• Title/Summary/Keyword: Pectobacterium carotovorum subsp. carotovorum

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Identification of Novel Bioactive Hexapeptides Against Phytopathogenic Bacteria Through Rapid Screening of a Synthetic Combinatorial Library

  • Choi, Jae-Hyuk;Moon, Eun-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.792-802
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    • 2009
  • Antimicrobial peptides (AMPs) are considered to be a promising alternative to conventional antibiotics for future generations. We identified four novel hexapeptides with antimicrobial activity: KCM11 (TWWRWW-$NH_2$), KCM12 (KWRWlW-$NH_2$), KCM21 (KWWWRW-$NH_2$), and KRS22 (WRWFIH-$NH_2$), through positional scanning of a synthetic peptide combinatorial library (PS-SCL). The ability of these peptides to inhibit the growth of a variety of bacteria and unicellular fungi was evaluated. KCM11 and KRS22 preferentially inhibited the normal growth of fungal strains, whereas KCM12 and KCM21 were more active against bacterial strains. Bactericidal activity was addressed in a clear zone assay against phytopathogenic bacteria, including Pectobacterium spp., Xanthomonas spp., Pseudomonas spp., etc. KCM21 showed the highest activity and was effective against a wide range of target organisms. Application of KCM21 with inoculation of Pectobacterium carotovorum subsp. carotovorum on detached cabbage leaves resulted in an immune phenotype or a significant reduction in symptom development, depending on the peptide concentration. Cytotoxicity of the four hexapeptides was evaluated in mouse and human epithelial cell lines using an MTT test. The results revealed a lack of cytotoxic effects.

Biological Control of Soft Rot on Chinese Cabbage Using Beneficial Bacterial Agents in Greenhouse and Field (유용세균(Beneficial Bacterial Agents)을 이용한 배추 무름병의 생물적 방제)

  • Shrestha, Anupama;Kim, Eun-Chang;Lim, Chuen-Keun;Cho, Sae-Youll;Hur, Jang-Hyun;Park, Duck-Hwan
    • The Korean Journal of Pesticide Science
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    • v.13 no.4
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    • pp.325-331
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    • 2009
  • Three beneficial bacterial agents, Lactobacillus strain KLF01, Lactococcus strain KLC02 and Paenibacillus strain KPB3 were showed clear zone against plated Pectobacterium carotovorum subsp. carotovorum (Pcc) soft rot pathogen. In greenhouse test, bio-control efficacy was more significantly effective in the treatments by KLC02 and KPB3 as 64%, 50%, 56% and 66%, 57%, 58% according to date of evaluation, respectively. In case of KLF01 control effect was relatively lower than treatments of KLC02 and KPB3 but its efficacy was still significantly observed when compared to control (only water treatment). Furthermore, KLF01, KLC02 and KPB3 showed 55%, 60% and 62% bio-control efficacy, respectively in field test from early March to late July of 2009. Thus, we suggest that these strains can be useful as bio-control agents against soft rot caused by Pcc.

Effect of Light-Induced ROS Generation Unit on Inactivation of Foodborne Pathogenic Bacteria in Water (광유도 ROS 발생장치의 세척용수 중 식중독 세균에 대한 불활성화 효과)

  • Choi, Jaehyuk;Kim, Dawoon;Jung, Kyu-Seok;Roh, Eunjung;Ryu, Kyoung-Yul;Ryu, Jae-Gee
    • 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.

Selection of a Susceptible Line (Susceptible to Pectobacterium 1, Atstp1) to Soft-rot Disease in T-DNA Insertion Mutants Pool of Arabidopsis (무름병에 감수성인 애기장대 돌연변이체 Atstp1 선발)

  • Choi, Chang-Hyun;Kim, Min-Gab;Ahn, Il-Pyung;Park, Sang-Ryeol;Bae, Shin-Chul;Hwang, Duk-Ju
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.312-315
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    • 2010
  • Pectobacterium carotovorum subsp. carotovorum (Pcc) causes soft rot disease in various plants. Although many studies about Pcc have been going on, little is known yet about the defense genes from plants. To identify defense associated genes in response to Pcc, we screened about 20 thousand Arabidopsis T-DNA knock out lines by inoculation with Pcc. We obtained a line (Atspt1) showing more susceptible symptom compared to WT (Col-0) on 1 day after the inoculation of Pcc on leaves of Arabidopsis with toothpicks. In this study, we optimized the system to select resistant and susceptible lines to Pcc from T-DNA inserted pool of Arabidopsis and expect the system and Atspt1 might be used for molecular breeding to produce resistant vegetables against Pcc.

A Synergistic Effect of Chitosan and Lactic Acid Bacteria on the Control of Cruciferous Vegetable Diseases

  • Lin, Yu-Chen;Chung, Kuang-Ren;Huang, Jenn-Wen
    • The Plant Pathology Journal
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    • v.36 no.2
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    • pp.157-169
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    • 2020
  • Two lactic acid bacteria (LAB) designated J02 and J13 were recovered from fermented vegetables based on their ability to suppress soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum (Pcc) on radish. J02 and J13 were identified as Lactobacillus pentosus and Leuconostoc fallax, respectively. The ability of J02 and J13 to suppress plant diseases is highly dependent on chitosan. LAB alone has no effect and chitosan alone has only a moderate effect on disease reduction. However, J02 or J13 broth cultures plus chitosan display a strong inhibitory effect against plant pathogens and significantly reduces disease severity. LAB strains after being cultured in fish surimi (agricultural waste) and glycerol or sucrose-containing medium and mixed with chitosan, reduce three cruciferous vegetable diseases, including cabbage black spot caused by Alternaria brassicicola, black rot caused by Xanthomonas campestris pv. campestris, and soft rot caused by Pcc. Experimental trials reveal that multiple applications are more effective than a single application. In-vitro assays also reveal the J02/chitosan mixture is antagonistic against Colletotrichum higginsianum, Sclerotium rolfsii, and Fusarium oxysporum f. sp. rapae, indicating a broad-spectrum activity of LAB/chitosan. Overall, our results indicate that a synergistic combination of LAB and chitosan offers a promising approach to biocontrol.

Water Extract from Spent Mushroom Substrate of Hericium erinaceus Suppresses Bacterial Wilt Disease of Tomato

  • Kwak, A Min;Min, Kyeong Jin;Lee, Sang Yeop;Kang, Hee Wan
    • Mycobiology
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    • v.43 no.3
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    • pp.311-318
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    • 2015
  • Culture filtrates of six different edible mushroom species were screened for antimicrobial activity against tomato wilt bacteria Ralstonia solanacearum B3. Hericium erinaceus, Lentinula edodes (Sanjo 701), Grifola frondosa, and Hypsizygus marmoreus showed antibacterial activity against the bacteria. Water, n-butanol, and ethyl acetate extracts of spent mushroom substrate (SMS) of H. erinaceus exhibited high antibacterial activity against different phytopathogenic bacteria: Pectobacterium carotovorum subsp. carotovorum, Agrobacterium tumefaciens, R. solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. axonopodis pv. citiri, and X. axonopodis pv. glycine. Quantitative real-time PCR revealed that water extracts of SMS (WESMS) of H. erinaceus induced expressions of plant defense genes encoding ${\beta}$-1,3-glucanase (GluA) and pathogenesis-related protein-1a (PR-1a), associated with systemic acquired resistance. Furthermore, WESMS also suppressed tomato wilt disease caused by R. solanacearum by 85% in seedlings and promoted growth (height, leaf number, and fresh weight of the root and shoot) of tomato plants. These findings suggest the WESMS of H. erinaceus has the potential to suppress bacterial wilt disease of tomato through multiple effects including antibacterial activity, plant growth promotion, and defense gene induction.

Evidence for Volatile Memory in Plants: Boosting Defence Priming through the Recurrent Application of Plant Volatiles

  • Song, Geun Cheol;Ryu, Choong-Min
    • Molecules and Cells
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    • v.41 no.8
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    • pp.724-732
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    • 2018
  • Plant defence responses to various biotic stresses via systemic acquired resistance (SAR) are induced by avirulent pathogens and chemical compounds, including certain plant hormones in volatile form, such as methyl salicylate and methyl jasmonate. SAR refers to the observation that, when a local part of a plant is exposed to elicitors, the entire plant exhibits a resistance response. In the natural environment, plants are continuously exposed to avirulent pathogens that induce SAR and volatile emissions affecting neighbouring plants as well as the plant itself. However, the underlying mechanism has not been intensively studied. In this study, we evaluated whether plants "memorise" the previous activation of plant immunity when exposed repeatedly to plant defensive volatiles such as methyl salicylate and methyl jasmonate. We hypothesised that stronger SAR responses would occur in plants treated with repeated applications of the volatile plant defence compound MeSA than in those exposed to a single or no treatment. Nicotiana benthamiana seedlings subjected to repeated applications of MeSA exhibited greater protection against Pseudomonas syringae pv. tabaci and Pectobacterium carotovorum subsp. carotovorum than the control. The increase in SAR capacity in response to repeated MeSA treatment was confirmed by analysing the defence priming of the expression of N. benthamiana Pathogenesis-Related 1a (NbPR1a) and NbPR2 by quantitative reverse-transcription PCR compared with the control. We propose the concept of plant memory of plant defence volatiles and suggest that SAR is strengthened by the repeated perception of volatile compounds in plants.

Development of an Efficient Bioassay Method for Testing Resistance to Bacterial Soft Rot of Chinese Cabbage (효율적인 배추 무름병 저항성 검정법 개발)

  • Lee, Soo Min;Choi, Yong Ho;Kim, Hun;Kim, Heung Tae;Choi, Gyung Ja
    • Research in Plant Disease
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    • v.26 no.3
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    • pp.159-169
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    • 2020
  • 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 Chinese cabbage to bacterial soft rot. Resistance degrees of 65 commercial Chinese cabbage cultivars to the Pcc KACC 10225 isolate were investigated. For further study, three Chinse cabbage cultivars (Taebong, Hadaejangkun, CR Alchan) 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 growth stage of Chinse cabbage seedling, inoculum concentration, and incubation temperature after inoculation. On the basis of the results, we suggest that an efficient screening method for resistant Chinses cabbage to Pcc is to inoculate twenty one-day-old seedlings with a bacterial suspension of Pcc at a concentration of 1×107 cfu/ml, and to incubate the plants in a dew chamber at 25℃ for 24 hr and then to cultivate in a growth room at 25℃ and 80% relative humidity with 12-hr light per day.

A New Screening Method for the Selection of Calla Lily Zantedeschia aethiopica Cultivars Resistant to Calla Lily Soft Rot (칼라의 무름병 저항성 품종 선발을 위한 검정 방법 개발)

  • Joung, Hyang Young;Choi, Mok Pil;Han, Kyung Sook;Kim, Su;Goo, Dae Hoe;Kang, Yun Im;Choi, Youn Jung;Park, Sang Kun
    • 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.

First Report of Soft Rot by Pectobacterium carotovorum subsp. brasiliense on Amaranth in Korea

  • Jee, Samnyu;Choi, Jang-Gyu;Hong, Suyoung;Lee, Young-Gyu;Kwon, Min
    • Research in Plant Disease
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    • v.24 no.4
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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.