• Title/Summary/Keyword: ITS region of rDNA

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An Endophytic Nodulisporium sp. from Central America Producing Volatile Organic Compounds with Both Biological and Fuel Potential

  • Syed, Riyaz-Ul-Hassan;Strobel, Gary;Geary, Brad;Sears, Joe
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.29-35
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    • 2013
  • A Nodulisporium sp. (Hypoxylon sp.) has been isolated as an endophyte of Thelypteris angustifolia (Broadleaf Leaf Maiden Fern) in a rainforest region of Central America. It has been identified both on the basis of its morphological characteristics and by scanning electron microscopy as well as ITS sequence analysis. The endophyte produces volatile organic compounds (VOCs) that have both fuel (mycodiesel) and use for biological control of plant disease. When grown on potato dextrose agar, the organism uniquely produces a series of ketones, including acetone; 2-pentanone; 3-hexanone, 4-methyl; 3-hexanone, 2,4-dimethyl; 2-hexanone, 4-methyl, and 5-hepten, 2-one and these account for about 25% of the total VOCs. The most abundant identified VOC was 1,8 cineole, which is commonly detected in this group of organisms. Other prominent VOCs produced by this endophyte include 1-butanol, 2-methyl, and phenylethanol alcohol. Moreover, of interest was the presence of cyclohexane, propyl, which is a common ingredient of diesel fuel. Furthermore, the VOCs of this isolate of Nodulisporium sp. were selectively active against a number of plant pathogens, and upon a 24 h exposure caused death to Phytophthora palmivora, Rhizoctonia solani, and Sclerotinia sclerotiorum and 100% inhibition to Phytophthora cinnamomi with only slight to no inhibition of the other pathogens that were tested. From this work, it is becoming increasingly apparent that each isolate of this endophytic Nodulisporium spp., including the Daldina sp. and Hypoxylon spp. teleomorphs, seems to produce its own unique set of VOCs.

Pink Mold Rot on Unishiu Orange (Citrus unshiu Mac.) Caused by Trichothecium roseum (Pers.) Link ex Gray in Korea (Trichothecium roseum에 의한 감귤 분홍빛열매썩음병 발생)

  • Kwon, Jin-Hyeuk;Kang, Dong-Wan;Choi, Okhee;Shim, Hong-Sik
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.226-228
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    • 2013
  • In 2012, a pink mold rot was observed on unishiu orange (Citrus unshiu Mac.) fruits at the Wholesale Market for Agricultural Products, Jinju, Korea. The symptom on unishiu orange was a water-soaked lesion on the surface of fruit, which later on enlarged to form softened brown rot lesions. The diseased fruits were covered with pink-colored mold, consisting of conidia and conidiophores of the pathogen. Optimum temperature for mycelial growth was $25^{\circ}C$. Conidia were hyaline, smooth, 2-celled, and thick-walled conidia with truncate bases, ellipsoidal to pyriform, characteristically held together zig-zag chains and $12-26{\times}8-12{\mu}m$ in size. Conidiophore was erect, colorless, unbranched, and 4-5 ${\mu}m$ wide. On the basis of mycological characteristics, pathogenicity test, and molecular analysis with complete ITS rDNA region, the causal fungus was identified as Trichothecium roseum (Pers.) Link ex Gray. This is the first report of pink mold rot caused by T. roseum on unishiu orange in Korea.

OPTHiS Identifies the Molecular Basis of the Direct Interaction between CSL and SMRT Corepressor

  • Kim, Gwang Sik;Park, Hee-Sae;Lee, Young Chul
    • Molecules and Cells
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    • v.41 no.9
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    • pp.842-852
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    • 2018
  • Notch signaling is an evolutionarily conserved pathway and involves in the regulation of various cellular and developmental processes. Ligand binding releases the intracellular domain of Notch receptor (NICD), which interacts with DNA-bound CSL [CBF1/Su(H)/Lag-1] to activate transcription of target genes. In the absence of NICD binding, CSL down-regulates target gene expression through the recruitment of various corepressor proteins including SMRT/NCoR (silencing mediator of retinoid and thyroid receptors/nuclear receptor corepressor), SHARP (SMRT/HDAC1-associated repressor protein), and KyoT2. Structural and functional studies revealed the molecular basis of these interactions, in which NICD coactivator and corepressor proteins competitively bind to ${\beta}-trefoil$ domain (BTD) of CSL using a conserved ${\varphi}W{\varphi}P$ motif (${\varphi}$ denotes any hydrophobic residues). To date, there are conflicting ideas regarding the molecular mechanism of SMRT-mediated repression of CSL as to whether CSL-SMRT interaction is direct or indirect (via the bridge factor SHARP). To solve this issue, we mapped the CSL-binding region of SMRT and employed a 'one- plus two-hybrid system' to obtain CSL interaction-defective mutants for this region. We identified the CSL-interaction module of SMRT (CIMS; amino acid 1816-1846) as the molecular determinant of its direct interaction with CSL. Notably, CIMS contains a canonical ${\varphi}W{\varphi}P$ sequence (APIWRP, amino acids 1832-1837) and directly interacts with CSL-BTD in a mode similar to other BTD-binding corepressors. Finally, we showed that CSL-interaction motif, rather than SHARP-interaction motif, of SMRT is involved in transcriptional repression of NICD in a cell-based assay. These results strongly suggest that SMRT participates in CSL-mediated repression via direct binding to CSL.

MAGED4 Expression in Glioma and Upregulation in Glioma Cell Lines with 5-Aza-2'-Deoxycytidine Treatment

  • Zhang, Qing-Mei;Shen, Ning;Xie, Sha;Bi, Shui-Qing;Luo, Bin;Lin, Yong-Da;Fu, Jun;Zhou, Su-Fang;Luo, Guo-Rong;Xie, Xiao-Xun;Xiao, Shao-Wen
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.8
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    • pp.3495-3501
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    • 2014
  • Melanoma-associated antigen (MAGE) family genes have been considered as potentially promising targets for anticancer immunotherapy. MAGED4 was originally identified as a glioma-specific antigen. Current knowledge about MAGED4 expression in glioma is only based on mRNA analysis and MAGED4 protein expression has not been elucidated. In the present study, we investigated this point and found that MAGED4 mRNA and protein were absent or very lowly expressed in various normal tissues and glioma cell line SHG44, but overexpressed in glioma cell lines A172,U251,U87-MG as well as glioma tissues, with significant heterogeneity. Furthermore, MAGED4 protein expression was positively correlated with the glioma type and grade. We also found that the expression of MAGED4 inversely correlated with the overall methylation status of the MAGED4 promoter CpG island. Furthermore, when SHG44 and A172 with higher methylation were treated with the DNA demethylating agent 5-aza-2'-deoxycytidine (5-AZA-CdR) reactivation of MAGED4 mRNA was mediated by significant demethylation in SHG44 instead of A172. However, 5-AZA-CdR treatment had no effect on MAGED4 protein in both SHG44 and A172 cells. In conclusion, MAGED4 is frequently and highly expressed in glioma and is partly regulated by DNA methylation. The results suggest that MAGED4 might be a promising target for glioma immunotherapy combined with 5-AZA-CdR to enhance its expression and eliminate intratumor heterogeneity.

Molecular Cloning and Function Analysis of an Anthocyanidin Synthase Gene from Ginkgo biloba, and Its Expression in Abiotic Stress Responses

  • Xu, Feng;Cheng, Hua;Cai, Rong;Li, Lin Ling;Chang, Jie;Zhu, Jun;Zhang, Feng Xia;Chen, Liu Ji;Wang, Yan;Cheng, Shu Han;Cheng, Shui Yuan
    • Molecules and Cells
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    • v.26 no.6
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    • pp.536-547
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    • 2008
  • Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

Crystal Structure of the Regulatory Domain of MexT, a Transcriptional Activator of the MexEF-OprN Efflux Pump in Pseudomonas aeruginosa

  • Kim, Suhyeon;Kim, Songhee H.;Ahn, Jinsook;Jo, Inseong;Lee, Zee-Won;Choi, Sang Ho;Ha, Nam-Chul
    • Molecules and Cells
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    • v.42 no.12
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    • pp.850-857
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    • 2019
  • The Gram-negative opportunistic pathogen, Pseudomonas aeruginosa, has multiple multidrug efflux pumps. MexT, a LysR-type transcriptional regulator, functions as a transcriptional activator of the MexEF-OprN efflux system. MexT consists of an N-terminal DNA-binding domain and a C-terminal regulatory domain (RD). Little is known regarding MexT ligands and its mechanism of activation. We elucidated the crystal structure of the MexT RD at 2.0 Å resolution. The structure comprised two protomer chains in a dimeric arrangement. MexT possessed an arginine-rich region and a hydrophobic patch lined by a variable loop, both of which are putative ligand-binding sites. The three-dimensional structure of MexT provided clues to the interacting ligand structure. A DNase I footprinting assay of full-length MexT identified two MexT-binding sequence in the mexEF-oprN promoter. Our findings enhance the understanding of the regulation of MexT-dependent activation of efflux pumps.

Development of Detection Method of Unapproved Genetically Modified Potato (EH92-527-1) in Korea using Duplex Polymerase Chain Reaction (Duplex PCR을 이용한 국내 미승인 유전자변형 감자(EH92-527-1)의 검사법 개발)

  • Yoo, Myung-Ryul;Kim, Jae-Hwan;Yea, Mi-Chi;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.156-160
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    • 2013
  • A duplex polymerase chain reaction (PCR) method was developed to detect unapproved genetically modified (GM) potato (EH92-527-1) in Korea. The UDP-glucose pyrophosphorylase (UGP) gene was selected as an endogenous reference gene for potato and used to validate the specificity for 14 different crops. The primer pair EH92-F/R was designed to amplify the junction sequence between the genome and transgenic region introduced in GM potato. Its specificity was also validated using several different GM events. The detection limit of the duplex PCR method is approximately 0.05%. This duplex PCR method could be useful for monitoring cultivation of unauthorized GM potato in Korea.

Screening for the 3' UTR Polymorphism of the PXR Gene in South Indian Breast Cancer Patients and its Potential role in Pharmacogenomics

  • Revathidevi, Sundaramoorthy;Sudesh, Ravi;Vaishnavi, Varadharajan;Kaliyanasundaram, Muthukrishnan;MaryHelen, Kilyara George;Sukanya, Ganesan;Munirajan, Arasambattu Kannan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3971-3977
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    • 2016
  • Background: Breast cancer, the commonest cancer among women in the world, ranks top in India with an incidence rate of 1,45,000 new cases and mortality rate of 70,000 women every year. Chemotherapy outcome for breast cancer is hampered due to poor response and irreversible dose-dependent cardiotoxicity which is determined by genetic variations in drug metabolizing enzymes and transporters. Pregnane X receptor (PXR), a member of the nuclear receptor superfamily, induces expression of drug metabolizing enzymes (DMEs) and transporters leading to regulation of xenobiotic metabolism. Materials and Methods: A genomic region spanning PXR 3' UTR was amplified and sequenced using genomic DNA isolated from 96 South Indian breast cancer patients. Genetic variants observed in our study subjects were queried in miRSNP to establish SNPs that alter miRNA binding sites in PXR 3' UTR. In addition, enrichment analysis was carried out to understand the network of miRNAs and PXR in drug metabolism using DIANA miRpath and miRwalk pathway prediction tools. Results: In this study, we identified SNPs rs3732359, rs3732360, rs1054190, rs1054191 and rs6438550 in the PXR 3; UTR region. The SNPs rs3732360, rs1054190 and rs1054191 were located in the binding site of miR-500a-3p, miR-532-3p and miR-374a-3p resulting in the altered PXR level due to the deregulation of post-transcriptional control and this leads to poor treatment response and toxicity. Conclusions: Genetic variants identified in PXR 3' UTR and their effects on PXR levels through post-transcriptional regulation provide a genetic basis for interindividual variability in treatment response and toxicity associated with chemotherapy.

Laboratory and Field Evaluations of Entomopathogenic Lecanicillium attenuatum CNU-23 for Control of Green Peach Aphid (Myzus persicae)

  • Kim, Hyang-Yeon;Lee, Hyang-Burm;Kim, Young-Cheol;Kim, In-Seon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1915-1918
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    • 2008
  • An entomopathogenic fungus was isolated from an infected aphid. The isolate conformed most closely to Lecanicillium attenuatum CBS 402.78 (AJ292434) based on the internal transcribed spacer (ITS) region of its 18S rDNA, and thus was designated L. attenuatum CNU-23. Laboratory and field evaluations of CNU-23 blastospores were carried out for the control of green peach aphids. The laboratory evaluations of CNU-23 revealed an aphid mortality of about 80% with an estimated $LT_{50}$ of 3.72 days after the application of CNU-23 at $1{\times}10^6$ blastospores/ml. Meanwhile, the field evaluations of CNU-23 performed on greenhouse pepper plants during the rainy season showed an aphid mortality ranging from 72% to 97%. Significant sporulation was observed in the aphids treated with CNU-23. Therefore, the results suggest that L. attenuatum CNU-23 can be used as a biocontrol agent for green peach aphids on greenhouse pepper plants.

Twig Blight on Chinese Magnolia Vain Caused by Botryosphaeria dothidea in Korea (Botryosphaeria dothidea에 의한 오미자 줄기마름병)

  • Park, Sangkyu;Kim, Seung-Han;Lee, Seung-Yeol;Back, Chang-Gi;Kang, In-Kyu;Jung, Hee-Young
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.44-49
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    • 2016
  • The twig blight symptoms were observed in Chinese magnolia vine (Schisandra chinensis) at Mungyeong city, Gyeongbuk province, Korea in June 2015. The typical symptoms of infected plant were shriveled and wilted in leaves which led to blight resulted in death. Based on the morphological characteristics, the isolate was suspected as Botryosphaeria sp. Inoculation of isolated pathogen was performed to identify its pathogenicity according to Koch's postulates. Re-isolated fungi from the inoculated stem was showed same morphological characteristics with original pathogen. Phylogenetic analysis was performed using combined sequence of rDNA internal transcribed spacer region, EF1-${\alpha}$ and ${\beta}$-tubulin gene. The isolated pathogen was identified to the B. dothidea by phylogenetic analysis. This is the first report of twig blight on S. chinensis caused by B. dothidea in Korea.