• 제목/요약/키워드: ${\beta}$-tubulin gene

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순창군 장류로부터 분리된 황국균의 동정 및 특성 (Identification and Characterization of Aspergillus oryzae Isolated from Soybean Products in Sunchang County)

  • 임은미;이지영;모하메드;한갑훈;이보순;조용식;김현영
    • 한국균학회지
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    • 제42권4호
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    • pp.282-288
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    • 2014
  • 본 연구에서는 순창지역에서 만들어지는 장류에서 곰팡이를 분리하고 동정하여 보다 안전하고 기능성이 높은 발효제품을 위한 균주를 확보하고자 하였다. 순창지역에서 생산되는 장류 제품으로부터 곰팡이를 분리하여 ${\beta}$-tubulin 유전자 분석 통해 10개의 균주가 Aspergillus oryzae/flavus complex임을 알 수 있었다. 보다 정확한 동정을 위하여 아플라톡신 클러스터 유전자 중에 하나인 omtA의 염기서열을 증폭하여 A. oryzae와 A. flavus 표준 균주의 omtA 서열과 함께 계통 분류한 결과, A. oryzae의 표준 균주와의 유연관계가 높음을 알 수 있었다. 또한 norB-cypA 사이의 염기서열을 증폭한 결과 500 bp이 증폭 산물이 확인되었는데 이는 표준 균주인 A. oryzae의 norB-cypA 사이의 염기서열 증폭 산물과 동일한 크기임을 확인할 수 있었다. A. oryzae로 확인된 10균주를 활용하여 코지를 제조하고 ${\alpha}$-amylase 활성과 protease 활성을 측정하였다. Protease 활성은 6, 13, 17, 27, 37, 그리고 38 균주로 제조된 코지는 대조구(시판되고 있는 종균으로 제작한 코지)보다 2배 정도 높은 protease 활성을 보였으며, ${\alpha}$-amylase 활성은 257~320 U/mL로 측정되었다. 식품안전성을 위한 아플라톡신 분비 확인 결과, 63번 균주로 제조된 코지를 제외한 모든 코지에서 아플라톡신을 만들지 않는 것으로 확인되어, 순창에서 분리된 A. oryzae는 추후 메주 접종균으로 개발할 수 있음을 보여주었다.

Benzimidazole계 살균제에 대한 배 검은별무늬병균 Venturia nashicola 의 저항성과 β-Tubulin 유전자 돌연변이와의 관계 (Relationship of Resistance to Benzimidazole Fungicides with Mutation of β-Tubulin Gene in Venturia nashicola)

  • 곽연수;민지영;송장훈;김명수;이한찬;김흥태
    • 식물병연구
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    • 제23권2호
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    • pp.150-158
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    • 2017
  • Benzimidazole계 살균제 저항성을 모니터링하면서, 저항성과 ${\beta}$-tubulin 유전자의 198번째 코돈의 돌연변이와의 관계를 조사하였다. Venturia nashicola의 benzimidazole계 살균제에 대한 반응은 $0.16{\mu}g/ml$ carbendazim 배지에서 전혀 생장하지 못하는 감수성과 $4.0{\mu}g/ml$ 배지에서 생장이 안 되는 저저항성, $100{\mu}g/ml$에서조차 생장이 가능한 고저항성으로 구분할 수 있었다. 국내 3개 지역의 과원에서 채집한 30균주의 V. nashicola는 carbendazim에 대해서 모두 고저항성이었다. V. nashicola의 ${\beta}$-tubulin 유전자의 염기서열을 분석한 결과 저저항성 균주에서는 감수성 균주와 염기서열에 차이가 없었지만, 고저항성 균주는 GAG인 glutamic acid가 GCG인 alanine으로 치환되어 있었다. 옥천의 과원에서 분리한 10균주 중에서 5균주는 GGG인 glycine으로 치환되어 있었다. 특히 glycine으로 치환될 경우 carbendazim에 대해서 저항성이 되면서 carbendazim과 diethofencarb 혼합제에 대해서는 감수성이 더 커졌다. 결국 ${\beta}$-tubulin의 198번째 코돈의 돌연변이 E198A와 E198G는 V. nashicola가 benzimidazole계 살균제에 대해서 저항성이 되는 중요한 기작임을 알 수 있었다.

Evaluation of ${\alpha}$-Tubulin as an Antigenic and Molecular Probe to Detect Giardia lamblia

  • Kim, Ju-Ri;Shin, Myeong-Heon;Song, Kyoung-Ju;Park, Soon-Jung
    • Parasites, Hosts and Diseases
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    • 제47권3호
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    • pp.287-291
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    • 2009
  • The ${\alpha}/{\beta}$-tubulin heterodimer is the basic subunit of microtubules in eukaryotes. Polyclonal antibodies specific to recombinant ${\alpha}$-tubulin of Giardia lamblia were made, and found effective as a probe to specifically detect G. lamblia by immunofluorescence assays. Nucleotide sequences of ${\alpha}$-tubulin genes were compared between G. lamblia WB and GS strains, prototypes of assemblage A and assemblage B, respectively. A set of primers was designed and used to amplify a portion of the ${\alpha}$-tubulin gene from G. lamblia. PCR-RFLP analysis of this ${\alpha}$-tubulin PCR product successfully differentiated G. lamblia into 2 distinct groups, assemblages A and B.Theresults indicate that ${\alpha}$-tubulin can be used as a molecular probe to detect G.lamblia.

Molecular Analysis of Botrytis cinerea Causing Ginseng Grey Mold Resistant to Carbendazim and the Mixture of Carbendazin Plus Diethofencarb

  • Kim, Joo-Hyung;Min, Ji-Young;Bae, Young-Seok;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • 제25권4호
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    • pp.322-327
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    • 2009
  • A total of 23 isolates of Botrytis cinerea causing the grey mold were collected from infected ginseng in several fields of Korea. The sensitivity to carbendazim and the mixture of carbendazim plus diethofencarb was determined through a mycelial inhibition test on PDA amended with or without fungicides. B. cinerea isolates were classified as 3 phenotypes, which were the first phenotype resistant to both of carbendazim and the mixture ($Car^RMix^R$), the second one resistant to carbendazim and sensitive to the mixture ($Car^RMix^S$), and the last one sensitive to both of them ($Car^RMix^S$). Carbendazim resistance correlated with a single mutation $\beta$-tubulin gene of B. cinerea amplified with primer pair tubkjhL and tubkjhR causing a change of glutamate to alanine at amino acid position 198. Furthermore, the substitution of valine for glutamate led the resistance to carbendazim and the mixture at the same position of amino acid. PCR-restriction fragment length polymorphism (PCR-RFLP) analysis using the restriction endonuclease, Tsp451 and BstUI allowed differentiation of the PCR fragment of $\beta$-tubulin gene of $Car^SMix^S$ isolates from that of $Car^RMix^R$ and $Car^RMix^S$ isolates. This method will aid in a fast detection of resistance of carbendazim and the mixture of carbendazim plus diethofencarb in B. cinerea in ginseng field.

Identification and Characterization of Eurotium rubrum Isolated from Mejuin Korea

  • Yun, Yeo-Hong;Hyun, Min-Woo;Suh, Dong-Yeon;Kim, Yong-Min;Kim, Seong-Hwan
    • Mycobiology
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    • 제37권4호
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    • pp.251-257
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    • 2009
  • We isolated and identified a strain of Eurotium rubrum from Meju that has not been reported in Korea. This fungus is yellowish brown; reverse dark brown on CYA and PDA while yellow on 2% MEA at $25{^{\circ}C}$. Cleistothecia are first bright yellow and gradually turned brown. Mycerial growth on CYA attained a diameter of 30 mm at $20{^{\circ}C}$, 37 mm at $25{^{\circ}C}$ and 32 mm at $30{^{\circ}C}$ after 15 days. The isolate grew slower on 2% MEA ($<$ 20 mm 15 days at $25{^{\circ}C}$) compared to CYA and PDA ($<$ 40 mm 15 days at $25{^{\circ}C}$). Cleistothecia are superficial, yellow to light brown, globose to subglobose, 40~75 ${\mu}m$ in diameter. Asci are 8-spored and globose to subglobose 8~11 ${\mu}m$. Ascospores are disciform, 4.0~5.0 ${\mu}m$ in length and 4.2~4.5 ${\mu}m$ in width. Conidia are ovate or bacillar, finely roughened to densely spinulose, 4.6~6.0 ${\mu}m$ in length and 3.0~4.3 ${\mu}m$ in width. Compared to known Eurotium rubrum, the Korean isolate showed 99% sequence similarity in ITS rDNA (554 bp) and calmodulin (750 bp) gene and 100% in $\beta$-tubulin (1016 bp) gene. The E. rubrum isolate also had weak $\beta$-glucosidase and protease activities.

Morphological and Genetic Characterization of Penicillium spp. associated with post - harvest decay of fruits. (oral)

  • Oh, S.Y.;Yu, S.H.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.115.1-115
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    • 2003
  • Post-harvest decay, caused by Penicillium spp. is a serious problem of fruits worldwide. Morphological characteristics and molecular markers were used to characterize 22 Penicillium isolates from apples, 18 isolates from pears, 60 from oranges and 18 from grapes and 23reference isolates representing related Penicillium spp. to assess their diversity and resolve their taxonomy. Based on morphological and physiological characteristics, the isolates were grouped as identical or very similar to P. digitatum, P. italicum, P. ulaiense or very similar to P. crustosum, P. expansum, P. solitum and unidentified Penicillium spp. Based on sequence comparisons of ITS region, variable site were presented within and among the species, but there variation were not correlated with the species. Cluster analyses of AP-PCR fragment patterns using UP and L45 primer and the -tubulin gene sequence, the Penicillium species were segregated into distinct groups. Particularly. the -tubulin partial sequence data provided support for species concepts based on morphological and physiological characteristics.

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Diagnosis of Benzimidazole Resistance in Haemonchus contortus of Sheep by Allele Specific PCR

  • Tiwari, J.;Kolte, A.P.;Kumar, S.;Swarnkar, C.P.;Singh, D.;Pathak, K.M.L.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권1호
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    • pp.7-11
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    • 2007
  • The study was conducted on 162 adult male Haemonchus contortus of sheep collected from Avikanagar, Jaipur and Bikaner regions to diagnose the benzimidazole (BZ) resistance in H. contortus. The BZ resistance is primarily linked with the mutation in ${\beta}$-tubulin isotype 1 gene which substitute phenylalanine (Phe) into tyrosine (Tyr) at the 200 codon of the gene. An allele specific polymerase chain reaction (AS-PCR) technique was used for diagnosis of BZ resistance in H. contortus. In AS-PCR, one reverse primer (TGG 312) was used in two separate reactions with each of 2 forward primers (resistant TGG 331 and susceptible CAW 106 primer) that differed only at 3' nucleotide position. Therefore, the amplified products from resistant and susceptible parasites were produced 267 and 266 bp, respectively. A total of 162 parasites were genotyped, of which 130 parasites found homozygous resistant 'rr', 22 heterozygous 'rS' and 10 homozygous susceptible 'SS' type. The prevalence of 'rr' individuals was higher in Jaipur (98%) followed by Avikanagar (93%) and Bikaner (50%) regions. Overall, the prevalence of BZ resistant allele (r) was higher (87%) as compared to 13% of BZ susceptible allele (S).

First Report of Sclerotinia White Rot Caused by Sclerotinia nivalis on Panax ginseng in Korea

  • Cho, Hye Sun;Shin, Jeong-Sup;Kim, Jae-Hyun;Hong, Tae-Kyun;Cho, Dae-Hui;Kang, Je Yong
    • 식물병연구
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    • 제19권1호
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    • pp.49-54
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    • 2013
  • Sclerotinia white rot disease was observed on 5 and 6-year-old ginseng (Panax ginseng) roots in Hongchun, Cheorwon, and Yanggu, Gangwon Province, Korea from 2006 to 2010. Symptoms included a brownish watery soft rot of the roots, and black sclerotia were often found on the rotten roots. The causal agent of the disease was identified as Sclerotinia nivalis based on cultural characteristics and sequence analyses of the internal transcribed spacer region of rDNA and ${\beta}$-tubulin gene with 100% sequence similarity. Pathogenicity tests were performed on 2-year-old ginseng roots with mycelium plugs without wounds. A watery soft rot of the roots and black sclerotia were observed 10 days after inoculation. These symptoms were identical to those observed on naturally infected roots. The same fungus was re-isolated from the lesions induced by artificial inoculation. This is the first report of sclerotinia white rot caused by S. nivalis on P. ginseng in Korea.

원목 및 톱밥배지 버섯 재배사 내 실내 공기서 분리한 미기록 진균 보고 (Unrecorded Fungal Species Isolated from Indoor Air in the Log Bed- and Sawdust Media-based Mushroom Cultivation Houses)

  • 안금란;김지은;김준영;김성환
    • 한국균학회지
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    • 제46권4호
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    • pp.495-503
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    • 2018
  • 2016년 4월부터 6월까지 표고 원목재배 임가 4곳과 표고 톱밥재배 임가 1곳의 재배사 내 실내 공기에서 진균을 포집 및 분리 동정하였다. 총 6개의 국내 미기록 진균(Cenangium acuum, Periconia macrospinosa, Neopestalotiopsissurinamensis, Metarhiziummarquandii, Trichoderma petersenii, Trichoderma paratroviride)을 분리하였으며 이들 균류에 대하여 형태학적 특성 및 internal transcribed spacer rDNA region, large subunitregion, ${\beta}-tubulin$ 또는 translation elongation factor $1{\alpha}$유전자 염기서열에 기반하여 계통학적 분석 결과를 기술하였다.