of Plants Responses and Adaptation to Freezing Stress

A. Sakai. W Larcher Frost Survival of Plants Responses and Adaptation to Freezing Stress With 200 Figures Springer -Verlag Berlin Heidelberg New York London Paris Tokyo

Professor em. Dr. AKIRA SAKAI The Institute of Low Temperature Science Hokkaido University Sapporo, Japan Professor Dr. WALTER LARCHER Institut flir Botanik der Universitiit Innsbruck StemwartestraBe 15 A-6020 Innsbruck, Austria ISBN-13:978-3-642-71747-5 e-isbn-13:978-3-642-71745-1 001: 10.1007/978-3-642-71745-1 Library of Congress Cataloging in Publication Data. Sakai, A. (Akira), 1920-. Frost survival of plants. (Ecological studies; v. 62). Bibliography: p. Includes indexes. 1. Plants, Effect of cold on. 2. Plants-Frost resistance. 3. Acclimatization (Plants) 4. Plants-Wounds and injuries. I. Larcher, W. (Walter), 1929-. II. Title. III. Series. QK156.S235 1981 581.19'165 8fr.31544. This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, re-use of illustrations, recitation, broadcasting, reproduction on microftlms or in other ways, and storage in data banks. Duplication of this publication or parts thereof is only permitted under the provisions of the German Copyright Law of September 9,1965, in its version of June 24,1985, and a copyright fee must always be paid. Violations fall under the prosecution act of the German Copyright Law. Springer-Verlag Berlin Heidelberg 1981. Softcover reprint of the hardcover 1 st edition 1981 The use of registered names, trademarks, 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. 2131/3130-543210

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Preface Low temperature represents, together with drought and salt stress, one of the most important environmental constraints limiting the productivity and the distribution of plants on the Earth. Winter survival, in particular, is a highly complex phenomenon, with regards to both stress factors and stress responses. The danger from winter cold is the result not only of its primary effect, i.e. the formation of ice in plant tissues; additional threats are presented by the freezing of water in and on the ground and by the load and duration ofthe snow cover. In recent years, a number of books and reviews on the subject of chilling and frost resistance in plants have appeared: all of these publications, however, concentrate principally on the mechanisms of injury and resistance to freezing at the cellular or molecular level. We are convinced that analysis of the ultrastructural and biochemical alterations in the cell and particularly in the plasma membrane during freezing is the key to understanding the limits of frost resistance and the mechanisms of cold acclimation. This is undoubtedly the immediate task facing those of us engaged in resistance research. It is nevertheless our opinion that, in addition to understanding the basic physiological events, we should be careful not to overlook the importance of the comparative aspects of the freezing processes, the components of stress avoidance and tolerance and the specific levels of resistance. Frost resistance is a genetically determined ecophysiological trait which is expressed under environmental constraints. The ability to harden is the basis for the differences between individuals, ecotypes, varieties and species with respect to their potential frost resistance. The gene pool of every population exhibits a range of variations both in hardening potential and seasonality timing, enabling it to survive a certain degree of change in the environment. The variability of frost resistance within a population is essential for the survival of a species following frost of unusual severity or of untimely occurrence, as well as for its adaptation to long-term fluctuations in climate. The degree of scatter of resistance within the progeny is characteristic for a species and provides a measure of its scope and selection. Thus, the quantitative analysis of phenomena over the full range of their variability among plant groups, species and genotypes provides important information from the viewpoint of ecology, phylogenetics and applied science. Climatic stress promotes evolutionary adaptation and accelerates the differentiation of ecotypes and species.

VI Preface Progression in the cold hardiness of plants along latitudinal and altitudinal gradients, and transitions between the various categories of cold hardiness have been hypothesized as being steps of evolutionary adaptation to low temperatures during the geological periods in which colder climates prevailed, as well as being connected with the spread of plants to colder regions. The book has been designed to include both the cellular and the comparative aspects of freezing stress and plant survival. With this special problem in mind, an attempt has been made to cover the entire range of ecophysiological research, from the biochemical to an ecological viewpoint. We have not attempted to produce a comprehensive compendium of plant cryobiology, but rather to present data on responses of plants to freezing stress, drawing to a large extent upon our personal experience gained in more than three decades of experimental research. Let us hope that the information presented here will make a significant contribution to the efforts of investigators engaged in planning research strategies for the study of plant hardiness in order to reduce crop losses resulting from frost and related stresses. Furthermore, it is our hope that more attention than hitherto will be given to the ecological interpretation of cold stress and cold resistance. Many colleagues have helped us by means of discussion, by providing us with unpublished data or by placing at our disposal original photographs and graphs: we gratefully acknowledge the help of members of the Institute of Low Temperature Science at the Hokkaido University in Sapporo: S. Yoshida, T. Niki, T. Sato, M. Ishikawa, M. Uemura, F. Yoshie; of the Hokkaido Forest Tree Breeding Institute: S. Eiga; and of the Institute of Botany at the University of Innsbruck: M. Bodner and U. Tappeiner. Further, we should like to express our thanks to C. J. Weiser and K. Tanino (Oregon State University), E. L. Proebsting (Washington State University), P. Wardle (DSIR, New Zealand), A. Kacperska (Warsaw University), M. J. Earnshaw (University of Manchester), M. Senser (University of Munich) and E. Beck (University of Bayreuth) for valuable suggestions and stimulating discussions. We should also like to take this opportunity to extend our gratitude to all persons and institutions who have helped us over the years by sending us their publications. We are most grateful to J. Wagner, A. Deutsch, R. Gapp and S. Hirn for their assistance in preparing the manuscript and with the compilation of indices. Our special thanks go to J. Wieser who has translated part of the text and carefully checked the remainder. It is a great pleasure to express our sincere thanks to Dr. K. F. Springer and the Editor responsible for the Ecological Studies, Prof. Dr. o. L. Lange, for including our book in their program, also to Dr. D. Czeschlik and his colleagues at Springer-Verlag for their cooperation and care in its preparation. Sapporo and Innsbruck, January 1987 AKIRA SAKAI WALTER LARCHER

Contents 1. Low Temperature and Frost as Environmental Factors 1.1 Low Temperature Hazards....... 1.2 Cold, Frost, and Snow.......... 1.3 The Occurrence of Cold, Frost, and Snow. 1.4 Frost and Snow in the Plant's Environment. 1.5 Time of Onset, Severity and Duration of Frost: The Freezing Risk............ 1.6 Temperature Fluctuations in Wintering Trees. 1 1 1 3 9 15 19 2. The Freezing Process in Plants...... 21 2.1 Freezing of Water and Aqueous Solutions 21 2.1.1 Ice Nucleation.......... 21 2.1.2 Freezing of Solutions....... 23 2.1.3 Sequence of Events in the Freezing of Liquids 24 2.1.4 Vitrification of Water and Solutes 24 2.2 Freezing of Plant Cells... 26 2.2.1 Extracellular Freezing.. 26 2.2.2 Intracellular Freezing... 29 2.2.3 Freezing of Intact Tissues. 31 2.2.4 Threshold Freezing Temperature and Supercooling of Plant Tissues............... 33 2.2.5 Initiation and Progress of Freezing in the Whole Plant. 36 3. Freezing Injuries in Plants..... 39 3.1 Typology of Freezing Mechanisms 39 3.2 Causes of Death by Freezing... 41 3.3 Phenomenology of Frost Damage. 47 3.3.1 Symptoms of Injury.... 47 3.3.2 Freezing Damage Peculiar to Trees. 48 3.4 Results of Freezing Injury and Chances of Recovery. 54 4. Mechanisms of Frost Survival.. 59 4.1 Components of Frost Survival 59 4.1.1 Frost Mitigation.... 59

VIII Contents 4.1.2 Frost Exclusion........ 62 4.1.3 Components of Frost Resistance. 65 4.2 Supercooling as a Survival Mechanism. 70 4.2.1 Persistent Supercooling of Leaves 70 4.2.2 Deep Supercooling of Xylem Ray Parenchyma Cells 72 4.3 Extraorgan and Extratissue Freezing....... 4.3.1 Deep Supercooling and Extraorgan Freezing of Hydrated Seeds............ 4.3.2 Deep Supercooling and Extraorgan Freezing of Floral Primordia of Woody Angiosperms. 4.3.3 Extraorgan Freezing of Conifer Buds.... 4.4 Comparison and Classification of Frost Survival Mechanisms 89 4.5 Survival at Ultralow Temperatures 91 4.5.1 Desiccated Plant Tissues 91 4.5.2 Hydrated Plant Tissues... 93 76 77 81 85 5. Cold Acclimation in Plants....... 97 5.1 The Annual Course of Frost Resistance 97 5.2 The Seasonal Cold Acclimation Process 99 5.2.1 Seasonality Timing....... 100 5.2.2 Induction of Cold Acclimation and the First Stage of the Hardening Process........... 102 5.2.3 The Second Stage of Frost Hardening..... 105 5.3 Environmental Control of the Level of Frost Resistance 107 5.3.1 Fluctuation of Frost Resistance During Winter 108 5.3.2 Water Content of the Plant and Frost Resistance. 112 5.4 Biochemical and Structural Changes During Cold Acclimation 113 5.4.1 Metabolic Pathway Shifts from the Growing State to the Dormant State..... 113 5.4.2 Sugars and Related Compounds 114 5.4.3 Amino Acids...... 117 5.4.4 Nucleic Acids and Proteins 118 5.4.5 Lipid Changes...... 124 5.4.6 Abscisic Acid...... 132 5.4.7 Cytological Changes During Cold Acclimation. 133 6. Frost Resistance in Plants............... 138 6.1 Genetic Variation in Frost Resistance......... 138 6.1.1 Differences in Frost Resistance Between Plant Taxa 138 6.1.2 Differences in Frost Resistance Within Populations. 142 6.1.3 Inheritance of Frost Resistance......... 153

Contents IX 6.2 Differences in Frost Resistance of Various Plant Organs and Tissues....... 157 6.2.1 Vegetative Organs........ 160 6.2.2 Reproductive Organs....... 167 6.3 Ontogenetic Variation in Frost Resistance 168 6.3.1 Variation of Frost Resistance During the Cell Cycle 169 6.3.2 Frost Resistance of Seeds............ 170 6.3.3 Frost Susceptibility During Germination and at the Early Seedling Stage.............. 171 6.3.4 Frost Resistance During Establishment and Ageing. 172 7. Regional Distribution of Plants and Their Adaptive Responses to Low Temperatures 174 7.1 Woody Plants Within the Tropics. 175 7.1.1 Mangroves 179 7.1.2 Palms 181 7.2 Evergreen Broad-Leaved Trees and Shrubs 185 7.2.1 Temperate Rain Forest Species 186 7.2.2 Mediterranean Sclerophylls 196 7.3 Transition from an Evergreen to a Deciduous Flora and Its Consequences for Frost Survival 200 7.3.1 Woody Plants..202 7.3.2 Ferns. 206 7.4 Conifers. 210 7.4.1 The Freezing Resistance of Various Conifer Families and Its Connection with Their Distribution Range.211 7.4.2 Characteristics of Freezing Resistance of Conifers.213 7.5 Mountain Plants 218 7.5.1 Dwarf Shrubs and Herbs of Temperate Mountains 218 7.5.2 Caulescent Giant Rosette Plants of Tropical High Mountains.224 7.6 Trends of Adaptive Improvement of Low Temperature Resistance in Vascular Plants. 229 8. Winter Damage as the Result of a Complexity of Constraints. 235 8.1 Winter Desiccation.................. 238 8.2 Damage Due to Ice Encasement and Compact Snow... 251 8.3 Harmful Effects of Heavy and Long-Lasting Snow Cover.. 252 8.4 Winter Survival: A Complex Response.......... 259

x Appendix Determination of Frost Resistance for Comparative Studies Field Survival......... Artificial Freezing Tests..... Analysis of the Freezing Process Frost Treatment of the Samples Identification of Frost Injuries. Quantification of Frost Resistance References........ Terminology and Definitions Taxonomic Index Subject Index.. Contents 260 260 260 261 262 263 264 266 267 305 307 317

Abbreviations DR DTA HTE K LN2 LT LT j LTo LTso LTE LTEso Tse Degree Celsius (centigrade); temperature level Directly recorded temperature ec) Differential thermal analysis (K) High temperature exotherm Kelvin; unit of temperature, temperature difference Liquid nitrogen (-196 C) Lowest temperature at a defined degree of lethality; quantification of frost resistance Initial injury temperature; incipient killing temperature Lowest temperature without injury Temperature at 50% lethality Low temperature exotherm Median temperature of L TEs Actual level of frost resistance corresponding to the state of hardening at a given time Maximum attainable level of frost resistance Threshold freezing temperature Threshold supercooling temperature For Terminology and Definitions see pp. 305-306.