Plant Aging Basic and Applied Approaches NATO ASI Series Advanced Science Institutes Series

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Volume 186-Plant Aging: Basic and Applied Approaches edited by Roberto Rodriguez, R. Sanchez Tames, and D. J. Durzan

Series A: Life Sciences Plant Aging Basic and Applied Approaches Edited by Roberto Rodriguez and R. Sanchez Tames University of Oviedo Oviedo, Spain

and D.J. Durzan University of California, Davis Davis, California

Plenum Press New York and London Published in cooperation with NATO Scientific Affairs Division Proceedings of a NATO Advanced Study Institute on Molecular Basis of Plant Aging, held July 2-15, 1989, in Ribadesella, Spain

Library of Congress Cataloging in Publication Data Plant aging: basic and applied approaches I edited by Roberto Rodriguez and R. Sanchez Tames, and D. J. Durzan. p. cm.-(NATO ASI series. Series A, Life sciences; vol. 186) "Published in cooperation with NATO Scientific Affairs Division." "Proceedings of a NATO Advanced Study Institute on Molecular Basis of Plant Aging, held July 2-15,1989 in Ribadeselia, Spain"-Copr. p. Includes bibliographical references. ISBN-13: 978-1-4684-5762-9 e-ISBN-13: 978-1-4684-5760-5 DOl: 10.1007/978-1-4684-5760-5 1. Plants-Aging-Congresses. 2. Plant propagation-Congresses. I. Rodriguez, Roberto. II. Sanchez, Tames, R. III. Durzan, D. J. IV. North Atlantic Treaty Organization. Scientific Affairs Division. V. NATO Advanced Study In• stitute on Molecular Basis of Plant Aging (1989: Ribadeselia, Spain) VI. Series: NATO ASI series. Series A, Life sciences; v. 186. QK762.5.P58 1990 90-6996 582'.0372-dc20 CIP

© 1990 Plenum Press, New York A Division of Plenum Publishing Corporation 233 Spring Street, New York, N.Y. 10013 All rights reserved No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher ORGANIZING AND SCIENTIFIC COMMITTEE

RODRIGUEZ, R. Lab. Fisiologia Vegetal, Dpto. B.O.S. Facultad de Biologia, Universidad de Oviedo. Arias de Velasco, sin. 33005 Oviedo. Spain

DURZAN, D. Department of Environmental Horti• culture. University of California. Davis, CA 95616 USA

PAIS, M.S. Departamento de Biologia Vegetal Facultade de Ciencias de Lisboa, Boco C2, Campo Grande 1700. Lisboa Portugal

WITHERS, L.A. International Board for Plant Gene• tic Resources. clo Food and Agricul• ture Organization of the United Na• tion. Via delle Terme di Caracalla 00100 Rome. Italy

TRAl.~ THANH-VAN, K. Institut de Physiologie V~g~tale. CNRS, 91190 Gif sur Yvette. France.

SANCHEZ TAMES, R. Lab. Fisiologia Vegetal, Dpto. B.O.S Facultad de Biologia, Universidad de Oviedo. Arias de Velasco, sin. 33005 Oviedo. Spain

SABATER, B. Dpto. de Biologfa Vegetal, Universi• dad de Alcal~ de Henares. Apdo. 20, Alcal~ de Henares. 28871-Madrid. Spain

NORTON, C.R. Faculty of Environmental Studies Meriot-Watt University Edinburgh, E43 9 DF. Scotland

PASQUALETO, P.L. Azienda Agricola Meristema SRL. Laboratorio di micropropagazione, Via Martiri della liberta, n. 13, 56030 Cascine di Buti (Pisa), Italy PREFACE

For many, the terms aging, maturation and are synonymous and used interchangeably, but they should not be. Whereas senescence represents an endogenously controlled degenerative programme leading to plant or organ , genetiC aging encompasses a wide array of passive degenerative genetiC processes driven primarily by exogenous factors (Leopold, 1975). Aging is therefore considered a consequence of genetiC lesions that accumulate over time, but by themselves do not necessarily cause death. These lesions are probably made more severe by the increase in size and complexity in trees and their attendant physiology. Thus while the withering of flower petals following pollination can be considered senescence, the loss of viability of stored more clearly represents aging (Norden, 1988). The very recent book "Senescence and Aging in Plants" does not discuss trees, the most dominant group of plants on the earth. Yet both angiospermic and gymnospermic trees also undergo the above phenomena but less is known about them. Do woody plants senesce or do they just age? What is phase change? Is this synonymous with maturation?

While it is now becoming recognized that there is no programmed senescence in trees, senescence of their parts, even in gymnosperms (e. g., needles of temperate conifers las t an average of 3.5 years), is common; but aging is a readily acknowledged phenomenon. In theory, at least, in the absence of any programmed senescence trees should -live forever, but in practice they do not. Is death simply a response to external environmental factors? As trees grow older, their growth rates decrease, and although they apparently never stop growing, very old trees increase in size rather insignificantly. Under natural conditions, a plot of the cumulative height and diameter growth by dominant trees in a stand exhibits a sigmoidal growth curve, with the maximum annual increment occurring relatively early in the age of the tree (Assma~n, 1970, quoted in Greenwood, 1988). The time at which the maximum increment occurs appears to be species-specific; and for the same species occurs later on relative poor soils, but at the same relative height and diameter. While there is little doubt that the maximum size a tree can attain is primarily a function of its genetics, is incremental growth more a function of size rather than chronological age? Is the phase-change process a consequence of the amount of growth that has occurred or is it the result of the physiological consequences of increased size (Greenwood, 1988)? There are other equally intriguing questions. Do the above changes arise from the accumulation of harmful metabolites with time, or from the physical, biophysical and physiological consequences of trees 'having to sustain such a large mass, the requeriments to move large volumes of water and nutrients, and photosynthates over great distances? Although the debate as to whether themselves age continues, more and more researchers have come to accept this idea, as first suggested by Shaffali tsky de flluckadell (1959); so much so that today we accept the fact that the oldest part of a tree chronologically is the youngest part developmentally, and attempt to utilize this fact in clonal propagation of woody plants. The organizers of the 1989 NATO Advanced Study Insti tute (ASI) on the "Molecular Basis of Plant Aging" recognized these and other dilemmas we face in undQrstanding the behaviour of woody plants. They explicitly state that "Aging is a phenomenon of considerable theoretical importance in relation to morphogenic control, differentiation and determination in plant development. It also has practical importance due to its implication in several economical areas such as flowering, fruit set, "in vitro" tree manipulation, genetic improvement, etc".

Al though many theories have been advanced to explain aging, the mechanisms involved in this process still are among the least understood factors in biology. Thus, to increase our knowledge in this scientific field will be very useful for a better understanding of plant biology. Due to the loss of morphogenic competence, aging is one of the most critical barriers to capture special genetic trai ts of selected trees. In order to fully exploit the possibili ties that tissue culture technologies provide for forest tree improvement, it is imperative to establish first the conditions to maintain plant tissues in their most reactive phase.

Thus I was delighted to accept the invitation of Prof. Roberto Rodriguez of the University of Oviedo, to participate in this ASI, which was held in Ribadesella, Asturias, Spain from June 25 to July 8, 1989. The objective of such ASI is to disseminate advanced knowledge not yet in University curricula and foster international scientific contacts through high level teaching courses. The organizers of this ASI accepted this mandate, and rose to the challenge of providing a detailed description of the process which occurs during aging. Thus, 'Some 70 participants from 15 countries, consisting of graduate students, post doctoral fellows, recently and long established researchers from Uni versi ties, Government Research Insti tutions and Private Industry assembled in ribadesella to address their charge.

The programme was divided into five sections, namely, 1. Juvenility, maturation and , 2. Vegetative propagation, advantages and limitations, 3. Ultrastructural, genetic and biochemical characteristics of aging and senescence, 4. Modulators of aging and maturation, and 5. Genetic manipulations. Information was presented in the form of major lectures (review and primary data), posters and short presentations. The work of each section ended with a round-table discussion. The results of these deliberations are captured in this volume. It is not easy to summarize in a few sentences the gist of a meeting with so wide a theme and with such a large number of presentations, both oral and by posters. Nevertheless, in a very superficial way, one can divide the meeting in two very broad areas. These were: 1) problems and practices of woody plant tissue culture, and 2) biochemical and molecular aspects of aging. Any such division is bound to be arbitrary and several papers in this volume fall into nei ther area, and perhaps this division structuralizes the ASI, which has led to the production of this very important volume. A number of papers dealt with the culture and manipulation of woody plants, particularly as these were affected by the maturation process. Some technology, such as use of the vibratome for explant production, thin layer methodology and micrografting techniques, was also presented. Factors affecting culture establishment and multiplication, problems of vitrification, polyphenol production, generation of somaclonal variation, the use of minimal culture media, including the possible harmful effects of , and the gEl.seous environment on the cultures all received attention. Because mature woody species are less responsive in culture, not only was the nature of the phase-change discussed, but empirical approaches to reactivate or reinvigorate woody explants for ease of manipulation in culture were also described. Some physiological and biochemical aspects of regeneration were also covered. The second major focus was on the genetic biochemical and molecular basis of phase change, and aging in woody species. The use of information from senescent herbaceous systems served as a model for understanding aging and its related phenomena, as many of the manifestations of aging are similar to senescence. Nitrogen metabolism, oxidative stress, free radical generation, and the role of phytohormones were among the topics covered. One view that seemed to emerge is that the rate of results from aging, a balance between oxidative stress damage and regenerative repair capacity. Lastly, the potential genetic manipulation of forest trees by traditional and molecular approaches to generate the forest tree of tomorrow highlighted the last session. The value of NATO ASIs, such as this one, cannot be overestimated. It brought together citizens of various countries, scientists from different disciplines, people of both sexes and of different ages and experiences for a period of two weeks to think on the common topic. Different perspectives were brought to bear on the subject and the format allowed for the free and unfettened exchange of ideas. Although many questions remain uncovered, we are all a little clearer in our understanding of aging and maturation in woody plants, of the role that stress plays in tissue cultures, etc. This volume' allows us to share our experiences with a much wider audience. A trust that all of you who read and study their articles will gain as much knowledge and pleasure in doing so, as we who participated in this ASI had during our two weeks in Ribadesella. The organizers and Scientific Committee under the Directorships of Roberto Rodriguez and Don Durzan deserve our sincere gratitude for job well done.

Trevor A. Thorpe Professor of Botany, Department of Biological Sciences; and Associate Dean (Research) Faculty of Science, Univ. of Calgary, Alberta, Canada.

REFERENCES GREENWOOD, M. 1988. The effect of phase change on annual growth increment in Eastern Larch. INRA-IUFRO International Symposium on Forest Tree Physiology, Nancy, France, Sept. 25-30. LEOPOLD, A.C. 1975. Aging, senescence and turnover in plants. BioScience 25: 654-662.

NORDEN, L.D •. 1988. The phenomena of senescence and aging. In: "Senescence and Aging in Plants" (Norden LD and Leopold, AC, eds.). pp. 1-50. Academic Press, New York. NORDEN, L.D. and LEOPOLD, A.C. 1988. Senescence and Aging in Plants. Academic Press, New York. 526 pp.

SCHAFFALITSKY DE MUCKADELL, M. 1959. Investigations on aging of apical meristems in woody plants its importance in silviculture. Torstl. Torsgsv. Danm. 25: 310-455. CONTENTS

SECTION I: AGING, MATURATION AND REJUVENATION

Aging of Meristems and Morphogenetic Potentialities ••••••••••••••• 3 V.S. Trippi

Maturation and Senescence: Types of Aging ••••••••••••••••••••••••• 11 V.S. Trippi

Adult vs. Juvenile Explants: Directed Totipotency ••••••••••••••••• 19 D.J. Durzan

Recovery of Transient Juvenile Capacities During Micropropagation of Filbert •••..••.••••••••••.••••••••••••• 27 C. Diaz-Sala, M. Rey and R. Rodriguez

Juvenility and Maturity of Woody Species in New Zealand ••••••••••• 37 J.L. Oliphant

Development of Rejuvenation Methods for "in vitro" Establishment, Multiplication and Rooting of Mature Trees...... 43 A. Ballester, M.C. Sanchez, F.C. San-Jose, F.J. Vieitez and A.M. Vieitez

Aging in Tree Species: Present Knowledge...... 51 A. Meier-Dinkel amd J. Kleinschmit

SECTION II: VEGETATIVE PROPAGATION: ADVANTAGES AND LIMITATIONS

Special Problems and Prospects in the Propagation of Woody Species...... 67 T.A. Thorpe and I.S. Harry

Factors Affecting Tissue Culture Success in Mass Propagation. • • • • • • . • • • • • . • . . • • • • • • • • • • • • • • • • • • . • • . • • • • • • • • • 75 P.L. Pasqualetto

Effective Handling of Plant Tissue Culture ••••••..••••••.•..•••.•• 81 R. Sanchez Tames, B. Fernandez Muniz and J.P. Majada

vii "In vitro" Micromul tiplication of Grapevine: Effect of Age, Genotype and Culture Conditions on Induction of Callus in Vitis spp. Leaf Segments •••••••••••••••••••••• 89 K.A. Roubelakis-Angelakis and K. Katsirdakis

Plant Cells and Protoplast Immovilization as Tools for Studies on Cell Function, Metabolism and Differentiation ...... ,e • • • • • • • • • • • • • • • • • • • • • • • 97 M.S. Pais and J.M.S. Cabral.

Recovery of Somatic Variation in Resistance of Populus to Septoria Musi va. • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •• 113 M.E. Ostry, D.D. Skilling, O.Y. Lee-Stadelmann W.P. Hackett

Shoot-Tip Grafting "in vitro" of Woody Species and its Influence on Plant Age...... 117 L. Navarro

The Control by Cryopreservation of Age-Related Changes in Plant Tissue Culture...... 125 E. Benson and K. Harding

Vitrification in Plant Tissue Culture ••••••••••••••••••••••••••••• 133 P.L. Pasqualetto

SECTION III: ULTRASTRUCTURAL, GENETIC AND BIOCHEMICAL CHARACTERISTIC OF AGING AND SENESCENCE

The Molecular Genetics of Maturation in Eastern Larch (Larix laricina (Du Roi) K. Koch) ••••••••••••••••••••••••••• 141 K. Hutchison, M. Greenwood, C. Sherman, J. Rebbeck and P. Singer

Cellular, Biochemical and Molecular Characteristics Related to Maturation and Rejuvenation in Woody Species...... • ...... 147 W.P. Hackett, J.R. Murray, H.H. Woo, R.E. Stapfer and R. Geneve

Uptake Characteristics of Sugars and Amino Acids by Vitis vinifera L. Protoplasts •••••••••••••••••••••••••••••• 153 K.A. Roubelakis-Angelakis and P.A. Theodoropoulos

Improvement of Micropropagat~ori Methods Linked to Biochemical Properties During "in vitro" CuI tures...... 161 A.M. Hirsch and D. Fortune

Age Dependence of Different Components of Variance •••••••••••••••• 169 M. Huehn and J. Kleinschmit

Interactions Between Genotype and Developmental Factors Modifying Peroxidase Expression •••••••••••••••••••• 183 A.M. Hirsch

viii Organogenesis: Structural, Physiological and Biochemical Aspects .•...... ,e • • • • • • • • • • • • • • • • • • • • • • • • • • • • •• 191 T.A. Thorpe

Changes in Carbohydrate Metabolism During Transient Senescence of Mustard (Sinapsis alba L.)· Cotyledons...... 199 H. 1. Kasemir

Study of Gene Expression During "in vitro" Culture of Tobacco Thin Cell Layers by Two-Dimensional Electrophoresis of Prote~ns •••••••••••••••••••••••••••••••• 207 K. Tran Thanh Van, M. Zivy, A. Cousson and H. Thiellement

An EXi)erimental Model for the Analysis of Plant Cell Differentiation: Thin Cell Layer Concept, Strategy, Methods, Records and Potantial ••••••••••••••••••• 215 K. Tran Thanh Van. L. Richard and G.A. Cyrille

Nutrient Remobilization, Nitrogen Metabolism and Chloroplast Gene Expression in Senescent Leaves. • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • . • • . • • • . • . • . • • • • • • • •• 225 B. Sabater, A. Vera, R. Tomas and M. Martin

Oxidative Stress and Senescence in Oat Leaves ••••••••••••••••••••• 231 V.S. Trippi

Structure and Function of Glutamate Dehydrogenase During Transient Senescence of Mustard (Sinapsis alba L.) Cotyledons •••••••••••••••••••••••••••••• 239 W. Lettgen, L. Britsch and H.I. Kasemir

SECTION IV: MODULATION OF AGING AND MADURATION

Effects of Vascular Tissue Containing a of Adjacent Pith: Hormone-Like Factors ••••••••••••••••••••••• 249 J. Lustinec

Hormonal Control of Senescence ••••••.••••••••••••••••••••••••••••• 257 B. Sabater, M. Martin, F.J. Sanchez and A. Vera

Modulation, Purification and Function of a Soluble Auxin Receptor...... 263 P.C.G. Van der Linde and A.M. Mennes

Free Radicals in Stressed and Aging Plant Tissue Cui tures. • • . • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •• 269 E. Benson

Po1yamines and Aging: Effect of Polyamine Biosynthetic Inhibitors on Plant Regeneration in Maize Callus CuI tured "in vi tro" ...... 277 A.F. Tiburcio, X. Figueras, I. Claparols, M. Santos and J.M. Torne

ix Hormone Action and Sensitivity: Possible Relation to Aging .••••••• 285 P.C.G. Van der Linde

SECTION V: GENETIC MANIPULATION

Cellular and Morphogenic Reorientation Induced by Inserting Foreing DNA...... 295 A. Boronat

Genetic Manipulation of Forest Trees: Implications for Physiological Processes •••••••••••••••••••••••••••••••• 303 D.J. Durzan

Biotechnology in Forest Tree Improvement: Trees of the Future...... 311 V. Chalupa

Biotechnology in Forest Tree Improvement: Trees of the Future...... 319 J. Kleinscdhmit and A. Meier-Dinkel

SELECTED POSTERS

Micropropagation of Vitis vinifera from "Vinho Verde" Region of Portugal: A Method for Grapevine Leafroll Virus Elimination .•••••••.•••..••.•.•••••••••••••• 327 M. Casal, and M.S. Pais

"In vitro" Culture of Pistacia vera L. Embryos and Aged Trees Explants...... 335 A. Gonzalez and D. Frutos

Tissues Culture and Regeneration in Jojoba •••••••••••••••••••••••• 339 S. Unique

Effect of Cold Temperature on Shoot Regeneration "in vitro" From Aged Cultures of GF-677 (Prunus persica x Prunus amygdalus) •••••••••••••••••••••••• 345 K. Dimasi-Theriou and A. Economou

Age and Meristem "in vitro" Culture Behaviour in Filbert. • • • • • • • • • • • • • • • • • • • • • • • . • • • • • • • • • • • • • • . • . • • • • • • • • •• 351 M~.T. Fernandez, M. Rey, C. Diaz-Sala and R. Rodriguez

Influence of Explant Source on "in vitro" Axillary Shoot Formation in Oak (Quercus robur L.) Seedlings. • • • • • • • • • . • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • . • • • • • . •• 357 H. Volkaert

Segmentation Effect of Immature Spike on Triticale Calli Induction...... 361 H. Guedes-Pinto, O. Pinto-Carnide and F. Leal

Cuticle Development in Dianthus caryophyllus Plantlets ••.••••••••• 367 M.A. Fal, P. Bernad, R. Obeso and R. Sanchez Tames

x Partial "Rejuvenation" of 20-Year-Old Sitka Spruce by Lower Crown Pruning...... 373 C. Selby, R. Lee and B.M.R. Harvey

Phase Change in Seqoiadendron giganteum ••••••••••••••••••••••••••• 377 O. Monteuuis and M.C. Bon

Rejuvenation of Adult Specimens of Castanea sativa Mill: Through "in vitro" Micropropagation •••••••••••••••••••••••• 383 J.A. Feijo and M.S. Pais

Embryogenic Cultures of Tobacco Pollen as a Model System To Study Plant Rejuvenation •••••••••••••••••••••••••••••••• 389 O. Vicente, D. Garrido, N. Eller, R.M. Benito Moreno, A. Alwen and E. Heberle-Bors

The Meristematic Calli of Maize: A Maintenance System of Tissue Juvenility...... 395 J.M. Torne and M.A. Santos

Growth and Development "in vitro" Long Term Cultures •••.•••••••••• 399 R.W. Joy IV and T.A. Thorpe

Protein Patterns on Corylus avellana L. Rooting Capacity •••••••••• 405 A. Gonzalez, R.S. Tames and R. Rodriguez

Inducing Artificial Tetraploids from Diploid Meadow Fescue (Festuca pratensis Huds.) Varieties and the Investigation of Some Meiotic Characteristics of Them •••••.•••••••••••.•••••••••••••••••• 411 B. Deniz

Inactivation of Peroxidase. Its Role in Plant Senescence •••••••••• 417 M. Acosta" M.B. Arnao, J .A. del Rio, J .L. Casas J. Sanchez-Bravo and F. Garcia-Canovas

Electron Microscope Observations and BA Effects in Apple Cells...... 423 R.J. Ordas, B. Fernandez and R. Rodriguez

Polyamine and Ethylene Metabolisms During Tomato Frui t Ripening...... 429 M. Acosta, J.L. Casas, J.A. del Rio, M.B. Arnao A. Ortufio and F. Sabater

Levels of Cytokinins in Aging and Rejuvenated Corylus avellana L. Tissues ••.••••••••••••••••••••••••••••• 435 B. Fernandez, R. Rodriguez, M.J. Canal, H. Andres and A. Rodriguez

Changes in Polyamines Related With Pruning As a Method for Rejuvenation in Filbert •••••••••••••••••••• 439 M. Rey, R. Astorga, C. Diaz-Sala, A.F. Tiburcio and R. Rodriguez

Early Flowering in Seedlings of Asparagus Promoted by Di uron and Atrazine. • • • • • • • • • • • • • • • . • • • • • • . • • • • • • • • • • •.• 445 M.L. Gonzalez Castanon

Index. . . . • • • • . • . • . • • • • • . . . • • . . • ...... • ...... • . • ...... •. 449

xi