DOCSLIB.ORG

Candidate Gene Identification from the Wheat QTL-2DL for Resistance Against Fusarium Head Blight Based on Metabolo-Genomics Approach

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Candidate Gene Identification from the Wheat QTL-2DL for Resistance Against Fusarium Head Blight Based on Metabolo-Genomics Approach Candidate gene identification from the wheat QTL-2DL for resistance against Fusarium head blight based on metabolo-genomics approach Udaykumar Kage Department of Plant Science McGill University, Montreal, Canada August 2016 A thesis submitted to the McGill University in partial fulfillment of the requirements of the degree of Doctor of Philosophy ©Udaykumar Kage (2016) i Dedicated to my beloved parents ii TABLE OF CONTENTS TABLE OF CONTENTS………………………………………………………………………...iii LIST OF TABLES……………………………………………………………………………...viii LIST OF FIGURES………………………………………………………………………………ix LIST OF APPENDICES…………………………………………………………………………xv LIST OF ABBREVIATIONS…………………………………………………………………...xvi ABSTRACT…………………………………………………………………………………...xviii ACKNOWLEDGEMENT……………………………………………………………………...xxi PREFACE AND CONTRIBUTION OF THE AUTHORS……………………………………..01 PREFACE……………………………………………………………………………………......01 CONTRIBUTION OF THE AUTHORS………………………………………………………...02 CHAPTER I: GENERAL INTRODUCTION…………………………………………………...03 GENERAL HYPOTHESIS……………………………………………………………………...07 GENERAL OBJECTIVES……………………………………………………………………....07 CHAPTER II: REVIEW OF THE LITERATURE……………………………………………...08 2.1 Fusarium head blight of wheat at a glance…………………………………………………...08 2.1.1 Fusarium head blight epidemiology and impact on wheat production…………………08 2.1.2 FHB management practices ……………………………………………………….……10 2.1.3 FHB management through breeding for disease resistance…………………………….11 2.1.3.1 Types of Fusarium head blight resistance………………………………………….12 2.1.3.2 Molecular markers and QTL for FHB resistance……………………………….…13 2.2 Functional genomics to identify candidate genes for FHB resistance…………………….....14 2.2.1 Metabolomics and analytical platforms for metabolite profiling……………………….14 2.2.2 Putative metabolite identification and databases………………………………………..16 2.2.3 Databases and bioinformatics tools to identify candidate genes……………………..…16 2.2.4 Application of metabolomics…………………………………………………………...17 2.2.5 Functional characterization of candidate genes………………………………………....18 CONNECTING STATEMENT FOR CHAPTER III……………………………………………20 CHAPETER III…………………………………………………………………………………..21 iii Identification and characterization of a Fusarium head blight resistance gene TaACT in wheat QTL-2DL……………………………………………………………………………………...21 3.1 Abstract………………………………………………………………………………………21 3.2 Introduction…………………………………………………………………………………..22 3.3 Material and Methods………………………………………………………………………..23 3.3.1 Plant material and experimental layout…………………………………………………23 3.3.2 Pathogen production and inoculation…………………………………………………...24 3.3.3 Sample collection, metabolite extraction and metabolite analysis using liquid chromatography-high resolution mass spectrometry (LC-HRMS)…………………………...24 3.3.4 Data processing using MZmine software and statistical analysis………………………24 3.3.5 Putative metabolite identification……………………………………………………….25 3.3.6 Disease severity and quantification of fungal biomass…………………………………25 3.3.7 Physical localization of QTL-2DL and identification of TaACT gene…………………26 3.3.8 Cloning, sequencing and sequence analysis of TaACT gene…………………………...26 3.3.9 RNA isolation and candidate gene expression based on semi-qPCR and qPCR……….27 3.3.10 Histochemical localization of HCAAs……………………………….………………..27 3.3.11 Expression and purification of recombinant protein in E. coli………………………...28 3.3.12 Virus-induced gene silencing of TaACT……………………...……………………….28 3.3.13 TaACT functional complementation study in Arabidopsis…………………………....29 3.4 Results………………………………………………………………………………………..30 3.4.1 Disease severity in NILs………………………………………………………………...30 3.4.2 Metabolite profiling in NILs……………………………………………………………31 3.4.3 Histochemical localization of HCAAs……………………………………………….…31 3.4.4 Identification of candidate gene TaACT in the QTL-2DL……………………………...31 3.4.5 Expression of gene TaACT based on semi-qPCR and qPCR…………………………...32 3.4.6 Functional characterization of TaACT using VIGS…………………………………….33 3.4.7 Functional characterization of TaACT based on complementation study in Arabidopsis……………………………………………………………………………………33 3.5 Discussion……………………………………………………………………………………33 3.5.1 FHB resistance in NIL-R is due to high fold induction of HCAAs…………………….34 3.5.2 TaACT induced high levels of coumaroylagmatine and coumaroylputrescine………....35 iv 3.5.3 Functional validation of TaACT………………………………………………………...35 CONNECTING STATEMENT FOR CHAPTER IV……………………………………………51 CHAPTER IV……………………………………………………………..…………………..…52 TaWRKY70 transcription factor in wheat QTL-2DL regulates downstream genes to biosynthesize metabolites to contain Fusarium graminearum spread………………………………………….52 4.1 Abstract………………………………………………………………………………………52 4.2 Introduction…………………………………………………………………………………..53 4.3 Materials and methods……………………………………………………………………….55 4.3.1 Plant production and experimental design……………………………………………...55 4.3.2 Pathogen production and inoculation…………………………………………………...56 4.3.3 Sample collection, metabolite analysis using liquid chromatography-high resolution mass spectrometry (LC-HRMS) and data processing………………………………………...56 4.3.4 Disease severity and fungal biomass assessment……………………………………….57 4.3.5 Candidate gene identification, based on high fold-change RR metabolites and their physical localization within QTL-2DL……………………………………………………….57 4.3.6 Gene cloning, sequencing and sequence analysis……………………………………....58 4.3.7 RNA isolation and gene expression based on qPCR……………………………………58 4.3.8 Nuclear localization assay………………………………………………………………59 4.3.9 Luciferase (LUC) transient expression assay…………………………………………...59 4.3.10 Construction of BSMV vectors and virus-induced gene silencing of TaWRKY70……59 4.3.11 Confirmation of gene silencing by qPCR, and estimation of fungal biomass and targeted metabolites…………………………………………………………………………...60 4.4 Results………………………………………………………………………………………..60 4.4.1 Fungal biomass quantification…………………………………………………………..60 4.4.2 Metabolite profiles of NILs……………………………………………………………..61 4.4.3 Identification of candidate genes in QTL-2DL…………………………………………61 4.4.4 TaWRKY gene sequencing and sequence analysis……………………………………...62 4.4.5 Sequence variation of TaWRKY70 between NILs and differential gene expression during Fg infection…………………………………………………………………………………...62 4.4.6 Gene expression, promoter analysis of RR metabolite biosynthetic genes and their physical interaction with RTaWRKY……………………………………………………………..63 v 4.4.7 Nuclear localization TaWRKY70 protein………………………………………………64 4.4.8 Response to Fg infection after knocking down of TaWRKY70 in wheat…………….…64 4.4.9 Silencing of TaWRKY70 affected transcriptional response of RRRM genes and RR metabolite accumulation………………………………………………………………………65 4.5 Discussion……………………………………………………………………………………65 CONNECTING STATEMENT FOR CHAPTER V…………………………………………….85 CHAPETER V…………………………………………………………………………………...86 Liquid chromatography and high resolution mass spectrometry‐based metabolomics to identify quantitative resistance‐related metabolites and genes, in spikelets of wheat NILs with QTL-2DL, against Fusarium head blight……………………………………………………..86 5.1 Abstract………………………………………………………………………………………86 5.2 Introduction…………………………………………………………………………………..87 5.3 Material and Methods………………………………………………………………………..89 5.3.1 Plant material and production…………………………………………………………..89 5.3.2 Pathogen production and inoculation…………………………………………………...89 5.3.3 Experiment layout, sample collection, metabolite analysis, and data processing………89 5.3.4 Putative metabolite identification……………………………………………………….90 5.3.5 Relative quantification of fungal biomass……………………………………………....91 5.3.6 Identification of candidate genes in FHB QTL-2DL……………………………...........91 5.3.7 RNA isolation, cDNA synthesis and qPCR analysis……………………………….…..91 5.4 Results………………………………………………………………………………………..92 5.4.1 Fungal biomass varied between NILs………………………………………………..…92 5.4.2 Clustering of observations based on multivariate analysis……………………………..92 5.4.3 Resistance related metabolites associated with spikelets with varying alleles at QTL- 2DL……………………………………………………………………………………………92 5.4.4 Identification of candidate genes in QTL-2DL…………………………………………93 5.4.5 Gene expression based on qPCR……………………………………………………….94 5.5 Discussion……………………………………………………………………………………94 5.5.1 Toxic phytochemicals slow down pathogen progress in plant…………………………95 5.5.2 Cell wall fortification…………………………………………………………………...95 5.5.3 Proposed model for role of QTL-2DL in spikelet resistance to FHB…………………..97 vi CHAPTER VI…………………………………………………………………………………..109 GENERAL DISCUSSION AND FUTURE STUDIES………………………………………...109 6.1 General discussion……………………………………………………………………….109 6.2. Future studies…………………………………………………………………………...112 REFERENCES…………………………………………………………………………………113 APPENDIX……………………………………………………………………………………..136 vii LIST OF TABLES Table 3.1: Fold change in abundance of resistance related (RR) metabolites detected in wheat rachis following F. graminearum inoculation. Table 3.2: List of primers used in the experiments. Table 4.1: List of high fold-change resistance related induced (RRI) metabolites identified in NILs with contrasting FHB resistance alleles at QTL-2DL. Table 4.2: Promoter analysis of RRRM genes regulated by TaWRKY70. Table 4.3: List of primers used in this study. Table 4.4: Predicted genes present in the QTL-2DL based on in-silico gene prediction and synteny with Brachypodium and Rice. Table 5.1: Resistance related (RR) metabolites identified in spikelets of NILs, harboring contrasting alleles of QTL-2DL, upon F. graminearum or mock inoculation. Table 5.2: List of primers used in the study for gene expression viii LIST OF FIGURES Figure 3.1: F. graminearum
Load more

Recommended publications
  • Astragalin: a Bioactive Phytochemical with Potential Therapeutic Activities
    Hindawi Advances in Pharmacological Sciences Volume 2018, Article ID 9794625, 15 pages https://doi.org/10.1155/2018/9794625 Review Article Astragalin: A Bioactive Phytochemical with Potential Therapeutic Activities Ammara Riaz,1 Azhar Rasul ,1 Ghulam Hussain,2 Muhammad Kashif Zahoor ,1 Farhat Jabeen,1 Zinayyera Subhani,3 Tahira Younis,1 Muhammad Ali,1 Iqra Sarfraz,1 and Zeliha Selamoglu4 1Department of Zoology, Faculty of Life Sciences, Government College University, Faisalabad 38000, Pakistan 2Department of Physiology, Faculty of Life Sciences, Government College University, Faisalabad 38000, Pakistan 3Department of Biochemistry, University of Agriculture, Faisalabad 38000, Pakistan 4Department of Medical Biology, Faculty of Medicine, Nigde O¨ mer Halisdemir University, Nigde 51240, Turkey Correspondence should be addressed to Azhar Rasul; [email protected] Received 8 January 2018; Revised 5 April 2018; Accepted 12 April 2018; Published 2 May 2018 Academic Editor: Paola Patrignani Copyright © 2018 Ammara Riaz et al. %is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Natural products, an infinite treasure of bioactive chemical entities, persist as an inexhaustible resource for discovery of drugs. %is review article intends to emphasize on one of the naturally occurring flavonoids, astragalin (kaempferol 3-glucoside), which is a bioactive constituent of various traditional
    [Show full text]
  • Shilin Yang Doctor of Philosophy
    PHYTOCHEMICAL STUDIES OF ARTEMISIA ANNUA L. THESIS Presented by SHILIN YANG For the Degree of DOCTOR OF PHILOSOPHY of the UNIVERSITY OF LONDON DEPARTMENT OF PHARMACOGNOSY THE SCHOOL OF PHARMACY THE UNIVERSITY OF LONDON BRUNSWICK SQUARE, LONDON WC1N 1AX ProQuest Number: U063742 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a com plete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest U063742 Published by ProQuest LLC(2017). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C ode Microform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106- 1346 ACKNOWLEDGEMENT I wish to express my sincere gratitude to Professor J.D. Phillipson and Dr. M.J.O’Neill for their supervision throughout the course of studies. I would especially like to thank Dr. M.F.Roberts for her great help. I like to thank Dr. K.C.S.C.Liu and B.C.Homeyer for their great help. My sincere thanks to Mrs.J.B.Hallsworth for her help. I am very grateful to the staff of the MS Spectroscopy Unit and NMR Unit of the School of Pharmacy, and the staff of the NMR Unit, King’s College, University of London, for running the MS and NMR spectra.
    [Show full text]
  • Polypodium Vulgare L.: Polyphenolic Profile, Cytotoxicity and Cytoprotective Properties in Different Cell Lines
    Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 14 May 2021 doi:10.20944/preprints202105.0351.v1 Article Polypodium vulgare L.: polyphenolic profile, cytotoxicity and cytoprotective properties in different cell lines Adrià Farràs1,2, Víctor López2,4, Filippo Maggi3, Giovani Caprioli3, M.P. Vinardell1, Montserrat Mitjans1* 1Department of Biochemistry and Physiology, Faculty of Pharmacy and Food Sciences, Universitat de Barcelona, 08028 Barcelona, Spain; [email protected] (A.F.); [email protected] (P.V.); [email protected] (M.M.) 2Department of Pharmacy, Faculty of Health Sciences, Universidad San Jorge, Villanueva de Gállego, Zaragoza, 50830 Spain; [email protected] (A.F.); [email protected] (V.L.) 3School of Pharmacy, Università di Camerino, 62032 Camerino, Italy; [email protected] (F.M.); [email protected] (G.C.) 4Instituto Agroalimentario de Aragón-IA2, CITA-Universidad de Zaragoza, 50013 Zaragoza, Spain *Correspondence: [email protected] Abstract: Pteridophytes, represented by ferns and allies, are an important phytogenetic bridge between lower and higher plants (gymnosperms and angiosperms). Ferns have evolved independently of any other species in the plant kingdom being its secondary metabolism a reservoir of phytoconstituents characteristic of this taxon. The study of the possible medicinal uses of Polypodium vulgare L. (Polypodiaceae), PV, has increased particularly when in 2008 the European Medicines Agency published a monograph about the rhizome of this species. Thus, our objective is to provide scientific knowledge on the methanolic extract from the fronds of P. vulgare L., one of the main ferns described in the Prades Mountains, to contribute to the validation of certain traditional uses. Specifically, we have characterized the methanolic extract of PV fronds (PVM) by HPLC-DAD and investigated its potential cytotoxicity, phototoxicity, ROS production and protective effects against oxidative stress by using in vitro methods.
    [Show full text]
  • Phytochem Referenzsubstanzen
    High pure reference substances Phytochem Hochreine Standardsubstanzen for research and quality für Forschung und management Referenzsubstanzen Qualitätssicherung Nummer Name Synonym CAS FW Formel Literatur 01.286. ABIETIC ACID Sylvic acid [514-10-3] 302.46 C20H30O2 01.030. L-ABRINE N-a-Methyl-L-tryptophan [526-31-8] 218.26 C12H14N2O2 Merck Index 11,5 01.031. (+)-ABSCISIC ACID [21293-29-8] 264.33 C15H20O4 Merck Index 11,6 01.032. (+/-)-ABSCISIC ACID ABA; Dormin [14375-45-2] 264.33 C15H20O4 Merck Index 11,6 01.002. ABSINTHIN Absinthiin, Absynthin [1362-42-1] 496,64 C30H40O6 Merck Index 12,8 01.033. ACACETIN 5,7-Dihydroxy-4'-methoxyflavone; Linarigenin [480-44-4] 284.28 C16H12O5 Merck Index 11,9 01.287. ACACETIN Apigenin-4´methylester [480-44-4] 284.28 C16H12O5 01.034. ACACETIN-7-NEOHESPERIDOSIDE Fortunellin [20633-93-6] 610.60 C28H32O14 01.035. ACACETIN-7-RUTINOSIDE Linarin [480-36-4] 592.57 C28H32O14 Merck Index 11,5376 01.036. 2-ACETAMIDO-2-DEOXY-1,3,4,6-TETRA-O- a-D-Glucosamine pentaacetate 389.37 C16H23NO10 ACETYL-a-D-GLUCOPYRANOSE 01.037. 2-ACETAMIDO-2-DEOXY-1,3,4,6-TETRA-O- b-D-Glucosamine pentaacetate [7772-79-4] 389.37 C16H23NO10 ACETYL-b-D-GLUCOPYRANOSE> 01.038. 2-ACETAMIDO-2-DEOXY-3,4,6-TRI-O-ACETYL- Acetochloro-a-D-glucosamine [3068-34-6] 365.77 C14H20ClNO8 a-D-GLUCOPYRANOSYLCHLORIDE - 1 - High pure reference substances Phytochem Hochreine Standardsubstanzen for research and quality für Forschung und management Referenzsubstanzen Qualitätssicherung Nummer Name Synonym CAS FW Formel Literatur 01.039.
    [Show full text]
  • Etude Du Potentiel Herbicide Des Extraits Végétaux Des Espèces Xero-Halophytes Tunisiennes Et Détermination De Leurs Modes D’Action
    Etude du potentiel herbicide des extraits végétaux des espèces xero-halophytes Tunisiennes et détermination de leurs modes d’action Promoteurs : Pr Haissam Jijakli – Pr Riadh Ksouri Année civile : 2020 COMMUNAUTÉ FRANÇAISE DE BELGIQUE UNIVERSITÉ DE LIÈGE - GEMBLOUX AGRO-BIO TECH Etude du potentiel herbicide des extraits végétaux des espèces xero-halophytes Tunisiennes et détermination de leurs modes d’action Sofiene Ben Kaab Dissertation originale présentée en vue de l’obtention du grade de docteur en Sciences agronomiques et ingénierie biologique Promoteurs : Pr Haissam Jijakli – Pr Riadh Ksouri Année civile : 2020 © Ben kaab sofiene, février 2020 Résumé Les adventices constituent une contrainte majeure pour l’agriculture. En effet, connues par leur pouvoir allélopathique important, ces adventices peuvent causer des dommages économiques sévères atteignant 34% dans les grandes cultures. Dans cette optique, suite à la forte limitation des herbicides chimiques sur le marché et plus spécifiquement le retrait progressif du glyphosate, la demande du public pour des herbicides alternatifs d'origine biologique et davantage respectueux de l’environnement, est croissante. Cette thèse s’inscrit dans la politique de minimiser l’utilisation des herbicides chimiques principalement le glyphosate. Les travaux de recherche effectué dans le cadre de cette thèse ont permis de sélectionner 2 extraits végétaux à forte activité herbicide à savoir, l’extrait phénoliques de Cynara cardunculus et l’huile essentielle de Rosmarinus officinalis. Par la suite nous avons procédé à un fractionnement bio-guidé afin de déterminer les composés bioactifs majoritaires. Ainsi, la myricitrine extraite de C. cardunculus a été identifiée pour la première fois comme molécule à potentiel herbicide. Ensuite, nous avons élaboré une nouvelle formulation à base de ces deux extraits végétaux sélectionnés qui pour la première fois améliore en même temps leurs efficacités et leurs stabilités.
    [Show full text]
  • Wine Phenolic Compounds Differently Affect the Host-Killing Activity of Two Lytic Bacteriophages Infecting the Lactic Acid Bacte
    viruses Article Wine Phenolic Compounds Differently Affect the Host-Killing Activity of Two Lytic Bacteriophages Infecting the Lactic Acid Bacterium Oenococcus oeni Cécile Philippe 1, Amel Chaïb 1, Fety Jaomanjaka 1 , Stéphanie Cluzet 1, Aurélie Lagarde 1, Patricia Ballestra 1, Alain Decendit 1,Mélina Petrel 2, Olivier Claisse 1,3 , Adeline Goulet 4,5, Christian Cambillau 4,5 and Claire Le Marrec 1,6,* 1 EA4577-USC1366 INRAE, Unité de Recherche OEnologie, Université de Bordeaux, Institut des Sciences de la Vigne et du Vin (ISVV), F-33140 Villenave d’Ornon, France; [email protected] (C.P.); [email protected] (A.C.); [email protected] (F.J.); [email protected] (S.C.); [email protected] (A.L.); [email protected] (P.B.); [email protected] (A.D.); [email protected] (O.C.) 2 Bordeaux Imaging Center, UMS3420 CNRS-INSERM, University Bordeaux, F-33000 Bordeaux, France; [email protected] 3 INRAE, ISVV, USC 1366 Oenologie, F-33140 Villenave d’Ornon, France 4 Architecture et Fonction des Macromolécules Biologiques, Aix-Marseille Université, Campus de Luminy, F-13020 Marseille, France; [email protected] (A.G.); [email protected] (C.C.) 5 Architecture et Fonction des Macromolécules Biologiques, Centre National de la Recherche Scientifique (CNRS), Campus de Luminy, F-13020 Marseille, France 6 Bordeaux INP, ISVV, EA4577 OEnologie, F-33140 Villenave d’Ornon, France * Correspondence: clehenaff@enscbp.fr; Tel.: +33-55-757-5831 Received: 29 October 2020; Accepted: 14 November 2020; Published: 17 November 2020 Abstract: To provide insights into phage-host interactions during winemaking, we assessed whether phenolic compounds modulate the phage predation of Oenococcus oeni.
    [Show full text]
  • DOI 10.22607IJACS.2020.802006.Pdf
    Indian Journal of Article Advances in Chemical Science Simultaneous Determination of Isoquercitrin and Astragalin in Plant (Leaf) Extract Using Liquid Chromatography with Tandem Mass Spectrometry Method for the Application of Toxicology Studies in Matrix Meet Patel, Padmaja Prabhu, Alpesh Patel, Purushottam Trivedi Department of Chemistry, Jai Research Foundation, Valvada, Vapi, Gujarat, India ABSTRACT An analytical approach has been developed and validated using liquid chromatography (LC)-mass Spectrometry (MS)/MS for the simultaneous determination of isoquercitrin and astragalin from the TGT Primaage (plant [leaf] extract), following the application of toxicology studies. The purpose of analytical method validation was established a sensitive data, which was investigated for chronic toxicology studies evaluation. The method validations were implemented for determining the individuality and concentration of the analytes, matrix match effects and provide an estimated analytical method validation. The method validation was carried out by performing different parameters using reference standards and test substance solutions of isoquercitrin and astragalin in the matrix and analyzed onto LC-MS/MS. This analytical validated method was successfully applied to the actual samples of the toxicology studies for the dose formulation analysis of TGT Primaage (plant [leaf] extract) samples which had the contents of isoquercitrin and astragalin. Key words: Flavonoid, Liquid chromatography-Mass spectrometry/mass spectrometry, Toxicology studies, International Conference on Harmonization Q2 (R1), Extraction technique, Matrix effect, Chemical compound. 1. INTRODUCTION The objectives of the present work are (i) to develop and validate the analytical method for the determination of isoquercitrin and astragalin Isoquercitrin is used as a flavonoid and also used as a chemical of plant (leaf) extract of TGT Primaage samples in the matrix, (ii) to compound [1].
    [Show full text]
  • Quercetagetin and Patuletin: Antiproliferative, Necrotic and Apoptotic Activity in Tumor Cell Lines
    molecules Article Quercetagetin and Patuletin: Antiproliferative, Necrotic and Apoptotic Activity in Tumor Cell Lines Jesús J. Alvarado-Sansininea 1, Luis Sánchez-Sánchez 2, Hugo López-Muñoz 2, María L. Escobar 3 , Fernando Flores-Guzmán 2, Rosario Tavera-Hernández 1 and Manuel Jiménez-Estrada 1,* 1 Laboratorio 2-10, Departamento de Productos Naturales, Instituto de Química, Universidad Nacional Autónoma de México, 04510 Ciudad de México, Mexico; [email protected] (J.J.A.-S.); [email protected] (R.T.-H.) 2 Laboratorio 6, 2do piso, UMIEZ, Facultad de Estudios Superiores Zaragoza, Universidad Nacional Autónoma de México, 09230 Ciudad de México, Mexico; [email protected] (L.S.-S.); [email protected] (H.L.-M.); [email protected] (F.F.-G.) 3 Laboratorio de Microscopía Electrónica, Departamento de Biología Celular, Facultad de Ciencias, Universidad Nacional Autónoma de México, 04510 Ciudad de México, Mexico; [email protected] * Correspondence: [email protected]; Tel.: +52-(55)-56-22-44-30 Received: 22 August 2018; Accepted: 4 October 2018; Published: 9 October 2018 Abstract: Quercetagetin and patuletin were extracted by the same method from two different Tagetes species that have multiple uses in folk medicine in Mexico and around the globe, one of which is as an anticancer agent. Their biological activity (IC50 and necrotic, apoptotic and selective activities of these flavonols) was evaluated and compared to that of quercetin, examining specifically the effects of C6 substitution among quercetin, quercetagetin and patuletin. We find that the presence of a methoxyl group in C6 enhances their potency. Keywords: necrotic; apoptosis; quercetin; quercetagetin; patuletin 1.
    [Show full text]
  • Developing Drug and Gene Therapies for Peroxisome Biogenesis Disorders of the Zellweger Spectrum
    Developing drug and gene therapies for peroxisome biogenesis disorders of the Zellweger Spectrum Catherine Argyriou Department of Human Genetics McGill University, Montréal, Canada June 2018 A thesis submitted to McGill University in partial fulfillment of the requirements of the degree of Doctor of Philosophy © Catherine Argyriou 2018 ABSTRACT Zellweger spectrum disorder (ZSD) usually results from biallelic mutations in PEX genes required for peroxisome biogenesis. PEX1-G843D is a common hypomorphic allele associated with milder disease. We previously showed that fibroblasts from patients with a PEX1-G843D allele recovered peroxisome functions when cultured with the nonspecific chaperone betaine and flavonoid acacetin diacetate. To identify more effective flavonoids for preclinical trials, we compared 54 flavonoids using our cell-based peroxisomal assays. Diosmetin showed the most promising combination of potency and efficacy; co-treatments of diosmetin and betaine showed the most robust additive effects. This was confirmed by 5 independent assays in primary PEX1-G843D patient cells. Neither agent was active in PEX1 null cells. I propose that diosmetin acts as a pharmacological chaperone to improve stability, conformation, and function of PEX1/PEX6 exportomer complexes. All individuals with a PEX1-G843D allele develop a retinopathy that progresses to blindness. To investigate pathophysiology and identify endpoints for experimental trials, I used the knock-in mouse model for the equivalent human mutation, PEX1-G844D. I characterized the progression of retinopathy and found reduced cone cell function and number early in life with more gradual deterioration of rod cell function. Electron microscopy at later stage retinopathy showed disorganization of photoreceptor inner segments and enlarged mitochondria. As retino-cortical function was relatively well-preserved, I propose that the vision defect in the Pex1-G844D mouse is primarily at the retinal level.
    [Show full text]
  • Asteraceae)§ Karin M.Valant-Vetscheraa and Eckhard Wollenweberb,*
    Chemodiversity of Exudate Flavonoids in Seven Tribes of Cichorioideae and Asteroideae (Asteraceae)§ Karin M.Valant-Vetscheraa and Eckhard Wollenweberb,* a Department of Plant Systematics and Evolution Ð Comparative and Ecological Phytochemistry, University of Vienna, Rennweg 14, A-1030 Wien, Austria b Institut für Botanik der TU Darmstadt, Schnittspahnstrasse 3, D-64287 Darmstadt, Germany. E-mail: [email protected] * Author for correspondence and reprint requests Z. Naturforsch. 62c, 155Ð163 (2007); received October 26/November 24, 2006 Members of several genera of Asteraceae, belonging to the tribes Mutisieae, Cardueae, Lactuceae (all subfamily Cichorioideae), and of Astereae, Senecioneae, Helenieae and Helian- theae (all subfamily Asteroideae) have been analyzed for chemodiversity of their exudate flavonoid profiles. The majority of structures found were flavones and flavonols, sometimes with 6- and/or 8-substitution, and with a varying degree of oxidation and methylation. Flava- nones were observed in exudates of some genera, and, in some cases, also flavonol- and flavone glycosides were detected. This was mostly the case when exudates were poor both in yield and chemical complexity. Structurally diverse profiles are found particularly within Astereae and Heliantheae. The tribes in the subfamily Cichorioideae exhibited less complex flavonoid profiles. Current results are compared to literature data, and botanical information is included on the studied taxa. Key words: Asteraceae, Exudates, Flavonoids Introduction comparison of accumulation trends in terms of The family of Asteraceae is distributed world- substitution patterns is more indicative for che- wide and comprises 17 tribes, of which Mutisieae, modiversity than single compounds. Cardueae, Lactuceae, Vernonieae, Liabeae, and Earlier, we have shown that some accumulation Arctoteae are grouped within subfamily Cichori- tendencies apparently exist in single tribes (Wol- oideae, whereas Inuleae, Plucheae, Gnaphalieae, lenweber and Valant-Vetschera, 1996).
    [Show full text]
  • Identifying Bioactive Phytochemicals in Spent Coffee Grounds for Cosmetic Application Through Global Metabolite Analysis
    IDENTIFYING BIOACTIVE PHYTOCHEMICALS IN SPENT COFFEE GROUNDS FOR COSMETIC APPLICATION THROUGH GLOBAL METABOLITE ANALYSIS _______________________________________ A Thesis Presented to the Faculty of the Graduate School University of Missouri-Columbia _______________________________________________________ In Partial Fulfillment of the Requirements for the Degree Master of Science _____________________________________________________ by JIHYUN PARK Dr. Chung-Ho Lin, Thesis Supervisor DECEMBER 2017 The undersigned, appointed by the dean of the Graduate School, have examined the thesis entitled: IDENTIFYING BIOACTIVE PHYTOCHEMICALS IN SPENT COFFEE GROUNDS FOR COSMETIC APPLICATION THROUGH GLOBAL METABOLITE ANALYSIS presented by JIHYUN PARK a candidate for the degree of Master of Science and hereby certify that in their opinion it is worthy of acceptance Professor Chung-Ho Lin Professor Shibu Jose Professor Gary Stacey Professor Minviluz Stacey ACKNOWLEDGEMENTS I would like to thank my advisor, Professor Chung-Ho Lin about his guidance with huge effort, and the Center for Agroforestry, without their considerable assistant, I could not have done this research. Conducting this project provided me a great opportunity to meet and work with many people who have a comprehensive mind and passion. It is very pleasure for me to offer thanks to them. In addition, my thesis committee members, Dr. Shibu Jose, Gary Stacey, and Bing Stacey, were the most responsible professors in their field and they leading me to focus on this research. They realized the novelty of my thesis topics paying attention to this project and encouraging me to accomplish the experiment on time. Also I would like to say thanks to Nahom Taddese Ghile who gave me the first help beginning this project and the laboratory members Van Ho, Danh Vu, Phuc Vo, who provided assistance in several steps of my experiment with familiarity.
    [Show full text]
  • In the Name of the Beneficent, the Merciful
    IN THE NAME OF ALLAH THE BENEFICENT, THE MERCIFUL In vitro and in vivo toxicological study of selective medicinal plants By Mazhar Abbas M. Phil. (UAF) A thesis submitted for partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY IN BIOCHEMISTRY DEPARTMENT OF BIOCHEMISTRY FACILITY OF SCIENCE UNIVERSITY OF AGRICULTURE FAISALABAD, PAKISTAN 2014 DECLARATION I hereby declare that the contents of the thesis," In vitro and in vivo Toxicological Study of Selective Medicinal Plants" is product of my own research and no part has been copied from any published source (expect the references, standard mathematical and geometrical models/ equations/ formulate/ protocols etc.). I further declare that this work has not been submitted for the award of any other diploma/ degree. The university may take actions if the information provided is found inaccurate at any stage. Mazhar Abbas 2004-ag-237 To The Controller of Examinations, University of Agriculture, Faisalabad. “We, the Supervisory Committee, certify that the contents and form of the thesis submitted by Mr. Mazhar Abbas 2004-ag-237, have been found satisfactory and recommend that it be processed for evaluation, by the External Examiner(s) for the award of Ph.D. degree”. Supervisory Committee 1. Chairman __________________________ Dr. Muhammad Shahid. 2. Member __________________________ Prof. Dr. Munir Ahmad Sheikh. 3. Member __________________________ Prof. Dr. Ghulam Muhammad. DEDICATED To My loving Parents Who always provide me compassion Throughout my life and my success is really the fruit of their prayers ACKNOWLEDGMENT In the name of Allah, the merciful, the beneficent All praises (belong) to Allah alone, the cherisher and Sustainer of the world.
    [Show full text]