• The Plant Pathology Journal
• /
• v.26 no.1
• /
• pp.49-56
• /
• 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.

• Food Science and Preservation
• /
• v.14 no.6
• /
• pp.688-694
• /
• 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.

• The Korean Journal of Mycology
• /
• v.47 no.4
• /
• pp.393-406
• /
• 2019

In August 2017, anthracnose symptoms were observed on the petioles and leaf veins of Korean radish (Rhaphanus sativus) in Hongcheon, Jeongseon, and Pyeongchang of the Gangwon province, Korea. Many grayish to dark-brown spots of 1-2 mm in diameter, appeared on the lower surface and leaf veins of the radish leaves. The spots gradually enlarged and coalesced to form dark-brown irregular lesions. Ten Colletotrichum isolates were obtained from the affected tissues of the Korean radish. Out of them, eight isolates were identified as C. higginsianum and two isolates were identified as C. truncatum based on morphological characteristics and multi-locus molecular phylogenetic analysis using the internal transcribed spacers and intervening 5.8S rDNA (ITS), partial beta-tubulin gene (TUB2), partial actin gene (ACT), and partial chitin synthase-1 gene (CHS1). The pathogenicity test was carried out on wounded and unwounded Korean radish (cv. Siraegimu and Osarimu), and Chinese cabbage (cv. Chuno and Smart) by inoculating with a spore suspension. All isolates except one C. truncatum isolate developed typical symptoms on both wounded and unwounded Korean radish. In Chinese cabbage, only the plants inoculated with C. higginsianum isolates developed symptoms regardless of the wound. This is the first report of anthracnose caused by C. truncatum on Korean radish in Korea.

• The Plant Pathology Journal
• /
• v.38 no.6
• /
• pp.629-636
• /
• 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
• /
• v.48 no.5
• /
• pp.390-399
• /
• 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.

• The Korean Journal of Mycology
• /
• v.50 no.3
• /
• pp.205-215
• /
• 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
• /
• v.3 no.2
• /
• pp.207-213
• /
• 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.

• The Korean Journal of Mycology
• /
• v.43 no.1
• /
• pp.58-63
• /
• 2015

Mold contamination is one of the detrimental factors affecting sawdust media-based shiitake cultivation in greenhouses. During mold monitoring of indoor air of greenhouses, several fungi were isolated. Among them, Aspergillus pulverulentus and Cosmospora butyri were found to be new in Korea and Lecanicillium psalliotae and L. antillanum were known in Korea without taxonomic validation. In this study the morphological characteristics and phylogenetic analysis based on the internal transcribed spacer (ITS) rDNA region or ${\beta}$-tubulin gene of the four identified species were described.

• The Korean Journal of Mycology
• /
• v.43 no.1
• /
• pp.71-76
• /
• 2015

Aquatic plant Hydrocharis dubia (Blume) Backer was collected from the Dalsung wetland in Daegu. Sixteen endophytic fungi with different colony morphologies were isolated from the roots of aquatic plants. Waito-c rice (WR) seedlings were treated with fungal culture filtrates (FCF) for screening plant growth-promoting activity. In the results, HD1008 strain isolated from aquatic plant showed highest plant growth-promoting activity. The FCF of HD1008 strain was analyzed using gas chromatography mass spectrometry (GC/MS) with selected ion monitoring (SIM). Analysis of the FCF of HD1008 strain found that it contained gibberellins (GA) ($GA_1$, 1.2 ng/100 mL; $GA_4$, 5 ng/100 mL). Phylogenetic tree of HD1008 strain was constructed by partial internal transcribed spacer (ITS) region and partial beta-tubulin gene sequences. Therefore, we describe HD1008 strain as a new gibberellin-producing Penicillium trzebinskii based on morphological and molecular characteristics.

• Journal of Applied Biological Chemistry
• /
• v.51 no.1
• /
• pp.1-6
• /
• 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.