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국화에 발생하는 반쪽시들음병균 Verticillium dahliae 검출용 등온 증폭법 개발
백창기 ( Chang-gi Back ),박미정 ( Mi-jeong Park ),한경숙 ( Kyung-sook Han ),박종한 ( Jong-han Park ) 한국균학회 2019 韓國菌學會誌 Vol.47 No.4
Verticillium wilt disease is caused by a fungal plant pathogen Verticillium dahliae, which attacks commercial crops such as chrysanthemum. The conventional methods so far used to identify this fungal pathogen require high expertise and are time-consuming. Therefore, in this study, we developed an assay for the rapid and specific detection of V. dahliae infection using loop-mediated isothermal amplification (LAMP) method. For this assay, four primers for LAMP were designed for targeting cellulose-growth-specific protein partial mRNA gene in Verticillium dahliae. Under standard condition, the optimum reaction temperature for amplification is around 60 ℃ within 60 minutes. This LAMP assay was designed to amplify only present in V. dahliae. When this LAMP assay applied to the DNAs for four other soil-borne fungi and host plants, no amplification was detected. Therefore, this LAMP assay we developed for V. dahliae is expected to do detection at the early stage of its infection. The fast and reliable detection method will allow us to develop effective management system to monitor and control infection of this pathogen in chrysanthemum plant.
우리나라에 발생하는 잿빛곰팡이병균 Botrytis cinerea의 분자계통학적 유연관계
백창기 ( Chang Gi Back ),이승열 ( Seung Yeol Lee ),정희영 ( Hee Young Jung ) 한국균학회 2014 Mycobiology Vol.42 No.2
잿빛곰팡이병의 전형적인 병징을 나타내는 병든 사과, 고추, 딸기, 오이, 토마토에서 곰팡이를 분리하고, 그들의 배양학적 특성과, 형태적 특성 및 PCR-RFLP을 통해 이 병원성 곰팡이를 모두 Botrytis cinerea로 동정하였다. 또한, 배양학적 특징에 따라 사과, 고추, 오이에서 분리한 잿빛곰팡이병균의 표현형은 균핵형이며, 딸기와 토마토에서 분리한잿빛곰팡이병균은 균사형이었다. 각각의 잿빛곰팡이병균의 ITS 영역 염기서열을 포함한 4종의 유전자(RPB2, HSP60, G3PDH)의 염기서열을 결정하고 분자계통학적 유연관계를 분석하였다. RPB2 유전자 염기서열을 제외한 ITS 영역, HSP60유전자 및 G3PDH 유전자의 염기서열은 Botrytis cinerea 종 내 뿐만 아니라 Botrytis 속 종간에도 매우 높은 상동성을 나타내어 계통학적 유연관계 분석이 어려웠다. 하지만, 3종의 유전자(RPB2, HSP60, G3PDH)를 결합한 유전자 염기서열을 이용한 분자 계통수 작성 결과, 본 연구에서 분리한 잿빛곰팡이병균은 Botrytis 속의 다른 종들과 구별되며, 사과, 고추, 오이, 토마토의 분리주는 아주 높은 근연관계에 있고, 딸기잿빛곰팡이병균은 다른 분리주와 달리 종내 다른 lineage를 형성하였다. Several isolates were collected from apple, pepper, strawberry, cucumber and tomato having typical gray mold symptoms. All the isolates were identified as Botrytis cinerea by using morphological characteristics and PCR-RFLP method. It was difficult to analyze the phylogenetic relationship of these isolates by using ITS region, HSP60 and G3PDH because these genes were highly homologous in their nucleotide in inter-species of B. cinerea and intra-species of genus Botrytis. However, phylogenetic analysis using combined sequences (RPB2, HSP60 and G3PDH genes) clearly showed that all isolate of B. cinerea were different from Botrytis spp. Furthermore, it was also confirmed that strawberry isolate was distantly related to apple, pepper, cucumber and tomato isolates that were closely related to each other in nucleotide level.
총설 : 사과나무를 가해하는 한국산 갈색무늬병균의 생물학적 특성
백창기 ( Chang Gi Back ),정희영 ( Hee Young Jung ) 한국균학회 2014 韓國菌學會誌 Vol.42 No.3
Apple blotch is a major apple disease and recently it causes critical economic losses in apple orchards in Korea. In this review, we described the history of apple blotch researches, ecology and life cycle of Marssonina coronaria, cultural and molecular characteristics and simple isolation method as well. Furthermore, characteristic of apple blotch like symptoms, its cause still unknown, occurs frequently in cv. “Fuji” was described in detail.
양파 노균병균 Peronospora destructor의 분자계통학적 유연관계 분석과 PCR 검출기술 개발
백창기 ( Chang-gi Back ),황선경 ( Sun-kyung Hwang ),박미정 ( Mi Jeong Park ),권영석 ( Young-seok Kwon ),정희영 ( Hee-young Jung ),박종한 ( Jong-han Park ) 한국균학회 2017 韓國菌學會誌 Vol.45 No.4
우리나라 양파 재배지의 주요 곰팡이병인 양파 노균병의 원인균 Peronospora destructor를 채집하여, internal transcribed spacer (ITS) 영역을 포함한 4개 유전자의 염기서열을 분석하여 상동성 비교와 분자계통학적 유연관계를 분석하였다. 그 결과, 경남 창녕군, 함안군, 합천군과 전남 무안군, 신안군, 해남군에 발생하는 P. destructor의 4종의 유전자 염기서열 모두 100% 상동성을 보였고, 유연관계 분석에서도 모두 동일한 계통으로 확인되었다. 본 연구에서는 ITS 영역 유전자 염기서열을 이용하여 P. destructor 검출용 PCR법을 개발하였고, 노균병균만을 특이적으로 검출하였다. 또한 식물체를 포함한 total genomic DNA로 검출한계를 검정한 결과, 0.07 ng/μL까지 가능하였다. 양파 노균병균 검출용 PCR법은 육안상 병징이 나타나지 않을 때도 충분히 감염유무가 확인되었다. Onion downy mildew, caused by Peronospora destructor, is a major disease in onion cultivation areas in Korea. The causal fungi were collected and analyzed based on sequence similarity and molecular phylogenetic relationships of multi-gene sequences, including the internal transcribed spacer (ITS) region. All isolates from Changnyeong-gun, Hamyang-gun, and Hapcheon-gun in Gyeongnam province, and Muan-gun, Haenam-gun, and Sinan-gun in Jeonnam province were identical in the four types of gene sequences, indicating they were genetically the same strains. In this study, a PCR method was developed based on the ITS gene sequences to amplify the specific DNA fragment for P. destructor only. The detection limit of was total genomic DNA of the P. destructor and the plant 0.7 ng/μL. Therefore, the developed PCR method could be used to detect P. destructor effectively from symptomless onion leaves.
가뭄저항성 GM벼(Agb0103)와 non-GM 일미벼 간 표면미생물의 변화 및 병 발생 양상 비교
이승열(Seung-Yeol Lee),백창기(Chang-Gi Back),박상규(Sangkyu Park),김경민(SangJae Suh),서상재(Kyung-Min Kim),정희영(Hee-Young Jung) 한국육종학회 2016 한국육종학회지 Vol.48 No.2
In this study, we compared disease incidence rate and phyllosphere microbial community between drought resistance ransgenic rice (Agb0103) and non-transgenic Ilmi (NGM) during 2011-2014 to examine an environmental risk assessment of drought resistance transgenic rice (Agb0103). As the results, major diseases such as sheath blight, brown spot, leaf blast and false nsmut were occurred, however, there were no significant disease incidence rate between Agb0103 and NGM. As the results of counting bacterial and fungal viable cell, the colonies were increased or decreased which affected by environmental conditions, however there were no differences between Agb0103 and NGM. Also unweighted pair-group method with arithmetic averaging(UPGMA) analysis based on polymerase chain reaction with denaturing gel electrophoresis (PCR-DGGE) revealed that DGGE band pattern of bacterial and fungal communities were clustered by each month and there were no differences between Agb0103 and NGM. Furthermore, isolated casual agents causing sheath blight and brown spot were collected from Agb0103 and NGM, and they revealed that each of pathogens were no differences in morphology and pathogenicity. Therefore, our results suggested that Agb0103 showed no differences in disease incidence rate, characteristic of pathogens and phyllosphere community with NGM. In this way, it can be assumed that transgenic rice Agb0103 could not influence phyllosphere microorganism community and environmental conditions.
토양에서 분리한 Diaporthe tectonae에 대한 보고
박상규,이승열,이재진,백창기,이향범,정희영,Park, Sangkyu,Lee, Seung-Yeol,Lee, Jae-Jin,Back, Chang-Gi,Lee, Hyang Burm,Jung, Hee-Young 한국균학회 2017 韓國菌學會誌 Vol.45 No.1
전북 전주시 완산구의 토양으로부터 국내 미기록종인 균류를 분리하였다. 형태학적 특징 및 ITS영역, ${\beta}-tubulin$, $EF1-{\alpha}$ 유전자를 이용한 계통학적 분석 결과, 토양에서 분리한 균류는 Diaporthe tectonae인 것으로 확인되었으며, 본 연구를 통해 D. tectonae가 국내에 존재함을 최초로 보고하였다. An unrecorded fungal species in Korea, Diaporthe tectonae was isolated from soil in Jeon-ju of Korea. The isolate was characterized morphologically, and a phylogenetic analysis using a combined dataset of internal transcribed spacer, ${\beta}-tubulin$, and elongation factor $1-{\alpha}$ sequences indicated its similarity to D. tectonae strains reported previously. To our knowledge, this is the first report of D. tectonae in Korea.