• 원예과학기술지
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• 제32권6호
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• pp.901-905
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• 2014

The efficacy of ozone micro-bubble water (OMBW) in reducing microbial populations on alfalfa seeds was investigated in this study. We observed the surface of alfalfa seeds using microscopy and found that many cracks and crevices existing on the surface could harbor pathogens. Alfalfa seeds were treated with tap water (TW), micro-bubble water (MBW), ozone water (OW), ozone micro-bubble water (OMBW), and chlorine water (CL) for 5 min, and total microbial population, E. coli and Salmonella spp. colonies were determined. Also, the sterilized seeds were germinated and cultivated for 5 d after sowing to investigate the percentage of germination and the growth of alfalfa sprouts. The treatments with OMBW and CL were most effective in reducing total microbial populations and E. coli was eliminated by OW, OMBW, and CL treatments. CL treatment reduced the percentage of germination and fresh weight of alfalfa sprouts, but OMBW did not cause any negative effects on the germination and growth of alfalfa sprouts. These results indicate that OMBW can be used as an effective sanitizer for eliminating seed-borne pathogens without detrimental effects on seed viability.

• The Plant Pathology Journal
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• 제19권3호
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• pp.184-187
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• 2003

Verticillium wilt was first observed in 2001 on potatoes (Solanum tuberosum) cv. Superior at Daegwallyong area, one of the major seed potato producing areas in Korea. The wilted potato plants showed typical symptoms including gradual yellowing and interveinal necrosis. There was discoloration in the vascular tissues of the infected stems which turned light brown. Fungal isolates from discolored vascular tissues were whitish to creamy with folding on potato dextrose agar medium, where they used to produce resting dark mycelia but no micro-sclerotia. Conidiophores were septate with side branches, swelled at the base, and arranged in a whorl. Conidia were 2.5-11.2$\times$2.0-4.5 $\mu\textrm{m}$ um in size and were borne in small clusters at the tips of phialides. Optimal temperature range for mycelial growth was $25-30^{\circ}C$. Based on these cultural and morphological characteristics, the fungus was identified as Verticillium albo-atrum Reink & Berth. Pathogenicity tests by root dipping method revealed that the fungus caused the same symptoms as observed in naturally infected potato plants. This is the first report of Verticillium wilt on potato caused by Verticillium albo-atrum in Korea.

• 한국약용작물학회지
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• 제8권1호
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• pp.79-82
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• 2000

본 연구는 율무 주산단지인 연천지역에서 율무 종자전염병인 잎마름병 및 깜부기병에 대한 종자 소독 실태와 종자소독제 처리에 의한 방제효과를 검정한 결과를 요약하면 아래와 같다. 율무의 잎마름병 종자감염율은 연천지역 농가에서 수집한 종자를 검사한 결과 $32.7{\sim}45.7%$였으며, 율무종자 소독기간은 율무 1호가 $0{\sim}7$일, 연천 재래종이 $0{\sim}20$일이었다. 농가별 종자소독기간 분포 비율은 무소독 20%, $1{\sim}5$일 20%, $6{\sim}10$일 55% 그리고 11일 이상이 15%였다. 종자소독처리제인 후루디옥소닐종자처리액상수화제의 잎마름병에 대한 종자소독효과는 97%, 깜부기병에 대한 종자 소독효과는 93.4% 로 높은 방제가를 나타냈다.

• 한국미생물·생명공학회지
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• 제46권4호
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• pp.346-359
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• 2018

Xanthomonas oryzae, a bacterial pathogen causing leaf blight disease (BLB) in rice, can cause widespread disease and has caused epidemics globally, resulting in severe crop losses of 50% in Asia. The pathogen is seed-borne and is transmitted through seeds. Thus, control of BLB requires the elimination of the pathogen from seeds. Concern about environment-friendly organic production has spurred improvements in a variety of biological disease control methods, including the use of bacteriophages, against bacterial plant pathogens. The present study explored the potential of bacteriophages isolated from diseased plant leaves and soil samples in killing the bacterial pathogen in rice seeds. Eight different phages were isolated and evaluated for their bacteriolytic activity against different pathogenic X. oryzae strains. Of these, a phage designated ${\varphi}XOF4$ killed all the pathogenic X. oryzae strains and showed the broadest host range. Transmission electron microscopy of ${\varphi}XOF4$ revealed it to be a tailed phage with an icosahedral head. The virus was assigned to the family Siphoviridae, order Caudovirales. Seedlings raised from the seeds treated with $1{\times}10^8pfu/ml$ of ${\varphi}XOF4$ phage displayed reduced incidence of BLB disease and complete bacterial growth inhibition. The findings indicate the potential of the ${\varphi}XOF4$ phage as a potential biological control agent against BLB disease in rice.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• The Plant Pathology Journal
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• 제34권6호
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• pp.490-498
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• 2018

Panicle blight and seed rot disease caused mainly by Burkholderia glumae and Burkholderia gladioli is threatening rice cultivation worldwide. The bacteria have been reported as seed-borne pathogens from rice. Accurate detection of both pathogens on the seeds is very important for limiting the disease dissemination. Novel primer pairs targeting specific molecular markers were developed for the robust detection of B. glumae and B. gladioli. The designed primers were specific in detecting the target species with no apparent cross-reactions with other related Burkholderia species at the expected product size. Both primer pairs displayed a high degree of sensitivity for detection of B. glumae and B. gladioli separately in monoplex PCR or simultaneously in duplex PCR from both extracted gDNA and directly preheated bacterial cell suspensions. Limit of detection was as low as 0.1 ng of gDNA of both species and $3.86{\times}10^2cells$ for B. glumae and $5.85{\times}10^2cells$ for B. gladioli. On inoculated rice seeds, the designed primers could separately or simultaneously detect B. glumae and B. gladioli with a detection limit as low as $1.86{\times}10^3cells$ per rice seed for B. glumae and $1.04{\times}10^4cells$ per rice seed of B. gladioli. The novel primers maybe valuable as a more sensitive, specific, and robust tool for the efficient simultaneous detection of B. glumae and B. gladioli on rice seeds, which is important in combating rice panicle blight and seed rot by early detection and confirmation of the dissemination of pathogen-free rice seeds.

• 식물병연구
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• 제17권1호
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• pp.52-58
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• 2011

우리나라에서 주로 재배되는 십자화과 작물(상추, 콜라비, 무, 배추, 양배추)의 종자 전염 병원균 중에서 세균성 병원균 Xanthomonns campestris pv. campestris(Xcc)와 바이러스 병원균 Lettuce Mosaic Virus(LMV)를 종자에서 동시검출하기 위한 One-step multiplex RT-PCR을 개발하였다. 각각의 병원균을 특이적으로 증폭시키는 병원균 검출용 프라이머 2종(Xcc-F/R, LMV-F/R)을 primerblast 프로그램을 이용하여 제작하였고 이들 프라이머 세트는 프라이머간 또는 병원균 cDNA간의 간섭없이 특이적으로 타겟 병원균만을 검출하였다. PCR을 이용한 병원균의 검출 최소 민감도는 1 ng이었다. 십자화과 작물중에서 유통 중인 콜라비 10품종, 상추 50품종, 무 20품종, 배추 20품종 그리고 양배추 20품종에 대한 종자 감염 병원균 검출을 위한 One-step multiplex RT-PCR 수행 결과, LMV는 전체 120품종 중에서 39품종에서 검출되었고, Xcc는 2개 품종에서 검출되었다. 그리고 50품종의 상추 종자 시료 중에 8품종의 시료에서 LMV와 Xcc가 동시 검출되었다.

• 한국식물병리학회지
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• 제11권3호
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• pp.252-257
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• 1995

Growth rates of the low temperature growing isolates, Pseudomonas fluorescens M45 and MC07, reached maximum stationary phase within 50 hrs at the low temperature, 4$^{\circ}C$. But an ordinary biocontrol agent P. putida Pf3 did not reach logarithmic growth phase until 80 hrs. The culture filtrates of M45 and MC07 significantly inhibited the mycelial growths of Pythium ultimum, Rhizoctonia solani and Phytophthora capsici. Detached cotyledons of cucumber grown on Murashige and Skoog agar medium were much enhanced in their growth, compared to those without the filtrates. Population densities of M45 and MC07 in the rhizosphere at 14$^{\circ}C$ were more stable than at 27$^{\circ}C$. When M45 and MC07 were treated into soil, the population density of MC07 continuously increased up to 9 days after treatment, and sustained the initial inoculum density up to 60 days. Cucumber damping-offs caused by P. ultimum and R. solani were significantly reduced by applying M45 as seed-inoculant and by soil treatment with MC07. The combined treatment of M45 and MC07 provided greater effect in reducing the disease incidence than that obtained by single treatments.

• 한국균학회지
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• 18권2호
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• pp.102-108
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• 1990

16과(科) 28종(種)의 작물추출액(植物抽出液)을 공시(供試)하여 Pythium ultimum에 대한 항균성(抗菌性)을 검정(檢定)하여 다음과 같은 결과를 얻었다. 목단, 자리공, 대황 등 9개의 작물추출액(植物抽出液)에서 유주자낭(遊走子囊) 발아억제효과(發芽抑制效果)가 큰 것으로 나타난다. 목단, 대황, 자리공에서 균사생장(菌絲生長) 억제효과(抑制效果)가 컸으며 그 중 목단이 가장 효과가 컸다. 참깨에서 목단 추출액(油出液)이 가장 강한 발병억제효과(發炳抑制效果)를 나타냈고, 오이에서는 억제효과(抑制效果)가 없었다. 자리공추출액(抽出液)에서 종자발아(種子發芽)에 약간의 약해가 나타났고 목단과 대황의 추출액(抽出液)에서는 약해가 없었다.

• The Plant Pathology Journal
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• 제36권1호
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• pp.29-42
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• 2020

Acidovorax citrulli causes bacterial fruit blotch in Cucurbitaceae, including watermelon. Although A. citrulli is a seed-borne pathogen, it can cause diverse symptoms in other plant organs like leaves, stems and fruits. To determine the infection routes of A. citrulli, we examined the virulence of six isolates (Ac0, Ac1, Ac2, Ac4, Ac8, and Ac11) on watermelon using several inoculation methods. Among six isolates, DNA polymorphism reveals that three isolates Ac0, Ac1, and Ac4 belong to Clonal Complex (CC) group II and the others do CC group I. Ac0, Ac4, and Ac8 isolates efficiently infected seeds during germination in soil, and Ac0 and Ac4 also infected the roots of watermelon seedlings wounded prior to inoculation. Infection through leaves was successful only by three isolates belonging to CC group II, and two of these also infected the mature watermelon fruits. Ac2 did not cause the disease in all assays. Interestingly, three putative type III effectors (Aave_2166, Aave_2708, and Aave_3062) with intact forms were only found in CC group II. Overall, our results indicate that A. citrulli can infect watermelons through diverse routes, and the CC grouping of A. citrulli was only correlated with virulence in leaf infection assays.