• Research in Plant Disease
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• v.29 no.4
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• pp.399-404
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• 2023

Agricultural activities and the number of farms in the subarctic regions have been increasing annually after the coronavirus disease 2019 pandemic to achieve food self-sufficiency. Potatoes are vulnerable to soft rot bacteria at all stages of production, storage, and transportation. A novel bacterium, Pseudomonas sp. N3-W, isolated from Alaska tundra soil, grows at 5-25℃ and produces extracellular protease(s). N3-W caused necrotic spots (hypersensitivity) in hot pepper leaves and soft rot disease (pathogenicity) in potato tubers. The psychrotolerant N3-W caused significant soft rot symptoms on potatoes at a broad temperature range (5℃, 15℃, and 25℃). In contrast, mesophilic Pectobacterium carotovorum KACC 16999 induced severe rotting symptoms in potatoes at their optimal growth temperature of 15℃ and 25℃. However, it barely produced symptoms at 5℃, which is the appropriate storage and transportation temperature for potatoes. The results of pathogenicity testing imply that psychrotolerant soft rot pathogens from polar regions may cause severe soft rot not only during the crop growing season but also during storage and transportation. Our study indicates the possibility of new plant pathogen emergence and transmission due to the expansion of crop cultivation areas caused by permafrost thawing in response to recent polar warming.

• Journal of Microbiology and Biotechnology
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• v.27 no.4
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• pp.791-807
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• 2017

The type II secretion system (T2SS), which transports selected periplasmic proteins across the outer membrane, has rarely been studied in nonpathogens or in organisms classified as Betaproteobacteria. Therefore, we studied Cupriavidus metallidurans (Cme), a facultative chemilithoautotroph. Gel analysis of extracellular proteins revealed no remarkable differences between the wild type and the T2SS mutants. However, enzyme assays revealed that native extracellular alkaline phosphatase is a T2SS substrate, because activity was 10-fold greater for the wild type than a T2SS mutant. In Cme engineered to produce three Ralstonia solanacearum (Rso) exoenzymes, at least 95% of their total activities were extracellular, but unexpectedly high percentages of these exoenzymes remained extracellular in T2SS mutants cultured in rich broth. These conditions appear to permit an alternative secretion process, because neither cell lysis nor periplasmic leakage was observed when Cme produced a Pectobacterium carotovorum exoenzyme, and wild-type Cme cultured in minimal medium secreted 98% of Rso polygalacturonase, but 92% of this exoenzyme remained intracellular in T2SS mutants. We concluded that Cme has a functional T2SS despite lacking any abundant native T2SS substrates. The efficient secretion of three foreign exoenzymes by Cme is remarkable, but so too is the indication of an alternative secretion process in rich culture conditions. When not transiting the T2SS, we suggest that Rso exoenzymes are probably selectively packaged into outer membrane vesicles. Phylogenetic analysis of T2SS proteins supports the existence of at least three T2SS subfamilies, and we propose that Cme, as a representative of the Betaproteobacteria, could become a new useful model system for studying T2SS substrate specificity.

• Research in Plant Disease
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• v.21 no.4
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• pp.255-260
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• 2015

We investigated influence of three commercial antibiotics viz., oxolinic acid, streptomycin, and validamycin A, on biocontrol and plant growth promoting activities of Bacillus vallismortis EXTN-1 and BS07M in Chinese cabbage. Plants were pre-drenched with these strains followed by antibiotics application at recommended and ten-fold diluted concentration to test the effect on biocontrol ability against soft rot caused by Pectobacterium carotovorum SCC1. The viability of the two biocontrol strains and bacterial pathogen SCC1 was significantly reduced by oxolinic acid and streptomycin in vitro assay, but not by validamycin A. In plant trials, strains EXTN-1 and BS07M controlled soft rot in Chinese cabbage, and there was a significant difference in disease severity when the antibiotics were applied to the plants drenched with the two biocontrol agents. Additional foliar applications of oxolinic acid and streptomycin reduced the disease irrespective of pre-drench treatment of the PGPRs. However, when the plants were pre-drenched with EXTN-1 followed by spray of validamycin A at recommended concentration, soft rot significantly reduced compared to untreated control. Similarly, strains EXTN-1 and BS07M significantly enhanced plant growth, but it did not show synergistic effect with additional spray of antibiotics. Populations of the EXTN-1 or BS07M in the rhizosphere of plants sprayed with antibiotics were significantly affected as compared to control. Taken together, our results suggest that the three antibiotics used for soft rot control in Chinese cabbage could affect bacterial mediated biocontrol and plant growth promoting activities. Therefore, combined treatment of the PGPRs and the commercial antibiotics should be carefully applied to sustain environmental friendly disease management.

• Journal of Microbiology and Biotechnology
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• v.26 no.9
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• pp.1542-1550
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• 2016

This is the first report that paromomycin, an antibiotic derived from Streptomyces sp. AG-P 1441 (AG-P 1441), controlled Phytophthora blight and soft rot diseases caused by Phytophthora capsici and Pectobacterium carotovorum, respectively, in chili pepper (Capsicum annum L.). Chili pepper plants treated with paromomycin by foliar spray or soil drenching 7 days prior to inoculation with P. capsici zoospores showed significant (p < 0.05) reduction in disease severity (%) when compared with untreated control plants. The disease severity of Phytophthora blight was recorded as 8% and 50% for foliar spray and soil drench, respectively, at 1.0 ppm of paromomycin, compared with untreated control, where disease severity was 83% and 100% by foliar spray and soil drench, respectively. A greater reduction of soft rot lesion areas per leaf disk was observed in treated plants using paromomycin (1.0 μg/ml) by infiltration or soil drench in comparison with untreated control plants. Paromomycin treatment did not negatively affect the growth of chili pepper. Furthermore, the treatment slightly promoted growth; this growth was supported by increased chlorophyll content in paromomycin-treated chili pepper plants. Additionally, paromomycin likely induced resistance as confirmed by the expression of pathogenesis-related (PR) genes: PR-1, β-1,3-glucanase, chitinase, PR-4, peroxidase, and PR-10, which enhanced plant defense against P. capsici in chili pepper. This finding indicates that AG-P 1441 plays a role in pathogen resistance upon the activation of defense genes, by secretion of the plant resistance elicitor, paromomycin.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.12-12
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• 2017

Genetic improvement in potato can be carried out through several approaches, as sexual crosses, somatic hybridization, mutation and genetic engineering. Although the approach is different, but the goal is the same, to get a superior cultivar. Mutation and genetic engineering are very interesting methods for genetic improvement of potato plants. Mutation by gamma-ray irradiation have been performed to get some new potato cultivars which are more resistant to disease and have higher productivity. We have carried out a mutation of some potato cultivars and obtained some excellent clones to be potentially released as new superior cultivars. By the mutation method, we have released one potato cultivar for the French fries industry, and we registered one cultivar of potato for chips, and two cultivar for vegetable potatoes. Actually we are doing multi-location trial for three clones to be released as new cultivars. Through genetic engineering, several genes have been introduced into the potato plant, and we obtained several clones of transgenic potato plants. Transgenic potato plants containing FBPase gene encoding for fructose bisphosphatase, have a higher rate of photosynthesis and higher tuber productivity than non-transgenic plants. This result suggests that FBPase plays an important role in increasing the rate of photosynthesis and potato tuber productivity. Some transgenic potatoes containing the Hd3a gene are currently being evaluated for their productivity. Over expression of the Hd3a gene is expected to increase tuber productivity and induce flowering in potatoes. Transgenic potato plants containing MmPMA gene encoding for plasma membrane ATPse are more tolerant to low pH than non-transgenic plants, indicating that plasma membrane ATPase plays an important role in the potato plant tolerance to low pH stress. Transgenic potato plants containing c-lysozyme genes, are highly tolerant of bacterial wilt diseases caused by Ralstonia solanacearum and bacterial soft rot disease caused by Pectobacterium carotovorum. Expression of c-lyzozyme gene plays an important role in increasing the resistance of potato plants to bacterial diseases.

• The Plant Pathology Journal
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• v.28 no.1
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• pp.68-74
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• 2012

Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

• Korean Journal of Soil Science and Fertilizer
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• v.34 no.5
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• pp.333-341
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• 2001

The Compost-decomposing-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. Cloning of CMCase encoding gene was accompanied by shotgun method. The pLK100 have yellow activity ring on CMC medium, that was carried 2.2 kb insert DNA in pBluescript II $SK^+$ vector, named BglC gene. The BglC was very similar to Pectobacterium carotovorum Gun_CLOAB(P15704) with score of 57% identity and 71% homology over 508 aa. The BglC was measured molecular weight 56 kDa by CMC-SDS-PAGE. Optimum cellulase activity Bacillus subtilis LYH201 was temperature $50^{\circ}C$ and pH 7.

• The Plant Pathology Journal
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• v.38 no.6
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• pp.656-664
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• 2022

Pectobacterium odoriferum is the primary causative agent in Kimchi cabbage soft-rot diseases. The pathogenic bacteria Pectobacterium genera are responsible for significant yield losses in crops. However, P. odoriferum shares a vast range of hosts with P. carotovorum, P. versatile, and P. brasiliense, and has similar biochemical, phenotypic, and genetic characteristics to these species. Therefore, it is essential to develop a P. odoriferumspecific diagnostic method for soft-rot disease because of the complicated diagnostic process and management as described above. Therefore, in this study, to select P. odoriferum-specific genes, species-specific genes were selected using the data of the P. odoriferum JK2.1 whole genome and similar bacterial species registered with NCBI. Thereafter, the specificity of the selected gene was tested through blast analysis. We identified novel species-specific genes to detect and quantify targeted P. odoriferum and designed specific primer sets targeting HAD family hydrolases. It was confirmed that the selected primer set formed a specific amplicon of 360 bp only in the DNA of P. odoriferum using 29 Pectobacterium species and related species. Furthermore, the population density of P. odoriferum can be estimated without genomic DNA extraction through SYBR Green-based real-time quantitative PCR using a primer set in plants. As a result, the newly developed diagnostic method enables rapid and accurate diagnosis and continuous monitoring of soft-rot disease in Kimchi cabbage without additional procedures from the plant tissue.

• Research in Plant Disease
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• v.25 no.3
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• pp.114-123
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• 2019

QD LED has an ideal light source for growing crops and can also be used to control plant pathogenic microorganisms. The mycelial growth inhibition effect of QD LED light on Rhizoctonia solani, Phytophthora drechsleri, Sclerotinia sclerotiorum, Sclerotinia minor, Botrytis cinerea, Fusarium oxysporum, Pectobacterium carotovorum, and Xanthomonas campestris were investigated. According to the results, BLUE (450 nm) light, suppressed S. sclerotiorum by 16.7% at 50 cm height from the light source, and 94.1% mycelial growth at 30 cm height. Mycelial growth of Sclerotinia minor was inhibited by 80.4% at 50 cm height and 36.3% at 50 cm height in B. cinerea. S. minor, and B. cinerea was inhibited by 100% mycelial growth at a height of 30 cm from the light source. At 15 cm height, all three pathogens (B. cinerea, S. minor, and S. sclerotiorum) was inhibited by 100%. QD RED (M1) and QD RED (M2) light suppressed mycelial growth of S. minor and B. cinerea by 100% at 30 cm and 15 cm height from the light source. For S. sclerotiorum, QD RED (M1) and QD RED (M2) showed 75.2% and 100% inhibition, respectively. Further experiment was conducted to know the suppression effect of lights after inoculating the fungal pathogens on lettuce crop. According to the results, QD RED (M2) suppressed the S. sclerotiorum by 59.9%. In addition, Blue (450 nm), QD RED (M1), and QD RED (M2) light reduce the infestation by 59.9%. In case of B. cinerea, disease reduction was found 84% by BLUE (450 nm) light. Results suggest that the growth inhibition of mycelium increases by Quantum dot LED light.

• The Korean Journal of Pesticide Science
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• v.17 no.4
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• pp.388-393
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• 2013

It has been well documented that Bacillus vallismortis strain EXTN-1, a beneficial rhizosphere bacterium, could enhance plant growth and induce systemic resistance to diverse pathogens in plants. However, the molecular mechanisms for how the EXTN-1 promote plant growth and induce resistances to diverse pathogens. Here, we show that 3-Hydroxy-2-butanone, a volatile organic compound (VOCs) emitted from the EXTN1, is a key factor for the bacteria-mediated beneficial effects on plant growth and defense systems. We found that the presence of volatile signals of EXTN-1 resulted in growth promotion of tobacco seedlings. The identification and analysis of EXTN-1-secreted volatile signals by solid-phase microextraction (SPME) and gas chromatography/mass spectrometry (GC/MS) indicated that a 3-hydroxy-2-butanone could provide not only the plant growth promotion, but also higher resistance against Pectobacterium carotovorum SCC1. These results suggest that a volatile compound released from EXTN-1 enhances the plant growth promotion and immunity of plants.