• The Plant Pathology Journal
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• 제26권1호
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• pp.49-56
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• 2010

Thirty-six isolates of Colletotrichum spp. were obtained from infected grapes in two different locations of Korea; 18 isolates from Cheonahn, where carbendazim (MBC) and the mixture of MBC and diethofencarb (NPC) had been applied to control grape ripe rot, and 18 isolates from Cheongju, where no fungicides had been used. Sequences analysis of the internal transcribed spacer (ITS) and the $\beta$-tubulin gene identified 34 of the 36 isolates as Colletotrichum gloeosporioides. The remaining two isolates from Cheongju were identified as C. acutatum. Of the 18 isolates from Cheonahn, 12 were resistant to both MBC and the mixture (MBC+NPC), and six were sensitive to them. All C. gloeosporioides isolates from Cheongju, but not the two C. acutatum isolates, were sensitive to these fungicides. Sequence analysis of the $\beta$-tubulin gene in all isolates revealed that C. gloeosporioides resistant to MBC and MBC+NPC had a tyrosine instead of phenylalanine at the amino acid position 200. The appearance of resistance to MBC and the mixture in C. gloeosporioides correlated with the history of fungicide application in Korea.

• 한국식품저장유통학회지
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• 제14권6호
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• pp.688-694
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• 2007

To evaluate the human risk of long-term intake of genetically modified (GM) rice, we carried out RT-PCR of housekeeping genes. Housekeeping genes, which show highly uniform expression in living organisms during various stages of development and under different environmental conditions, were normalized by RT-PCR. We assessed the expression of 10 common housekeeping genes (18s rRNA, 25S rRNA, UBC, UBQ5, UBQ10, ACT11, GAPDH, eEF-$1{\alpha}$, ${\beta}$-TUB, GAPDH, ${\beta}$-actin, B2m, G6pd2, Gyk, Gus, Hprt, Cyclophlin A, Tfrc, ${\alpha}$-tubulin and RPL13A) in the liver, stomach, small intestine, large intestine, kidney and spleen of mice fed GM or non-GM rice. We found no significant differences in the expression of housekeeping genes between the two groups of mice.

• 한국균학회지
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• 제47권4호
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• pp.393-406
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• 2019

2017년 8월경, 강원도 홍천군, 정선군, 평창군 소재 무 재배 포장에서 잎자루와 잎의 주맥에 형성된 탄저병 병징이 관찰되었다. 회색 내지 짙은 갈색을 띤 1-2 mm 크기의 점무늬가 아래 잎과 잎의 주맥에 형성되었다. 이러한 점무늬는 점차 확대되고 합쳐지면서 갈색을 띤 불규칙한 병반으로 나타났다. 병든 무 잎에서 10개의 Colletotrichum균을 분리하였다. 분리 균주의 형태적 특성과 internal transcribed spacers and intervening 5.8S rDNA (ITS), partial beta-tubulin gene (TUB2), partial actin gene (ACT) 및 partial chitin synthase-1 gene (CHS-1) 부위를 사용한 다자위 분자 계통 분석(multilocus molecular phylogenetic analysis)에 의해 8개 균주는 Colletotrichum higginsianum, 2개 균주는C. truncatum으로 동정되었다. 무(시래기무, 오사리무)와 배추(품종: 추노, 스마트)를 대상으로 상처와 무상처로 구분하여 포자현탁액을 떨어뜨려서 접종하는 방법을 사용하여 병원성 검정을 수행하였다. 무에서는C. truncatum 1개 균주를 제외하고 상처와 무상처 접종에서 모두 병징을 나타냈고, 배추의 경우는C. higginsianum 접종구에서만 상처의 유무에 관계없이 병이 발생하였다. 국내에는 기존에 C. higginsianum은 무와 배추의 탄저병균으로 보고되었으며, C. truncatum에 의한 무 탄저병의 발생은 본 연구에서 국내 최초로 보고한다.

• The Plant Pathology Journal
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• 제38권6호
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• pp.629-636
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• 2022

Colletotrichum species is known as the major causal pathogen of red pepper anthracnose in Korea and various groups of fungicides are registered for the management of the disease. However, the consistent use of fungicides has resulted in the development of resistance in many red pepper-growing areas of Korea. Effective management of the occurrence of fungicide resistance depends on constant monitoring and early detection. Thus, in this study, various methods such as agar dilution method (ADM), gene sequencing, allele-specific polymerase chain reaction (PCR), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were applied for the detection of benzimidazole resistance among 24 isolates of Colletotrichum acutatum s. lat. and Colletotrichum gloeosporioides s. lat. The result of the ADM showed that C. gloeosporioides s. lat. was classified into sensitive and resistant isolates to benomyl while C. acutatum s. lat. was insensitive at ≥1 ㎍/ml of benomyl. The sequence analysis of the β-tubulin gene showed the presence of a single nucleotide mutation at the 198th amino acid position of five isolates (16CACY14, 16CAYY19, 15HN5, 15KJ1, and 16CAYY7) of C. gloeosporioides s. lat. Allele-specific PCR and PCR-RFLP were used to detect point mutation at 198th amino acid position and this was done within a day unlike ADM which usually takes more than one week and thus saving time and resources that are essential in the fungicide resistance management in the field. Therefore, the molecular techniques established in this study can warrant early detection of benzimidazole fungicide resistance for the adoption of management strategies that can prevent yield losses among farmers.

• Entomological Research
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• 제48권5호
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• pp.390-399
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• 2018

In order to precisely assess gene expression levels, the suitable internal reference genes must be served to quantify real-time reverse transcription polymerase chain reaction (RT-qPCR) data. For armyworm, Mythimna separata, which reference genes are suitable for assessing the level of transcriptional expression of target genes have yet to be explored. In this study, eight common reference genes, including ${\beta}$-actin (${\beta}$-ACT), 18 s ribosomal (18S), 28S ribosomal (28S), glyceraldehyde-3-phosphate (GAPDH), elongation fator-alpha ($EF1{\alpha}$), TATA box binding protein (TBP), ribosomal protein L7 (RPL7), and alpha-tubulin (${\alpha}$-TUB) that in different developmental stages, tissues and insecticide treatments of M. separata were evaluated. To further explore whether these genes were suitable to serve as endogenous controls, three software-based approaches (geNorm, BestKeeper, and NormFinder), the delta Ct method, and one web-based comprehensive tool (RefFinder) were employed to analyze and rank the tested genes. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated according to normalized HSP70, and MsepCYP321A10 gene expression data. We found that the most suitable reference genes for the different experimental conditions. For developmental stages, 28S/RPL7 were the optimal reference genes, both $RPL7/EF1{\alpha}$ were suitable for experiments of different tissues, whereas for insecticide treatments, $28S/{\alpha}-TUB$ were suitable for normalizations of expression data. In addition, $28S/{\alpha}-TUB$ were the suitable reference genes because they have the most stable expression among different developmental stages, tissues and insecticide treatments. Our work is the first report on reference gene selection in M. separata, and might serve as a precedent for future gene expression studies.

• 한국균학회지
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• 제50권3호
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• pp.205-215
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• 2022

Penicillium pimiteouiense was discovered in South Korea during an investigation of fungal communities in soil collected from the Gyeongsangbuk-do province. In this study, we performed molecular analysis of this fungal isolate using internal transcribed spacer rDNA, β-tubulin, and Calmodulin gene sequences. We also performed morphological analysis using five agar media, potato dextrose, oatmeal, malt extract, czapek yeast extract, and yeast extract sucrose. In this study, the molecular and morphological analyses of P. pimiteouiense with detailed descriptions and figures has been carried out.

• Animal cells and systems
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• 제3권2호
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• pp.207-213
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• 1999

During the differentiation of Naegleria gruberi amoebas into flagellates, the amoebas undergo sequential changes in cell shape and form new cellular organelles. To understand the nature of the signal which initiates this differentiation and the signal transduction pathway, we treated cells with four agents, PMA, retinoic acid (RA), okadaic acid, and cAMP. Retinoic acid and cAMP had specific effects on the differentiation of N. gruberi depending on the time of the drug treatment. Addition of (100$\mu$M) retinoic acid at the initiation of differentiation inhibited differentiation by blockinq the transcription of differentiation specific genes (e.g., $\beta$-tubulin). This inhibition of differentiation by retinoic acid was overcome by co-treatment with cAMP (or dbcAMP, 20 $\mu$M). Addition of retinoic acid at later stages (30 and 70 min) had no effect on the transcriptional regulation of the $\beta$-tubulin gene, however the differentiation was inhibited by different degrees. Co-treatment of cAMP at these stages did not overcome the inhibitory effect of retinoic acid. These results suggest that the role of retinoic acid as a transcriptional regulator might be conserved throughout the evolution of eukaryotes.

• 한국균학회지
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• 제43권1호
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• pp.58-63
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• 2015

진균에 의한 오염은 재배사에서 톱밥 배지를 이용한 표고 재배에 중대한 영향을 미치기 때문에 표고 재배사 실내 공기에 존재하는 진균에 대한 모니터링을 수행하던 중 여러 진균을 분리하였다. 분리된 진균 중에는 국내 미기록인 Aspergillus pulverulentus와 Cosmospora butyri 등 2종을 비롯하여 기록은 있으나 균학적 확증이 부족한 Lecanicillium psalliotae와 L. antillanum 등 2종이 존재하였다. 본 논문에서는 이들 균류에 대한 형태적 특성과 더불어 internal transcribed spacer (ITS) rDNA region 또는 ${\beta}$-tubulin 유전자 염기서열에 기반한 계통학적 분석 결과를 기술하였다.

• 한국균학회지
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• 제43권1호
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• pp.71-76
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• 2015

수생식물 샘플은 낙동강과 금호강이 만나는 달성습지에서 자라풀을 채집하였다. 자라풀의 뿌리에서 내생균류를 분리하고 형태가 다른 균주를 관찰하여 최종적으로 16개 균주를 선발하였다. 내생균류의 배양여과액은 식물생장촉진활성 검정을 위하여 난장이벼에 처리하여 스크리닝하였으며, HD1008 균주가 식물생장촉진활성이 가장 높은 것으로 확인되었다. HD1008 균주의 배양여과액을 HPLC와 GC/MS-SIM을 이용하여 분석하였고, HD1008 균주가 식물호르몬인 지베렐린 $GA_1$ (1.2 ng/100 mL)과 $GA_4$ (5 ng/100 mL) 를 생산하는 것을 정량분석을 통하여 확인하였다. 또한, HD1008균주의 beta-tubulin 유전자 염기서열을 이용하여 동정에 이용하였으며, 분자적인 방법과 형태적인 방법으로 관찰하였을 때, 지베렐린을 생산하는 새로운 P. trzebinskii로 동정되었다.

• Journal of Applied Biological Chemistry
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• 제51권1호
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• pp.1-6
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• 2008

Estrogens, a group of steroid compounds functioning as the primary female sex hormone, play an important role in the development and progression of breast cancer. In this study, using a novel annealing control primer-based GeneFishing PCR technology, five differentially expressed genes (DEGs), expressed using 10nM mitogenic estrogens, $17{\beta}$-estradiol (E2) and $16{\alpha}$-hydroxyestrone ($16{\alpha}$-OHE1), were selected from the estrogen receptor (ER)-positive MCF-7 human breast cancer cells. The DEGs, MRPL42, TUBA1B, SSBP1, KNCT2, and RUVBL1, were identified by comparison with the known genes via direct sequencing and sequence homology search in BLAST. Quantitative real-time PCR data showed that two DEGs, tubulin ${\alpha}1b$ and kinetochore associated 2, were greater than 2-fold upregulated by E2 or $16{\alpha}$-OHE1. Both genes could be new biomarkers for the treatment and prognosis of cancers, and further study may provide insights into the molecular mechanisms underlying development and progression of breast cancer.