• 식물병연구
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• 제24권4호
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• pp.339-341
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• 2018

Amaranth has the potential for good materials related to nutrients and health benefits. There are several diseases of amaranth such as leaf blight, damping-off, and root rot. As a causal agent of soft rot disease, Pectobacterium spp. could infect various plant species. In this study, we isolated the bacterial pathogen causing soft rot of amaranth in South Korea. In Gangneung, Gangwon province during 2017, amaranth plants showed typical soft rot symptoms such as wilting, defoliation and odd smell. To isolate pathogen, the macerated tissues of contaminated amaranth were spread onto LB agar plates and purified by a single colony subculture. One ml bacterial suspension of a representative isolate was injected to the stem of five seedlings of 2-week-old amaranth with a needle. Ten mM magnesium sulfate solution was used as a negative control. 16S rDNA gene and recA gene were sequenced and compared with the reference sequences using the BLAST. In the phylogenetic tree based on 16S rDNA gene and recA gene, GSA1 strain was grouped in Pcb.

• 식물병연구
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• 제29권4호
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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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• 제27권4호
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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.

• 식물병연구
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• 제21권4호
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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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• 제26권9호
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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.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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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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• 제28권1호
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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.

• 한국토양비료학회지
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• 제34권5호
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• pp.333-341
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• 2001

퇴비화 촉진 미생물인 Bacillus subtilis LYH201균주가 분비하는 섬유소 분해효소를 분자생물학적으로 연구한 결과는 다음과 같다. 섬유소를 분해하는 유전자는 유전자은행에 의해 구한 약 5,000개의 clone 중 CMC 배지 상에서 활성을 가지는 clone을 선발하여 bglC(pLYH7-39)로 명명하였다. 섬유소를 분해하는 bglC 유전자는 Pvu II, EcoRI, SspI의 제한효소 site를 가지고 있었으며, BglC는 Clostridium acetobutylicum GUN_CLOAB(P15704)와 57%의 identity와 71%의 homology를 나타내어 상동성이 가장 높았으며, CMC-SDS-PAGE 분석으로 56 kDa의 분자량을 나타냈고, 온도는 $50^{\circ}C$, pH는 7에서 활성이 가장 높았다.

• The Plant Pathology Journal
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• 제38권6호
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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.

• 식물병연구
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• 제25권3호
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• pp.114-123
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• 2019

Quantum Dot LED (QD LED) 조명은 소형의 크기, 좁은 대역파장, 긴 수명, 전자 시스템을 통한 제어가 용이하여 현재 시설재배에 이용되는 형광등, 할로겐램프, HID, HSP 램프의 단점을 보완할 수 있는 작물생육에 이상적인 광원으로서 잠재력을 가지고 있다. QD LED 조명을 이용하여 식물 병원성 미생물의 방제가 가능하다면 작물재배에 사용되는 인력 및 비용을 절감하고 화학적 방제제를 사용하지 않은 안전성 높은 생산물을 얻을 수 있다는 장점이 있다. 본 연구에서는 식물공장 및 온실에서 많이 재배되고 있는 상추에 큰 피해를 입히는 주요 식물 병원성 곰팡이에 대한 QD LED 조명의 영향과 생장억제효과를 확인하기 위해 시행하였다. 상추에 주로 발생하여 작물에 피해를 입히는 Rhizoctonia solani, Phytophthora drechsleri, Sclerotinia sclerotiorum, Sclerotinia minor, Botrytis cinerea, Fusarium oxysporum, Pectobacterium carotovorum, Xanthomonas campestris균을대상으로 QD LED 조명에 의한 균사생장억제 효과를 조사하였으며 처리한 6종류의 조명 중 BLUE (450 nm) 조명은 Sclerotinia sclerotiorum는 50 cm 거리에서 16.7%의 억제율을 보였으며 30 cm 거리에서 94.1%의 균사생장억제율을 보였다. S. minor는 50 cm 거리에서 80.4%, B. cinerea는 50 cm 거리에서 36.3%의 균사생장이 억제되었으며 30 cm 거리에서 S. minor와 B. cinerea는 100%의 균사생장억제율을 보였다. 15 cm 거리에서는 3종의 병원균 모두 100%의 억제율을 보였다. QD RED (M1), QD RED (M2)조명은 30 cm와 15 cm 거리에서 Sclerotinia minor와 Botrytis cinerea의 균사생장을 100% 억제했으며 Sclerotinia sclerotiorum의 경우 30 cm 거리에서 QD RED (M1)과 QD RED (M2)조명에 대해 각각 75.2%, 100%의 억제율을 보였으나 15 cm 거리에서는 각각 5.8%, 36.3%의 억제율을 보였다. 상추에 병원균을 접종하여 LED 광원 하에 생장을 확인한 결과 QD RED (M2)광원에서 S. sclerotiorum의 감염을 59.9% 억제하였고 S. minor는 BLUE (450 nm), QD RED (M1), QD RED (M2) 광원에서 59.9%의 억제율을 보였다. B. cinerea의 경우 BLUE (450 nm) 광원에서 84%의 높은 억제율을 보였다.