• Title/Summary/Keyword: RNA polymerase II

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First Report of Pyrenophora tritici-repentis in Wheat (Triticum aestivum L.) in Korea and in vitro Selection of an Effective Fungicide

  • Min-Hye Jeong;Eu Ddeum Choi;Seol-Hwa Jang;Sang-Min Kim;Sook-Young Park
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.277-286
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    • 2023
  • Tan spot, caused by Pyrenophora tritici-repentis, is a major foliar disease in wheat worldwide. In April 2021, tan spot symptoms were observed in a commercial wheat field in Suncheon, Jeonnam Province, Korea, with over 5% of the wheat leaves exhibiting symptoms. These symptoms included oval-shaped tan necrosis surrounded by a bright halo. The three representative isolates exhibited irregular mycelial growth on V8-potato dextrose agar and produced pseudothecia. Based on the concatenated sequence datasets of four multi-genes, including the internal transcribed spacer, large subunit ribosomal RNA, glyceraldehyde-3-phosphate dehydrogenase, and RNA polymerase II second-largest subunit genes, phylogenetic analysis revealed that these three isolates clustered in the same clade as P. tritici-repentis. Results of pathogenicity test indicated that the initial symptoms appeared 5 days post-inoculation (dpi), with typical tan spot symptoms developing at 7 dpi. The pathogen was successfully re-isolated from the symptomatic tissues, thus fulfilling Koch's postulates. Furthermore, we selected three fungicides that effectively inhibited the mycelial growth of P. tritici-repentis by more than 90% in vitro. To the best of our knowledge, this is the first report of tan spot disease in wheat in Korea.

Generation of Transgenic Mice Overexpression Mouse RESISTIN

  • J. R. Chun;S. J. Song;J. T. Do;K. S. Chung;Lee, H. T.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.99-99
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    • 2002
  • The hormone resistin is associated with typeII diabetes mellitus in rodent model. Resistin impairs glucose tolerance and insulin action. A new class of anti-diabetic drugs were called thiazolidinediones (TZDs) downregulates a resistin which is induced during adipocyte differentiation. But the connection between increased adiposity and resistin remains unknown. The objectives of this study was to clone a mouse resistin cDNA and to generate transgenic mice overexpressing mouse resistin gene. The 555 bp of mouse resistin was amplified from mob cDNAS by polymerase chain reaction (PCR) and cloned into pCR$\^$(R)/ 2.1 TOPO T-vector. Mouse resistin mRNA on the basis of Genbank sequence (acession no. AF323080). Then, the PCR product was cloned into pTargeT$\^$TM/ mammalian expression vector that has pCMV promoter and chimeric intron. Restriction enzyme analysis with BamH I and Not I was carried out to determine an orientation of the insert in the vector. The pCMV-mus/resistin gene was prepared from previous recombinant pTargeT$\^$TM/-mus/resistin by digestion of Bgl II, and has used for microinjection into pronuclei of one cell embryos. The microinjected embryos were transfered to pseudopregnant foster-mother. Mouse resistin expression was detected in transgenic F1 mice by Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR). Resistin gene expression mouse has heavier body weight which was measured higher level of plasma glucose than that of normal mouse. And in diet-induced experiments, the abdominal fat pads were isolated from each 24h starvation and re-feeding after fasting group mice that were assessed by RT-PCR analysis. In fasting group mice, resistin expression was higher than that of re-feeding group mice. This result suggests that the resistin gene overexpressing mice may be became to obesity and be useful as an animal disease model to be diabetes mellitus caused by insulin resistance of resistin.

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Melittin induces autophagy to alleviate chronic renal failure in 5/6-nephrectomized rats and angiotensin II-induced damage in podocytes

  • Yufan Zhang;Huaping Xu;Hongwei Qiao;Ya Zhao;Minmin Jiang
    • Nutrition Research and Practice
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    • v.18 no.2
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    • pp.210-222
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    • 2024
  • BACKGROUND/OBJECTIVES: Chronic renal failure (CRF) is a complex pathological condition that lacks a cure. Certain Chinese medicines, such as melittin, a major component in bee venom, have shown efficacy in treating CRF patients. On the other hand, the mechanisms underlying the therapeutic effects of melittin are unclear. MATERIALS/METHODS: A 5/6 nephrectomy model (5/6 Nx) of renal failure was established on rats for in vivo assays, and mouse podocyte clone 5 (MPC5) mouse podocyte cells were treated with angiotensin II (AngII) to establish an in vitro podocyte damage model. The 24-h urine protein, serum creatinine, and blood urea nitrogen levels were evaluated after one, 2, and 4 weeks. Hematoxylin and eosin staining, Masson staining, and periodic acid-Schiff staining were used to examine the pathological changes in kidney tissues. A cell counting kit 8 assay was used to assess the cell viability. Reverse transcription polymerase chain reaction and Western blot were used to assess the mRNA and protein levels in the cells, respectively. RESULTS: In the rat 5/6 Nx, melittin reduced the 24-h urinary protein excretion and the serum creatinine and blood urea nitrogen levels. Furthermore, the renal pathology was improved in the melittin-treated 5/6 Nx rats. Melittin promoted podocin, nephrin, Beclin 1, and the LC3II/LC3I ratio and inhibited phosphorylated mammalian target of rapamycin (mTOR)/mTOR in 5/6 Nx-induced rats and AngII-induced MPC5 mouse podocyte cells. Moreover, inhibiting autophagy with 3-MA weakened the effects of melittin on podocin, nephrin, and the LC3II/LC3I ratio in podocytes. CONCLUSION: Melittin may offer protection against kidney injury, probably by regulating podocyte autophagy. These results provide the theoretical basis for applying melittin in CRF therapy.

A systematic study of nuclear interactome of C-terminal domain small phosphatase-like 2 using inducible expression system and shotgun proteomics

  • Kang, NaNa;Koo, JaeHyung;Wang, Sen;Hur, Sun Jin;Bahk, Young Yil
    • BMB Reports
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    • v.49 no.6
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    • pp.319-324
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    • 2016
  • RNA polymerase II C-terminal domain phosphatases are newly emerging family of phosphatases that contain FCPH domain with Mg+2-binding DXDX(T/V) signature motif. Its subfamily includes small CTD phosphatases (SCPs). Recently, we identified several interacting partners of human SCP1 with appearance of dephosphorylation and O-GlcNAcylation. In this study, using an established cell line with inducible CTDSPL2 protein (a member of the new phosphatase family), proteomic screening was conducted to identify binding partners of CTDSPL2 in nuclear extract through immunoprecipitation of CTDSPL2 with its associated. This approach led to the identification of several interacting partners of CTDSPL2. This will provide a better understanding on CTDSPL2.

DNA Profiling of Leuconostoc citreum Strains in Fermented Foods by Repetitive Element Polymerase Chain Reaction

  • Kaur, Jasmine;Sharma, Anshul;Lee, Sulhee;Park, Young-Seo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1778-1782
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    • 2017
  • To identify and discriminate the bacterial species at the subspecific level, rep-PCR is a reliable genomic fingerprinting tool. Fourteen strains of bacteria were isolated from different food sources, identified as Leuconostoc citreum using 16S rRNA gene sequencing, and amplified using rep-primers (REP, ERIC, and $(GTG)_5$). Fingerprinting patterns generated bands in the range of 300-6,000 bp with REP, 150-6,000 bp with ERIC, and 200-1,700 bp with $(GTG)_5$ primers. In UPGMA dendrogram analysis, 14 strains were clustered into three clades (I, II, and III) with all the primers, thus differentiating them at the molecular level. The present study revealed the differentiation of L. citreum strains using rep-PCR.

Amino Acid Sequence Homology of Hybrid Poplar O-methyltransferuse Involved in Lignin Biosynthesis

  • Park, Young-Goo;Sul, Ill-Whan;Shin, Dong-Ill;Park, Jang-Won;Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.3 no.3
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    • pp.131-134
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    • 2001
  • In $\lambda$-Zap II vector system, a cDNA library was constructed for the developing secondary xylem mRNA from hybrid poplar, Populus nigra x maximowiczii. A cDNA clone of 1.5 kb in size, pOMTB1.4 encoding a lignin-bispecific O-methyltransferase was screened by plaque hybridization using a probe of 540 bp cDNA amplified by polymerase chain reaction from the cDNA library and identified by nucleotide sequencing. Its nucleotide sequence contains one open reading frame of 366 amino acids. The deduced amino acid sequence in comparison with that of Populus tremuloides showed the differences of 9 amino acids and revealed 85-99% homology among alfalfa, poplar and aspen.

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First Report of Leaf Spot of Datura metel Caused by Alternaria tenuissima in Korea

  • Aktaruzzaman, Md.;Kim, Joon-Young;Afroz, Tania;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.330-333
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    • 2015
  • In June 2013, we collected leaf spot disease samples of Datura metel from Gangneung, Gangwon Province, Korea. The symptoms observed were small circular to oval dark brown spots with irregular in shape or remained circular with concentric rings. We isolated the pathogen from infected leaves and cultured the fungus on potato dextrose agar. We examined the fungus morphologically and confirmed its pathogenicity according to Koch's postulates. The results of morphological examinations, pathogenicity tests, and the rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4), glycerol-3-phosphate dehydrogenase (G3PDH) and the RNA polymerase II second largest subunit (RPB2) gene sequence revealed that the causal agent was Alternaria tenuissima. To the best of our knowledge, this is the first report of leaf spot of D. metel caused by A. tenuissima in Korea as well as worldwide.

Characterization and Pathogenicity of Alternaria vanuatuensis, a New Record from Allium Plants in Korea and China

  • Li, Mei Jia;Deng, Jian Xin;Paul, Narayan Chandra;Lee, Hyang Burm;Yu, Seung Hun
    • Mycobiology
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    • v.42 no.4
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    • pp.412-415
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    • 2014
  • Alternaria from different Allium plants was characterized by multilocus sequence analysis. Based on sequences of the ${\beta}$-tubulin (BT2b), the Alternaria allergen a1 (Alt a1), and the RNA polymerase II second largest subunit (RPB2) genes and phylogenetic data analysis, isolates were divided into two groups. The two groups were identical to representative isolates of A. porri (EGS48-147) and A. vanuatuensis (EGS45-018). The conidial characteristics and pathogenicity of A. vanuatuensis also well supported the molecular characteristics. This is the first record of A. vanuatuensis E. G. Simmons & C. F. Hill from Korea and China.

Talaromyces halophytorum sp. nov. Isolated from Roots of Limonium tetragonum in Korea

  • You, Young-Hyun;Aktaruzzaman, Md.;Heo, Inbeom;Park, Jong Myong;Hong, Ji Won;Hong, Seung-Beom
    • Mycobiology
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    • v.48 no.2
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    • pp.133-138
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    • 2020
  • Talaromyces halophytorum sp. nov. was isolated from the roots of halophyte Limonium tetragonum collected from Seocheon-gun, Korea in November 2015. It showed a slow growth on yeast extract sucrose agar at 25 ℃, no growth at 4 ℃ or 37 ℃ and produced smooth-walled and globose to sub-globose conidia. T. halophytorum is phylogenetically distinct from the other reported Talaromyces species of section Trachyspermi based on multi-locus sequence typing results using partial fragments of β-tubulin, calmodulin, ITS, and RNA polymerase II genes.

Characterization of Paecilomyces variotii and Talaromyces amestolkiae in Korea Based on the Morphological Characteristics and Multigene Phylogenetic Analyses

  • Nguyen, Thi Thuong Thuong;Paul, Narayan Chandra;Lee, Hyang Burm
    • Mycobiology
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    • v.44 no.4
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    • pp.248-259
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    • 2016
  • During fungal diversity surveys of the order Eurotiales in Korea, two fungal strains, EML-DG33-1 and EML-NCP50, were isolated from samples of rat dung and fig tree leaf collected at a garden located in Gwangju in 2014. To complete the National Species List of Korea, it is a prerequisite to verify whether many questionable species, which were previously recorded but not confirmed, indeed present in Korea. Herein, the isolates were confirmed as undescribed species, Paecilomyces variotii and Talaromyces amestolkiae based on the combination of morphological and phylogenetic analyses of multigenes including the rDNA internal transcribed spacer, ${\beta}-tubulin$, and RNA polymerase II subunit 2.