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The Physiological Effects of Phycobilisome Antenna Modification on the Cyanobacterium Synechocystis Sp. PCC 6803

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The Physiological Effects of Phycobilisome Antenna Modification on the Cyanobacterium Synechocystis Sp. PCC 6803 Washington University in St. Louis Washington University Open Scholarship All Theses and Dissertations (ETDs) Winter 1-1-2012 The hP ysiological Effects of Phycobilisome Antenna Modification on the Cyanobacterium Synechocystis sp. PCC 6803 Lawrence Edward Page Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/etd Recommended Citation Page, Lawrence Edward, "The hP ysiological Effects of Phycobilisome Antenna Modification on the Cyanobacterium Synechocystis sp. PCC 6803" (2012). All Theses and Dissertations (ETDs). 1015. https://openscholarship.wustl.edu/etd/1015 This Dissertation is brought to you for free and open access by Washington University Open Scholarship. It has been accepted for inclusion in All Theses and Dissertations (ETDs) by an authorized administrator of Washington University Open Scholarship. For more information, please contact [email protected]. WASHINGTON UNIVERSITY IN ST. LOUIS Division of Biology & Biomedical Sciences Biochemistry Dissertation Examination Committee: Himadri Pakrasi, Chair Tuan-Hua David Ho Joseph Jez Tom Smith Gary Stormo Yinjie Tang The Physiological Effects of Phycobilisome Antenna Modification on the Cyanobacterium Synechocystis sp. PCC 6803 by Lawrence Edward Page II A dissertation presented to the Graduate School of Arts and Sciences of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy December 2012 St. Louis, Missouri © Copyright 2012 by Lawrence Edward Page II. All rights reserved. Table of Contents List of Figures .........................................................................................................................................x List of Tables....................................................................................................................................... xiv List of Abbreviations..........................................................................................................................xv Acknowledgements.........................................................................................................................xvii Dedication .........................................................................................................................................xxiv List of Publications ....................................................................................................................xxv Abstract of the Dissertation ........................................................................................................xxvi Chapter 1: Introduction to Photosynthetic Light Harvesting Antenna Complexes and The Cyanobacterial Phycobilisome ................................................................................................1 Introduction..............................................................................................................................................2 The role of cyanobacteria in the evolution of life on earth ...................................................................2 Synechocystis sp. PCC 6803 as a model organism for the study of photosynthesis .............................2 Oxygenic Photosynthesis ......................................................................................................................3 Light harvesting antenna complexes .....................................................................................................4 The phycobilisome of cyanobacteria.....................................................................................................6 Phycobilisome structure in Synechocystis 6803....................................................................................8 Antenna mutants and photoautotrophic productivity ............................................................................9 This Work...............................................................................................................................................11 References...............................................................................................................................................13 ii Chapter 2: Physiological consequences of phycobilisome antenna truncation on expression and function of Photosystem II............................................................................... 21 Introduction............................................................................................................................................22 Materials and Methods..........................................................................................................................25 Strains and growth conditions .............................................................................................................25 Western blots.......................................................................................................................................25 Oxygen saturation curves ....................................................................................................................26 Low temperature fluorescence spectra................................................................................................26 Flash-induced oxygen evolution .........................................................................................................26 Flash-induced Fluorescence ................................................................................................................27 Results.....................................................................................................................................................28 Examination of active PSII expression ...............................................................................................28 Low temperature fluorescence ............................................................................................................28 Determination of S-state turnover .......................................................................................................29 Discussion ...............................................................................................................................................31 References...............................................................................................................................................34 Chapter 3: Organization and Flexibility of Cyanobacterial Thylakoid Membranes Examined by Neutron Scattering.................................................................................................. 48 Introduction............................................................................................................................................49 Materials and Methods..........................................................................................................................52 Electron microscopy............................................................................................................................52 Small angle neutron scattering ............................................................................................................52 Data analysis........................................................................................................................................53 Quasi-elastic neutron scattering ..........................................................................................................53 Results.....................................................................................................................................................55 iii Discussion ...............................................................................................................................................61 References...............................................................................................................................................67 Chapter 4: Reduction of Photoautotrophic Productivity in the Cyanobacterium Synechocystis sp. PCC 6803 by Phycobilisome Antenna Reduction .................................. 86 Introduction............................................................................................................................................87 Materials and Methods..........................................................................................................................90 Growth and maintenance of bacterial strains ......................................................................................90 Absorption spectra...............................................................................................................................90 Growth curves .....................................................................................................................................90 Photobioreactor specifications ............................................................................................................90 Growth in photobioreactors.................................................................................................................91 Sample measurements .........................................................................................................................92 Results.....................................................................................................................................................93 Growth comparison of WT and antenna mutants................................................................................93
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