• The Plant Pathology Journal
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• v.40 no.2
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• pp.151-159
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• 2024

Bacterial soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc) is one of the most severe diseases in radish cultivation. To control this plant disease, the most effective method has been known to cultivate resistant cultivars. Previously, we developed an efficient bioassay method for investigating resistance levels with 21 resistant and moderately resistant cultivars of radish against a strain Pcc KACC 10421. In this study, our research expanded to investigate the resistance of radish cultivars against six Pcc strains, KACC 10225, KACC 10421, ATCC 12312, ATCC 15713, LY34, and ECC 301365. To this end, the virulence of the six Pcc strains was determined based on the development of bacterial soft rot in seedlings of four susceptible radish cultivars. The results showed that the Pcc strains exhibited different virulence in the susceptible cultivars. To explore the race differentiation of Pcc strains corresponding to the resistance in radish cultivars, we investigated the occurrence of bacterial soft rot caused by the six Pcc strains on the 21 resistant and moderate resistant cultivars. Our results showed that the average values of the area under the disease progress curve were positively correlated with the virulence of the strains and the number of resistant cultivars decreased as the virulence of Pcc strains increased. Taken together, our results suggest that the resistance to Pcc of the radish cultivars commercialized in Korea is more likely affected by the virulence of Pcc strains rather than by race differentiation of Pcc.

• The Plant Pathology Journal
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• v.16 no.5
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• pp.252-256
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• 2000

The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

• The Plant Pathology Journal
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• v.29 no.2
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• pp.174-181
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• 2013

The plant growth-promoting rhizobacterium Ochrobactrum lupini KUDC1013 elicited induced systemic resistance (ISR) in tobacco against soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum. We investigated of its factors involved in ISR elicitation. To characterize the ISR determinants, KUDC1013 cell suspension, heat-treated cells, supernatant from a culture medium, crude bacterial lipopolysaccharide (LPS) and flagella were tested for their ISR activities. Both LPS and flagella from KUDC1013 were effective in ISR elicitation. Crude cell free supernatant elicited ISR and factors with the highest ISR activity were retained in the n-butanol fraction. Analysis of the ISR-active fraction revealed the metabolites, phenylacetic acid (PAA), 1-hexadecene and linoleic acid (LA), as elicitors of ISR. Treatment of tobacco with these compounds significantly decreased the soft rot disease symptoms. This is the first report on the ISR determinants by plant growth-promoting rhizobacteria (PGPR) KUDC1013 and identifying PAA, 1-hexadecene and LA as ISR-related compounds. This study shows that KUDC1013 has a great potential as biological control agent because of its multiple factors involved in induction of systemic resistance against phytopathogens.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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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.

• Research in Plant Disease
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• v.17 no.2
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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.

• The Plant Pathology Journal
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• v.25 no.3
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• pp.286-290
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• 2009

The stability of three different kinds of pUC-derived plasmids, pDsRed, pZsYellow, and pGFPuv, was investigated in Pectobacterium strains to utilize those plasmids as tracers. All three plasmids pDsRed, pZsYellow and pGFPuv showed their specific colors in Pectobacterium strains. Especially, the plasmid pDsRed conferred bright pink colonies on the Pectobacterium strains. When the bacteria lost the plasmid pDsRed, the colonies turned white, suggesting that the plasmid could be a good marker system for Pectobacterium strains on different environmental conditions. The effect of the antibiotic pressure on the stability of the plasmid was different depending on the host bacteria. P. carotovorum subsp. betavasculorum was more sensitive to the antibiotic pressure than P. carotovorum subsp. carotovorum Pcc21. However, temperature change significantly affected plasmid stability on both Pectobacterium strains. Almost all strains lost the plasmids with the shift in temperature from $28^{\circ}C$ to $37^{\circ}C$. Presence of the plasmids did not affect bacterial pathogenicity on their own host plants. Among three plasmids, pZsYellow was not useful as a marker because the yellow fluorescent proteins from pZs Yellow were interfered with the yellow natural fluorescence of the plant tissues induced by the defense system. Since the red color of DsRed can be seen with naked eyes, plasmid pDsRed was applicable as a marker. However, the color change was slow so that additional manipulation to increase the expression speed was necessary. Plasmid pGFPuv could serve as a perfect marker without any problem, tracing the reproduction and spread of the plant pathogens perfectly.

• Research in Plant Disease
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• v.24 no.3
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• pp.193-201
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• 2018

Pectobacterium carotovorum subsp. carotovorum (Pcc) causes bacterial soft rot on a wide range of crops worldwide, especially in countries with warm and humid climates. This study was conducted to establish an efficient screening method for resistant cultivars of radish (Raphanus sativus) to bacterial soft rot. Resistance degrees of 60 commercial radish cultivars to the Pcc KACC 10421 isolate were investigated. For further study, six radish cultivars (Awooriwoldong, YR Championyeolmu, Jeonmuhumu, Bitgoeunyeolmu, Sunbongaltari, Housecheongok) showing different level of resistance to the bacterium were selected. The development of bacterial soft rot on the cultivars was tested according to several conditions such as incubation temperature, seedling stage of radish, inoculum concentration to develop the disease. On the basis of the results, we suggest that an efficient screening method for resistant radish to Pcc is to inoculate twenty-day-old seedlings with a bacterial suspension of Pcc at a concentration of $8{\times}10^5cfu/ml$ and then to cultivate the plants in a growth room at $25^{\circ}C$ and 80% RH with 12-hour light per day.

• Horticultural Science & Technology
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• v.30 no.2
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• pp.137-143
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• 2012

Chinese cabbage plant was grown hydroponically for 4 weeks in order to examine the temporal relationship of calcium concentration of the nutrient solution with calcium content in the leaf tissue and susceptibility of the tissue to soft rot disease by $Pectobacterium$ $carotovorum$ ssp. $carotovorum$ (Pcc). Calcium concentration from 0.5 to 32.0 mM was maintained for 1 week using Hoagland & Arnon solution. The calcium content of the leaf was proportionally increased to the concentration of the nutrient in the solution (r = 0.912). In contrast, the severity of soft rot symptom in the young leaves was inversely related with the amount of calcium supplied to the nutrient solution (r = 0.899). Water-soluble chitosan, prepared by hollow fiber filtration (> 100 kDa) was applied into the nutrient solution from 0.0 to 5,000 ppm. The chitosan of 10 ppm was the most effective to promote calcium uptake of the leaf, showing 155% of the control. The same chitosan solution prohibited most soft rot development of the leaf by Pcc, exhibiting only 53% of the control. Among different molecular weight fractions, chitosan fraction obtained from 30-100 kDa molecular weight cut-off promoted calcium uptake the most up to 163% of the control, and reduced the development of soft rot disease recording merely 36% of the control of the leaf tissue. The results obtained in the present study suggest that large scale production of water-soluble chitosan with an optimum molecular weight and its commercial application to Chinese cabbage production will be important to improve yield and quality of the crop.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.302-309
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• 2005

Phytopathogenic Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 secretes multiple isozymes of the plant cell wall degrading enzyme endoglucanases. We have cloned a third cel gene encoding CMCase from Pcc LY34. The structural organization of the celC gene (AY188753) consisted of an open reading frame (ORP) of 1,116 bp encoding 371 amino acid residues with a signal peptide of 22 amino acids within the NH$_2$-terminal region of pre-CelC. The predicted amino acid sequence of CelC was similar to that of Peetobaeterium ehrysanthemi Cel8Y (AF282321). The CelC has the conserved region of the glycoside hydrolase family 8. The apparent molecular mass of CelC was calculated to be 39 kDa by CMC-SDS-PAGE. The cellulase­minus mutant of Pee LY34 was as virulent as the wild-type in pathogenicity tests on tubers of potato. The results suggest that the CelC of Pce LY34 is a minor factor for the pathogenesis of soft-rot.

• Korean Journal of Organic Agriculture
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• v.26 no.1
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• pp.129-140
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• 2018

Pectobacterium carotovorum subsp. carotovorum was found to be highly virulent to various vegetables, including Chinese cabbage. The antibacterial isolate CAB12243-2 was tested in a field bioassay for suppressing soft rot disease. The nucleotide sequencing of the 16S rRNA gene identified, the CAB12243-2 strain used in this study as Bacillus toyonensis. B. toyonensis CAB12243-2 inhibited the pectate lyase process by soft rot pathogens, and used trehalose and glucose as carbon sources. In field tests, the antibacterial isolate B. toyonensis CAB12243-2 suppressed soft rot disease with 73.0% control efficacy on the spring cultivar "Norangbom" and with 68.9% efficacy on the fall cultivar "Bulam 3". These results suggest that B. toyonensis CAB12243-2 can be used as a biological control agent for the control of soft rot diseases on vegetables.