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Methods in Molecular Biology 1744 Yongfeng Guo Editor Plant Senescence Methods and Protocols M M B ethods in olecular iology Series Editor John M. Walker School of Life and Medical Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For further volumes: http://www.springer.com/series/7651 Plant Senescence Methods and Protocols Edited by Yongfeng Guo Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, Shandong, China Editor Yongfeng Guo Tobacco Research Institute Chinese Academy of Agricultural Sciences Qingdao, Shandong, China ISSN 1064-3745 ISSN 1940-6029 (electronic) Methods in Molecular Biology ISBN 978-1-4939-7670-6 ISBN 978-1-4939-7672-0 (eBook) https://doi.org/10.1007/978-1-4939-7672-0 Library of Congress Control Number: 2017964498 © Springer Science+Business Media, LLC 2018 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Printed on acid-free paper This Humana Press imprint is published by Springer Nature The registered company is Springer Science+Business Media, LLC The registered company address is: 233 Spring Street, New York, NY 10013, U.S.A. Preface As the final phase of plant development, senescence is of great significance for the life cycle of plants as well as for agriculture. Plant senescence is a type of senescence distinct from senescence processes in animals and other systems (Chapter 1), mainly because of the nutri- ent remobilization process which, as a result of evolutionary fitness, recycles nutrients in senescing tissues to sink organs, a critical step for yield formation in agriculture. Nutrients in senescing organs are released through massive degradation of macromolecules such as proteins, polysaccharides, lipids, and nucleotides. The macromolecule degradation and nutrient remobilization processes are driven by active expression of thousands of senes- cence-associated genes (SAGs) (Chapter 2), which makes plant senescence distinct from many other plant developmental processes. Besides age, senescence of plants can be induced by a great number of factors including reproductive growth, phytohormones, and environmental cues. There are also many factors which function to inhibit plant senescence (Chapter 2). Thus, a complex regulatory net- work involving transduction of various signals, gene expression, and metabolism is respon- sible for the initiation of plant senescence. Protocols related to plant senescence induced by ethylene (Chapter 8), abscisic acid (Chapters 9 and 11), jasmonic acid, salicylic acid, strigo- lactones, peptide hormones (Chapter 11), shade/darkness (Chapter 12), salt (Chapter 13), pathogen (Chapter 15), oxidative stress (Chapter 16), nutritional status (Chapter 14), and postharvest conditions (Chapter 17) are summarized in this volume. Once senescence is initiated, plants undergo a similar senescence execution process, the so-called senescence syndrome. Protocols commonly used in characterizing molecular, cellular, and biochemical aspects of the senescence syndrome are included in this volume. These are methods in studying chlorophyll catabolism (Chapter 18), programmed cell death (Chapter 21), cell wall polysaccharides (Chapter 25), vacuolar proteases (Chapter 22), chloroplast protein degradation (Chapter 24), and autophagy (Chapter 23). Methods for studying different types of senescence, including leaf senescence (Chapter 3), flower senescence (Chapter 4), fruit ripening (Chapter 7), nodule senescence (Chapter 5), and floral organ abscission (Chapters 6 and 26), are described. Also included are techniques for studying plant senescence in general, such as gene identification and functional analysis (Chapter 19), MicroRNAs (Chapter 20), map-based cloning (Chapter 17), virus-induced gene silencing (Chapter 4), electron microscopy (Chapter 26), transcriptome (Chapter 27), and metabolome (Chapter 28). The techniques covered in this volume are either developed specially for plant senes- cence studies or optimized for studying senescing plants. Every method includes not only a detailed description of the procedures but also thoughtful notes for correct interpreta- tion, special cautions, or alternative options. The aim is to provide a useful handbook of standard protocols for plant molecular biologists working on senescence. Qingdao, Shandong, China Yongfeng Guo v Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix Part I IntroductIon of Plant SeneScence 1 Concepts and Types of Senescence in Plants .............................. 3 Susheng Gan 2 Initiation, Progression, and Genetic Manipulation of Leaf Senescence........... 9 Akhtar Ali, Xiaoming Gao, and Yongfeng Guo Part II P henotyPIc analySIS and Molecular MarkerS of Plant organ SeneScence 3 Phenotypic Analysis and Molecular Markers of Leaf Senescence................ 35 Liming Zhao, Yan Xia, Xiao-Yuan Wu, Jos H.M. Schippers, and Hai-Chun Jing 4 Investigation of Petal Senescence by TRV-Mediated Virus-Induced Gene Silencing in Rose.............................................. 49 Chenxia Cheng, Junping Gao, and Nan Ma 5 Phenotypic Analysis and Molecular Markers of Plant Nodule Senescence......... 65 Xiuying Xia 6 Quantitative Analysis of Floral Organ Abscission in Arabidopsis Via a Petal Breakstrength Assay........................................ 81 Chun-Lin Shi and Melinka A. Butenko 7 Characterization of Climacteric and Non-Climacteric Fruit Ripening ........... 89 Xiaohong Kou and Mengshi Wu Part III horMonal control of Plant SeneScence 8 Ethylene Treatment in Studying Leaf Senescence in Arabidopsis ............... 105 Zhonghai Li and Hongwei Guo 9 The Assay of Abscisic Acid-Induced Stomatal Movement in Leaf Senescence...... 113 Yanyan Zhang and Kewei Zhang 10 The EPR Method for Detecting Nitric Oxide in Plant Senescence.............. 119 Aizhen Sun 11 Hormone Treatments in Studying Leaf Senescence......................... 125 Zenglin Zhang and Yongfeng Guo Part IV StreSS-Induced SeneScence In PlantS 12 Methods to Study Darkness-Induced Leaf Senescence....................... 135 Yi Song and Lin Li vii viii Contents 13 Salt Treatments and Induction of Senescence ............................. 141 Yasuhito Sakuraba, Dami Kim, and Nam-Chon Paek 14 Methods for Elucidation of Plant Senescence in Response to C/N-Nutrient Balance............................................ 151 Shoki Aoyama, Junji Yamaguchi, and Takeo Sato 15 Study of Cotton Leaf Senescence Induced by Alternaria alternata Infection ..... 161 Wei Liu, Wenwei Zhang, Na Zheng, Weibo Zhai, and Fangjun Qi 16 Study of Hydrogen Peroxide as a Senescence-Inducing Signal................. 173 Stefan Bieker, Maren Potschin, and Ulrike Zentgraf 17 Identification of Postharvest Senescence Regulators Through Map-Based Cloning Using Detached Arabidopsis Inflorescences as a Model Tissue .......... 195 Donald A. Hunter, Rubina Jibran, Paul Dijkwel, David Chagné, Kerry Sullivan, Aakansha Kanojia, and Ross Crowhurst Part V Molecular and cellular ProceSSeS In Plant SeneScence 18 Chlorophyll and Chlorophyll Catabolite Analysis by HPLC................... 223 Aditi Das, Luzia Guyer, and Stefan Hörtensteiner 19 Identification and Functional Analysis of Senescence-Associated Genes in Wheat... 237 Geng Wang, Ke Li, and Chunjiang Zhou 20 Isolation, Purification, and Detection of Micro RNAs in Plant Senescence........ 247 Yujun Ren and Ying Miao 21 In Situ Detection of Programmed Cell Death in Senescing Nicotiana tabacum Leaves Using TUNEL Assay........................... 267 Branka Uzelac, Dušica Janošević, and Snežana Budimir 22 Activities of Vacuolar Cysteine Proteases in Plant Senescence.................. 283 Dana E. Martínez, Lorenza Costa, and Juan José Guiamét 23 Study of Autophagy in Plant Senescence................................. 299 Xuefei Cui, Jing Zheng, Jinxin Zheng, and Qingqiu Gong 24 Isolation of Chloroplasts for In Organelle Protein Degradation Assay ........... 307 Scott W. Vande Wetering and Judy A. Brusslan 25 Plant Cell Walls: Isolation and Monosaccharide Composition Analysis........... 313 Yingzhen Kong, Malcolm O’Neill, and Gongke Zhou 26 Visualizing Morphological Changes of Abscission Zone Cells in Arabidopsis by Scanning Electron Microscope........................... 321 Chun-Lin Shi and Melinka A. Butenko Part VI SySteMS BIology In Plant SeneScence 27 RNA-Seq Analysis of the Transcriptome of Leaf Senescence in Tobacco ......... 331 Wei Li and Yongfeng Guo 28 Comprehensive Metabolomics Studies of Plant Developmental Senescence....... 339 Mutsumi Watanabe, Takayuki Tohge, Salma Balazadeh, Alexander Erban, Patrick Giavalisco, Joachim Kopka, Bernd Mueller-Roeber, Alisdair R. Fernie, and Rainer Hoefgen Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359 Contributors akhtar alI • Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, Shandong, China; Nuclear Institute for Food and Agriculture, Peshawar, Pakistan ShokI aoyaMa • Faculty of Science and Graduate School of Life Science, Hokkaido University, Sapporo, Japan SalMa Balazadeh • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany; Institute of Biochemistry and Biology, University of Potsdam, Potsdam-Golm, Germany Stefan BIeker • ZMBP, Department of General Genetics, University Tuebingen, Tuebingen, Germany Judy a. BruSSlan • Department of Biological Sciences, California State University, Long Beach, CA, USA Snežana BudIMIr • Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia MelInka a. Butenko • Department of Molecular Biosciences, University of Oslo, Oslo, Norway daVId chagné • The New Zealand Institute for Plant & Food Research Limited, Food Industry Science Centre, Palmerston North, New Zealand chenxIa cheng • Department of Ornamental Horticulture, China Agricultural University, Beijing, China; Beijing Key Laboratory of Development and Quality Control of Ornamental Crops, China Agricultural University, Beijing, China lorenza coSta • Instituto de Fisiología Vegetal (INFIVE)—Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de La Plata, Buenos Aires, Argentina roSS crowhurSt • The New Zealand Institute for Plant & Food Research Limited, Mt. Albert Research Centre, Auckland, New Zealand xuefeI cuI • Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, Tianjin, China; Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin, China adItI daS • Institute of Plant and Microbial Biology, University of Zurich, Zurich, Switzerland Paul dIJkwel • Institute of Fundamental Sciences, Massey University, Palmerston North, New Zealand alexander erBan • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany alISdaIr r. fernIe • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany SuSheng gan • Plant Biology Section, School of Integrative Plant Science, College of Agriculture and Life Sciences, Cornell University, Ithaca, NY, USA xIaoMIng gao • Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, Shandong, China ix x Contributors JunPIng gao • Department of Ornamental Horticulture, China Agricultural University, Beijing, China; Beijing Key Laboratory of Development and Quality Control of Ornamental Crops, China Agricultural University, Beijing, China PatrIck gIaValISco • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany QIngQIu gong • Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, Tianjin, China; Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin, China Juan JoSé guIaMét • Instituto de Fisiología Vegetal (INFIVE)—Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de La Plata, Buenos Aires, Argentina yongfeng guo • Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, Shandong, China hongweI guo • The State Key Laboratory of Protein and Plant Gene Research, College of Life Sciences, Peking-Tsinghua Center of Life Sciences, Peking University, Beijing, China; Department of Biology, South University of Science and Technology of China, Shenzhen, Guangdong, China luzIa guyer • Institute of Plant and Microbial Biology, University of Zurich, Zurich, Switzerland raIner hoefgen • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany Stefan hörtenSteIner • Institute of Plant and Microbial Biology, University of Zurich, Zurich, Switzerland donald a. hunter • The New Zealand Institute for Plant & Food Research Limited, Food Industry Science Centre, Palmerston North, New Zealand dušIca JanošeVIć • Institute of Botany and Botanical Garden “Jevremovac”, Faculty of Biology, University of Belgrade, Belgrade, Serbia ruBIna JIBran • The New Zealand Institute for Plant & Food Research Limited, Food Industry Science Centre, Palmerston North, New Zealand; Institute of Fundamental Sciences, Massey University, Palmerston North, New Zealand haI-chun JIng • Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing, China aakanSha kanoJIa • Institute of Fundamental Sciences, Massey University, Palmerston North, New Zealand daMI kIM • Department of Plant Science, Plant Genomics and Breeding Institute, Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea yIngzhen kong • Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, Shandong, China JoachIM koPka • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany xIaohong kou • School of Chemical Engineering and Technology, Tianjin University, Tianjin, China lIn lI • Institute of Plant Biology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China ke lI • College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei, China weI lI • Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, Shandong, China Contributors xi zhonghaI lI • The State Key Laboratory of Protein and Plant Gene Research, College of Life Sciences, Peking-Tsinghua Center of Life Sciences, Peking University, Beijing, China weI lIu • State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China nan Ma • Department of Ornamental Horticulture, China Agricultural University, Beijing, China; Beijing Key Laboratory of Development and Quality Control of Ornamental Crops, China Agricultural University, Beijing, China dana e. Martínez • Instituto de Fisiología Vegetal (INFIVE)—Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de La Plata, Buenos Aires, Argentina yIng MIao • Center for Molecular Cell and Systems Biology, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, Fujian, China Bernd Mueller-roeBer • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany; Institute of Biochemistry and Biology, University of Potsdam, Potsdam-Golm, Germany MalcolM o’neIll • Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA naM-chon Paek • Department of Plant Science, Plant Genomics and Breeding Institute, Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea Maren PotSchIn • ZMBP, Department of General Genetics, University Tuebingen, Tuebingen, Germany fangJun QI • State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China yuJun ren • Center for Molecular Cell and Systems Biology, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, Fujian, China yaSuhIto SakuraBa • Department of Plant Science, Plant Genomics and Breeding Institute, Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea takeo Sato • Faculty of Science and Graduate School of Life Science, Hokkaido University, Sapporo, Japan JoS h.M. SchIPPerS • Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing, China; Molecular Ecology, Institute for Biology I, RWTH Aachen University, Aachen, Germany chun-lIn ShI • Department of Molecular Biosciences, University of Oslo, Oslo, Norway yI Song • Institute of Plant Biology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China kerry SullIVan • The New Zealand Institute for Plant & Food Research Limited, Food Industry Science Centre, Palmerston North, New Zealand aIzhen Sun • The National Key Laboratory of Plant Molecular Genetics and National Center for Plant Gene Research (Shanghai), CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China takayukI tohge • Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany Branka uzelac • Institute for Biological Research “Siniša Stanković”, University of Belgrade, Belgrade, Serbia

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