Plant Biochemistry

1049 Pages·1965·29.755 MB·English

by  James Bonner and J. E. Varner (Eds.)

PLANT BIOCHEMISTRY
EDITED BY
JAMES BONNER
DIVISION OF BIOLOGY
CALIFORNIA INSTITUTE OF TECHNOLOGY
PASADENA, CALIFORNIA
J. E. VARNER
MSU/AEC PLANT RESEARCH LABORATORY
MICHIGAN STATE UNIVERSITY
EAST LANSING, MICHIGAN
1965
@
ACADEMIC PRESS New York and London
COPYRIGHT© 1965, BY ACADEMIC PRESS INC.
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LIST OF CONTRIBUTORS
Numbers in parentheses indicate the pages on which the authors' contributions begin.
T. AKAZAWA,* The International Rice Research Institute, Los Banos,
Laguna, Philippines (258)
PETER ALBERSHEIM, Department of Chemistry, University of Colorado,
Boulder, Colorado (151, 298)
B. AXELROD, Department of Biochemistry, Purdue University, Lafayette,
Indiana (231)
ROBERT S. BANDURSKI, Department of Botany and Plant Pathology, Mich
igan State University, East Lansing, Michigan (467)
J. A. BASSHAM, Lawrence Radiation Laboratory, University of California,
Berkeley, California (875)
LAWRENCE BOGORAD, Department of Botany, University of Chicago, Chi
cago, Illinois (717)
JAMES BONNER, Division of Biology, California Institute of Technology,
Pasadena, California (3, 21, 38, 213, 665, 850)
WALTER D. BONNER, JR., Johnson Foundation, University of Pennsyl
vania, Philadelphia, Pennsylvania (89)
R. H. BURRIS, Department of Biochemistry, University of Wisconsin,
Madison, Wisconsin (961)
EMANUEL EPSTEIN, Department of Soils and Plant Nutrition, University
of California, Davis, California (438)
L. FOWDEN, Botany Department, University College, London, England
(361)
J. B. HARBORNE, John Innes Institute, Hertford, England (618)
ERICH HEFTMANN, WURDD, ARS, U. S. Department of Agriculture,
Division of Biology, California Institute of Technology, Pasadena,
California (693)
* Present address: Nagoya University School of Agriculture, Anjo, Aichi, Japan.
v
Vi LIST OF CONTRIBUTORS
ROBERT W. HOLLEY, U. S. Plant, Soil, and Nutrition Laboratory, U.S.D.A.,
and Department of Biochemistry, Cornell University, Ithaca, New
York (346)
EUGENE F. JANSEN, Western Regional Research Laboratory, Albany, Cali
fornia (641)
BESSEL KOK, Bioscience Department, RIAS, Baltimore, Maryland (903)
J. A. LOCKHART, Department of Plant Physiology, University of Hawaii,
Honolulu, Hawaii (826)
A. C. NEISH, Atlantic Regional Laboratory, National Research Council
of Canada, Halifax, Nova Scotia, Canada (581)
RODERIC B. PARK, Botany Department and Lawrence Radiation Labora
tory, University of California, Berkeley, California (124)
GERHARD W. E. PLAUT, Laboratory for the Study of Hereditary and
Metabolic Disorders, and Departments of Biological Chemistry and
Medicine, University of Utah College of Medicine, Salt Lake City,
Utah (391)
S. L. RANSON, University of Newcastle upon Tyne, Newcastle upon
Tyne, England (493)
JOHN H. RICHARDS, Gates and Crellin Laboratories of Chemistry, Cali
fornia Institute of Technology, Pasadena, California (526)
MARY SPENCER, Departments of Plant Science and Biochemistry, Uni
versity of Alberta, Edmonton, Alberta, Canada (793)
P. K. STUMPF, Department of Biochemistry and Biophysics, University
of California, Davis, California (322)
T. SWAIN, LOW Temperature Research Station, Cambridge, England
(552)
GUY A. THOMPSON, JR., Department of Biochemistry, University of Wash
ington, Seattle, Washington (64)
J. E. VARNER, MSU/AEC Plant Research Laboratory, Michigan State
University, East Lansing, Michigan (14, 189, 213, 763, 867)
CLAIRE HUMMEL WINESTOCK, Laboratory for the Study of Hereditary
and Metabolic Disorders, and Departments of Biological Chemistry
and Medicine, University of Utah College of Medicine, Salt Lake
City, Utah (391)
PREFACE
This treatise is intended for the advanced student or professional
worker in the plant sciences. It is directed to the biochemist who wishes
to become informed about areas of biochemistry which are unique to
plants, for example, cell wall matters, photosynthesis, or nitrogen fixation,
or who may wish to discover to what degree plants share biochemical
pathways found in other organisms. This work will also be valuable to
plant biologists in general. Biochemistry can and does contribute to the
understanding and solution of the problems involved in many of the
more specialized aspects of plant biology—taxonomy, morphology, ecol­
ogy, horticulture, agronomy, phytopathology, to name a few. We believe
this book can help students and research workers in these diverse fields
by providing them with a ready source of biochemical information
directly applicable to plants. Finally, we feel that it can be used suc­
cessfully as a text in plant biochemistry courses. The student in such a
course would need some background in organic chemistry, but previous
study of biochemistry would not necessarily be required.
We have tried to present each topic comprehensively in the sense
that we have started with general principles and ended with the current
state of the subject. We hope that the reader, after having studied a
topic in this book, will find himself qualified to go into his laboratory
and start investigations possessing the latest knowledge available in that
field. To assist the research worker we have included references per­
tinent to the original literature; to assist the student we have also
included suggestions for more general reading on each topic. The
student without previous knowledge of biochemistry will find such read­
ing desirable—perhaps necessary.
In the fifteen years since the appearance of our first "Plant Biochem­
istry" we have seen a knowledge explosion in the fields of modern
biochemistry and molecular biology. Plant biochemistry has participated
in this vast increase of information and insight. The format of the present
volume reflects these welcome changes in our science. No longer is it
possible for one person to retain the knowledge required for the pres­
entation of all of plant biochemistry at a high level. We have therefore
vii
Vili PREFACE
asked our colleagues to assist us with contributions in their specialized
fields, while we have contented ourselves with covering those topics
in which we are competent.
We wish to make a special acknowledgment not only of the enthusi­
asm with which our collaborators, the contributors to this treatise, agreed
to accept their tasks, but of the speed and energy with which they have
completed them. We are deeply indebted to them. We are indebted also
to the staff of Academic Press, both for having encouraged us to begin
this volume and for their continuous and skillful help during its prepara­
tion. One of us (J.E.V.) gratefully acknowledges the support of RIAS,
a division of the Martin Company, Baltimore, Maryland, during the early
period in the preparation of this volume. A final note of appreciation is
due to the directors of the Herman Frasch Foundation for Agricultural
Chemistry who by their policy of supporting basic work in plant bio­
chemistry have done so much to promote our present understanding of
this field.
November, 1965 JAMES BONNER
J. E. VARNER
Chapter 1
CELL AND SUBCELL
JAMES BONNER
I. Introduction 3
II. The Subcellukr Components of the Plant Cell 3
III. The Logic of Cell Life 6
IV. Methods of Cell Fractionation 7
V. Assignment of Enzyme Activities to Individual Subnuclear Fractions . 11
VI. Cell as Community 12
References 13
I. INTRODUCTION
One of the most powerful generalizations of biochemistry is that cells
of all kinds and of all creatures possess the same, rather small, number
of kinds of subcellular components. These subcellular entities are similar
as between the different kinds of cells not only in their morphology and
submicroscopic structure, but also in chemical composition and, most
importantly, in chemical function, each kind contributing its own mite
to the overall functioning of the cell. The untangling of the biochemical
pathways of metabolism, and the development of our understanding of
the strategy of life, has been due very largely to the technology which
has developed since approximately 1950 by means of which it has
become possible to separate the several subcellular components from
one another and to identify the enzyme systems associated with each.
We turn our attention therefore first to the subcellular components of
the plant cell.
II. THE SUBCELLULAR COMPONENTS OF THE PLANT CELL
The principal subcellular components of the plant cell, and those
with which biochemistry in its present stage is principally concerned, are
the nucleus, the chloroplasts, the mitochondria, the ribosomes, messenger
ribonucleic acid (RNA), and the individual soluble enzymes. Table I
summarizes the number of each of these kinds of entities to be found
in a typical or average cell. The vast majority of plant cells contain, of
3
4 J. BONNER
course, one nucleus although many instances of multinucleate cells are
known—even in higher plants, as, for example, the latex vessels or the
sieve tubes, cells which are multinucleate by virtue of dissolution of
transverse cell walls. Chloroplasts in the photosynthetic portion of the
plant number in general a few tens, fifty as a rough average. To this
number should perhaps be added the proplastids, from which mature
chloroplasts arise, but since we have still today no good estimate of the
number of proplastids to be found in a typical cell, we will not further
consider them. The proplastids are found in the nonphotosynthetic as
well as in the photosynthetic portions of the plant—e.g., in roots. Mito­
chondria characteristically occur in the plant cell in the order of hun­
dreds, five hundred to one thousand being a typical number. Ribosomes,
the next smaller category of particle, occur in vastly greater numbers
TABLE I
NUMBERS AND SIZES OF SUBCELLULAR PARTICLES OF VARIOUS
CLASSES PRESENT IN A TYPICAL PLANT CELL
Subcellular Number
particle Diameter per cell
Nucleus 5-20 1
μ
Chloroplasts 5-20 μ 50-200
Mitochondria 1-5 μ 500-2000
Ribosomes 250 A 5 to 50 X IO6
Enzyme molecules 20-100 Â 5 to 50 X IO8
than chloroplasts or mitochondria. A growing functional plant cell might
perhaps contain a few hundred thousand ribosomes, although this num­
ber varies greatly with age, state of senescence, and so on. The bulk of
the cytoplasmic protein, the portion to which we refer in general as the
nonparticulate cytoplasm is, of course, composed of enzyme molecules—
in fact, of a great number of kinds of enzyme molecules. The total
number of enzyme molecules in a typical cell will be of the order of one
thousand million. These consist of several thousand, perhaps one to ten
thousand, different species of enzyme molecules, each qualified to
catalyze a specific kind of chemical reaction. A typical plant cell might
then contain one thousand million enzyme molecules of ten thousand
different kinds, one hundred thousand representatives of each of the ten
thousand species being present. The proportion which any individual
kind of enzyme molecule constitutes of the total soluble cytoplasmic
protein often departs widely, however, from the average one-thousandth
of one per cent which would be expected on the basis of the above
calculation. Thus we know that particular kinds of enzyme molecules in
1. Cell and Subcell 5
particular kinds of cells may constitute from a few tenths to as much
as several per cent of total soluble protein. As a general rule, however,
we must expect, because there are so many kinds of enzyme molecules,
that each one will constitute but a small portion of the total, and it is
not surprising therefore, that in the purification of enzymes enrichments
of ten thousandfold or more are not uncommonly needed to achieve
pure material.
We have referred above to messenger RNA as a typical component
of the plant cell. Messenger RNA may indeed be isolated and char­
acterized by methods to be considered in a later chapter. For the time
being, it may be noted that messenger RNA may most easily and char­
acteristically be detected in the plant cell by virtue of its interaction with
the ribosomes. Ribosomes interact with, and attach to, messenger RNA,
and since a single messenger RNA strand may simultaneously bind many
ribosomes, the great bulk of the ribosomes of the plant cell are often
detected as large aggregates—the so-called poly somes. Transfer RNA,
characterized by its small molecular size, like the soluble enzymes is a
component of the nonparticulate, cytoplasmic material and for this
reason is often known in the literature of the cell as soluble RNA.
The entire assemblage of subcellular structures outlined above is, of
course, contained within a membrane system, and we might properly
include the membrane system as one of the most characteristic of cellular
components. This membrane system comprises not only the protoplasmic
membrane itself, but also the membranes which surround nucleus, chloro-
plast, and vacuole as well as the membranous elements of the cytoplasm,
the endoplasmic reticulum, discussed in detail in Chapter 5. The plant
cell is characterized also by the cell wall external to the protoplasmic
membrane, and in a sense, the wall too might be considered as a sub­
cellular component characteristic of the plant cell. The plant cell contains
still further subcellular systems with which we cannot, at present, be con­
cerned in detail because of lack of knowledge of their function or
biochemistry. Among these subcellular components whose study remains
a challenge for the future are, for example, the Golgi bodies, mem­
branous elements of characteristic structure known from electron micros­
copy. In this category also we may place the spindle fibers, which are
responsible for the movements of chromosomes at mitosis and meiosis,
as well as the spindle fiber-generating organelles. Further subcellular
components of more evident function are the starch grains, considered
in Chapter 12; fat droplets, considered in Chapter 14; the calcium oxalate
crystals characteristic of many cells, considered in Chapter 20; and the
aleurone grains, those dense protein bodies in which the reserve protein
of seeds are characteristically deposited, considered in Chapter 29.
6 J. BONNER
III. THE LOGIC OF CELL LIFE
Before we consider in detail the operation of individual subcellular
systems, it will be helpful to consider the overall logic or strategy which
the cell uses in the conduct of its affairs. The enzyme molecules conduct,
of course, the transformation of available substrates into the kinds of
molecules, the building blocks, from which further cell components are
to be made. It is a basic law of biology that, for each kind of chemical
reaction conducted in a cell, there is a kind of enzyme molecule which
catalyzes that reaction. It is by this means that the living organism
selects from all thermodynamically possible chemical reactions those
which it will use in cellular metabolism. The enzyme molecules of varied
kinds are thus the basic elements of cellular transformations, and their
presence is the basic requirement for life and growth. All the other
subsystems of the cell are associated directly or indirectly with the
production of enzyme molecules. Since enzyme molecules are not alive
and cannot reproduce themselves, they must be synthesized, that is,
assembled from their constituent amino acids. The function of enzyme
synthesis is shared by ribosomes and messenger RNA. The long-chain
molecules of messenger RNA may be likened to punched tapes contain­
ing information about the sequence in which amino acids are to be
assembled to make a particular kind of enzyme molecule. Ribosomes
decode this information and, with the assistance of transfer RNA and
appropriate specialized kinds of enzyme molecules, considered in Chap­
ter 15, assemble enzyme molecules. The production of ribosomes then,
is an important task, and it is one function of the nucleolus to produce
the ribosomes (Chapter 4). The generation of messenger RNA is the
function of the chromosomes of the nucleus. We know, for plant as for
all cells, that the formation of each kind of enzyme is controlled by a
gene or genes of the genetic material. The genetic material, made of
DNA, possesses the ability to print off copies of itself. These copies, each
containing the information of one or a few genes are the messenger RNA
molecules, which are then available for decoding by the ribosomes.
Finally, it is the function of the nucleus not only to produce ribo­
somes and messenger RNA, but in addition to replicate the genetic
material. Such replication of the DNA is requisite to cell division, and
by means of such replication it is assured that each daughter cell gets
a complete copy of the genetic information, the information about how
to make all of the kinds of enzyme molecules required in the cellular
economy (Fig. 1).
Where, in the logic of cell life, do chloroplasts and mitochondria fit
in? Chloroplasts and mitochondria are both relatively large bodies sur-