• Journal of Sericultural and Entomological Science
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• v.44 no.2
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• pp.64-68
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• 2002

The antibacterial protein gene, nuecin was expressed in Sf9 cells using baculovirus expression vector system (BEVS). The antibactetial activity of mature nuecin against Pectobacterium carotovorum subsp. carotovorum, Ralstonia solanacearum and Pseudomonas tolaasii was significantly high, demonstrating that nuecin had a wider antibacterial spectrum on gram negative and positive bacteria. The result appears to be superior to other antibacterial peptide, attacin. The nuecin was purified by SP-sepharose and Mono Q HR ion-exchange chromatography, and then by Superdex 200 HR 10/30 column. The purified nuecin is quite stable at 80$\^{C}$ and 100$\^{C}$ for several hours of incubation and in a wide pH range (pH 2-12).

• 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.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.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.306-311
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• 2009

A new PCR method was developed to detect Pectobacterium carotovorum which is the causative agent of soft rot in Brassica pekinensis. A specific detection primer set based on Lytic murein transglycolase gene was designed and evaluated. Using ERB3_F (5'-TGC GAC ACC TCC TCA TCA CG-3') and ERB3_R (5'-CTT ATC ACG CTG TAA CCA GC-3') primers, 437 nucleotides long fragment was specifically amplified. The amplified products were observed in 52 out of 55 strains of P. carotovorum or Pectobacterium carotovorum subsp. carotovorum. On the other hand, no amplification was observed in 8 organisms including Chinese cabbage and potato. The optimal PCR condition for the ERB3_F/ERB3_R primer set was $58^{\circ}C$ for annealing and 15 mM for $MgCl_2$. With serially diluted templates, the specific PCR sensitivitie limit was $2\times10^3$ copies. Also, this method can be applied not only to DNA but also to field samples. This PCR method may be expected to be useful for specific detection of P. carotovorum.

• Korean Journal of Environmental Agriculture
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• v.35 no.3
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• pp.191-201
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• 2016

BACKGROUND: The aim of this study was to investigate the antimicrobial effects of the ethanol extracts from various medicinal herbs against plant pathogens to understand the possible the crop protection agents.METHODS AND RESULTS: Among the tested medicinal herbs, Zizyphus jujuba ethanol extract had the potent antimicrobial activity against Phytophthora capsici, Erwinia carotovorum subsp. carotovora, Pseudomonas syringae pv. syringae and Ralstonia solanacearum. The major constituents of Z. jujuba were identified to eugenol(40.45%), dodecanoic acid(18.40%), β-caryophyllene (10.05%) and isoeugenol(9.85%) by GC/MS. Eugenol and isoeugenol had strong inhibitory activity on spore germination against P. capsici and growth against E. carotovorum subsp. carotovora, P. syringae pv. syringae and R. solanacearum.CONCLUSION: In this regard, eugenol and isoeugenol were found to be responsible for the antimicrobial activity of Z. jujuba ethanol extract against plant pathogens. In addition, Z. jujuba ethanol extract, eugenol and isoeugenol can be used the potent antimicrobial agents.

• Journal of Food Hygiene and Safety
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• v.34 no.6
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• pp.583-590
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• 2019

As the consumption of fresh fruits and vegetables increases, food poisoning caused by foodborne pathogen contamination is not decreasing. To prevent the contamination of produce, a quick and easy, low-cost, environmentally-safe disinfection method that does not affect produce freshness or quality is needed. This study demonstrates a new-concept, circulating-water disinfection system that purifies water by using newly developed 'LED-PS (photosensitizer)-induced ROS generation unit'. Using various types of LED-PS induced ROS generation units, we investigated the conditions for reducing the density of various pathogenic bacteria by more than 3 log CFU / mL in 1 hour. The major operational factors affecting the density reduction of the LED-PS-induced ROS generation unit were analyzed. Depending on bacteria species, the density reduction rate was varied. The effect of the units on reducing the density of Bacillus cereus and Pectobacterium carotovorum subsp. carotovorum was high, but the effect on foodborne bacteria such as Escherichia coli was relatively low. In this circulating water disinfection system, the density reduction effect tended to increase as the flow rate increased and the initial bacterial density decreased. As the amount of PS absorbed beads increased, the density reduction effect increased exponentially in some bacteria. Model 3280, a double cylindrical unit connecting two single cylindrical units, could completely sterilize more than 3 log CFU/mL of B. cereus and P. carotovorum subsp. carotovorum in 30 minutes of LED irradiation.

• Research in Plant Disease
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• v.29 no.2
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• pp.168-173
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• 2023

Agricultural antibiotics are widely used to inhibit the growth of phytopathogenic bacteria involved in plant diseases. However, continuous antibiotic overuse in crop production may lead to the development of antibiotic resistance in phytopathogenic bacteria. This study was conducted to evaluate the resistance to three different agricultural antibiotics (oxytetracycline+streptomycin, streptomycin, and validamycin A) in 91 strains of phytopathogenic bacteria including Pectobacterium carotovorum, Pseudomonas syringae pv. actinidiae, Clavibacter michiganensis subsp. michiganensis, C. michiganensis subsp. capsici, and Xanthomonas arboricola pv. pruni. Bacterial growth in the presence of various concentrations of validamycin A was also assessed spectrophotometrically by analyzing the optical density. All strains did not grow when the cells were exposed to oxytetracycline+streptomycin or 100× of streptomycin. However, among the 91 strains, 4% and 2% strains showed bacterial growth at the concentrations of 1× and 10× of streptomycin, respectively. Furthermore, 97%, 93%, and 73% strains were resistant to the 1×, 10×, and 100× of validamycin A, respectively, and especially, P. carotovorum contained the highest resistance to the validamycin A. Minimum bactericidal concentration values of validamycin A did not correlate with the patterns of agricultural antibiotic resistance. Further studies are needed to understand the incidence and development of antibiotic resistance in phytopathogenic bacteria.

• Journal of Life Science
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• v.19 no.8
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• pp.1177-1182
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• 2009

In order to establish in vitro infection model for research of plant pathogen based on tissue softening disease in napa cabbage, eighty independent bacterial strains were isolated from the softened napa cabbage tissues. Eight bacterial isolates were primarily screened with the generation of reproducible tissue softening disease to fresh napa cabbages within 24${\sim}$48 hours after inoculation. Through various microbiological biochemical and morphological examinations, three Gram (-) isolates which harbor independent biological properties were finally chosen, and named as RBI, RB2 and RB6. Collective results obtained from API 20E test and analyses of VITEK 2 COMPACT and nucleotide sequences of 165 rRNA of each isolate proposed that isolates RBI and RB2 are close to the Erwinia carotovora subsp. odorifera, and RB6 is close to the Erwinia carotovora subsp. carotovora. These isolates grew optimally at $30^{\circ}C$ with neutral pH culture condition. The isolates caused softening tissue disease with dose-dependent manner regardless of pre-surface damages of napa cabbage. Minimum dose to cause soft rot disease for RBI, RB2 or RB6 were $8.0{\times}10^8$ CFU/mt $10^9$ CFU/ml or $4.7{\times}10^6$ CFU/ml respectively. These isolates caused tissue softening disease to eggplant, paprika and napa cabbage out of 14 different tested vegetables, indicating that these isolates damages specific plant tissues. The bacterial isolates obtained in this research and in vitro plant infection model will be adapted in the understanding of the mechanism of pathogenesis by plant pathogen.

• Horticultural Science & Technology
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• v.31 no.3
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• pp.366-370
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• 2013

This experiment was carried out to establish the screening methods for the selection of Zantedeschia aethiopica cultivars resistant to calla lily soft rot caused by Pectobacterium carotovorum subsp. carotovorum. A center of leaf and petiole tissues of four Z. aethiopica cultivars were wounded and inoculated with three different inoculum concentrations, $1{\times}10^7cfu/mL$, $1{\times}10^8cfu/mL$, and $1{\times}10^9cfu/mL$, of EccNHRI-21 isolate. And they were evaluated at 4, 18, and 26 hours after inoculation. The lesion of maceration was developed well in the leaf tissues inoculated with $1{\times}10^9cfu/mL$ concentration. And evaluation of resistance in 18 hours after inoculation was correlated with field resistance positively. Using this method, ten Z. aethiopica commercial cultivars and four wild types were screened. 'Crowbrough' and 'White Cutie' were selected as highly resistant genotypes and 'Mont Blanc' and 'Silky White' as resistant genotypes. 'Wedding March' and 'Kiwi Blush' were evaluated as moderate resistant and the others including 'Childsiana' were susceptible. And all of four wild types were evaluated as more than moderate resistant. In spite of control through cultural, biological and chemical methods, the use of resistant cultivars is most efficient to overcome calla lily soft rot. This newly developed screening method was helpful for breeding new varieties resistant to calla lily soft rot.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.346-354
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• 2020

Pectobacterium, which causes soft rot disease, is divided into 18 species based on the current classification. A total of 225 Pectobacterium strains were isolated from 10 main cultivation regions of potato (Solanum tuberosum), napa cabbage (Brassica rapa subsp. pekinensis), and radish (Raphanus sativus) in South Korea; 202 isolates (90%) were from potato, 18 from napa cabbage, and five from radish. Strains were identified using the Biolog test and phylogenetic analysis. The pathogenicity and swimming motility were tested at four different temperatures. Pectolytic activity and plant cell-wall degrading enzyme (PCWDE) activity were evaluated for six species (P. carotovorum subsp. carotovorum, Pcc; P. odoriferum, Pod; P. brasiliense, Pbr; P. versatile, Pve; P. polaris, Ppo; P. parmentieri, Ppa). Pod, Pcc, Pbr, and Pve were the most prevalent species. Although P. atrosepticum is a widespread pathogen in other countries, it was not found here. This is the first report of Ppo, Ppa, and Pve in South Korea. Pectobacterium species showed stronger activity at 28℃ and 32℃ than at 24℃, and showed weak activity at 37℃. Pectolytic activity decreased with increasing temperature. Activity of pectate lyase was not significantly affected by temperature. Activity of protease, cellulase, and polygalacturonase decreased with increasing temperature. The inability of isolated Pectobacterium to soften host tissues at 37℃ may be a consequence of decreased motility and PCWDE activity. These data suggest that future increases in temperature as a result of climate change may affect the population dynamics of Pectobacterium.