• The Plant Pathology Journal
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• 제30권2호
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• pp.117-124
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• 2014

The plant pathogenic bacterial genus Pectobacteirum consists of heterogeneous strains. The P. carotovorum species is a complex strain showing divergent characteristics, and a new subspecies named P. carotovorum subsp. brasiliensis has been identified recently. In this paper, we re-identified the P. carotovorum subsp. brasiliensis isolates from those classified under the subspecies carotovorum and newly isolated P. carotovorum subsp. brasiliensis strains. All isolates were able to produce plant cell-wall degrading enzymes such as pectate lyase, polygalacturonase, cellulase and protease. We used genetic and biochemical methods to examine the diversity of P. carotovorum subsp. brasiliensis isolates, and found genetic diversity within the brasiliensis subsp. isolates in Korea. The restriction fragment length polymorphism analysis based on the recA gene revealed a unique pattern for the brasiliensis subspecies. The Korean brasiliensis subsp. isolates were divided into four clades based on pulsed-field gel electrophoresis. However, correlations between clades and isolated hosts or year could not be found, suggesting that diverse brasiliensis subsp. isolates existed.

• The Plant Pathology Journal
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• 제38권2호
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• pp.102-114
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• 2022

Pectobacterium carotovorum subsp. carotovorum (Pcc) is a gram-negative, broad host range bacterial pathogen which causes soft rot disease in potatoes as well as other vegetables worldwide. While Pectobacterium infection relies on the production of major cell wall degrading enzymes, other virulence factors and the mechanism of genetic adaptation of this pathogen is not yet clear. In the present study, we have performed an in-depth genome-wide characterization of Pcc strain ICMP5702 isolated from potato and compared it with other pathogenic bacteria from the Pectobacterium genus to identify key virulent determinants. The draft genome of Pcc ICMP5702 contains 4,774,457 bp with a G + C content of 51.90% and 4,520 open reading frames. Genome annotation revealed prominent genes encoding key virulence factors such as plant cell wall degrading enzymes, flagella-based motility, phage proteins, cell membrane structures, and secretion systems. Whereas, a majority of determinants were conserved among the Pectobacterium strains, few notable genes encoding AvrE-family type III secretion system effectors, pectate lyase and metalloprotease in addition to the CRISPR-Cas based adaptive immune system were uniquely represented. Overall, the information generated through this study will contribute to decipher the mechanism of infection and adaptive immunity in Pcc.

• 한국식물병리학회지
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• 제13권3호
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• pp.179-183
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• 1997

십자화과 식물에 검은썩음병을 일으키는 Xanthomonas campestris pv. campestris에 대하여 담배 18품종 중 가장 과민반응을 강하게 나타내는 품종(Nicotiana tabacum cv. TC500)을 과민반응 비기주식물로 선발하였다. NTG 돌연변이를 실시하여 얻은 약 4,500개의 mutant들을 $10^{8}$ cells/ml의 농도로 주사 접종하여 비기주식물인 담배 잎에서 과민반응을 유도하지 않는 HR-mutans(XHN 514-774, XHN 620-831)를 선발하였고, 기주식물인 양배추 잎에서 병원성을 나타내지 않는 비병원성변이주(XPN 1001)를 선발하였다. HR mutant를 기주식물인 양배추에 접종한 결과 병원성을 나타내었고, nonpathogenic mutant를 비기주식물인 담배잎에 접종한 결과 과민반응을 유도하였다. 담배 잎에서 HR-mutant와 wild type의 population를 조사한 결과 wild type은 24시간 이후 급격히 감소하는 반면, HR mutant는 일정수준을 유지하였으나 72시간 이후에 감소하였다. 그리고 mutant들의 protease, cellulase, amylase, polygalacturonate lyase 분비여부를 조사한 결과 wild type과 비슷하게 분비하였다.

• 식물병연구
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• 제18권3호
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• pp.240-244
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• 2012

양파 무름병 병원균인 Burkholder gladioli pv. allicola CH1을 양파에 접종했을 때 $20^{\circ}C$ 이상에서는 전형적인 양파 무름병 증상을 보이면서 심하게 부패하였지만, $10^{\circ}C$ 이하에서는 일부 조직이 무름증상을 보였지만 겉으로는 건전하게 보였다. B. gladioli pv. allicola CH1은 oxolinic acid WP, streptomycin + copper hydroxide WP와 Streptomycin WP에 강한 감수성을 보였다. 분비효소 중 펙틴질 분해효소의 활성은 검출할 수 있었지만, 섬유소 분해효소인 carboxymethylcellulase 활성은 검출되지 않았다. 그러나 펙틴질 분해활성 중에서 pectin(pectate) lyase의 활성은 검출되지 않고 polygalacturonase 활성만 검출할 수 있었다. PGA 액체배지에서 체외 효소는 18시간까지 급속한 효소반응을 보였으며, 24시간 후에 최고치를 가진 후 계속적인 활성을 가졌으며 효소의 생성은 균의 생육과 거의 일치하는 것으로 나타났다. PGA-SDS-PAGE에 의한 섬유소분해효소 직접활성 염색법을 이용하여 3종류의 polygalacturonase isozyme으로 추정되는 45 kDa, 35 kDa, 29 kDa 크기의 활성밴드를 관찰할 수 있었다.

• 농업과학연구
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• 제41권4호
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• pp.265-274
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• 2014

Physiological characteristics of two Korean golden kiwifruit cultivars, 'Halla Gold' and 'Haehyang', were compared to determine the storage potential of fruit. The soluble solid levels of the fruit were 8.9 and 6.9 oBrix in 'Halla Gold' and 'Haehyang' at harvest, respectively but increased up to 15.4 in 'Halla Gold' and 17.5 oBrix in 'Haehyang' after 2 months of storage. Major sugars were fructose and glucose, and sucrose content was relatively low regardless of cultivar. The edible quality of 'Haehyang' was better than 'Halla Gold' because of higher amount of sugars. Firmness of the fruits gradually decreased as the increase of storage period in 'Halla Gold' in both flesh and core tissue. Th firmness loss of 'Haehyang' fruit was faster in the first 2 months and then became slow. After 75 days of storage, the firmness of 'Haehyang' fruit was only 5.2% at harvest. Core tissue was soften enough to eat at ripe stage. Wall modifying enzyme activities including xylanase, ${\alpha}$-L-arabinofuranosi-dase and ${\beta}$-galactosidase were consistently higher in 'Haehyang' and the activity of pectate lyase was more increased than 'Halla Gold' after 2 months of storage. Respiration rate of 'Haehyang' was higher than 'Halla Gold' and further increased after 2 months of storage. Weight loss was much higher in 'Haehyang' which showed higher rate of the firmness loss. The storage potential of golden kiwifruit was estimated to be about 2 months for 'Haehyang' and 3 months for 'Halla Gold' when determined on the basis of the fruit firmness.

• 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.

• The Plant Pathology Journal
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• 제36권4호
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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.

• 한국유기농업학회지
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• 제26권1호
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• pp.129-140
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• 2018

Pectobacterium carotovorum subsp. carotovorum은 배추를 포함한 대부분의 채소작물에 강한 병원성을 나타낸다. 포장 방제 효과 검정을 통하여 배추 무름병 억제인자로 선발된 CAB12243-2 균주는 16S rRNA gene의 염기서열 상동성 분석으로 Bacillus toyonensis로 확인되었다. B. toyonensis CAB12243-2 균주는 무름병균의 펙틴분해 작용을 억제하였고 탄소원으로 glucose와 trehalose를 이용하였다. 또한 이 균주는 '노랑봄' 품종으로 봄배추 재배 시 73%, '불암3호' 품종으로 가을배추 재배 시 68.9%의 포장 방제 효과를 보였다. 이러한 결과로 B. toyonensis CAB12243-2 균주는 채소류의 무름병에 대한 효과적인 생물적 방제제로 활용이 가능한 것을 확인할 수 있었다.

• 농약과학회지
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• 제20권4호
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• pp.380-387
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• 2016

인삼 모잘록병을 일으키는 병원균의 생물적 방제제 개발을 위하여 인삼 근권 토양으로 부터 500종의 미생물을 분리하였다. 인삼모잘록병원균을 대상으로 항균활성을 검정한 결과, 항균활성이 우수한 균주 3종을 선발하였다. 선발한 균주를 대상으로 항생물질과 세포벽분해효소 생성능력을 조사하였으며, fengycin, bacillomycin D, surfactin, iturin A와 zwittermicin A와 같은 리포펩타이드 생합성 유전자 유무를 조사하였다. ES1과 ES3 균주에서 iturin A와 surfactin 생합성 유전자를 확인하였으며, 세포벽 분해효소 생성능을 확인한 결과 선발한 모든 균주에서 cellulase, pectate lyase, protease를 생성하였다. 16S rRNA 염기서열 분석에 기반하여 계통도를 분석한 결과 ES1 균주와 ES3 균주는 Bacillus methylotrophucus로 확인되었으며, ES2는 B. amyloliquefaciens로 동정되었다. 포장실험 결과 ES1, ES2, ES3 처리구에서 각각 32.4%, 46.8%, 36.7%의 방제효과를 보였다. 이러한 결과로부터 항균활성과 세포벽 분해 효소 생성능이 우수한 길항균주를 선발하였으며, 향후 포장실험과 제형화 개발 등을 통해 인삼모잘록병 방제를 위한 생물적 방제제로 활용할 수 있을 것으로 기대된다.