• 한국식품위생안전성학회지
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• 제37권2호
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• pp.55-62
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• 2022

세균성 무름병은 배추와 같은 채소의 주요 질병중의 하나이며, 특히 김치의 주원료인 배추의 저장에 있어서 발생할 경우, 품질 저하로 인한 경제적 손실을 유발한다. 우리는 무름병이 발생한 배추로부터 원인이 되는 세균들을 분리하였고, 해당 세균이 신선한 배추 조직에 인위적으로 접종하였을 경우, 무름병이 발생하는 것을 확인하였다. 해당 원인균 2종을 동정한 결과, Pseudomonas kribbensis와 Pantoea vagans로 확인되었다. 이러한 원인 세균을 제어하기 위하여 차아염소산수 처리와 항균활성 유산균의 배양 상등액을 이용하여 제어하는 테스트를 진행하였다. 본 테스트에 사용한 항균활성 유산균은 리스테리아와 같은 병원성 미생물과 Pseudomonas 속 계열을 저해하는 Lactobacillus plantarum PL203 유산균을 선별하여 사용하였다. 제어 테스트는 정상 배추 조직에 무름병 원인균 2종을 각각 10⁵ cfu/g 이 되도록 접종한 후, 차아염소산수를 농도별로 분무하거나 항균활성 유산균 배양상등액을 0.5 mL 분무한 후, 25℃에서 5일간 배양한 결과, 차아염소산수 120 ppm 이상 첨가한 구간과 항균활성 유산균 배양 상등액을 처리한 구간에서 대조구와 비교하여 배추 무름 원인세균을 완전하게 제어하는 것을 확인하였다.

• 식물병연구
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• 제29권1호
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• pp.72-78
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• 2023

In September 2021, gray-to-brown discoloration and expanding water-soaked lesions were observed on the outer and inner layers and the core of kimchi cabbage (Brassica rapa subsp. pekinensis) in fields located in Samcheok, Gangwondo, Korea. A bacterial strain designated as KNUB-02-21 was isolated from infected cabbage samples. Phylogenetic analysis based on the sequences of the 16S rRNA region and the dnaX, leuS, and recA genes confirmed that the strain was affiliated with Pectobacterium versatile. Additionally, the biochemical and morphological profiles of the isolate were similar to those of P. versatile. Based on these results, the isolate was identified as a novel strain of P. versatile. Healthy kimchi cabbage slices developed soft rot upon inoculation with P. versatile KNUB-02-21 and exhibited symptoms similar to those observed in the diseased plants in fields. The re-isolated strains were similar to those of P. versatile. Prior to our study, P. versatile as the causative pathogen of kimchi cabbage soft rot had not been reported in Korea.

• The Plant Pathology Journal
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• 제39권1호
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• pp.62-74
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• 2023

Plant pathogenic Pectobacterium species cause severe soft rot/blackleg diseases in many economically important crops worldwide. Pectobacterium utilizes plant cell wall degrading enzymes (PCWDEs) as the main virulence determinants for its pathogenicity. In this study, we screened a random mutant, M29 is a transposon insertion mutation in the metC gene encoding cystathionine β-lyase that catalyzes cystathionine to homocysteine at the penultimate step in methionine biosynthesis. M29 became a methionine auxotroph and resulted in growth defects in methionine-limited conditions. Impaired growth was restored with exogenous methionine or homocysteine rather than cystathionine. The mutant exhibited reduced soft rot symptoms in Chinese cabbages and potato tubers, maintaining activities of PCWDEs and swimming motility. The mutant was unable to proliferate in both Chinese cabbages and potato tubers. The reduced virulence was partially restored by a complemented strain or 100 µM of methionine, whereas it was fully restored by the extremely high concentration (1 mM). Our transcriptomic analysis showed that genes involved in methionine biosynthesis or transporter were downregulated in the mutant. Our results demonstrate that MetC is important for methionine biosynthesis and transporter and influences its virulence through Pcc21 multiplication in plant hosts.

• 식물병연구
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• 제17권2호
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• pp.111-120
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• 2011

A bacterial artificial chromosome library of Pectobacterium carotovorum 35 was constructed to characterize the genome and to sequence its hrp region. The hrp cluster of P. carotovorum 35 consisted of 26 open reading frames in five operons. A promoter-based green fluorescent protein technology was used to identify the genes regulated by the alternative sigma factor, HrpL, in P. carotovorum 35. The majority of the selected clones contained the hrpJ operon promoter sequence, which harbors a hrp box, but no putative hrp boxes were detected within the promoter sequences of two other hrpL-regulated genes encoding for pectate lyase and large repetitive protein. Although the promoters of five other hrp operons also contained hrp boxes, their expression was not HrpL-dependent in the promoter-based selection in E. coli. However, transcriptional analysis showed that expression from all operons harboring hrp boxes, except for the hrpN operon, was reduced significantly in the hrpL mutant. The severity of soft-rot symptoms when the hrpL mutant was applied to the surface of tobacco leaves, mimicking natural infection, was greatly attenuated. These results indicate that the hrpL gene of P. carotovorum 35 may be involved in the development of soft-rot symptoms.

• 식물병연구
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• 제24권3호
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• pp.193-201
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• 2018

Pactobacterium carotovorum subsp. carotovorum (Pcc)에 의한 무름병은 전세계적으로 문제가 되고 있고 특히 온난 다습한 지역에서 문제가 심각하다. 본 연구는 Pcc에 의해 발생하는 무 무름병에 대한 효율적인 저항성 검정법을 확립하기 위하여 수행되었다. 시판 무 품종 60개의 Pcc에 대한 저항성 정도를 조사하고, 추후 실험을 위해 저항성에 차이를 보이는 6개 품종을 선발하였다. 이들 6개 품종의 접종하는 무의 생육 시기, 접종원 농도, 접종 후 재배 온도 등의 발병 조건에 따른 무름병 발생을 조사하였다. 이들 실험의 결과로부터 무의 무름병에 대한 저항성을 검정하기 위해서는, 무 종자를 파종하고 온실($25^{\circ}C{\pm}5^{\circ}C$)에서 20일 동안 재배한 유묘에 Pcc 균주의 세균현탁액($8{\times}10^5cfu/ml$)을 분무하여 접종하고, 접종한 식물은 $25^{\circ}C$ 습실상에 24시간 동안 배양한 후에 $25^{\circ}C$, 상대습도 80%의 생육상으로 이동하여 재배하는 것을 제안하고자 한다.

• The Plant Pathology Journal
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• 제16권5호
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• pp.286-289
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• 2000

Mechanol extracts of three cold-tolerant eucalyptus trees-Eucalyptus darlympleana, E. gunnii and E. unigera were screened for antimicrobial activity against twenty two phyto-pathogenic fungi and six food-borne bacterial pathogens. E. unigera showed the antagonistic activity against all the tested pathogens. Among the tested fungal pathogens, Pythium species were highly sensitive to the leaf extracts. Especially, P. vanterpoolii, a causal agent of leaf blight in creeping bentgrass (Agrostis palustris), was completely inhibited by the extracts. The eucalyptus extracts were also effective in inhibiting the fungal growth of Botrytis cinerea and Phomopsis sp. isolated from the lesions of kiwifruit soft rot during post-harvest storage. Escherichia coli O-157 was less sensitive to the inhibition than the other bacterial pathogens tested. It was likely that Gram positive bacteria-Bacillus subtilis and Streptococcus mutans were more sensitive to the eucalyptus extracts than Gram negative bacteria-Escherichia coli, Salmonella enteritidis and Pseudomonas aeruginosa. Our findings suggest that the cold-tolerant eucalyptus species have antimicrobial properties that can serve the development of novel fungitoxic agents or food preservatives.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2016년도 춘계학술대회 및 임시총회
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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.

• Journal of Microbiology and Biotechnology
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• 제31권1호
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• pp.70-78
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• 2021

Identifying the extracellular metabolites of microorganisms in fresh vegetables is industrially useful for assessing the quality of processed foods. Pectobacterium carotovorum subsp. carotovorum (PCC) is a plant pathogenic bacterium that causes soft rot disease in cabbages. This microbial species in plant tissues can emit specific volatile molecules with odors that are characteristic of the host cell tissues and PCC species. In this study, we used headspace solid-phase microextraction followed by gas chromatography coupled with mass spectrometry (HS-SPME-GC-MS) to identify volatile compounds (VCs) in PCC-inoculated cabbage at different storage temperatures. HS-SPME-GC-MS allowed for recognition of extracellular metabolites in PCC-infected cabbages by identifying specific volatile metabolic markers. We identified 4-ethyl-5-methylthiazole and 3-butenyl isothiocyanate as markers of fresh cabbages, whereas 2,3-butanediol and ethyl acetate were identified as markers of soft rot in PCC-infected cabbages. These analytical results demonstrate a suitable approach for establishing non-destructive plant pathogen-diagnosis techniques as alternatives to standard methods, within the framework of developing rapid and efficient analytical techniques for monitoring plant-borne bacterial pathogens. Moreover, our techniques could have promising applications in managing the freshness and quality control of cabbages.

• The Plant Pathology Journal
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• 제38권1호
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• pp.33-45
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• 2022

To screen antagonistic fungi against plant pathogens, dual culture assay (DCA) and culture filtrate assay (CFA) were performed with unknown soil-born fungi. Among the different fungi isolated and screened from the soil, fungal isolate ANU-301 successfully inhibited growth of different plant pathogenic fungi, Colletotrichum acutatum, Alternaria alternata, and Fusarium oxysporum, in DCA and CFA. Morphological characteristics and rDNA internal transcribed spacer sequence analysis identified ANU-301 as Aspergillus terreus. Inoculation of tomato plants with Fusarium oxysporum f. sp. lycopersici (FOL) induced severe wilting symptom; however, co-inoculation with ANU-301 significantly enhanced resistance of tomato plants against FOL. In addition, culture filtrate (CF) of ANU-301 not only showed bacterial growth inhibition activity against Dickeya chrysanthemi (Dc), but also demonstrated protective effect in potato tuber against soft rot disease. Gas chromatography-tandem mass spectrometry analysis of CF of ANU-301 identified 2,4-bis(1-methyl-1-phenylethyl)-phenol (MPP) as the most abundant compound. MPP inhibited growth of Dc, but not of FOL, in a dose-dependent manner, and protected potato tuber from the soft rot disease induced by Dc. In conclusion, Aspergillus terreus ANU-301 could be used and further tested as a potential biological control agent.

• 식물병연구
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• 제29권3호
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• pp.304-309
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• 2023

Wilted and water-soaked lesion symptoms were observed on cucumbers in greenhouses located in Daejeon, Chungcheongnam-do, Korea, in June 2021. A bacterial strain, designated KNUB-04-21, was isolated from the cucumbers, which was subsequently identified as Pectobacterium brasiliense through a phylogenetic analysis based on sequences of the 16S rRNA region, dnaX, leuS, and recA genes. The biochemical characteristics of KNUB-04-21 were also similar to those of P. brasiliense through investigation using the API ID 32 GN system. The pathogenicity of KNUB-04-21 was confirmed by inoculating it into healthy cucumber plants. The reisolated strains were also found to be same to the original strain. To our knowledge, this is the first report of P. brasiliense being identified as the causative agent of cucumber soft rot in Korea.