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The Interaction Between Pseudomonas Syringae Pv. Tomato and the Lectin from Tomato (Lycopersicon Esculentum) a Possible Role in Determining Pathogen Specificity

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The Interaction Between Pseudomonas Syringae Pv. Tomato and the Lectin from Tomato (Lycopersicon Esculentum) a Possible Role in Determining Pathogen Specificity University of Windsor Scholarship at UWindsor Electronic Theses and Dissertations Theses, Dissertations, and Major Papers 1-1-1985 The interaction between pseudomonas syringae pv. tomato and the lectin from tomato (Lycopersicon esculentum) a possible role in determining pathogen specificity. Jamie Seymour Pitts University of Windsor Follow this and additional works at: https://scholar.uwindsor.ca/etd Recommended Citation Pitts, Jamie Seymour, "The interaction between pseudomonas syringae pv. tomato and the lectin from tomato (Lycopersicon esculentum) a possible role in determining pathogen specificity." (1985). Electronic Theses and Dissertations. 6903. https://scholar.uwindsor.ca/etd/6903 This online database contains the full-text of PhD dissertations and Masters’ theses of University of Windsor students from 1954 forward. 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CANADIAN THESES ON MICROFICHE THESES CANADIENNES SUB MICROFICHE National Library of Canada BibliothPque rationale du Canada • I* Collections Development Bran,ch Direction dti dPveioppement des collections Canadian Theses on : Service des theses canadiennes Microfiche Service • ' sur microfiche .■ ■ Ottawa/Canada K1A0N4 . ■ NOTICE AVIS The quality of this microfiche is heavily dependent upon the La quality de cette microfiche depend grandement de la quality quality of the' original thesis submitted for microfilming. Every de la thPse soumise aiimicrofilmage. 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Reproduction in full or in part of this film is governed by^he La reproduction, mPme partielle', de ce microfilm'est soumise Canadian Copyright Act, R.S.C. 1970, c. C-30. Please read P la Loi canadienne sur le droit d’auteur, SRC 1970, c. C-30. the authorization forms which accompany this thesis. Veuillez prendre connaissance d es fdrmules d’autorisation qui • accompagnent cette thPse. THIS DISSERTATION LA THESE A ETE HAS BEEN MICROFILMED MICROFILM IrE TELLE QUE EXACTLY AS RECEIVED NOUS L*AVONS REQUE Canada NL 339 (r. 65/09) National Library Bibliothfcqua nationale CANADIAN THESES'- THtSES CANADIENNES i * of Canada du Canada ON MICROFICHE \ SUR UICRQFWHE 67995 -\ NAME OF AUTHOR//VOM DE L'AUTEUR— — JaEie Seymour Pitts TITLE OF THESIS/TSTRE DE LA ru rsF ? h e Interaction Bettjeen Psfeitdomonn’-s S-grinyrio P i t . T rm ntn and the lectin from Tomato (Lycobersicon. EsculentumT; A .Possible Role in Determining Pathogen Specificity. UN IVERSlTi/UNIVERSITim, DEGREE FOR WHICH THESIS WAS PRESENTED/. „ ■ GRADE POUR LEQUEL CETTE THESE FUT PpFrfnt/ f H. S c , YEAR THIS DEGREE C O N F E R R E D //* fl'ORTENTION DE CE GRADE. 1 9 8 5 ' NAME OF SUPERVISOR/ATOM DU DIRECTEUR DE THtSE^_________ Dr. Park?-pH Permission is hereby granted to the NATIONAL LIBRARY OF L’autorisation est, par la prSsente, accordde d la BIBUOTHl o CANADA to microfilm this thesis and to lend or sell copies QUE NATIONALE DU CANADA de microfilmer cette thSse t 1! of the film. j de prSter ou de vendre des exemplaires du film. The author reserves other publication rights, and neither the V.auteur se rdserve les autres droits de publication?ni, thesis nor extensive extracts from it may be printed or other­ th ise n i de longs extraits de celle-ci m doivent etre imprinu wise reproduced without the author's written permission. ou autrement reprpduits sans I'autorisation dcrite de I'auteui "V DATED/0A TE, SIGNED/S/dWf. PERMANENT ADDRESS/R/fSIDENCE F IX t. NU-*I U-J4) THE INTERACTION BETWEEN PSmBOMOMS SYSMGAE PV. TOMA TO AND THE LECTIN FROM TOMATO \LYGOPERSWON ESCULENTVM$. A POSSIBLE ROLE IN DETERMINING PATHOGEN SPECIFICITY by . Jamie Seymour Pitts A thesis . submitted to the Faculty of Graduate Studies through the Department of Biology in Partial Fulfillment " of the requirements for the Degree ^ of Masters, of Science at The University of Windsor » Windsor, Ontario, Canada © Jamie Seymour Pitts 1985 AH Rights Reserved. 3 829405 Approved: This thesis' is dedicated to my .parents • and the memory of the late Mrs. Stephanie Brown. J * ■o: j c ' j ABSTRACT • / .4 . : . A leetin isolated frojn the. tomato fruit using a 60S ammonium sulphate precipitation, was the sanjie as that previously isolated by Naehbar ei tf/(198Q) and ■L Kilpatrick {1980). The tomato leotin had a molecular 'Weight of .140,000, as "determined'% gel filtration, and was specific for oligomers of N - aeetylglueosamine. As well as being a hemagglutinin, the lectin was a bacterial agglutinin, which agglutinated a wide- range of bacterial species, including Pseudomonas syringa'e py .tomato ( P.tomato). The specific activity of the lectin was 80 times greater for bacterial agglutination than hemagglutination. Bacterial agglutination titres were dramatically increased when divalent cations were present, and mimiaked the prozone phenomenon, known to occur between an antig£fr and antibody. * P.tomato produced an exopolysaocharide in addition to fevan, which completely inhibited its agglutination by the tomato (lectin. A lipopolysacoharide extract of P tomato also inhibited bacterial agglutination, to the same degree. 7 In consideration of the ledft recognition model, developed by Sequbira and Graham (1977), it seems likely that P.tomato is pathogenic to the tomato plant at least partly because it avoids recognition (lectin binding to the bacterial lipopolysacoharide). The ability to avoid recognition is imparted by thgC^ exopolysaccharide, which competitively inhibits the lectin from binding the, bacterial lipopolysacoharide. r v a ACKNOWLEDGEMENTS I would like to express my deep appreciation to Dr. ILB. Faeferell for the advice and endless encouragement he lias offerred me throughout the entire course \o f this study. Thanks also go to the other members of my committee, Dr. B.A. Cotter and Dr. N.F. Taylor. , Iwould also like to thank my good friends, Mr.’ Lou Passador, Mr. John Austin and Ms. Renu Sarao For making those long nights on the third floor - bearable. • v . Last, but certainly not least, thank-you Sandra, for your devotion and understanding during the most difficult of times. «/ x TABLE OF CONTENTS PAGE Ab d i c a t i o n . • IT ABSTRACT.'....\.. ........... ¥ 'f ACKNOWLEDGEMENTS... ¥1 LIST OF ILLUSTRATIONS. ix o % LIST OF TABLBSi. • y ; •■••••• x ■ ' x LIST OP ABBREVIATIONS .!...•....................... T. '................ .xi INTRODUCTION.:...'........ ............ .......................,. .1 ? LITERATURE REVIEW . ; ; 9 • » ; * * * V : -Bacterial Speck of Tomato.'... .......................v .................5- ■ Plant lectins....... ’. ?................ '..14 MATERIALS AND METHODS r ' ”0' ' ' ' ' *" ' ' * Bacterial Cultures. .......... ..........„ . ............;. .21 Isolation of a le ctin from tomato, fruit.............................*.. ____ 22 • . Cel Filtration.... ............ ..............................cr ' . •'............22 ti . Lectin Concentration in Orud? Extract ..................................... ....23 ; Hemagglutination (HA) Assay .................. ..............................'.................. .24 1 ■ 1 Carbohydrate S p e c ific ity ............ .25 ;* Bacterial Agglutination (BA) Assay....... ...... ............. ; .................25 Ex^polysaccharide Extraction ..................... .26 Lipopolyshccharide Extraction.. ......... f ......................26 e ' I * inhibition of Bacterial Agglutination ;j ■ 4 A) Exopolysaccharide InhibitiW x>i__^f .......... 27 B) Lipopolysaccharide Inhibition.............. , v ..........., .28 RESULTS ' - - : . - ■ ?■ . '• • - ■■ •. - jft * c> , V .Chapter- 1. .Characterization of Tomato'.Lectin '■-'a . _o * 5 * . J " Erythrocyte Specificity, r, ,.J. .„.......... .;.30 g. ' * ’* .V - Molecular Weight Determination.".; ..........» . ......... .80. - - '' ; . ' ''» . * Carbohydrate Specificity............. ■...........; ............................35 Bacterial Agglutination;.. v..- ............. .38 . - .Lectin Concentration in Crpde. E x t r a c t . ...................... '..42 - i Is the Bacterial Agglutinin the- 9 f \ . Hemagglutinin. ...... ................ .-a
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