• 한국균학회지
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• 제46권4호
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• pp.467-477
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• 2018

탄저병균인 Colletotrichum circinans는 세계적으로 양파에 심각한 피해를 주는 병원균이다. 본 연구에서는 일반 PCR방법과 real-time PCR방법으로 C. circinan를 정확하면서도 쉽고 빠르게 검출이 가능한 특이 마커를 개발하였다. $tef-1{\alpha}$ 유전자와 ${\beta}-tubulin$ 유전자를 분석하여 C. circinan를 특이적으로 검출할 수 있는 cirTef-F/cirTef-R set와 cirTu-F/cirTu-R set를 제작하였다. 일반 PCR 방법으로 cirTef-F/cirTef-Rset는 100pg, cirTu-F/cirTu-Rset는 1ng까지 검출이 가능하였고 real-time PCR 방법으로는 각각 10 pg, 100 pg까지 검출이 가능하였다. C. circinans에 인공적으로 감염된 양파 종자에서도 cirTef-F/cirTef-Rset를 사용하여 일반 PCR방법과 real-time PCR 방법 모두 C. circinans검출이 가능하였다. 본 연구에서 개발한 C. circinans 특이 검출마커는 수출입 되는 채소 및 종자에서 빠르고 정확하게 탄저병균인 C. circinans를 검출하는데 사용될 수 있을 것이다.

• Current Research on Agriculture and Life Sciences
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• 제31권4호
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• pp.245-249
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• 2013

In 2008, leaf blight symptoms were observed on several Chinese chive farms in Sangju. The Pathogenicity of the isolate was confirmed by artificial inoculation, where the pathogen exhibited a strong pathogenicity toward healthy plants. Morphological classification identified the isolate as from the Fusarium genus. For further analysis, PCR and phylogenetic classification were performed with ITS region and 28S rRNA gene which are commonly used for fungal identification. However, the results provided a poor resolution. To solve this problem, we analyzed translation elongation factor 1-alpha (TEF-$1{\alpha}$) gene. The analyzed results using TEF-$1{\alpha}$ gene indicated that the isolate was F. proliferatum. Therefore, it is assumed that TEF-$1{\alpha}$ gene is important when Fusarium sp. was identified using molecular classification method.

• 한국균학회지
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• 제46권2호
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• pp.186-192
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• 2018

PCR 기술을 이용하여 고추와 토마토의 탄저병을 일으키는 Colletotrichum coccodes 균을 빠르고 정확하게 검출하는 방법을 개발하였다. Colletotrichum 13종 22개 균주로 부터 translation elongation factor 1 alpha 유전자를 분석하여 C. coccodes에 특이적인 coccoTef-F/cocco Tef-R primer set를 제작하였다. 제작된 primer를 사용한 결과 일반 PCR 방법으로 10 ng, real-time PCR 방법으로는 10 pg 수준에서 C. coccodes가 특이적으로 검출이 가능하였다. 인공적으로 C. coccodes에 감염시킨 고추와 토마토 종자에서도 일반 PCR 방법과 real-time PCR 방법 모두 C. coccodes검출이 가능하였다. 본 연구에서 개발한 PCR 방법은 수출입되는 종자에서 신속하고 정확하게 탄저병균 C. coccodes를 특이적으로 검출하는데 활용될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1563-1570
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• 2010

To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

• Mycobiology
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• 제47권1호
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• pp.76-86
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• 2019

Scab disease caused by Venturia nashicola is of agroeconomic importance in cultivation of Asian pear. However, little is known about the degree of genetic diversity in the populations of this pathogen. In this study, we collected 55 isolates from pear scab lesions in 13 major cultivation areas in Korea and examined the diversity using sequences of internal transcribed spacer (ITS) region, ${\beta}$-tubulin (TUB2), and translation elongation factor-$1{\alpha}$ ($TEF-1{\alpha}$) genes as molecular markers. Despite a low level of overall sequence variation, we found three distinctive subgroups from phylogenetic analysis of combined ITS, TUB2, and $TEF-1{\alpha}$ sequences. Among the three subgroups, subgroup 1 (60% of isolates collected) was predominant compared to subgroup 2 (23.6%) or subgroup 3 (16.4%) and was distributed throughout Korea. To understand the genetic diversity among the subgroups, RAPD analysis was performed. The isolates yielded highly diverse amplicon patterns and none of the defined subgroups within the dendrogram were supported by bootstrap values greater than 30%. Moreover, there is no significant correlation between the geographical distribution and the subgroups defined by molecular phylogeny. Our data suggest a low level of genetic diversification among the populations of V. nashicola in Korea.

• The Plant Pathology Journal
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• 제26권4호
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• pp.409-416
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• 2010

Fusarium species, a large group of plant pathogens, potentially pose quarantine concerns worldwide. Here, we focus on the development of a method for detecting four Fusarium species in quarantined plants in Korea: F. solani f. sp. cucurbitae, F. stilboides, F. redolens, and F. semitectum var. majus. Species-specific primers were designed from the nucleotide sequences of either the translation elongation factor-1 alpha (TEF1) gene or RNA polymerase II subunit (RPB2) gene. Two different primer sets derived from TEF1, all specific to F. solani f. sp. cucurbitae, were able to differentiate the two races (1 and 2) of this species. A set of nested primers for each race was designed to confirm the PCR results. Similarly, two primer sets derived from RPB2 successfully amplified specific fragments from five F. stilboides isolates grouped within a single phylogenetic clade. A specific TEF1 primer set amplified a DNA fragment from only four of the 12 F. redolens strains examined, which were grouped within a single phylogenetic clade. All of the F. semitectum var. majus isolates could be specifically detected with a single RPB2 primer set. The specificity of the primer sets developed here was confirmed using a total of 130 Fusarium isolates.

• 한국균학회지
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• 제45권1호
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• pp.74-82
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• 2017

꽃도라지(Eustoma grandiflorum)는 국내에서 장식용 화훼류로 널리 이용되는 절화류이다. 2015년 경기도 여주와 경남 김해 지역에서 꽃도라지가 시들고, 지제부가 잘록해지면서 위축되고, 뿌리가 썩는 증상이 나타났다. 병든 식물체는 포장이나 육묘상자에서 황화되고, 뿌리 발육이 저해되었고, 진전되면 전체적으로 황화되면서 시들고, 위축되며, 결국 2~3개월 안에 완전히 말라 죽는다. 병든 식물체 지제부에서 Fusarium균이 분리되었고, 9개 균주를 단포자 분리하여 형태적 특성을 조사한 결과, Fusarium solani로 동정되었다. 대형포자는 통통하고, 직선형이거나 약간 굽은 초승달 모양이고, 소형분생포자는 긴mono형태의 분생포자원세포에서 false head상으로 형성되었다. 후벽포자는 균사 중간 혹은 끝부분에 풍부하게 형성되었다. 이와 같은 동정 결과는 translation elongation factor 1 alpha (TEF)와 RNA polymerase II subunit (RPB2) 유전자의 염기서열 분석으로 재확인되었다. 그 결과, 분리 균주는 NCBI GenBank에 등록된 F. solani와 TEF 유전자는 99.2~99.9%, RPB2 유전자는 98.0~98.1%의 상동성을 나타내었다. 건전한 꽃도라지 유묘의 뿌리를 포자현탁액에 침지 접종하여 병원성 검정을 수행한 결과, 접종 7일 이내에 접종한 식물체에서만 병징이 관찰되었다. 따라서 이 병을 F. solani에 의한 꽃도라지 뿌리썩음병으로 명명하며, 병의 발생을 국내에서 처음으로 보고한다.

• 한국균학회지
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• 제46권1호
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• pp.58-68
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• 2018

클로탈라리아는 국내에서 토양개량을 위한 질소고정, 토양침식 감소, 잡초 및 선충 억제를 위해 풋거름작물로 이용하고 있다. 2014년, 클로탈라리아를 풋거름작물로 재배하는 완주 지역의 포장에서 시들음 증상이 관찰되었다. 감염된 식물체의 잎은 아래잎부터 황화 되면서 시들기 시작하였고, 위쪽 잎도 황화 되었으며, 결국 식물체는 완전히 고사하였다. 감염된 식물체 줄기에서 어두운 색의 자낭각이 다수 관찰되었고, 이 자낭각으로부터 자낭포자를 단포자 분리하여 6개 균주를 분리하였다. 균학적 특성에 의해 분리균은 Fusarium udum(완전세대: Gibberella indica)로 동정되었다. 대형포자의 정단세포는 대부분 갈고리 모양으로 굽어 있고, 소형포자는 단경자에서 false head 상으로 형성되었다. 후벽포자는 균사에서 단일 혹은 무리지어 풍부하게 형성되었다. 이와 같은 동정 결과는 translation elongation factor 1 alpha(TEF), calmodulin (CAL), histone 3 (HIS3) 유전자 염기서열 분석으로 재확인되었다. 그 결과, 분리 균주는 NCBI GanBank에 등록된 F. udum과 TEF 유전자는 94.4~96.2%, CAL 유전자는 99.7%, HIS3 유전자는 99.6~99.8%의 상동성을 보였다. 클로탈라리아와 두 품종의 콩을 대상으로 포자현탁액을 토양에 관주 접종하여 병원성 검정을 수행한 결과, 접종 14~21일 이내에 접종한 클로탈라리아와 태광콩에서 병징이 관찰되었다. 따라서 이 병을 F. udum에 의한 클로탈라리아 시들음병으로 명명하고자 제안하며, 국내에서 처음으로 보고한다.

• Mycobiology
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• 제47권4호
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• pp.408-414
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• 2019

Crinum asiaticum and Hymenocallis littoralis, commonly known as spider lilies are bulbous perennial and herbaceous plants that widely planted in Malaysia as ornamental. During 2015-2016, symptom of leaf blight was noticed on the hosts from several locations in Penang. The symptom appeared as irregular brown to reddish lesions surrounded by yellow halos. As the disease progressed, the infected leaves became blighted, dried, and fell off with the presence of black microsclerotia and pycnidia on the lesions parts. The present study was conducted to investigate the causal pathogen of leaf blight on C. asiaticum and H. littoralis. Based on morphological characteristics and DNA sequences of internal transcribed spacer (ITS) region and translation elongation factor 1-alpha (TEF1-α) gene, the causal pathogen was identified as Macrophomina phaseolina. Phylogenetic analysis of combined dataset of ITS and TEF1-α grouped the isolates studied with other isolates of M. phaseolina from GenBank. The grouping of the isolates was supported by 96% bootstrap value. Pathogenicity test proved the role of the fungus in causing leaf blight on both hosts.

• 한국균학회지
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• 제46권3호
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• pp.213-225
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• 2018

Trichoderma (Hypocreaceae) is one of the most ubiquitous genera worldwide. This genus has an excellent ability to adapt to diverse environments, even under poor nutritional conditions, such as in freshwater. However, little is known about the diversity of Trichoderma species in freshwater environments. In this study, we isolated diverse fungal strains from algae, plant litter, and soil sediment in streams in Korea. The strains were identified based on molecular phylogenetic analyses of internal transcribed spacer (ITS) rDNA and translation elongation factor 1 ($TEF1{\alpha}$) sequences. We also investigated their morphological characteristics by microscopic observation and determination of cultural features on different media. As a result, six Trichoderma species were found in Korea: T. afroharzianum, T. capillare, T. guizhouense, T. paraviridescens, T. reesei, and T. saturnisporum. Interestingly, T. paraviridescens showed both cellulose activity and hypoxia stress tolerance phenotypes, indicating its role as a decomposer in freshwater ecosystems. Our study revealed that freshwater environment could be a good candidate for investigating the species diversity of Trichoderma.