oryzicola YC7007 was used as a biological control agent against bakanae disease of rice caused by Fusarium fujikuroi, and its mechanism of interaction with the pathogen and the rice was further elucidated. Root drenching with B. oryzicola YC7007 suspension reduced the disease severity of bakanae significantly when compared with the untreated controls. The treatments of B. oryzicola YC7007 suspension ($2.0{\times}10^7cfu/ml$) to the rice rhizosphere reduced bakanae severity by 46-78% in pots and nursery box tests containing autoclaved and non-autoclaved soils. Moreover, in the detached rice leaves bioassay, the development of necrotic lesion and mycelial expansion of F. fujikuroi were inhibited significantly by spraying the culture filtrate of B. oryzicola YC7007. Drenching of ethyl acetate extracts of the culture filtrate to the rhizosphere of rice seedlings also reduced the bakanae disease severity in the plant culture dish tests. With the root drenching of B. oryzicola YC7007 suspension, the accumulation of hydrogen peroxide was observed at an early stage of rice seedlings, and a hormonal defense was elicited with and without pathogen inoculation. Our results showed that the strain B. oryzicola YC7007 had a good biocontrol activity against the bakanae disease of rice by direct inhibition, and was also capable of inducing systemic resistance against the pathogen via primed induction of the jasmonic acid pathway.

• Microbiology and Biotechnology Letters
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• v.39 no.3
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• pp.252-258
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• 2011

A bacterial strain capable of hydrolyzing xylan and locust bean gum (LBG) was isolated from farm soil by enrichment culture using mixture of palm kernel meal (PKM) and wheat bran as carbon source. Nucleotide sequence of 16S rDNA amplified from the isolate YB-1107 showed high similarity with those of genus Cellulosimicrobium strains. Xylanase productivity was increased when the Cellulosimicrobium sp. YB-1107 was grown in the presence of wheat bran or oat spelt xylan, while mannanase productivity was increased drastically when grown in the presence of PKM or LBG. Particularly, maximum mannanase and xylanase activities were obtained in the culture filtrate of media containing 0.7% PKM or 1% wheat bran, respectively. Both enzyme activities were produced at stationary growth phase. Mannanase from the culture filtrate showed the highest activity at $55^{\circ}C$ and pH 6.5. Xylanase activity was optimal at $65^{\circ}C$ and pH 5.5. The predominant products resulting from the mannanase or xylanase hydrolysis were oligosaccharides for LBG or xylan, respectively. In addition, the enzymes could hydrolyze wheat bran and rice bran into oligosaccharides.

• The Plant Pathology Journal
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• v.36 no.3
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• pp.244-254
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• 2020

Gom-chwi (Ligularia fischeri) is severely infected with Phytophthora drechsleri, the causal organism of Phytophthora root rot, an economically important crop disease that needs management throughout the cultivation period. In the present study, Phytophthora root rot was controlled by using bacterial isolates from rhizosphere soils collected from various plants and screened for antagonistic activity against P. drechsleri. A total of 172 bacterial strains were isolated, of which, 49 strains showed antagonistic activities by dual culture assay. In the seedling assay, six out of the 49 strains showed a predominant effect on suppressing P. drechsleri. Among the six strains, the ObRS-5 strain showed remarkable against P. drechsleri when treated with seed dipping or soil drenching. The ObRS-5 strain was identified as Enterobacter asburiae based on 16S ribosomal RNA gene sequences analysis. The bacterial cells of E. asburiae ObRS-5 significantly suppressed sporangium formation and zoospore germination in P. drechsleri by 87.4% and 66.7%, respectively. In addition, culture filtrate of E. asburiae ObRS-5 also significantly inhibited sporangium formation and zoospore germination by 97.0% and 67.6%, respectively. Soil drenched bacterial cells, filtrate, and culture solution of E. asburiae ObRS-5 effectively suppressed Phytophthora root rot by 63.2%, 57.9%, and 81.1%, respectively. Thus, E. asburiae ObRS-5 could be used as a potential agent for the biological control of Phytophthora root rot infecting gom-chwi.

• The Plant Pathology Journal
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• v.38 no.5
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• pp.461-471
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• 2022

Erwinia amylovora is a causative pathogen of fire blight disease, affecting apple, pear, and other rosaceous plants. Currently, management of fire blight relies on cultural and chemical practices, whereas it has been known that few biological resources exhibit disease control efficacy against the fire blight. In the current study, we found that an SFC20201208-M01 fungal isolate exhibits antibacterial activity against E. amylovora TS3128, and the isolate was identified as a Penicillium brasilianum based on the 𝛽-tubulin (BenA) gene sequence. To identify active compounds from the P. brasilianum culture, the culture filtrate was partitioned with ethyl acetate and n-butanol sequentially. From the ethyl acetate layer, we identified two new compounds (compounds 3-4) and two known compounds (compounds 1-2) based on spectroscopic analyses and comparison with literature data. Of these active compounds, penicillic acid (1) exhibited promising antibacterial activity against E. amylovora TS3128 with a minimal inhibitory concentration value of 25 ㎍/ml. When culture filtrate and penicillic acid (125 ㎍/ml) were applied onto Chinese pearleaf crab apple seedlings prior to inoculation of E. amylovora TS3128, the development of fire blight disease was effectively suppressed in the treated plants. Our results provide new insight into the biocontrol potential of P. brasilianum SFC20201208-M01 with an active ingredient to control fire blight.

• The Korean Journal of Mycology
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• v.22 no.1
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• pp.50-61
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• 1994

From soil samples, 380 antagonistic microorgnisms were isolated. Among the isolates, 42 strains had mycelia growing inhibition ability against Fusariun solani, ginseng root rot causing pathogen. Isolates CHA 1 and S-PFHR 6 were proposed as antagonists for this study and they were identified as Promicromonospora sp. and Pseudomonas pseudoalcaligenes respectively. As an antagonism against hyphae of F. solani in dual culture test, CHA 1 and S-PFHR 6 inhibited linear growing, caused abnormal branching, and the membrane projection which formed by cell wall destruction. The secondary metabolites contained in the culture filtrates which prepared from PD broth and Nutrient broth inhibited the spore germination to 14.3%. The culture filtrate of S-PFHR 6 which prepared by a little amount of soil extract addition to nutrient rich medium had more strongly. inhibited the spore germination and spore germination decreased to less than 4.0% in it. The soil used in this study had fungistasis and the germination rate of macroconidia and chlamydospore of F.solani was 19.4% and 17.7% respectively. The steam sterilized soil lost fungistasis and germination rate of conidia increased to more than 97.9%. The soils amended with the propagule of CHA 1 and S-PFHR 6 increased fungistasis and the germination rate of macroconidia decreased to 14.7% and 11.7% respectively in each treatments. But the soil ammended with glucose and asparagine annulled fungistatic ability and the germination rate of macroconidia increased to more than 48.0%. As an antagonistic activity of the secondary metabolites of two antagonistic isolates in soil, the germination rate of macroconidia of F. solani was 9.3% in the soil amended with the culture filtrate of CHA 1 but the culture filtrate of S-PFHR 6 had no such activity. In the soil which treated with antagonist propagule or culture filtrate, the chlamydospore germination rate was lower than that in natural soil. The addition of glucose and asparagine to antagonist propagule treated soil did not enhanced the chlamydospore germination.