Plant Cell Biology: From Astronomy to Zoology

By Randy O. Wayne

Plant Cell Biology, Second Edition: From Astronomy to Zoology connects the fundamentals of plant anatomy, plant physiology, plant growth and development, plant taxonomy, plant biochemistry, plant molecular biology, and plant cell biology. It covers all aspects of plant cell biology without emphasizing any one plant, organelle, molecule, or technique. Although most examples are biased towards plants, basic similarities between all living eukaryotic cells (animal and plant) are recognized and used to best illustrate cell processes. This is a must-have reference for scientists with a background in plant anatomy, plant physiology, plant growth and development, plant taxonomy, and more.

• Includes chapter on using mutants and genetic approaches to plant cell biology research and a chapter on -omic technologies
• Explains the physiological underpinnings of biological processes to bring original insights relating to plants
• Includes examples throughout from physics, chemistry, geology, and biology to bring understanding on plant cell development, growth, chemistry and diseases
• Provides the essential tools for students to be able to evaluate and assess the mechanisms involved in cell growth, chromosome motion, membrane trafficking and energy exchange

• Language: English
• Publisher: Academic Press
• Release date: Nov 13, 2018
• ISBN: 9780128143728

Randy O. Wayne

Randy O. Wayne is a plant cell biologist at Cornell University notable for his work on plant development. In particular, along with his colleague Peter K. Hepler, Wayne established the powerful role of calcium in regulating plant growth; accordingly, their 1985 article, Calcium and plant development, was cited by at least 405 subsequent articles to earn the "Citation Classic" award from Current Contents magazine and has been cited by hundreds more since 1993. He is an authority on how plant cells sense gravity through pressure, on the water permeability of plant membranes, light microscopy, as well as the effects of calcium on plant development. He has published over 50 articles and is the author of another book, Light and Video Microscopy.

Book preview

Plant Cell Biology

From Astronomy to Zoology

Second Edition

Randy Wayne

Table of Contents

Cover image

Title page

Copyright

Dedication

Preface to the First Edition

Preface to the Second Edition

Chapter 1. On the Nature of Cells

1.1. Introduction: What Is a Cell?

1.2. The Basic Unit of Life

1.3. The Chemical Composition of Cells

1.4. A Sense of Cellular Scale

1.5. The Energetics of Cells

1.6. Are There Limits to the Mechanistic View?

1.7. The Mechanistic Viewpoint and God

1.8. What Is Cell Biology?

1.9. Summary

1.10. Questions

Chapter 2. Plasma Membrane

2.1. The Cell Boundary

2.2. Topology of the Cell

2.3. Evidence for the Existence of a Plasma Membrane

2.4. Structure of the Plasma Membrane

2.5. Isolation of the Plasma Membrane

2.6. Chemical Composition of the Plasma Membrane

2.7. Transport Physiology

2.8. Electrical Properties of the Plasma Membrane

2.9. Characterization of Two Transport Proteins of the Plasma Membrane

2.10. Plasma Membrane–Localized Physiological Responses

2.11. Structural Specializations of the Plasma Membrane

2.12. The Cytoskeleton–Plasma Membrane–Extracellular Matrix Continuum

2.13. Summary

2.14. Questions

Chapter 3. Plasmodesmata

3.1. The Relationship Between Cells and the Organism

3.2. Discovery and Occurrence of Plasmodesmata

3.3. Structure of Plasmodesmata

3.4. Isolation and Composition of Plasmodesmata

3.5. Permeability of Plasmodesmata

3.6. Summary

3.7. Questions

Chapter 4. Endoplasmic Reticulum

4.1. Significance and Evolution of the Endoplasmic Reticulum

4.2. Discovery of the Endoplasmic Reticulum

4.3. Structure of the Endoplasmic Reticulum

4.4. Structural Specializations That Relate to Function

4.5. Isolation of RER and SER

4.6. Composition of the Endoplasmic Reticulum

4.7. Function of the Endoplasmic Reticulum

4.8. Summary

4.9. Questions

Chapter 5. Peroxisomes

5.1. Discovery of Microbodies

5.2. Isolation of Peroxisomes

5.3. Composition of Peroxisomes

5.4. Function of Peroxisomes

5.5. Relationship Between Glyoxysomes and Peroxisomes

5.6. Metabolite Channeling

5.7. Other Functions

5.8. Biogenesis of Peroxisomes

5.9. Evolution of Peroxisomes

5.10. Summary

5.11. Questions

Chapter 6. Golgi Apparatus

6.1. Discovery and Structure of the Golgi Apparatus

6.2. Polarity of the Golgi Stack

6.3. Isolation of the Golgi Apparatus

6.4. Composition of the Golgi Apparatus

6.5. Function of the Golgi Apparatus

6.6. The Mechanism of Movement From Cisterna to Cisterna

6.7. Positioning of the Golgi Apparatus

6.8. Summary

6.9. Questions

Chapter 7. The Vacuole

7.1. Discovery of the Vacuole

7.2. Structure, Biogenesis, and Dynamic Aspects of Vacuoles

7.3. Isolation of Vacuoles

7.4. Composition of Vacuoles

7.5. Transport Across the Vacuolar Membrane

7.6. Functions of the Vacuole

7.7. Biotechnology

7.8. Summary

7.9. Questions

Chapter 8. Movement Within the Endomembrane System

8.1. Discovery of the Secretory Pathway

8.2. Movement to the Plasma Membrane and the Extracellular Matrix

8.3. Movement From the Endoplasmic Reticulum to the Golgi Apparatus to the Vacuole

8.4. Movement From the Endoplasmic Reticulum to the Vacuole

8.5. Movement From the Plasma Membrane to the Endomembranes

8.6. Disruption of Intracellular Secretory and Endocytotic Pathways

8.7. Summary

8.8. Questions

Chapter 9. Cytoplasmic Structure

9.1. Historical Survey of the Study of Cytoplasmic Structure

9.2. Chemical Composition of Protoplasm

9.3. Physical Properties of Cytoplasm

9.4. Microtrabecular Lattice

9.5. Summary

9.6. Questions

Chapter 10. Actin- and Microfilament-Mediated Processes

10.1. Discovery of Actomyosin and the Mechanism of Muscle Movement

10.2. Actin in Nonmuscle Cells

10.3. Force-Generating Reactions Involving Actin

10.4. Actin-Based Motility

10.5. Role of Actin in Membrane Transport

10.6. Summary

10.7. Questions

Chapter 11. Tubulin and Microtubule-Mediated Processes

11.1. Discovery of Microtubules in Cilia and Flagella and the Mechanism of Movement

11.2. Microtubules in Nonflagellated or Nonciliated Cells and the Discovery of Tubulin

11.3. Force-Generating Reactions Involving Tubulin

11.4. Tubulin-Based Motility

11.5. Microtubules and Cell Shape

11.6. Various Stimuli Affect That Microtubule Orientation

11.7. Microtubules and Cytoplasmic Structure

11.8. Intermediate Filaments

11.9. Centrin-Based Motility

11.10. Tensegrity in Cells

11.11. Summary

11.12. Questions

Chapter 12. Cell Signaling

12.1. The Scope of Cell Regulation

12.2. What Is Stimulus-Response Coupling?

12.3. Receptors

12.4. Cardiac Muscle as a Paradigm for Understanding the Basics of Stimulus-Response Coupling

12.5. A Kinetic Description of Regulation

12.6. Ca2+ Signaling System

12.7. Mechanics of Doing Experiments to Test the Importance of Ca2+ as a Second Messenger

12.8. Specific Signaling Systems in Plants Involving Ca2+

12.9. Phosphatidylinositol Signaling System

12.10. The Role of Ions in Cells

12.11. Summary

12.12. Questions

Chapter 13. Chloroplasts

13.1. Discovery of Chloroplasts and Photosynthesis

13.2. Isolation of Chloroplasts

13.3. Composition of the Chloroplasts

13.4. Thermodynamics and Bioenergetics in Photosynthesis

13.5. Organization of the Thylakoid Membrane and the Light Reactions of Photosynthesis

13.6. Physiological, Biochemical, and Structural Adaptations of the Light Reactions

13.7. Fixation of Carbon

13.8. Reduction of Nitrate and the Activation of Sulfate

13.9. Chloroplast Movements and Photosynthesis

13.10. Genetic System of Plastids

13.11. Biogenesis of Plastids

13.12. Summary

13.13. Questions

Chapter 14. Mitochondria

14.1. Discovery of the Mitochondria and Their Function

14.2. Isolation of Mitochondria

14.3. Composition of Mitochondria

14.4. Cellular Geography of Mitochondria

14.5. Chemical Foundation of Respiration

14.6. Other Functions of the Mitochondria

14.7. Genetic System in Mitochondria

14.8. Biogenesis of Mitochondria

14.9. Summary

14.10. Questions

Chapter 15. Origin of Organelles

15.1. Autogenous Origin of Organelles

15.2. Endosymbiotic Origin of Chloroplasts and Mitochondria

15.3. Origin of Peroxisomes, Centrioles, and Cilia

15.4. Ongoing Process of Endosymbiosis

15.5. Primordial Host Cell

15.6. Symbiotic DNA

15.7. Summary

15.8. Questions

Chapter 16. The Nucleus

16.1. The Discovery of the Nucleus and Its Role in Heredity, Systematics, and Development

16.2. Isolation of Nuclei

16.3. Structure of the Nuclear Envelope and Matrix

16.4. Chemistry of Chromatin

16.5. Morphology of Chromatin

16.6. Cell Cycle

16.7. Chromosomal Replication

16.8. Transcription

16.9. Nucleolus and Ribosome Formation

16.10. Summary

16.11. Questions

Chapter 17. Ribosomes and Proteins

17.1. Nucleic Acids and Protein Synthesis

17.2. Protein Synthesis

17.3. Protein Activity

17.4. Protein Targeting

17.5. Protein–Protein Interactions

17.6. Protein Degradation

17.7. Structure of Proteins

17.8. Functions of Proteins

17.9. Techniques of Protein Purification

17.10. Plants as Bioreactors to Produce Proteins for Vaccines

17.11. Summary

17.12. Questions

Chapter 18. The Origin of Life

18.1. Spontaneous Generation

18.2. Concept of Vitalism

18.3. The Origin of the Universe

18.4. Geochemistry of the Early Earth

18.5. Prebiotic Evolution

18.6. The Earliest Darwinian Ancestor and the Last Common Ancestor

18.7. Diversity in the Biological World

18.8. The Origin of Consciousness

18.9. Concept of Time

18.10. Summary

18.11. Questions

Chapter 19. Cell Division

19.1. Mitosis

19.2. Regulation of Mitosis

19.3. Energetics of Mitosis

19.4. Division of Organelles

19.5. Cytokinesis

19.6. Summary

19.7. Questions

Chapter 20. Extracellular Matrix

20.1. Relationship of the Extracellular Matrix of Plant and Animal Cells

20.2. Isolation of the Extracellular Matrix of Plants

20.3. Chemical Composition and Architecture of the Extracellular Matrix

20.4. Extracellular Matrix–Plasma Membrane–Cytoskeletal Continuum

20.5. Biogenesis of the Extracellular Matrix

20.6. Permeability of the Extracellular Matrix

20.7. Mechanical Properties of the Extracellular Matrix

20.8. Cell Expansion

20.9. Summary

20.10. Epilog

20.11. Questions

Chapter 21. Omic Science: Platforms and Pipelines

21.1. One, Two, Three, Infinity

21.2. Genomics

21.3. Transcriptomics

21.4. Proteomics

21.5. Interactomics

21.6. Metabolomics

21.7. Phenomics

21.8. Pan-Omics

21.9. Single-Cell Omics

21.10. One, Two, Three…Infinity Revisited

21.11. Summary

21.12. Questions

Appendix 1. SI Units, Constants, Variables, and Geometric Formulae

Appendix 2. A Cell Biologist's View of Non-Newtonian Physics

Appendix 3. Calculation of the Total Transverse Force and Its Relation to Stress

Appendix 4. Laboratory Exercises

Bibliography

Index

Copyright

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Cover pictures: Photomicrograph of diatoms arranged on a microscope slide taken with a differential interference contrast microscope (top, left); Photomicrograph of an onion epidermal cell taken with a phase contrast microscope (top, center); Photomicrograph of Paramecium bursaria taken with a differential interference microscope (top, right); Photomicrograph of cotton hair cells taken with a polarized light microscope (bottom).

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Dedication

This book is dedicated to Amy, my wife and soulmate, and to Erwin Chargaff, an Apollonian and Dionysian scientific hero who first isolated and characterized DNA as an informational macromolecule, suggested that we should sequence it, and understood where science was going.

Preface to the First Edition

This book is in essence the lectures I give in my plant cell biology course at Cornell University. Heretofore, the lecture notes have gone by various titles, including Cell La Vie, The Book Formerly Known as Cell La Vie, Molecular Theology of the Cell, Know Thy Cell (with apologies to Socrates), Cell This Book (with apologies to Abbie Hoffman), and Impressionistic Plant Cell Biology. I would like to take this opportunity to describe this course. It is a semester-long course for undergraduate and graduate students. Because the undergraduate biology majors are required to take genetics, biochemistry, and evolution as well as 1  year each of mathematics and physics and 2  years of chemistry, I have done my best to integrate these disciplines into my teaching. Moreover, many of the students also take plant anatomy, plant physiology, plant growth and development, plant taxonomy, plant biochemistry, plant molecular biology, and a variety of courses that end with the suffix -omics; I have tried to show the connections between these courses and plant cell biology. Nonbotanists can find a good introduction to plant biology in Mauseth (2009) and Taiz and Zeiger (2006).

Much of the content has grown over the past 20  years from the questions and insights of the students and teaching assistants who have participated in the class. The students' interest has been sparked by the imaginative and insightful studies done by the worldwide community of cell biologists, which I had the honor of presenting.

I have taken the approach that real divisions not only exist between subject areas taught in a university but only in the state of mind of the teachers and researchers. With this approach, I hope that my students do not see plant cell biology as an isolated subject area, but as an entry into every aspect of human endeavor. One of the goals of my course is to try to reestablish the connections that once existed between mathematics, astronomy, physics, chemistry, geology, philosophy, and biology. It is my own personal attempt, and it is an ongoing process. Consequently, it is far from complete. Even so, I try to provide the motivation and resources for my students to weave together the threads of these disciplines to create their own personal tapestry of the cell from the various lines of research.

Recognizing the basic similarities between all living eukaryotic cells (Quekett, 1852, 1854; Huxley, 1893), I discuss both animal and plant cells in my course. Although the examples are biased toward plants (as they should be in a plant cell biology course), I try to present the best example to illustrate a process and sometimes the best examples are from animal cells. I take the approach used by August Krogh (1929), that is, there are many organisms in the treasure house of nature and if one respects this treasure, one can find an organism created to best illuminate each principle! I try to present my course in a balanced manner, covering all aspects of plant cell biology without emphasizing any one plant, organelle, molecule, or technique. I realize, however, that the majority of papers in plant cell biology today are using a few model organisms and -omic techniques. My students can learn about the successes gained though this approach in a multitude of other courses. I teach them that there are other approaches.

Pythagoras believed in the power of numbers, and I believe that the power of numbers is useful for understanding the nature of the cell. In my class, I apply the power of numbers to help relate quantities that one wishes to know to things that can be easily measured (Hobson, 1923; Whitehead, 1925; Hardy, 1940; Synge, 1951, 1970; Feynman, 1965a; Schrödinger, 1996). For example, the area of a rectangle is difficult to measure. However, if one knows its length and width, and the relation that area is the product of length and width, the area can be calculated from the easily measurable quantities. Likewise, the circumference or area of a circle is relatively difficult to measure. However, if one measures the diameter and multiplies it by π, or the square of the diameter by π/4, one can easily obtain the circumference and area, respectively. In the same way, one can easily estimate the height of a tree from easily measurable quantities if one understands trigonometry and the definition of tangent.

My teaching was greatly influenced by a story that Hans Bethe told at a meeting at Cornell University commemorating the 50th anniversary of the chain reaction produced by Enrico Fermi. Bethe spoke about the difference between his graduate adviser, Arnold Sommerfeld, and his postdoctoral adviser, Enrico Fermi. He said that, in the field of atomic physics, Sommerfeld was a genius at creating a mathematical theory to describe the available data. Sommerfeld's skill, however, depended on the presence of data. Fermi, on the other hand, could come up with theories even if the relevant data were not apparent. He would make estimates of the data from first principles. For example, he estimated the force of the first atomic bomb by measuring the distance small pieces of paper flew as they fell to the ground during the blast in Alamogordo. Knowing that the force of the blast diminished with the square of the distance from the bomb, Fermi estimated the force of the bomb relative to the force of gravity. Within seconds of the blast, he calculated the force of the bomb to be approximately 20 kilotons, similar to which the expensive machines recorded (Fermi, 1954; Lamont, 1965).

To train his students to estimate things that they did not know, Fermi would ask them, How many piano tuners are there in Los Angeles? After they looked befuddled, he would say, You can estimate the number of piano tuners from first principles! For example, how many people are there in Los Angeles? One million? What percentage has pianos? Five percent? Then there are 50,000 pianos in Los Angeles. How often does a piano need to be tuned? About once a year? Then 50,000 pianos need to be tuned in a year. How many pianos can a piano tuner tune in a day? Three? Then one tuner must spend 16,667  days a year tuning pianos. But since there are not that many days in a year, and he or she probably only works 250  days a year, then there must be around 67 piano tuners in Los Angeles.

My students apply the power of numbers to the study of cellular processes, including membrane transport, photosynthesis, and respiration, to get a feel for these processes and the interconversions that occur during these processes between different forms of energy. My students apply the power of numbers to the study of cell growth, chromosome motion, and membrane trafficking to postulate and evaluate the potential mechanisms involved in these processes, and the relationships between these processes and the bioenergetic events that power them. Becoming facile with numbers allows the students to understand, develop, and critique theories. As the Greek origin of the word [theory] implies, the Theory is the true seeing of things—the insight that should come with healthy sight (Adams and Whicher, 1949).

Using the power of numbers to relate seemingly unrelated processes, my students are able to try to analyze all their conclusions in terms of first principles. They also learn to make predictions based on first principles. The students must be explicit in terms of what they are considering to be facts, what they are considering to be the relationship between facts, and where they are making assumptions. This provides a good entry into research because the facts must be refined and the assumptions must be tested (East, 1923).

I do not try to introduce any more terminology in my class than is necessary, and I try to explain the origin of each term. Some specialized terms are essential for precise communication in science just as it is in describing love and beauty. However, some terms are created to hide our ignorance, and consequently prevent further inquiry, because something with an official-sounding name seems well understood (Locke, 1824; Hayakawa, 1941; Rapoport, 1975). In Goethe's (1808) Faust Part One, Mephistopheles says: For at the point where concepts fail. At the right time a word is thrust in there. With words we fitly can our foes assail. Francis Bacon (1620) referred to this problem as the Idols of the Marketplace. Often we think we are great thinkers when we answer a question with a Greek or Latin word. For example, if I am asked, Why are leaves green? I quickly retort, Because they have chlorophyll. The questioner is satisfied, and says Oh. The conversation ends. However, chlorophyll is just the Greek word for green leaf. Thus, I really answered the question with a tautology. I really said Leaves are green because leaves are green and did not answer the question at all. It was as if I was reciting a sentence from scripture, which I had committed to memory without giving it much thought. However, I gave the answer in Greek, and with authority … so it was a scientific answer.

In An Essay Concerning Human Understanding, John Locke (1824) admonished that words are often used in a nonintellectual manner. He wrote,

… he would not be much better than the Indian before-mentioned, who, saying that the world was supported by a great elephant, was asked what the elephant rested on; to which his answer was, a great tortoise. But being again pressed to know what gave support to the broad-backed tortoise, replied, something he knew not what. And thus here, as in all other cases where we use words without having clear and distinct ideas, we talk like children; who being questioned what such a thing is, which they know not, readily give the satisfactory answer, that it is something; which in truth signifies no more, when so used either by children or men, but that they know not what; and that the thing that they pretend to know and talk of is what they have no distinct idea of at all, and so are perfectly ignorant of it, and in the dark.

Sometimes terms are created to become the shibboleths of a field, and sometimes they are created for political reasons, financial reasons, or to transfer credit from someone who discovers something to someone who renames it (Agre et al., 1995). Joseph Fruton (1992) recounted (and translated) a story of a conversation with a famous chemist in Honoré de Balzac's La Peau de Chagrin:

Well, my old friend, said Planchette upon seeing Japhet seated in an armchair and examining a precipitate, How goes it in chemistry?

It is asleep. Nothing new. The Académie has in the meantime recognized the existence of salicine. But salicine, asparagine, vauqueline, digitaline are not new discoveries.

If one is unable to produce new things, said Raphael, it seems that you are reduced to inventing new names.

That is indeed true, young man.

I teach plant cell biology with a historical approach and teach not only of the fruits but also of the trees which have borne them, and of those who planted these trees (Lenard, 1906). This approach also allows them to understand the origins and meanings of terms; to capture the excitement of the moment of discovery; to elucidate how we, as a scientific community, know what we know; and it emphasizes the unity and continuity of human thought (Haldane, 1985). I want my students to become familiar with the great innovators in science and to learn their way of doing science (Wayne and Staves, 1998, 2008). I want my students to learn how the scientists we learn about choose and pose questions, and how they go about solving them. I do not want my students to know just the results and regurgitate those results on a test (Szent-Györgyi, 1964; Farber, 1969). I do not want my students to become scientists who merely repeat on another organism the work of others. I want my students to become like the citizens of Athens, who according to Pericles do not imitate—but are a model to others. Whether or not my students become professional cell biologists, I hope they forever remain amateurs and dilettantes in terms of cell biology. That is, I hope that I have helped them become one who loves cell biology and one who delights in cell biology (Chargaff, 1986)—not someone who cannot recognize the difference between a pile of bricks and an edifice (Forscher, 1963), not someone who sells buyology (Wayne and Staves, 2008), and not someone who sells his or her academic freedom (Rabounski, 2006; Apostol, 2007).

Often people think that a science course should teach what is new, but I answer this with an amusing anecdote said by Erwin Chargaff (1986): Kaiser Wilhelm I of Germany, Bismark's old emperor, visited the Bonn Observatory and asked the director: ‘Well, dear Argelander, what's new in the starry sky?’ The director answered promptly: ‘Does your Majesty already know the old?’ The emperor reportedly shook with laughter every time he retold the story.

According to R. John Ellis (1996),

It is useful to consider the origins of a new subject for two reasons. First, it can be instructive; the history of science provides sobering take-home messages about the importance of not ignoring observations that do not fit the prevailing conceptual paradigm, and about the value of thinking laterally, in case apparently unrelated phenomena conceal common principles. Second, once a new idea has become accepted there is often a tendency to believe that it was obvious all along—hindsight is a wonderful thing, but the problem is that it is never around when you need it!

The historical approach is necessary, in the words of George Palade (1963), to indicate that recent findings and present concepts are only the last approximation in a long series of similar attempts which, of course, is not ended.

I teach my students that it is important to be skeptical when considering old as well as new ideas. According to Thomas Gold (1989),

New ideas in science are not always right just because they are new. Nor are the old ideas always wrong just because they are old. A critical attitude is clearly required of every scientist. But what is required is to be equally critical to the old ideas as to the new. Whenever the established ideas are accepted uncritically, but conflicting new evidence is brushed aside and not reported because it does not fit, then that particular science is in deep trouble—and it has happened quite often in the historical past.

To emphasize the problem of scientists unquestioningly accepting the conventional wisdom, Conrad H. Waddington (1977) proposed the acronym COWDUNG to signify the Conventional Wisdom of the Dominant Group.

In teaching in a historical manner, I recognize the importance of Thomas H. Huxley's (1853b) warnings that Truth often has more than one Avatar, and whatever the forgetfulness of men, history should be just, and not allow those who had the misfortune to be before their time to pass for that reason into oblivion and The world, always too happy to join in toadying the rich, and taking away the ‘one ewe lamb’ from the poor. Indeed, it is often difficult to determine who makes a discovery (Djerassi and Hoffmann, 2001). I try to the best of my ability to give a fair and accurate account of the historical aspects of cell biology.

My course includes a laboratory section and my students perform experiments to acquire personal experience in understanding the living cell and how it works (Hume, 1748; Wilson, 1952; Ramón y Cajal, 1999). Justus von Liebig (1840) described the importance of the experimental approach this way:

Nature speaks to us in a peculiar language, in the language of phenomena; she answers at all times the questions which are put to her; and such questions are experiments. An experiment is the expression of a thought: we are near the truth when the phenomenon, elicited by the experiment, corresponds to the thought; while the opposite result shows that the question was falsely stated, and that the conception was erroneous.

My students cannot wait to get into the laboratory. In fact, they often come in on nights and weekends to use the microscopes to take photomicrographs. At the end of the semester, the students come over to my house for dinner (I worked my way through college as a cook) and bring their best photomicrographs. After dinner, they vote on the 12 best, and those are incorporated into a class calendar. The calendars are beautiful and the students often make extra to give as gifts.

In 1952, Edgar Bright Wilson Jr. wrote in An Introduction to Scientific Research, There is no excuse for doing a given job in an expensive way when it can be carried through equally effectively with less expenditure. Today, with an emphasis on research that can garner significant money for a college or university through indirect costs, there is an emphasis on the first use of expensive techniques to answer cell biological questions and often questions that have already been answered. However, the very expense of the techniques often prevents one from performing the preliminary experiments necessary to learn how to do the experiment so that meaningful and valuable data and not just lists are generated. Unfortunately, the lists generated with expensive techniques often require statisticians and computer programmers, who are far removed from experiencing the living cells through observation and measurement, to tell the scientist which entries on the list are meaningful. Thus, there is a potential for the distinction between meaningful science and meaningless science to become a blur. I use John Synge's (1951) essay on vicious circles to help my students realize that there is a need to distinguish for themselves what is fundamental and what is derived.

By contrast, this book emphasizes the importance of the scientists who have made the great discoveries in cell biology using relatively low-tech quantitative and observational methods. But—and this is a big but—these scientists also treated their brains, eyes, and hands as highly developed scientific instruments. I want my students to have the ability to get to know these great scientists. I ask them to name who they think are the 10 best scientists who ever lived. Then I ask if they have ever read any of their original work. In the majority of the cases, they have never read a single work by the people who they consider to be the best scientists. This is a shame. They read the work of others … but not the best. Interestingly, they usually are well read when it comes to reading the best writers (e.g., Shakespeare, Faulkner, etc.).

Typically, the people on my students' lists of best scientists have written books for the layperson or an autobiography (Wayne and Staves, 1998). Even Isaac Newton wrote a book for the layperson! I give my class these references and encourage them to become familiar with their favorite scientists first hand. The goal of my lectures and this book is to facilitate my students' personal and continual journey in the study of life.

My goal in teaching plant cell biology is not only to help my students understand the mechanisms of the cell and its organelles in converting energy and material matter into a living organism that performs all the functions we ascribe to life. I also hope to deepen my students' ideas of the meaning, beauty, and value of life and the value in searching for meaning and understanding in all processes involved in living.

I thank Mark Staves and my family, Michelle, Katherine, Zack, Beth, Scott, my mother and father, and aunts and uncles for their support over the years. I also thank my colleagues at Cornell University and teachers at the Universities of Massachusetts, Georgia, and California at Los Angeles, and especially Peter Hepler and Masashi Tazawa, who taught me how to see the universe in a living cell.

Randy Wayne,     Department of Plant Biology, Cornell University

Preface to the Second Edition

The first edition of this book emphasized the great ideas regarding what makes life possible on the cellular level and the great observations and experiments that stimulated, supported, and challenged those ideas. That is, the first edition emphasized the questions asked by great thinkers to understand what life is and the observations and experiments performed by great observers and experimentalists to provide answers to those questions. It was an inquiry into the origin, certainty, and extent of our cell biological knowledge and left room for further questioning and wonder. Scientific inquiry not only begins with wonder but also usually ends with it too. The questions are eternal and the answers are provisional—becoming fleshed out and sometimes changed over time. Extensive references were given so that the reader could read the original papers and develop a personal relationship with the scientists who came up with the great observations, experiments, and ideas. My goal was to facilitate the prospect that the reader would be inspired to join a century-long collaboration with the scientists who wrote the original papers. Even if the reader did not join a century-long collaboration, he or she would not only be informed from reading the original references, meaning that the reader would know that something was the answer; but also, the reader would be enlightened, knowing why something was the answer. This goal was the scientific equivalent of Mortimer Adler's (See Adler and van Doren, 1967) philosophy that drove him to edit the Great Books of the Western World series and write How to Read a Book.

Biology has now become systems biology. The approach of systems biology (Cullis, 2008) "is to accumulate information and biological resources relating to as much of the genome as possible and then determining which parts are of importance. Moreover, the questions and answers obtained in the past are often considered biased and perhaps unworthy of study. According to Sheth and Thaker (2014), the main challenge confronting the field is not to look back (incorporating previous findings is critical but will be comparatively easy) but to look forward to how one might plan and interpret the enormous new data that soon will be generated." Technology is being developed to obtain big data at the systems biology level that is unbiased. However, recognizing what bias is and what wisdom and knowledge are depends on an ability to read the relevant papers, the ability to work with one's eyes and hands to make observations and do experiments, and the ability to understand the ideas, observations, and experiments. This edition maintains and updates the philosophical outlook of the first edition, emphasizing how we know what we know, so that the reader can come to his or her own conclusion about the value of the work.

This edition also includes a new chapter entitled, Omic Science: Platforms and Pipelines, which emphasizes the great ideas and experiments that provided the basis for the technologies used in systems biology. I emphasize the development of the technologies rather than the results obtained from the technologies, as a study of the development of the technologies reveals real scientific achievement and creativity, and the explanation of their development has real pedagogical value. On the other hand, the results produced by the technologies are by necessity wanting given that the goal is to characterize everything in an unbiased manner (Chory et al., 2000), and, at the present time, we are as close to knowing nothing new from the systems approach as we are to knowing everything.

According to Bruce Alberts, Marc Kirschner, Shirley Tilghman, and Harold Varmus (2012) "the system now favors those who can guarantee results rather than those with potentially path-breaking ideas that, by definition, cannot promise success. Young investigators are discouraged from departing too far from their postdoctoral work, when they should instead be posing new questions and inventing new approaches. Seasoned investigators are inclined to stick to their tried-and-true formulas for success rather than explore new fields." Where is the freedom to think? Not only are trained scientists working like automatons but also being replaced by automatons. According to Steve Strogatz (Manjoo, 2011), "Our time is limited. As thinking machines, they have a lot of advantages over us—this is obvious…We're not going to be the best players in town. I do think we'll be put out of business. This is really going to happen."

More importantly, I'd like to ask the question: What are the goals of science? Are the goals best done by computers? Or should the goals be based on the proposition that science is a field of human endeavor that promotes freedom by training people to think. John Dewey (1910) realized, "Genuine freedom, in short, is intellectual; it rests in the trained power ofthought, in ability to ‘turn things over,’ to look at matters deliberately, to judge whether the amount and kind of evidence requisite for decision is at hand, and if not, to tell where and how to seek such evidence. If a man's actions are not guided by thoughtful conclusions, then they are guided by inconsiderate impulse, unbalanced appetite, caprice, or the circumstances of the moment. To cultivate unhindered, unreflective external activity is to foster enslavement, for it leaves the person at the mercy of appetite, sense, and circumstance. We have to take seriously what Marcus Garvey (1938) said in a speech given in Nova Scotia, We are going to emancipate ourselves from mental slavery because whilst others might free the body, none but ourselves can free the mind." Bob Marley immortalized these words in Redemption Song.

The culture of scientific research is in danger of eliminating what is necessary to cultivate the intelligent and creative use of technology. Both Jacobus van't Hoff (1967), who viewed chemicals in three-dimensional space, and Peter Mitchell (1980), who viewed metabolism in three-dimensional space as well as time, quoted Henry Thomas Buckle's (1872) description of the type of imagination needed to make great scientific discoveries: "there is a spiritual, a poetic, and for aught we know a spontaneous and uncaused element in the human mind, which ever and anon, suddenly and without warning, gives us a glimpse and a forecast of the future, and urges us to seize the truth as it were by anticipation." Yes, serendipity favors a prepared mind.

This book is written for the students who are craving to be free to do curiosity-driven science that requires thinking, imagination, and skill. It is also written for the taxpayer who expects their hard-earned money to be invested wisely. It is my hope that this book will help turn the scientific pendulum back so that science can once again encourage a way of understanding nature that is fueled by the intelligent use of technology and the scientific imagination (Holton, 1978).

I wrote the following paragraph in the Preface to the First Edition:

In 1952, Edgar Bright Wilson Jr. wrote in An Introduction to Scientific Research, "There is no excuse for doing a given job in an expensive way when it can be carried through equally effectively with less expenditure." Today, with an emphasis on research that can garner significant money for a college or university through indirect costs, there is an emphasis on the first use of expensive techniques to answer cell biological questions and often questions that have already been answered. However, the very expense of the techniques often prevents one from performing the preliminary experiments necessary to learn how to do the experiment so that meaningful and valuable data and not just lists are generated. Unfortunately, the lists generated with expensive techniques often require statisticians and computer programmers, who are far removed from experiencing the living cells through observation and measurement, to tell the scientist which entries on the list are meaningful. Thus, there is a potential for the distinction between meaningful science and meaningless science to become a blur.

In this preface, I would like to update the paragraph by substituting Facilities & Administrative (F&A) costs for indirect costs.

I would like to heartily thank Mohanapriyan Rajendran for finding so many of my mistakes, typos, and omissions. Lastly, I would like to thank Karl Niklas, a scholar and a gentleman, for the fascinating and thought-provoking conversations we have about biology and teaching every morning.

Chapter 1

On the Nature of Cells

Abstract

Life consists of the ability to move and generate electricity; to take up nutrients and expel wastes; to perform chemical syntheses of organic molecules at ambient temperatures and pressures, and therefore grow; to reproduce itself with near-perfect fidelity; and to sense and respond to changes in the external environment to maintain itself. The cell is the lowest level of organization that has the ability to perform all these processes and thus is the basic unit of life. To get an idea of the nature of cells based on first principles, I describe the discovery of cells, their dimensions, their chemical composition, and the energetics of cells.

Keywords

Edmund B. Wilson; Henri Dutrochet; Hugo von Mohl; Jean Baptiste Carnoy; Matthias Schleiden; Nehemiah Grew; Robert Hooke; Thomas H. Huxley

The world globes itself in a drop of dew. The microscope cannot find the animalcule which is less perfect for being little. Eyes, ears, taste, smell, motion, resistance, appetite, and organs of reproduction that take hold on eternity—all find room to consist in the small creature. So do we put our life into every act. The true doctrine of omnipresence is that God reappears with all His parts in every moss and cobweb.

Ralph Waldo Emerson, Compensation.

The path of modern organ physiology is straight and clear, and we are not far from a complete understanding of life as an association of organs. But the organ is an assembly of cells and its properties and activities are dependent on the properties and activities of its component cells. Organ physiology has therefore, so to speak, begun its study from the midst of life; the beginning, the basis of life is in the cell.

Ivan Pavlov (cited in Heilbrunn, 1952).

1.1. Introduction: What Is a Cell?

In the introduction to his book, Grundzüge der Botanik, Matthias Schleiden (1842), often considered the cofounder of the cell theory, admonished, Anyone who has an idea of learning botany from the present book, may just as well put it at once aside unread; for from books botany is not learnt (quoted in Goebel, 1926). Likewise, I would like to stress that an understanding of plant cell biology, and what a plant cell is, comes from direct experience. I hope that this book helps facilitate your own personal journey into the world of the cell.

Dom Pérignon, according to André Simon (1934), did not discover, invent or create sparkling Champagne. He never claimed to have done so, nor did any of his contemporaries claim any such honour for him. In fact, Champagne was already being made in England where the oiled hemp rag that was traditionally used to stopper the bottle had just been replaced by cork (see history of Vintners' Hall at http://www.vintnershall.co.uk/?page=history_of_the_hall). While the oiled hemp rag kept the dust out, the cork allowed the build up of carbon dioxide in the wine. Exploring the world made accessible by the invention of the microscope, Robert Hooke (1665) took a look at the cork that may have been used by William Russell, the first Duke and fifth Earl of Bedford to stopper champagne bottles (Simon, 1934; Taber, 2007, 2009). Serendipitously, Hooke discovered a regular, repeating structure in cork that he called a cell. The word cell comes from the Latin celle, which in Hooke's time meant "a small apartment, esp. one of several such in the same building, used e.g., for a store-closet, slave's room, prison cell; also cell of a honeycomb; … also a monk's or hermit's cell" (Oxford English Dictionary, 1933). Hooke used the word cell to denote the stark appearance of the air-filled pores he saw in the honeycomb-like pattern in the cork that he viewed with his microscope (Fig. 1.1). Hooke's perspective of the emptiness of cells was propagated by Nehemiah Grew (1682), who compared the cells of the pith of asparagus to the froth of beer (Fig. 1.2), and is still implied in words with the prefix cytos, which in Greek means hollow place. Hooke, however, did realize that there might be more to a cell than he could see. He wrote,

Now, though I have with great diligence endeavoured to find whether there be any such thing in those microscopical pores of wood or piths, as the valves in the heart, veins and other passages of animals, that open and give passage to the contained fluid juices one way, and shut themselves, and impede the passage of such liquors back again, yet have I not hitherto been able to say anything positive in it; … but … some diligent observer, if helped with better microscopes, may in time, detect [them].

Hugo von Mohl (1852) pointed out in Principles of the Anatomy and Physiology of the Vegetable Cell, the first textbook devoted to plant cell biology, that indeed plant cells are not vacuous when viewed with optically corrected microscopes but contain a nucleus and an opake, viscid fluid of a white colour, having granules intermingled in it, which fluid I call protoplasm. Von Mohl, echoing the conclusions of Henri Dutrochet (1824) and John Quekett (1852), further revealed through his developmental studies that cells have a variety of shapes (Fig. 1.3) and give rise to all structures in the plant including the phloem and xylem. This was contrary to the earlier opinions of de Candolle and Sprengel (1821), who believed that there were three elementary forms in plants—dodecahedral-shaped cells, noncellular tubes, and noncellular spirals (Fig. 1.4). By focusing on mature plants, de Candolle and Sprengel had not realized that the tubelike vessels and the spiral-like protoxylem developed from dodecahedral-shaped cells. To further emphasize the vitality of cells, von Mohl also stressed that cells were endowed with the ability to perform all kinds of movements.

Figure 1.1  Cells of cork. 

From Hooke, R., [1665], 1961. Micrographia or Some Physiological Descriptions of Minute Bodies Made by Manifying Glasses with Observations and Inquiries thereupon. Dover Publications, New York.

Figure 1.2  The cortical cells of a small root of asparagus. 

From Grew, N., [1682], 1965. The Anatomy of Plants with an Idea of a Philosophical History of Plants and Several other Lectures Read before The Royal Society. Johnson Reprint Corp., New York.

In the world of the living cell, the only thing that is certain is change—movement occurs at all levels, from the molecular to the whole cell. While I was taught that plants, unlike animals, do not move, some plants can constantly change their position. Get a drop of pond water and look at it under the microscope. Watch a single-celled alga such as Dunaliella under the microscope (Fig. 1.5). See it swim? These plant cells are Olympic-class swimmers: they swim about 50  μm/s—equivalent to five body lengths per second. Not only can the cells swim but also change their motile behavior in response to external stimuli. When a bright flash of light (from the sun or a photographic flash) strikes swimming Dunaliella cells, like synchronous swimmers, they all swim backward for about a half second. From this observation, even a casual observer will conclude that individual cells have well-developed sensory systems that can sense and respond to external stimuli (Wayne et al., 1991).

Figure 1.3  Stellate cells from the petiole of a banana. 

From von Mohl, H., 1852. Principles of the Anatomy and Physiology of the Vegetable Cell. Translated by Henfrey, A. John van Voorst, London.

Figure 1.4  Spiral vessels, sap tubes, and cells of Maranatha lutea . 

From deCandolle, A.P., Sprengel, K., 1821. Elements of the Philosophy of Plants. William Blackwood, Edinburgh.

In contrast to Dunaliella, some cells, particularly those of higher plants, remain static within an immobile cell wall. Yet, if you look inside the cell, you are again faced with movement. You see that the protoplasm dramatically flows throughout a plant cell, a phenomenon known as cytoplasmic streaming (Kamiya, 1959). Look at the giant internodal cell of Chara (Fig. 1.6). The cytoplasm rotates around the cell at about 100  μm/s. If you electrically stimulate the cell, the cytoplasmic streaming ceases instantly. As the neurobiologists say, the cell is excitable and responds to external stimuli. In fact, action potentials were observed in characean internodal cells before they were observed in the nerve cells of animals (Cole and Curtis, 1938, 1939). The events that occur between electrical stimulation and the cessation of streaming are relatively well understood, and I discuss these throughout the book.

Figure 1.5  Photomicrograph of a swimming Dunaliella cell taken with Nomarski differential interference contrast optics.

Figure 1.6  Photomicrograph of a portion of a giant internodal cell of Chara showing several nuclei being carried by cytoplasmic streaming.

Lastly, take a look at the large single-celled plasmodium of the slime mold Physarum (Fig. 1.7; Coman, 1940; Kamiya, 1959; Carlisle, 1970; Konijn and Koevenig, 1971; Ueda et al., 1975; Durham and Ridgway, 1976; Chet et al., 1977; Kincaid and Mansour, 1978a,b; Hato, 1979; Dove and Rusch, 1980; Sauer, 1982; Dove et al., 1986; Bailey, 1997; Bozzone and Martin, 1998). Its cytoplasm streams at about 2000  μm/s. The force exerted by the streaming causes the plasmodium to migrate about 0.1  μm/s. Why does it move so slowly when streaming is so rapid? Notice that the cytoplasmic streaming changes direction in a rhythmic manner. The velocity in one direction is slightly greater than the velocity in the opposite direction. This causes the cell to migrate in the direction of the more rapid streaming. Because the plasmodium migrates toward food, the velocity of cytoplasmic streaming in each direction is probably affected by the gradient of nutrients. Nobody knows how this cell perceives the direction of food and how this signal is converted into directions for migration. Will you find out?

Figure 1.7  Dark-field photomicrograph of the slime mold Physarum polycephalum .

While looking at Physarum, notice that the protoplasm is not homogeneous but is full of relatively large round bodies rushing through the cell (Fig. 1.8). Is what you see the true nature of protoplasm, or are there smaller entities, which are invisible in a light microscope, that are also important in the understanding of cells? Edmund B. Wilson (1923) describes the power and the limitations of the light microscope in studying protoplasm:

When viewed under a relatively low magnification … only the larger bodies are seen; but as … we increase the magnification … we see smaller and smaller bodies coming into view, at every stage graduating down to the limit of vision … which in round numbers is not less than 200 submicrons. … Such an order of magnitude seems to be far greater than that of the molecules of proteins and other inorganic substances. … Therefore an immense gap remains between the smallest bodies visible with the microscope and the molecules of even the most complex organic substances. For these reasons alone … we should be certain that below the horizon of our present high-power microscopes there exists an invisible realm peopled by a multitude of suspended or dispersed particles, and one that is perhaps quite as complex as the visible region of the system with which the cytologist is directly occupied.

Figure 1.8  Bright-field photomicrograph of the streaming cytoplasm of the slime mold Physarum polycephalum .

We have now arrived at a borderland, where the cytologist and the colloidal chemist are almost within hailing distance of each other—a region, it must be added, where both are treading on dangerous ground. Some of our friends seem disposed to think that the cytologist should halt at the artificial boundary set by the existing limits of microscopical vision and hand over his inquiry to the biochemist and biophysicist with a farewell greeting. The cytologist views the matter somewhat differently. Unless he is afflicted with complete paralysis of his cerebral protoplasm he can not stop at the artificial boundary set up by the existing limits of microscopical vision.

Looking at the streaming plasmodium of Physarum inspires a sense of awe and wonder about life. How is that single cell able to sense the presence of the oatmeal flake and move toward it? How does it generate the force to move from within? What kind of endogenous timekeeper is in the cell that allows the streaming cytoplasm to move back and forth with the rhythm and regularity of a beating heart (Time, 1937, 1940)? We will explore these and other questions about living cells. However, to cross the artificial boundaries and comprehend the nature of the living cell, it is necessary to develop knowledge of mathematics, chemistry, and physics as well as cytology, anatomy, physiology, genetics, and developmental biology. The practice of cell biology that incorporates these various disciplines is still in its adolescent period and is treading on dangerous ground. As in any developing science, observations and measurements contain a given amount of uncertainty or probable error, and the exactness of the measurements, and thus the science, evolves (Hubble, 1954). Perhaps cell biology is at the stage thermodynamics was a century ago. Gilbert Newton Lewis and Merle Randall described the growth and development of thermodynamics in the Preface to their 1923 book, Thermodynamics and the Free Energy of Chemical Substances:

There are ancient cathedrals which, apart from their consecrated purpose, inspire solemnity and awe. Even the curious visitor speaks of serious things, with hushed voice, and as each whisper reverberates through the vaulted nave, the returning echo seems to bear a message of mystery. The labor of generations of architects and artisans has been forgotten, the scaffolding erected for their toil has long since been removed, their mistakes have been erased, or have become hidden by the dust of centuries. Seeing only the perfection of the completed whole, we are impressed as by some superhuman agency. But sometimes we enter such an edifice that is still partly under construction; then the sound of hammers, the reek of tobacco, the trivial jests bandied from workman to workman, enable us to realize that these great structures are but the result of giving to ordinary human effort a direction and a purpose.

Science has its cathedrals.

Cell biology is a young, vibrant, growing science, the beginnings of which took place in the early part of the 19th century when scientists, including Schleiden (1853), pondered what regular element may underlie the vast array of plant forms from the slender palm, waving its elegant crown in the refreshing breezes … to the delicate moss, barely an inch in length, which clothes our damp grottos with its phosphorescent verdue. Schleiden felt that we may never expect to be enabled to spy into the mysteries of nature, until we are guided by our researches to very simple relations … the simple element, the regular basis of all the various forms.

1.2. The Basic Unit of Life

Prior to 1824, organic particles or a vegetative force that organized organic particles were considered by some prominent scientists including Gottfried Leibniz, Comte de Buffon, and John Needham to be the basic unit of life (Roger, 1997). In fact, John Needham (1749) and John Bywater (1817, 1824) observed these living particles in infusions of plant and animal material that they placed under the microscope. Bywater observed that they writhed about in a very active manner and conjectured that the immediate source of the movement was thermal energy, which originated from the particles of [sun]light which come in contact with the earth, and have lost their rapid momentum. Bywater considered sunlight to carry the vital force, and concluded that the particles of which bodies are composed, are not merely inert matter, but have received from the Deity certain qualities, which render them actively instrumental in promoting the physical economy of the world.¹

Henri Dutrochet (1824) emphasized the importance of the cell, as opposed to living particles or the whole organism, as the basic unit of life. Dutrochet came to this conclusion from his microscopical observations, by which he observed plants are derived entirely from cells, or of organs which are obviously derived from cells. He extended his observations to animals and concluded that all organic beings are composed of an infinite number of microscopic parts, which are only related by their proximity (quoted in Rich, 1926). More than a decade later, Dutrochet's cell theory was promoted by Schleiden and Schwann. Schleiden (1838), a botanist, wrote

Every plant developed in any higher degree, is an aggregate of fully individualized, independent, separate beings, even the cells themselves. Each cell leads a double life: an independent one pertaining to its own development alone, and another incidental, in so far as it has become an integral part of a plant. It is, however, easy to perceive that the vital process of the individual cells must form the very first, absolutely indispensable fundamental basis.

Likewise, Schwann (1838), a zoologist, concluded that the whole animal body, like that of plants, is thus composed of cells and does not differ fundamentally in its structure from plant tissue. Thanks to the extensive research, and active promotion by Schleiden and Schwann; by the end of the 1830s, Dutrochet's concept that the cell is the basic unit of all life became well established, accepted, and extended to emphasize the interrelationships between cells. The expanded cell theory provided a framework to understand the nature of life and its origin and continuity.

We often divide various objects on Earth into two categories: the living and the lifeless. Therefore, the investigation of cells may provide us with a method to understand the question, What is life? We often characterize life as something that possesses attributes that the lifeless lack (Beale, 1892; Blackman, 1906; Tashiro, 1917; Osterhout, 1924; Harold, 2001). The power of movement is a distinctive aspect of living matter, where the movement has an internal rather than an external origin. Living matter generates electricity. Living matter also takes up nutrients from the external environment and, by performing synthetic reactions at ambient temperatures, converts the inorganic elements into living matter. Living matter also expels the matter that would be toxic to it. The ability to synthesize macromolecules from inorganic elements allows growth, another characteristic of living matter. Living matter also contains information, and thus has the ability to reproduce itself, with near-perfect fidelity. Lastly, living matter is self-regulating. It is capable of sensing and responding to environmental signals to maintain a homeostasis (Cannon, 1932, 1941) or to adjust to new conditions by entering metastable states, or other states, in a process known as allostasis (Spencer, 1864; Emerson, 1954; Sapolsky, 1998).

The above-mentioned properties are characteristic of living things and their possession defines a living thing. Mathews (1916) notes, When we speak of life we mean this peculiar group of phenomena; and when we speak of explaining life, we mean the explanation of these phenomena in the terms of better known processes in the non-living. There are entities like viruses that exhibit some, but not all, of the characteristics of life. Are viruses the smallest living organism as the botanist Martinus Beijerinck thought when he isolated the tobacco mosaic virus in 1898 or are they the largest molecules as the chemist Wendell Stanley thought when he crystallized the tobacco mosaic virus in 1935 (Stanley and Valens, 1961)? While the distinction between nonliving and living is truly blurred (Anonymous, 1905; Twain, 1923; Pirie, 1938; Baitsell, 1940), the cell in general is the smallest unit capable of performing all the processes associated with life.

For centuries, people believed that the difference between living and nonliving matter arises from the fact that living matter possesses a vital force, also known as the vis vitalis, a purpose, a soul, Maxwell's demon, a spirit, an archeus, or an entelechy (Reil, 1796; Loew, 1896; Lovejoy, 1911; Ritter, 1911; Driesch, 1914, 1929; Frankl, 1973; Waddington, 1977). According to the view of the vitalists and dualists, the laws of physics and chemistry used to describe inorganic nature are, in principle, incapable of describing living things. By contrast, mechanists, materialists, mechanical materialists, and monists believe that there is a unity of nature and a continuum between the nonliving and the living—and all things, whether living or not, are made of the same material and are subject to the same physical laws and mechanisms (Bernard, 1865; Dutrochet, 1824; von Helmholtz, 1903; Koenigsberger, 1906; Rich, 1926; Brooks and Cranefield, 1959). We will ask along with Erwin Schrödinger (1946), How can the events in space and time which take place within the spatial boundary of a living organism be accounted for by physics and chemistry?

Mary Shelley (1818) wrote about the potential of the materialistic/mechanical view and the ethics involved in experimentation on the nature of life when she described how Victor Frankenstein discovered that life could emerge spontaneously when he puts together the right combination of matter and activated it with electrical energy. In the materialist/mechanical view, living matter is merely a complex arrangement of atoms and molecules, performing chemical reactions and following physical laws. Thus, according to this view, the laws of chemistry and physics are not only applicable but also essential to the understanding of life (Belfast Address, Tyndall, 1898). Claude Bernard (1865) believed that "the term ‘vital properties’ is only provisional; because we call properties vital which we have not yet been able to reduce to physico-chemical terms; but in