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Protocells
Bridging Nonliving and Living Matter
Mark A. Bedau,
Liaohai Chen,
David Deamer,
David C. Krakauer,
Norman H. Packard,
and Peter F. Stadler

Protocells

Protocells
Bridging Nonliving and Living Matter
edited by Steen Rasmussen, Mark A. Bedau, Liaohai Chen, David Deamer,
David C. Krakauer, Norman H. Packard, and Peter F. Stadler
The MIT Press
Cambridge, Massachusetts
London, England

62009 Massachusetts Institute of Technology
All rights reserved. No part of this book may be reproduced in any form by any electronic or mechanical
means (including photocopying, recording, or information storage and retrieval) without permission in
writing from the publisher.
For information about special quantity discounts, please email special [email protected]
This book was set in Times New Roman and Syntax on 3B2 by Asco Typesetters, Hong Kong.
Printed and bound in the United States of America.
Library of Congress Cataloging in Publication Data
Protocells : bridging nonliving and living matter / edited by Steen Rasmussen . . . [et al.].
p. ; cm.
Includes bibliographical references and index.
ISBN 978 0 262 18268 3 (hardcover : alk. paper) 1. Artiﬁcial cells. 2. Life (Biology) I. Rasmussen,
Steen.
[DNLM: 1. Cells. 2. Biogenesis. 3. Cell Physiology. 4. Models, Biological. QU 300 P967 2008]
QH501.P76 2008
576.8
0
3 dc22 2007049243
10987654321

Contents
Preface ix
Acknowledgments xi
Introduction xiii
I Overview: Bridging Nonliving and Living Matter 1
1 The Early History of Protocells: The Search for the Recipe of Life 3
Martin M. Hanczyc
2 Experimental Approaches to Fabricating Artificial Cellular Life 19
David Deamer
3 Semisynthetic Minimal Cells: New Advancements and Perspectives 39
Pasquale Stano, Giovanni Murtas, and Pier Luigi Luisi
4 A Roadmap to Protocells 71
Steen Rasmussen, Mark A. Bedau, John S. McCaskill, and Norman H. Packard
II Integration 101
5 Steps Toward a Synthetic Protocell 107
Martin M. Hanczyc, Irene A. Chen, Peter Sazani, and Jack W. Szostak
6 Assembly of a Minimal Protocell 125
Steen Rasmussen, James Bailey, James Boncella, Liaohai Chen, Gavin Collis,
Stirling Colgate, Michael DeClue, Harold Fellermann, Goran Goranovic´, Yi Jiang,
Chad Knutson, Pierre-Alain Monnard, Fouzi Mouffouk, Peter E. Nielsen, Anjana Sen,
Andy Shreve, Arvydas Tamulis, Bryan Travis, Pawel Weronski, William H. Woodruff,
Jinsuo Zhang, Xin Zhou, and Hans Ziock

7 Population Analysis of Liposomes with Protein Synthesis and a Cascading Genetic
Network 157
Takeshi Sunami, Kanetomo Sato, Keitaro Ishikawa, and Tetsuya Yomo
8 Constructive Approach to Protocells: Theory and Experiments 169
Kunihiko Kaneko
9 Origin of Life and Lattice Artificial Chemistry 197
Naoaki Ono, Duraid Madina, and Takashi Ikegami
10 Models of Protocell Replication 213
Ricard V. Sole´, Javier Macı´a, Harold Fellermann, Andreea Munteanu, Josep Sardanye´s,
and Sergi Valverde
11 Compositional Lipid Protocells: Reproduction without Polynucleotides 233
Doron Lancet and Barak Shenhav
12 Evolutionary Microfluidic Complementation Toward Artificial Cells 253
John S. McCaskill
III Components 295
13 Self-Replication and Autocatalysis 299
Volker Patzke and Gu¨ nter von Kiedrowski
14 Replicator Dynamics in Protocells 317
Peter F. Stadler and Ba¨rbel M. R. Stadler
15 Peptide Nucleic Acids as Prebiotic and Abiotic Genetic Material 337
Peter E. Nielsen
16 The Core of a Minimal Gene Set: Insights from Natural Reduced Genomes 347
Toni Gabaldo´n, Rosario Gil, Juli Pereto´, Amparo Latorre, and Andre´s Moya
17 Parasitism and Protocells: Tragedy of the Molecular Commons 367
Jeffrey J. Tabor, Matthew Levy, Zachary Booth Simpson, and Andrew D. Ellington
18 Forming the Essential Template for Life: The Physics of Lipid Self-Assembly 385
Ole G. Mouritsen and Ask F. Jakobsen
19 Numerical Methods for Protocell Simulations 407
Yi Jiang, Bryan Travis, Chad Knutson, Jinsuo Zhang, and Pawel Weronski
vi Contents

20 Core Metabolism as a Self-Organized System 433
Eric Smith, Harold J. Morowitz, and Shelley D. Copley
21 Energetics, Energy Flow, and Scaling in Life 461
William H. Woodruff
IV Broader Context 475
22 Ga´nti’s Chemoton Model and Life Criteria 481
James Griesemer and Eo¨rs Szathma´ry
23 Viral Individuality and Limitations of the Life Concept 513
David C. Krakauer and Paolo Zanotto
24 Nonlinear Chemical Dynamics and the Origin of Life: The Inorganic-Physical Chemist
Point of View 537
Jerzy Maselko and Maciej Maselko
25 Early Ancestors of Existing Cells 563
Andrew Pohorille
26 Prebiotic Chemistry, the Primordial Replicator, and Modern Protocells 583
Henderson James Cleaves II
27 Cell-like Entities: Scientific Challenges and Future Applications 615
John M. Frazier, Nancy Kelley-Loughnane, Sandra Trott, Oleg Paliy, Mauricio
Rodriguez Rodriguez, Leamon Viveros, and Melanie Tomczak
28 Social and Ethical Issues Concerning Protocells 641
Mark A. Bedau and Emily C. Parke
Glossary 655
About the Authors 667
Index 679
Contents vii

Preface
The idea for this book grew out of two international protocell workshops in Sep-
tember 2003. One meeting, at Los Alamos National Laboratory and the Santa Fe
Institute, was organized by Steen Rasmussen, Liaohai Chen, David Deamer, David
Krakauer, Norman Packard, and Peter Stadler. The other meeting, at the European
Conference on Artiﬁcial Life (ECAL) in Dortmund, Germany, was organized by
Steen Rasmussen and Mark Bedau. We published a short summary of the state of
the art of protocell research as reﬂected in those workshops in early 2004 (Rasmussen
et al., 2004), and we planned to collect more details about this research in a longer
volume. That plan was the seed for this book. But a series of events intervened,
changing and delaying the book.
Those events grew out of the Seventh Artiﬁcial Life Conference in Portland, Ore-
gon, organized by Mark Bedau, John McCaskill, Norman Packard, and Steen Ras-
mussen in August 2000. Coinciding with the millennium, the conference aimed to
take stock of the young ﬁeld of artiﬁcial life. Out of the Oregon meeting came a com-
munity consensus of speciﬁc grand challenges in artiﬁcial life. One of these challenges
is to create wet artiﬁcial life from scratch.
Over the next three years, our activities were a portfolio of projects, most involv-
ing, in one way or another, the creation of life from scratch. In 2001 we coined the
termliving technologyas an umbrella for our activities. The next year we realized
how computer-controlled microﬂuidics could act as life support for the evolution of
minimal chemical systems, and two months later we started creating a new roadmap
to protocells. Our meetings led to a proposal for a new Center for Living Technology
at which scientiﬁc developments in this area could be nurtured and developed along
the way to producing practical applications.
The European Commission’s program on complex systems funded the ﬁrst phase
of these plans. Just before the ﬁrst protocell workshops in 2003, we learned that our
EC proposal on Programmable Artiﬁcial Cell Evolution (PACE) was funded. John
McCaskill led the PACE project, which consisted of fourteen European and U.S.
partners and included plans for a European Center for Living Technology in Venice.

One of the members of the PACE consortium was the new startup company, Proto-
Life Srl., of which Norman Packard and Mark Bedau became CEO and COO. Soon
afterwards Los Alamos National Laboratory awarded a complementary grant for a
project on Protocell Assembly, led by Steen Rasmussen. While these new activities
absorbed our time for a couple of years, this book was on the back burner.
In 2005 Emily Parke agreed to manage the editorial process of producing the
book. Our vision of the book had grown in the intervening years, so we solicited
chapters from many who had missed the original workshops.
Though it had a convoluted gestation, we hope this book will be both a resource
and an inspiration for the exciting and important quest to create life from scratch.
Mark Bedau
Norman Packard
Steen Rasmussen
Venice, Italy, June 2007
Reference
Rasmussen, S., Chen, L., Deamer, D., Krakauer, D., Packard, N., Stadler, P., & Bedau, M. (2004). Tran
sitions between nonliving and living matter.Science,303, 963.
x Preface

Acknowledgments
This book has been produced only because of the time and energy of a large group of
people. The editors are extremely grateful for the various kinds of help provided, and
we are pleased to have the opportunity here to acknowledge them.
Editorial Manager: Emily C. Parke
Editorial Board
James Bailey
James Boncella
James Cleaves
Stirling Colgate
Gavin Collis
Michael DeClue
Andrew Ellington
Goran Goranovic´
Martin M. Hanczyc
Takeshi Ikegami
Martin Nilsson Jacobi
Yi Jiang
Gu¨nter von Kiedrowski
Chad Knutson
Natalio Krasnegor
James La Clair
Doron Lancet
Pierre-Alain Monnard
Harold Morowitz
Ole Mouritsen
Andreea Munteanu
Peter Nielsen
Andrew Shreve
Eric Smith
Ricard Sole´
Pasquale Stano
Eo¨rs Szathma´ry
Bryan Travis
Woody Woodru¤
Hanz Ziock
Jinsuo Zhang

First, we want to convey special thanks to Emily Parke. As editorial manager,
Emily coordinated and oversaw the complex process of soliciting chapters, coordi-
nating reviews of the chapters, checking revisions, copy editing the ﬁnal manuscripts,
and generally keeping the project on track. None of the editors wants even to imag-
ine what producing this book would have been like without Emily’s unﬂagging ex-
pert assistance.
We are also especially grateful to the thoughtful critical attention devoted to early
drafts of the chapters by our Editorial Board.
We want to single out John McCaskill for thanks. John has been a key intellectual
partner and friend in the creation of the diverse portfolio of protocell-related activ-
ities, especially the European Commission’s Programmable Artiﬁcial Cell Evolution
(PACE) project, the Los Alamos Protocell Assembly project, the European Center
for Living Technology, and ProtoLife Srl. All of those projects, and so indirectly
this book, show the beneﬁt of John’s wit, intelligence, and vision.
We would also like to thank the MIT Press for critical early support that helped
launch this project and critical later patience when the timeline slipped. We thank
the Santa Fe Institute and the European Center for Living Technology for hospital-
ity while some of the work on this volume was completed. For ﬁnancial support for
some of this work, we thank EU-supported PACE integrated project, EC-FP6-IST-
FET-IP-002035, and the Los Alamos National Laboratory LDRD-DR Protocell As-
sembly project.
xii Acknowledgments

Introduction
I.1 What Are Protocells?
All life forms are composed of molecules that are not themselves alive. But how
exactly do living and nonliving matter di¤er? Is there a fundamental di¤erence at
all? How could we possibly turn a collection of nonliving materials into something
that is, at least operationally speaking, alive? This book is a compendium of the state
of the art on attempts to answer these questions by building bridges between non-
living chemistry and emergent living states of matter. It focuses on attempts to create
very simple life forms in the laboratory. These new forms of life might be quite
unfamiliar.
In this book a living system is operationally deﬁned as a system that integrates
three critical functionalities (ﬁgure I.1, outer ring): First, it maintains an identity
over time by localizing all its components. Second, it uses free energy from its envi-
ronment to digest environmental resources in order to maintain itself, grow, and
ultimately reproduce. Third, these processes are under the control of inheritable in-
formation that can be modiﬁed during reproduction. These properties enable selec-
tion and thus evolution as part of the reproduction process. Living systems are
sometimes said to include various further essential properties, such as autonomous
information processing, sensitivity to the environment, self-organization, and pur-
poseful behavior. We agree that these are central properties of living systems, but
we hold that they are derivative in the sense that they are e¤ectively implied by the
functionally integrated triad described previously.
This book aims to provide a very general understanding of how one could obtain
entities with these lifelike properties from experiments that begin with nonliving
materials. The book generally reﬂects the perspective that chemical instances of
such forms of life must embody the three operational functionalities in three inte-
grated chemical systems: ametabolismthat extracts usable energy and resources
from the environment,genesthat chemically realize informational control of liv-
ing functionalities, and acontainerthat keeps them all together (see ﬁgure I.1, inner

triangle). We will use the termprotocellto refer to any realization of these three func-
tional components. Our usage is close to the engineering wordprototype, that is, an
artiﬁcial structure that represents the ﬁrst simple working model of a designed sys-
tem. The importance of cooperative structures for minimal life was initially stressed
in Eigen’s hypercycle concept (Eigen, 1971), which focused on the cooperation be-
tween informational structures (genes). Ga´nti (1975) ﬁrst identiﬁed the minimal co-
operative structure capable of forming a cell as the triad of genes, metabolism, and
container.
We use the termscontainer,metabolism, andgenequite generally, with minimal
presupposition concerning their chemical details. In most contexts, the container
will be an amphiphilic structure such as a vesicle or micelle, but immobilizing chem-
icals on a surface can also achieve the required spatial localization. Similarly, metab-
olism could harvest redox energy or light, it could work with more or less complex
material precursors, and it might or might not use adenosine triphosphate (ATP)
and complex enzymes. Furthermore, genes might achieve informational control
and inheritance with some nucleotide other than DNA, or even without using any
biopolymers.
Figure I.1
The three essential operational functionalities of living systems (outer circle), together with the correspond
ing realizations of these functionalities in protocell architectures (inner triangle).
xiv Introduction

One way to produce a protocell is to combine an appropriate mix of nonliving
molecules and then let them react and self-assemble into a living protocell. This
process begins with molecular constituents and so may be considered abottom-up
approach to protocells. Most of the work in this book falls within this bottom-up
approach. Complementing this is atop-downapproach based on the recent success
of genomic research. The top-down approach begins with an existing contemporary
living cell, typically a very simple one, and reduces its genome by successive removal
of genes, to arrive at a minimal cell with just enough genes to maintain itself and
reproduce (Hutchison et al., 1999). One long-range aim of this top-down research is
to make an artiﬁcial cell by destroying a cell’s original genome and inserting a new
minimal genome that is synthesized externally from nucleotides using genomic tech-
nology (Glass et al., 2006). A spectrum of intermediate levels of functional organiza-
tion is spanned by top-down approaches beginning with living matter (contemporary
cells) and bottom-up approaches beginning with nonliving matter, as illustrated in
ﬁgure I.2.
Once a population of protocells exists, their functional e¤ectiveness might cause
some to be selected over others. If there is a combinatorially large family of possible
forms of informational control, and if information inheritance is neither too perfect
Figure I.2
The bottom up approach focuses on assembling a minimal protocell from simple inorganic and organic
components. The top down approach focuses on the simpliﬁcation of modern cells. Eventually these two
approaches will meet in the middle.
Introduction xv

nor too imperfect, then the process of evolution might be able to improve those func-
tionalities over time. Evolvability is often thought to be an essential property of life
(e.g., Maynard Smith, 1975), but it remains elusive in existing protocell realizations.
In fact, it is an open question under which conditions such a system will be able to
exhibit open-ended evolution, by which we mean a system’s ability to create new
properties in an open-ended manner over time as a result of selection and environ-
mental pressures (Bedau et al., 2000). Statistical analysis of evolutionary systems,
including artiﬁcial life models, indicates that the only systems known to exhibit
open-ended evolution are natural life (the biosphere) and human technology, which
is itself generated by living entities (Bedau et al., 1997; Skusa and Bedau, 2002). One
key to achieving open-ended evolvability is the ability of evolving systems to gener-
ate novel properties. Novel properties in molecular systems can be obtained not only
by genetic rearrangement but also by aggregation of existing systems. For example,
combining a lipid membrane system with a photosynthesizer might produce a simple
light-driven metabolic system. Aggregation-generated novelty could be as important
as genetic variation in the evolution of protocells.
Most of the protocell architectures presented in this book rely crucially on the pro-
cess of self-assembly. This means that the resulting protocell objects are not designed
or constructed piece by piece, as happens with the products of traditional engineer-
ing; rather, the requisite materials are brought together under the appropriate labo-
ratory conditions and the protocells spontaneously form. Protocells are themselves
emergent structures, so designing them and controlling their functionality will require
the development of new techniques of emergent engineering.
I.2 Scientific Roots of Protocell Research
Contemporary protocell research has a number of scientiﬁc roots. One is the long
tradition of work on the origin of life. The question of how life might be obtained
from nonliving materials is similar to that of how contemporary life originated, since
contemporary life presumably arose from nonliving materials, but these two ques-
tions lead us in very di¤erent directions. The methods and conditions used to make
protocells might be radically unlike those actually involved in life’s origins, and it is
an entirely open question whether such new forms of life would or could ever evolve
into anything like naturally existing life-forms. The understanding of life obtained
from making protocells, however, should contribute to research on the origin of life
as well as beneﬁt from it.
Research on the origin of life naturally spawned several of the research threads
that comprise current protocell e¤orts. A particularly important thread is the concept
of the RNA world, which concentrates on RNA as the primary element in origin of
life scenarios (see, e.g., Gilbert, 1986; Orgel, 1994). The connection with protocell re-
xvi Introduction

search comes when RNA chemistry is integrated with container structures (cf. Szos-
tak, Bartel, and Luisi, 2001; see also chapters 2 and 5).
Protocell research may also be seen as an endeavor within the ﬁeld of artiﬁcial life.
The phrase ‘‘artiﬁcial life’’ is much broader and refers to any attempt to synthesize
the essential features of living systems. Artiﬁcial life traditionally falls into three
branches, corresponding to three synthesis methods. ‘‘Soft’’ artiﬁcial life creates com-
puter simulations or other purely digital constructions that exhibit lifelike behavior.
‘‘Hard’’ artiﬁcial life produces hardware implementations of lifelike systems, usually
in robotics. ‘‘Wet’’ artiﬁcial life involves the creation of lifelike systems in a wet lab,
in most cases based on carbon chemistry in water. The holy grail of wet artiﬁcial life
is the construction of protocells, which is the focus of this volume.
Another strand of human activity that produces living cells that are not found in
nature is represented by the recent success of synthetic biology e¤orts to alter meta-
bolic pathways of existing contemporary cells by genetic manipulation (for an over-
view see Baker et al., 2006). These techniques may produce a range of more or less
artiﬁcial cells, depending on the extent of genetic modiﬁcation and the resulting dis-
tance from naturally existing life. If synthetic biology is characterized as the attempt
to engineer new biological systems, then protocell research can be viewed as a branch
of synthetic biology (see the introduction to part IV).
Astrobiology is concerned with the search for life elsewhere in the universe, so it
and protocell research share an interest in the fundamental properties of living sys-
tems. While the artiﬁcial life community asks what is minimal life and how can it be
useful, the astrobiology community asks where life comes from and whether it exists
only on the Earth. In contrast with astrobiology, protocell research does not need to
justify the cosmological or geochemical origins of its starting materials. However, the
possibility of detecting alien life on other planets or moons adds many intriguing
questions, some of which we will touch on in the last section of this book.
I.3 Purposes and Organization of This Book
This volume is a comprehensive general resource on protocell research intended to
serve a number of functions. The ﬁrst, of course, is simply to convey the state of the
art in contemporary protocell research. Doing this comprehensively involves a rich
multiplicity of perspectives: di¤erent disciplines (e.g., physics, chemistry, biology,
biochemistry, geochemistry, materials science, computer science, biophysics, evolu-
tionary theory, engineering, philosophy), di¤erent approaches to creating protocells,
the employment of di¤erent scientiﬁc methods (observation, experiment, simulation,
theory), and the understanding of processes happening on di¤erent spatial and tem-
poral scales. A resource that collects key protocell research in one place can promote
mutually beneﬁcial crosstalk with related and overlapping scientiﬁc and engineering
Introduction xvii

projects, such as studies of the origin of life and extraterrestrial life, e¤orts in syn-
thetic and systems biology, investigations in material science and nanotechnology,
as well as the study of artiﬁcial life and artiﬁcial chemistry in general.
This snapshot of protocell research today can also promote, inform, and construc-
tively focus future work in this area in at least two ways: by enhancing synergies
among the various perspectives and approaches currently being pursued within pro-
tocell research, and by identifying and calling attention to milestones that represent
the key tractable open questions that will most likely propel scientiﬁc progress.
Finally, being able to bridge the gap between the nonliving and the living will raise
a number of new practical issues for society to face. The capacity to engineer truly
living technology will galvanize a wealth of practical projects and surely spawn a
diverse ecology of revolutionary applications. But in the wrong hands, or with the
wrong designs, protocell technology could generate new kinds of risks to human
health and the environment. In addition, the practice of creating novel forms of life
from scratch could shock existing cultural norms, with the potential to fundamen-
tally reshape our sense of who we are and where we come from. A comprehensive
compendium on protocells could help those whether scientists, journalists, public
o‰cials, or the general public interested in gaining a scientiﬁcally informed per-
spective on this rapidly expanding new ﬁeld.
These purposes for this volume have informed its overall organization. Part I con-
tains four overviews of protocell science. These range from histories of experimen-
tal e¤orts to a conceptual framework for comparing experimental and theoretical
protocell achievements and proposals. Together the chapters provide complemen-
tary views on how to evaluate and categorize the ﬁeld. The eight chapters in part II
present some of the leading approaches to integrating the core complementary func-
tionalities of containment, metabolism, and informational control and transfer (genet-
ics) into a fully functioning protocell. The work presented here includes experimental
e¤orts, simulation of theoretical schemes, attempts to combine both approaches, and
a method of achieving integration with the help of computer-controlled microﬂuidic
and microelectrical devices.
Part III focuses on the foundational functional components of protocells provided
by containment, metabolism, and genetics. These nine chapters dive into experimen-
tal and simulation details of replicating informational molecules, lipid membranes
and compartments, and metabolic processes as well as energetics. This section also
includes a chapter on protocell simulation methods.
Part IV situates protocell science in the broader contexts of theories of life, mini-
mal forms of life, the related scientiﬁc investigation into the origin of life, and inves-
tigations of some practical applications and social and ethical implications. The book
closes with a glossary of technical terms found in the chapters.
xviii Introduction

I.4 Why Now?
Modern studies of simple protocells began to appear in the 1980s, typically in the
context of origin of life research. (For a historical account of earlier work, see chap-
ter 1.) This modern research was inspired by the discovery of Bangham and co-
workers in (1965) that phospholipids could not only assemble into the vesicular
structures now called liposomes, but also could trap concentration gradients of ions
and other solutes. From this work it immediately became apparent that the lipid
bilayer was the primary permeability barrier to the free di¤usion of ions and metab-
olites, and that specialized proteins must be present in the bilayer in the form of
channels and pumps to allow communication between the intracellular volume and
the external environment.
In the 1990s, other laboratories began to investigate encapsulated systems of mac-
romolecules. The early goals were to develop e‰cient encapsulation methods, design
lipid compositions and techniques that would permit external substrates to supply
encapsulated enzymes, establish conditions in which lipid vesicles could undergo
growth and division, and demonstrate catalyzed synthesis of polymers such as RNA,
DNA, and peptides within the liposome volume. As these goals were achieved and
reported in the literature, other laboratories began to think seriously about develop-
ing a variety of further protocell systems.
Much of the theoretical research on minimal life has its intellectual origin from the
ideas presented by the experimentalist Manfred Eigen back in 1971, when the notions
of quasispecies and hypercycle were created (Eigen, 1971). Cooperative structures
that provide mutual support of genes, metabolism, and containers can be seen as
generalizations of Eigen’s ideas. Also, Ilya Prigogine’s creation of the concept of dis-
sipative structures in the late 1970s gripped the imagination of many theorists trying
to understand the origins of life. The notion from the late 1980s of the RNA world
inspired both theoretical and experimental investigations of minimal life, and even
today this concept is probably still the most prevalent hypothesis about the bridge
between early minimal life and contemporary life.
With the turn of the millennium, ever-increasing numbers of publications explicitly
related to functional protocells began to appear. An Internet search in 2006 for the
termsprotocellandartiﬁcial cellproduced approximately 50,000 references to each
term. A similar search in the scientiﬁc literature revealed 435 citations forprotocell
and 1,310 citations forartiﬁcial cell. These numbers reveal exploding interest in these
topics since the ﬁrst modern publications about 25 years ago.
What might have given rise to this remarkable increase? First, of course, is the in-
trinsic interest in bridging the gap between nonliving and living matter. To truly un-
derstand what it would take to create a minimal living cell, the obvious challenge is
Introduction xix

to verify this understanding by assembling a minimal cell from its parts list. The ﬁrst
fabrication of a functioning protocell will be a major scientiﬁc breakthrough. It is
also clear that protocell technology will have signiﬁcant applications, particularly in
the biotechnology, material science, computer and information technology, and envi-
ronmental science industries.
All of these considerations make this book timely and of real use to the increasing
number of investigators who see research on protocells as their primary scientiﬁc
quest. We expect that this book will serve as a focus for such activity, calling atten-
tion to the progress that has been made so far, and identifying milestones to guide
future research and development.
In an environment in which basic scientiﬁc research su¤ers from decreasing re-
sources, society owes great thanks for the foresight and vision of national funding
sources, such as the Department of Energy’s support for Los Alamos National Lab-
oratory and NASA programs in Astrobiology and Exobiology, as well as for the Eu-
ropean Commission’s program on Future and Emerging Technologies, the United
Kingdom’s Engineering and Physical Sciences Research Council, the Japan Society
for the Promotion of Science, and the Japanese Minister of Education, Culture,
Sports, Science, and Technology, all of which have provided signiﬁcant ﬁnancial sup-
port for protocell research well in advance of any proven applications. Without these
kinds of resources, the research reported in this book would have been impossible.
The eventual applications of protocell research will come through the ﬂowering of
living technology. Living technology is one of the ﬁrst concrete realizations of what
the U.S. National Science Foundation (NSF) and the European Commission (EC)
have termedconvergent technologies. Convergent technologies are the emerging syn-
theses of nano-bio-info-cognitive (NBIC) knowledge production, and the NSF and
EC both believe that convergent technologies will have a very large socioeconomic
impact in the next 25 years. We hope that this volume will help forge an interna-
tional community of interested stakeholders working with protocell scientists to ex-
plore the broader global opportunities and challenges of protocell research and the
emergence of living technology.
References
Baker, D., Church, G., Collins, J., Endy, D., Jacobson, J., Keasling, J., & Modrich, P. (2006). Engineering
life: Building a fab for biology.Scientiﬁc American,294, 44 51.
Bangham, A. D., Standish, M. M., & Watkins, J. C. (1965). Di¤usion of univalent ions across the lamellae
of swollen phospholipids.Journal of Molecular Biology,13, 238 252.
Bedau, M. A., McCaskill, J. S., Packard, N. H., Rasmussen, S., Adami, C., Green, D. G., et al. (2000).
Open problems in artiﬁcial life.Artiﬁcial Life,6, 363 376.
Bedau, M. A., Snyder, E., Brown, C. T., & Packard, N. H. (1997). A comparison of evolutionary activity
in artiﬁcial evolving systems and the biosphere. In P. Husbands & I. Harvey (Eds.),Proceedings of the
fourth European conference on artiﬁcial life, ECAL97(pp. 125 134). Cambridge, MA: MIT Press.
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Eigen, M. (1971). Self organization of matter and the evolution of biological macromolecules.Naturwis
senschaften,58, 465 523.
Ga´nti, T. (1975). Organization of chemical reactions into dividing and metabolizing units: The chemotons.
Biosystems,7, 15 21.
Gilbert, W. (1986). The RNA world.Nature,319, 618.
Glass, J. I., Smith, H. O., Hutchinson III, C. A., Alperovich, N. Y., & Assad Garcia, N. (2007). Minimal
bacterial genome. U.S. Patent Application Publication 2007/0122826.
Hutchison, C. A. III, Peterson, S. N., Gill, S. R., Cline, R. T., White, O., Fraser, C. M., et al. (1999).
Global transposon mutagenesis and a minimal mycoplasma genome.Nature,286, 2165 2169.
Maynard Smith, J. (1975).The theory of evolution, 3rd ed. New York: Penguin.
Orgel, L. E. (1994). The origin of life on the Earth.Scientiﬁc American,271, 76 83.
Skusa, A., & Bedau, M. A. (2002). Towards a comparison of evolutionary creativity in biological and cul
tural evolution. In R. Standish, M. A. Bedau, & H. A. Abbass (Eds.),Artiﬁcial life VIII(pp. 233 242).
Cambridge, MA: MIT Press.
Szostak, J. W., Bartel, D. P., & Luisi, P. L. (2001). Synthesizing life.Nature,409, 387 390.
Introduction xxi

I
OVERVIEW: BRIDGING NONLIVING AND LIVING MATTER

1
The Early History of Protocells: The Search for the Recipe of Life
Martin M. Hanczyc
1.1 Introduction
Vitalism is an old ideology that makes a clear distinction between chemicals compris-
ing a living organism and chemicals from inanimate matter. The synthesis of urea by
Friedrich Wo¨hler (1828) challenged the philosophical distinction between nonliving
and living matter by demonstrating that a biological organic molecule can be synthe-
sized in a laboratory from nonliving chemical precursors. Further development of
organic chemistry demonstrated how most of the organic molecules that comprise a
cell could be synthesized in a laboratory under detailed reaction conditions. Subse-
quent experiments such as the Buchner brothers’ fermentation of sugar into ethanol
(a biological process) by the nonliving yeast extracts in 1897 opened the door further
to exploration of the conceptual gray space between chemistry and biology (Fried-
mann, 1997). The following brief historical introduction recounts the principal exper-
imental endeavors to create a protocell that began more than 100 years ago.
1.2 The First ‘‘Cell Models’’
The ﬁrst studies from the latter half of the nineteenth century that reported lifelike
behaviors from nonliving systems were not based on cells or even cell extracts. Begin-
ning in 1867, Moritz Traube described a system that demonstrated the formation and
growth of semipermeable membranes composed of copper ferrocyanide surrounding
a seed crystal of copper sulfate. Such an artiﬁcial membrane superﬁcially resembled a
cell membrane in that it exhibited selective permeability to di¤erent solutes and
responded to osmotic pressure (Traube, 1867). In 1892, Bu¨tschli described structures
with an amoebalike movement formed by combining olive oil and potash. The dy-
namic structures appeared to form pseudopodia and engulf other particles, behaviors
seen with living amoebae (Bu¨tschli, 1892). Leduc (1907) also produced ‘‘osmotic
cells’’ similar to Traube’s by placing a seed crystal of calcium chloride in a saturated

potassium salt solution. As the crystal dissolved, a membrane of calcium phosphate
appeared and formed a structure similar in morphological appearance to a cell. He
reported the shape change of the cell-like structures in response to environmental
changes in osmotic pressure that presumably modulated the surface of the structures.
These early cell models showed that simple, nonliving materials such as salt solutions
could organize into structures at least superﬁcially resembling living cells (Oparin,
1965a). Even though the relevance of such models to actual living cells was not
clearly established at that time, the tendency of nonliving matter to adopt some char-
acteristics of life fascinated researchers interested in not only the origin of but also
the synthesis of life.
1.3 Herrera’s Sulphobes
In 1897 Alfonzo L. Herrera published a book,Recueil des lois de ta Biologie Generale
(Collection of Laws of General Biology), in which he introduced the idea that the
functions and structures of living cells can be attributed to physicochemical laws
(Negro´n-Mendoza, 1994). In 1904, Herrera founded a new branch of science, plas-
mogeny, which he deﬁned as the study of the origin of protoplasm as an experimen-
tal science. Protoplasm was a term used to describe the living material inside a cell. It
had the observable properties of ﬂow and movement, was the site of metabolism, and
was able to self-replicate. Herrera began to explore how such properties could be the
result of physicochemical forces acting and interacting within living cells. His ap-
proach was to reconstitute such properties using nonbiological chemicals. By mixing
substances with di¤erent chemical potentials and di¤erent phases, he observed life-
like patterns emerging. He is most famous for producing ‘‘sulphobes’’ by exposing
thin ﬁlms of formaldehyde in water to fumes of ammonium sulﬁde (Herrera, 1942).
Through microscopy (ﬁgure 1.1), Herrera observed populations of many diverse
structures reminiscent of protoplasm, including structures that appeared to undergo
mitotic division (Herrera, 1912). Apparently Herrera’s sulphobes were so lifelike that
when presented to an ‘‘eminent microscopist,’’ they were determined to be living
creatures and were then classiﬁed as such (Young 1965, p. 357).
Plasmogeny o¤ered a view of biology seen through the eyes of a synthetic chemist.
With his sulphobes, Herrera demonstrated that the superﬁcial structure and organi-
zation of the protoplasm could be recapitulated through purely chemical means. Fur-
thermore, such structures produced chemistry that resulted in the fortuitous synthesis
of two amino acids and dyes. Therefore the sulphobes not only resembled lifelike
structures but also were capable of limited biologically relevant synthesis. Most nota-
bly Herrera’s synthesis of protoplasm-like properties in the laboratory was demon-
strated starting from simple, nonbiological constituents.
4 Martin M. Hanczyc

1.4 Bungenberg de Jong’s Coacervates
Although Herrera began an extensive experimental research program to recapitulate
and understand protoplasm through chemistry, the origin of the idea that the proto-
plasm is made up of chemicals subject to physicochemical laws can be found in early
research on colloidal systems. Thomas Graham (1805 1869), an English chemist,
studied the di¤usion of nonbiological materials and biological extracts such as
starch, gum, and gelatin. He termed the class of substances with very low di¤usion
ratescolloids. In 1861, he wrote that the plastic parts of the animal body also behave
like colloids (Graham, 1861).
The Dutch biochemist H. G. Bungenberg de Jong (1893 1977) coined the term
coacervateas a special from of colloid. When certain organic substances such as gel-
atin and gum arabic are mixed with an aqueous solution, the ﬂuids separate into two
distinct phases. Upon agitation, the bottom, organic-rich layer breaks up into a pop-
ulation of small microscopic spherical structures or coacervates (see ﬁgure 1.2). These
structures of a few microns in diameter do not readily dissolve because of a tight
layer of water molecules that interacts with the charged organics at the surface sur-
rounding the coacervate. This in e¤ect delays the di¤usion of concentrated material
in the coacervate sphere into the surrounding media. Bungenberg de Jong (1932)
noted the similarities between coacervates and living protoplasm, and like Graham
before, held the idea that the physical conditions that produced coacervates may
also explain something about the organization of the protoplasm.
Figure 1.1
Sample micrographs from the work of Herrera showing various cell like morphologies (800 magniﬁca
tion) from Herrera (1912).
The Early History of Protocells 5

Coacervates are interesting as model systems of simple cells in that they form
spontaneously, are rich in organic material (as opposed to the aqueous environment
around the coacervate), and provide a locally segregated environment with bounda-
ries that allow selective absorption of exogenous organic molecules. Coacervates are
most stable when formed from a mixture of positively and negatively charged sub-
stances resulting from an interplay of hydration and electrostatic forces. As a result,
coacervates are dynamic, can respond to external manipulation, and at the same
time are quite unstable, especially when the conditions under which they form are
changed. As a result of such investigations into colloidal and coacervate systems, it
was thought that living protoplasm consisted of di¤erent coacervate systems acting
and interacting together to produce the various dynamical properties observed in liv-
ing cells.
1.5 Crile’s Autosynthetic Cells
At the Cleveland Clinic Foundation in the 1930s, Doctor George Crile and col-
leagues experimented with the reconstitution of artiﬁcial cells from living material.
Two fractions were prepared from animal brain: one containing the lipid (lipoid) ma-
Figure 1.2
Coacervates containing bacterial polynucleotide phosphorylase enzyme able to synthesize RNA polymers,
from Oparin, 1965b (magniﬁcation not speciﬁed).
6 Martin M. Hanczyc

terial and another containing sterilized protein material. When these fractions were
mixed together with a salt solution representing the salinity found in the animal
brain, structures resembling cells self-assembled. These structures, typically 50 to
150 microns in diameter, were termedautosynthetic cells(Crile, Telkes, and Row-
land, 1932) (ﬁgure 1.3) and were scrutinized for lifelike behavior. Beyond their mor-
phological similarities to living protozoa, Crile determined that the autocatalytic cell
preparations consumed a certain amount of oxygen and produced carbon dioxide (in
comparison with controls), and concluded that a certain amount of metabolism was
taking place. However, it was never shown what this metabolism-like activity was, or
if it was localized to the cell-like structures. In addition, they observed budding and
division of the structures by microscopy. The autosynthetic cells could be maintained
for months by feeding the structures with the sterile protein fraction, or destroyed by
heat, poisons, radiation, lack of oxygen, or lack of food.
Crile and colleagues relied heavily on self-organized processes in living matter to
reconstitute some of the properties of life from biological extracts. They then used
currently available techniques to test for signs of life. The work of Noireaux and Lib-
chaber (2004) recreated a similar approach of artiﬁcially reconstituting cells from
extracts under more controlled conditions and using modern techniques.
1.6 Oparin and Coacervates
Although A. I. Oparin’s interests were focused on understanding the origin of life, he
made many contributions to the ﬁeld of protocell research, primarily by introducing
metabolism to coacervates. Oparin and colleagues continued the pioneering work of
Bungenberg de Jong and made coacervates from gum arabic (an extract from the
Figure 1.3
Crile’s autosynthetic cells (400 magniﬁcation; reproduced from Crile, Telkes, and Rowland, 1932).
The Early History of Protocells 7

Acaciatree, a complex and variable mixture of arabinogalactan oligosaccharides,
polysaccharides, and glycoproteins) and gelatin (a protein product formed by partial
hydrolysis of collagen extracted from animal skin, bones, cartilage, and ligaments).
Such coacervates were prepared containing phosphorylase enzymes capable of poly-
merizing the substrate glucose-1-phosphate into starch. Once the coacervates con-
taining the enzyme were prepared, the substrate was added to the external medium
and adsorbed by the coacervates. It was found under such conditions that starch pro-
duction proceeded within the coacervate. Taking this experimental design a step fur-
ther, Oparin prepared coacervates containing two di¤erent enzymes, phosphorylase
andb-amylase, which together could form a simple two-step enzymatic pathway.
When the substrate glucose-1-phosphate was added exogenously, it was found that
not only did starch form in the coacervate droplet but also maltose, the product of
b-amylase acting on the starch, was found in the external media (ﬁgure 1.4). Report-
edly Oparin and colleagues succeeded in encapsulating polyadenine synthesis (see
ﬁgure 1.2), NAD oxidative-reductive reactions, and ascorbic acid oxidation within
coacervates (Oparin, 1966). Oparin also understood that the rates of reaction for
each step determine whether the coacervate assembly would grow by incorporating
and converting the incoming substrates into retainable products or diminish by con-
verting all available molecules into highly di¤usible product (Oparin, 1965b).
This work with coacervates demonstrated a few essential points. First, structures
with a superﬁcial semblance to living protoplasm can be formed in the laboratory
from organic material. Such structures can trap functional molecules such as
enzymes. Molecules added to the external media can be absorbed by the coacervate
structure. The absorption is selective and depends on the composition of the coacer-
vate. Once absorbed, the molecules can be acted on by the trapped enzymes and the
product accrues within the structure. Finally, the products of the reaction can also
di¤use out of the structure into the environment. The essential role of selective per-
meability is clearly demonstrated. As Young stated in 1965, ‘‘It [a selectively perme-
able barrier] permits the cell to create its own internal environment necessary for its
metabolic and reproductive activities’’ (Young 1965, p. 348). On the importance of
metabolism, Oparin stated that ‘‘individual chemical reactions in living beings are
strictly coordinated and proceed in a certain sequence, which as a whole forms a
network of biological metabolism directed toward the perpetual self-preservation,
growth, and self-reproduction of the entire system under the given environmental
conditions’’ (Oparin 1965b, p. 331).
By engineering supramolecular structures capable of housing a simple metabolic
pathway, Oparin demonstrated that the reconstitution of a population of structures
with simple lifelike metabolism was possible. Although Oparin’s main objective for
this work was to understand fundamental processes related to chemical evolution
and the origin of cellular life, his work with coacervates followed an approach more
8 Martin M. Hanczyc

Figure 1.4
(A) Representation of molecules involved in the coacervate enzymatic pathway. (B) Schematic of the
encapsulated enzymatic pathway. The enzymes, phosphorylase andbamylase, are entrapped within
the coacervate structure. The substrate, glucose 1 phosphate, is supplied exogenously and absorbed by the
coacervate. The substrate is converted to starch within the coacervate droplet by phosphorylase. Then
thebamylase converts the starch to maltose, which di¤uses out of the coacervate structure (Crile, Telkes,
and Rowland, 1932).
The Early History of Protocells 9

akin to synthetic biology. This is because his work was based on organic molecules
and enzymes extracted and puriﬁed from already living matter. (In contrast, Her-
rera’s work involved the generation of structures directly from nonbiological chemi-
cals.) Because of his success in reconstituting lifelike microscale structures in the
1950s and 1960s, Oparin inspired many to pursue the fundamental questions of the
origin of life and to understand the basic physicochemical forces that underlie cellu-
lar life.
1.7 Jeewanu
In the 1960s Krishna Bahadur from the Department of Chemistry at the University
of Allahabad reported on the synthesis of protocells calledJeewanu(Bahadur, 1966).
The name given to these structures, Jeewanu, comes from a Sanskrit work for par-
ticles of life. Jeewanu were made using various protocols and components. Some
were made by combining peptides, ascorbic acid, ammonium molybdate, and inor-
ganic minerals. Others were made by mixing paraformaldehyde, molybdic acid, and
ferric chloride in water and then placing this mixture on a nutrient-rich agar. After
some time (hours to weeks) under exposure to a source of light (sunlight or artiﬁcial
ultraviolet light), globular structures of about 0.5 to 15 microns in diameter ap-
peared. Bahadur claimed that the Jeewanu had a similar morphology to living cells,
grew over time, possessed weak metabolic activity, and could reproduce by budding.
He also made the claim that Jeewanu were living cells. The creation of Jeewanu was
an attempt to synthesize actual living cells using simple precursors with the goal of
understanding the origin of life. The inadequate detail provided in his publications
led to criticisms (see Caren and Ponnamperuma, 1967) and doubt was cast on his
claims.
1.8 Fox’s Microspheres
With the development of molecular biology in the 1950s through the 1970s, the fun-
damental role of self-organization or self-assembly in biological systems became in-
creasingly apparent (Fox, 1968; Wald, 1954). It is this type of interaction among
molecules that gives rise to higher-order structures of complexity in modern-day cells
and is a main factor in the formation of sulphobes, autosynthetic cells, Jeewanu, and
coacervates as described earlier. In addition, self-organization provides a tool that
can be used to construct protocells without relying excessively on precise and often
untenable manipulation on the molecular level.
In the 1960s and 1970s, Sidney Fox (1912 1998) and colleagues contributed to the
pursuit of the synthesis of life by creating populations of peptide-based spherical
structures in the laboratory. Speciﬁcally, by heating amino acids, mixing in water or
10 Martin M. Hanczyc

salt solution, and then allowing the solution to cool slowly, Fox found that spherical
structures formed spontaneously (ﬁgure 1.5). These structures were similar in diame-
ter (typically a few microns), with each microsphere containing an estimated 10
10
proteinoid molecules. The proteinoid material formed during the heating step by
linking and crosslinking of the amino acid monomers to form more elaborate and
complex peptides. These peptides would then interact and self-assemble within
minutes into larger macromolecular structures called microspheres (Fox and Dose,
1972). Fox’s microspheres were found to be generally more stable than coacervates.
Fox extensively characterized the dynamics of his microspheres, reporting that he
observed many interesting phenomena including di¤usion of material from the inside
of the sphere to the outside, partial ﬁssion (due to dissolution of proteinoid and
change in surface tension), generation of double-layer membrane, shrinkage or swell-
ing resulting from osmotic changes, sensitivity to pH (depending on amino acid com-
position), selective permeation of polysaccharides and monosaccharides, motility
(with zinc and ATP), budding, growth by accretion, and formation of junctions
between microspheres. In addition, Fox and his colleagues reported some weak
enzymelike activity in the proteinoid products (Fox, 1965, 1968). It is not clear
whether microspheres exhibiting such behaviors were common or the conclusions
were based on a few rare events in the population.
Like Oparin, Fox was searching for abiotic processes that could have led to the
formation of cellular life. In much the same way, microspheres showed encapsulated
metabolism and supramolecular dynamics. One fundamental di¤erence between
Figure 1.5
Microspheres of proteinoid in water from Young, 1965 (900 magniﬁcation).
The Early History of Protocells 11

coacervates and microspheres is that Fox demonstrated the self-assembly of a popu-
lation of complex microstructures from simple precursors.
1.9 Toward the Modern Concept of the Protoplasm
The cell model systems presented here were simplistic conceptualizations of biologi-
cal cells and may not have been accurate representations of the important forces that
deﬁne a living cell. Concurrent research into properties of living cells painted a di¤er-
ent picture of the vital physicochemical properties of life. Although coacervates and
microspheres were shown to have some type of selectively permeable barriers, other
research has shown that all natural living cells use a particular kind of barrier com-
posed of a lipid membrane. In 1900, Charles E. Overton showed that nonpolar, oily
chemical substances were selectively absorbed by plant cells. He hypothesized that
the cell membrane barrier is similar in some way to olive oil and is lipoid in com-
position (Overton, 1900). Further investigation into the physicochemical nature of
lipids by Irving Langmuir (1917) led to the idea that fatty acid molecules form a
monolayer at the air-water interface with the fatty hydrocarbon chains oriented
away from the water. Gorter and Grendel (1925) then performed experiments similar
to Langmuir’s, but with lipid extracts from erythrocytes. They were able to calculate
that enough lipid was present to cover approximately twice the surface area of the
cells, concluding that the cells were covered in a layer of lipids ‘‘two molecules
thick.’’ This research may have inspired Crile and colleagues to use two fractions to
construct their autosynthetic cells: the protein fraction to reconstitute the cytosol and
the lipoid fraction to reconstitute the membrane.
Danielli and Davson (1935) proposed a bilayer model of lipid cell membrane in
which lipids were arranged such that the hydrocarbon chains formed the interior of
the membrane sandwich, and proteins covered the lipid headgroups on both sides
of the membrane. This hypothetical structure was further supported by electron mi-
croscopy studies in the 1950s, and renamed theunit membrane model(Robertson,
1957). Further evolution in the conceptualization of the cell membrane led to the cur-
rent ﬂuid mosaic model of the cell membrane (Singer and Nicholson, 1972). In this
model the cell is enclosed in a bilayer of lipids as before, but now the lipids as well as
integral proteins move within the membrane. The ﬂuid mosaic model allows for a
detailed understanding of processes in real cells such as osmotic response, selective
permeability, and speciﬁc cell-cell and cell-environment communication. Bangham
demonstrated in 1965 that dispersions of phospholipid extracts self-assemble into
spherical structures reminiscent of cell membranes, and these ‘‘liposomes’’ like cell
membranes are osmotically active (Bangham, Standish, and Watkins, 1965). This
provided the ﬁrst in vitro model of a real cell membrane. Since then, liposomes have
12 Martin M. Hanczyc

been developed extensively as models of natural cell membranes and used in con-
structing protocells.
In sulphobes, autosynthetic cells, coacervates, Jeewanu, and microspheres, the
morphology of the structures bears a superﬁcial resemblance to living cells. The elec-
trostatic forces of interacting polymers and the crosslinking of peptides may explain
their shape and consistency. However, the real structure of the cytoplasm is of a
much di¤erent complexity. While Graham, Bungenberg de Jong, and others were
investigating the properties of coacervates, other contemporaneous models of the
protoplasm were being developed. One included a view in which the internal contents
of the cell are physically organized in three dimensions by a protein network of
ﬁbers. In the eighteenth and nineteenth centuries, an intracellular ﬁber network was
proposed to explain contraction of animal muscles (see Frixione, 2000). Sigmund
Freud, the father of psychoanalysis, was one of the original proponents of the ﬁbril-
lary theory of protoplasm structure as it applied to his research into nerve cells
(Freud, 1882; Triarhou and Del Cerro, 1987). During the early part of the twentieth
century, similar conceptualizations were applied to explain the intracellular organiza-
tion and movement in many types of animal cells. Such hypotheses were supported in
the 1950s through the visualization of protein ﬁber networks by electron microscopy.
Today, we understand that the consistency of the eukaryotic cytoplasm is not due to
colloidal properties of the constituents after all, but rather to an elaborate protein
cytoskeleton that is responsible for the cell shape as well as internal organization
and cell movement. Evidence also exists for similar protein networks in bacteria (see
Carballido-Lopez, 2006).
Returning now to the attempts to synthesize protocells, those cell models did not,
after all, represent the true complexity of living protoplasm beyond superﬁcial mor-
phological similarity. Nevertheless, the early research into the recipe of life has
shown that much simpler aggregate constructions can exhibit some lifelike behaviors.
Furthermore, some structures have been engineered to contain primitive metabo-
lisms. So although such works may not have successfully explained the physico-
chemical forces of cellular life, they can be seen as useful attempts to synthesize a
simplistic form of artiﬁcial life.
1.10 Summary
The ideology of vitalism has been challenged, and its once-deﬁned lines blurred by
the research programs that ambitiously tried to synthesize life and lifelike behavior
from nonliving components. These early attempts at synthesizing life from nonlife fu-
eled the debate about what life essentially is. Such research projects attracted the in-
terest of many researchers and thinkers from a wide range of disciplines, and also of
The Early History of Protocells 13

religious leaders. Indeed, Sidney Fox was invited on multiple occasions to the Vati-
can for discussions about the origin and synthesis of life with the Pope and his scien-
tists (Fox, 1997). Self-organization of structure, response to environmental changes,
uptake of nutrients, internal metabolism, expulsion of waste, movement, and self-
replication: These concepts persist in modern-day attempts to synthesize protocells,
as described in the following chapters of this book. Although these early attempts
did not satisfactorily represent the real physicochemical forces that shape a modern
living cell, they did show the tendency of matter, under the proper conditions, to or-
ganize itself into structures possessing similar qualities to living cells. And in that
way, they imply that there may be many paths toward synthesizing primitive life.
In 1966, at the national meeting of the American Chemical Society (ACS), a sym-
posium on the synthesis of living systems was held to present the major steps in the
developing ﬁeld of molecular biology. The topics ranged from the laboratory synthe-
sis of bovine insulin and other proteins to the synthesis and replication of nucleic
acids. The symposium prompted the publication of a paper in 1967 suggesting that
‘‘[i]t is the accomplishments of the past decade and the conﬁdence in the creative
powers of scientists which have led to speculation that these accomplishments may
lead, before the end of the century to the synthesis of new types of rudimentary living
systems’’ (Price, 1967, p. 144).
In an earlier optimistic view published inSciencein 1960, Simpson noted that ‘‘[a]t
a recent meeting in Chicago, a highly distinguished international panel of experts was
polled. All considered the experimental production of life in the laboratory immi-
nent, and one maintained that this has already been done . . .’’ (Simpson 1960, p.
969).
Despite predictions, most researchers would agree that the goal of a synthetic cell
has not yet been achieved. The question remains: When will science create synthetic
life? This is di‰cult to answer. Recently, Jack Szostak remarked that $20 million dol-
lars and three more years of research would produce a living and evolving protocell
(Zimmer, 2004). Indeed, a well-funded, consistent research program may reach the
desired goal in a relatively short time. Protocell history captured in manuscripts and
texts over the past hundred years shows us that progress in chemistry, physics, biol-
ogy, biochemistry, and other areas inspires an enthusiasm that makes the goal of a
synthetic cell seem closer. Descriptive and exploratory research in many ﬁelds, along
with technological advancements, is intimately linked with the vision of turning our
knowledge of lifetoward thesynthesis of life.
Looking back over the history of protocell development, we can see that there are
many often related avenues that one can take to create a living cell through chemis-
try. We need not only a fundamental knowledge of a variety of subjects, but also our
creativity. A few scientists spent a career exploring the beneﬁts and limitations of the
14 Martin M. Hanczyc

chosen path described here. In the end, none of these e¤orts has produced a living
cell. But they all asked the important questions, highlighted the challenges, and
shaped our thinking. These attempts to synthesize living cells from chemistry show
that a viable experimental model of a living cell will depend on more than the co-
localization of various essential characters such as the type of molecule, source of in-
formation, and functional metabolism. Higher-order interactive networks must also
be in place to coordinate the form and function of all components in a living cell
with high reliance on the self-organizing properties of the system.
The creation of a self-replicating protocell is only one step toward the synthesis of
life. Our predecessors were interested in not only creation of a self-replicating proto-
cell through chemistry and physics but also the synthesis of a ‘‘protoplasm’’ capable
of evolving into a diverse array of cell types and a communal network of living enti-
ties. Although current e¤orts often focus on the creation of a single synthetic cell, our
history envisions an even larger goal of an interwoven community of synthetic struc-
tures acting and interacting to produce a rich environment suitable not only for the
creation but for the evolution of synthetic life.
Acknowledgments
My sincere thanks and gratitude to Christopher Barbour, Coordinator of Special
Collections, at the Tisch Library Special Collections of Tufts University, and Patricia
Killiard, Head of Electronic Services and Systems, of the Cambridge University
Library, for their assistance in acquiring many of the older references cited in this
chapter.
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The Early History of Protocells 17

2
Experimental Approaches to Fabricating Artificial Cellular Life
David Deamer
2.1 Introduction
Recent scientiﬁc advances suggest that it may be possible to fabricate an artiﬁcial cell
(i.e., a protocell) with most, if not all, of the properties associated with the living
state. At ﬁrst glance, this might seem to be an impossible task, but life appears to
have arisen spontaneously on the early Earth, so perhaps we can be optimistic. Yet
life did not spring into existence with a full complement of genes, ribosomes, mem-
brane transport systems, metabolism and the DNA!RNA!protein information
transfer that dominates all life today. There must have been something simpler, a
kind of sca¤old-life that was left behind in the evolutionary rubble. Can we repro-
duce that sca¤old? One possible approach is to incorporate one or a few genes into
artiﬁcial vesicles to produce molecular systems that display the properties of life. The
properties of the system may then provide clues to the process by which life began in
a natural setting of the early Earth, and perhaps lead to a second origin of life, but
this time in a laboratory setting.
What would such a system do? We can answer this question by listing the basic
functions that would be required of artiﬁcial cellular life:
Self-assembled lipidlike molecules form membrane-bounded compartments that en-
capsulate internal molecular machinery.
Macromolecules are encapsulated, but smaller nutrient molecules cross the mem-
brane barrier by di¤usion or through polymer pores.
Energy is captured by a pigment system (light energy), or from chemical energy or
oxidation-reduction reactions.
A primitive version of metabolism is initiated within the boundary membrane.
The energy is coupled to the synthesis of activated monomers, and macromolecules
grow by polymerization of the monomers.

The membrane-bounded compartment grows by addition of amphiphilic
molecules.
Macromolecular catalysts speed the growth process, and primitive regulatory
mechanisms evolve to control metabolism and polymerization processes.
Information is captured in the sequence of monomers in one set of polymers, and
used to direct the synthesis of a second set of catalytic polymers, thereby reproducing
the catalysts during growth.
The membrane-bounded system of macromolecules can divide into smaller
structures.
Genetic information is passed between generations by duplicating the informa-
tional polymers and sharing them between daughter cells.
Occasional mistakes (mutations) are made during replication or transmission of in-
formation so that the system can evolve through natural selection.
Looking at this list, one is struck by the complexity of even the simplest form of
cellular life. This is why it has been so di‰cult to ‘‘deﬁne’’ life in the usual sense of
a deﬁnition, that is, boiled down to a few sentences in a dictionary. Cellular life is a
complex system that cannot be captured in a few sentences, so perhaps listing its
observed properties is the best we can ever hope to do. Despite the apparent com-
plexity, it is also signiﬁcant that most of the functions have been reproduced in the
laboratory. We can now describe how this set of functions might be integrated into
artiﬁcial cellular life.
2.2 Self-Assembly Processes in Cellular Life
All modern cellular life incorporates two processes, which we will refer to as self-
assembly and directed assembly. Spontaneous self-assembly occurs when certain
compounds associate through noncovalent hydrogen bonds, electrostatic forces, and
nonpolar interactions that stabilize orderly arrangements of small and large mole-
cules. A classic example is the manner by which amphiphilic molecules in aqueous
phases form micelles and bimolecular structures (ﬁgure 2.1, top panel). Another ver-
sion of self-assembly requires the formation of covalent bonds between similar mo-
lecular species, such as the random polymers of amino acids that can be produced
by energy-dependent condensation reactions (ﬁgure 2.1, center). In contrast, the
directed assembly processes characteristic of today involve the formation of covalent
bonds by energy-dependent synthetic reactions, but also require that a coded se-
quence in one type of polymer in some way directs the sequence of monomer addi-
tion in a second polymeric species (ﬁgure 2.1, lower panel).
20 David Deamer

Spontaneous self-assembly of organic compounds in aqueous phases was presum-
ably common on the prebiotic Earth, and likely involved certain compounds that can
form closed membrane-bound microenvironments. Such boundary structures, and
the compartments they produce, have the potential to make energy available in the
form of ion gradients, and can provide a selective inward transport of nutrients. Fur-
thermore, membranous compartments, in principle, are capable of containing unique
systems of macromolecules. If a yet unknown macromolecular replicating system of
polymers could be encapsulated within a membrane-bounded compartment, the
components of the system would share the same microenvironment (ﬁgure 2.2) and
the result would be a major step toward cellularity, speciation, and true cellular func-
tion (Cavalier-Smith, 1987; Conde-Frieboes and Blochliger, 2001; Deamer et al.,
2002; Dyson, 1999; Hutchison et al., 1999; Kock and Schmidt, 1991; Morowitz,
Figure 2.1
Cellular life today uses both self assembly and directed assembly processes to grow. Self assembly (upper
panel) is essential for synthesis and stability of membrane structures and protein folding, whereas directed
assembly (lower panel) underlies the synthesis of proteins according to the base sequences in DNA and
mRNA. We assume that on the early Earth, random polymers similar to peptides and nucleic acids were
produced by a yet unknown synthetic pathway (center). The random polymers, if capable of growth in
a membrane bounded microenvironment, would be subjected to selection and thereby begin biological
evolution.
Experimental Approaches to Fabricating Artificial Cellular Life 21

1992; Ourisson and Nakatani, 1994; Segre´, Deamer, and Lancet, 2001; Szostak, Bar-
tel, and Luisi, 2001).
2.2.1 Self-Assembly Processes in Organic Mixtures
What physical properties are required for a molecule to become incorporated into a
stable bilayer? All bilayer-forming molecules are amphiphiles, with a hydrophilic
‘‘head’’ and a hydrophobic ‘‘tail’’ on the same molecule. If amphiphilic molecules
were present in the mixture of organic compounds available on the early Earth, it is
not di‰cult to imagine that their self-assembly into molecular aggregates was a com-
mon process.
It is reasonable to conclude that a variety of simpler amphiphilic molecules can
participate in the formation of membrane structures. The long-chain fatty acids and
alcohols that contribute the amphiphilic property of contemporary membrane lipids
are possible components. Signiﬁcantly, such simple amphiphiles readily form vesicles
(Apel, Deamer, and Mautner, 2002; Monnard et al., 2002). Stability of the vesicles is
strongly dependent on chain length, pH, ionic strength, amphiphile composition
and concentration, temperature, and head group characteristics. For example, even
a 9-carbon monocarboxylic acid nonanoic acid can form vesicles at concen-
trations of 85 mM and pH 7.0, which is the pK of the acid in bilayers. At this pH,
half of the carboxylic acid groups are protonated, half are anions, and hydrogen
bonding between the protonated and anionic head groups stabilizes the bilayer con-
ﬁguration. Addition of small amounts of a nonanol can also stabilize the bilayers
as a result of hydrogen bonding between the alcohol and acid head groups, so
that vesicles form at lower concentrations (@20 mM) over a pH range from 7 to
Figure 2.2
A protocell would have a minimal set of functional properties, including self assembly of boundary mem
branes, transport of monomers, capture of energy to drive polymerization reactions, and encapsulation of
polymer systems capable of growth.
22 David Deamer

11. The vesicles provide a selective permeability barrier, as indicated by osmotic
activity and entrapment of polar dyes. As chain length increases, stability also in-
creases, and vesicles form at lower concentrations (Apel et al., 2002; Monnard et al.,
2002).
2.2.2 What Amphiphiles Self-Assemble into Membranes?
Amphiphilic molecules are among the simplest of life’s molecular components, and
are readily synthesized by nonbiological processes. Virtually any normal alkane hav-
ing 10 or more carbons in its chain takes on amphiphilic properties if one end of the
molecule incorporates a polar or ionic group (see below). The simplest common
amphiphiles are therefore molecules such as monocarboxylic acids (anions), mono-
amines (cations), and alcohols (neutral polar groups).
CH
3-ðCH2Þn-COOH!H
þ
þCH 3-ðCH2Þn-COOðanionÞ
CH
3-ðCH2Þn-NH2þH
þ
!CH 3-ðCH2Þn-NH3
þðcationÞ
CH
3-ðCH2Þn-OHðneutral amphiphileÞ
Lipids are far more diverse chemically than other typical biomolecules such as
amino acids, carbohydrates, and nucleotides. The deﬁnition of lipids includes simple
fatty acids and their glycerol esters, sterols such as cholesterol, and phospholipids,
sphingolipids, and cerebrosides. Lipids are generally deﬁned by their common hydro-
phobic character, which makes them soluble in organic solvents such as chloroform.
Virtually all lipids also have a hydrophilic group, which makes them surface-active.
Eukaryotic phospholipids typically have two fatty acid chains esteriﬁed to a glyc-
erol, with the third position of the glycerol esteriﬁed to a phosphate group. Most
phospholipids also have a head group such as choline, ethanolamine, or serine
attached to the phosphate. One such lipid is shown in ﬁgure 2.3. The precise function
of the variable head groups has not yet been established.
The other lipid commonly present in eukaryotic membranes is cholesterol, a poly-
cyclic structure produced from isoprene by a complex biosynthetic pathway.
Figure 2.3
A phospholipid (1 palmitoyl, 2 oleoyl phosphatidylcholine). This molecule is amphiphilic as it has a
‘‘water hating’’ (hydrophobic) tail and a ‘‘water loving’’ (hydrophilic) head.
Experimental Approaches to Fabricating Artificial Cellular Life 23

2.3 The Fluid Mosaic Model of Membrane Structure
In the 1970s, the ﬂuid mosaic concept emerged as the most plausible model to ac-
count for the known structure and properties of biological membranes (Singer and
Nicolson, 1972). The fact that membranes exist as two-dimensional ﬂuids (liquid-
disordered), rather than in a gel state (solid-ordered), was clearly demonstrated by
Frye and Edidin (1970), who showed that the lipid and protein components of two
separate membranes di¤used into each other when two di¤erent cells were fused.
Since that time, numerous studies have measured the di¤usion coe‰cient of lipids
and proteins in membranes, and the di¤usion rates were found to correspond to
those expected of a ﬂuid with the viscosity of olive oil, rather than a gel phase resem-
bling wax.
The lipid components of membranes must be in a ﬂuid state to function as mem-
branes in living cells. Straight-chain fatty acids have relatively high melting points
because of the ease with which van der Waals interactions can occur along the hy-
drocarbon chains. Any discontinuity in the chains interrupts these interactions and
markedly decreases the melting point. As an example, stearic acid contains 18 car-
bons in its alkane chain and melts at 68

C, while oleic acid, with acis-double bond
between carbons 9 and 10, has a melting point of 16

C. If cellular life requires ﬂuid
membranes, it is reasonable to assume that the membranes of artiﬁcial cells could
also be composed of amphiphilic molecules in a ﬂuid state. However, alternative
boundary structures could also be incorporated, such as a self-assembling protein
coat resembling that of viruses.
The idea that the proteins of biological membranes are embedded in a ﬂuid sea of
lipids arose from our increasing understanding of membrane structure. It has been
demonstrated in numerous ways that most of the proteins associated with mem-
branes are embedded in the lipid bilayer phase, rather than simply adhering to the
surface. As a general rule, membrane proteins have stretches of hydrophobic amino
acids in their sequences, and these are threaded back and forth through the bilayer
multiple times, thereby anchoring the protein to the membrane. The hydrophobic
proteins often are involved in production of pores, or transmembrane channels, that
are essential for ion and nutrient transport processes.
Could channels capable of nutrient transport be produced in the bilayer mem-
branes of artiﬁcial cells? In fact, channel-like defects do appear when nonpolar pep-
tides interact with a lipid bilayer. For instance, polyleucine or polyalanine have been
induced to fuse with planar lipid membranes, and the bilayers exhibited transient
bursts of ionic conductance (Oliver and Deamer, 1994). More complex synthetic pep-
tides have also been demonstrated to produce ion-conducting channels in lipid
bilayers (Lear, Wasserman, and DeGrado, 1988). It is fair to expect that a variety
of polymers are likely to be able to penetrate bilayer membranes and produce chan-
24 David Deamer

nels that bypass the permeability barrier. This is an area that is ripe for further inves-
tigations, as described in a recent review by Pohorille, Schweighofer, and Wilson
(2005).
2.4 Function of Membranes in Artificial Cells
Membranes have many functions in addition to acting as containers for the macro-
molecular polymers of life. Three primary membrane functions associated with an
artiﬁcial cell might include selective inward transport of nutrients from the environ-
ment, capture of the energy available in light or oxidation-reduction potentials, and
coupling of that energy to some form of energy currency such as ATP to drive poly-
mer synthesis.
The simplest of these functions is that of a permeability barrier, which limits free
di¤usion of solutes between the cytoplasm and the external environment. Although
such barriers are essential for cellular life to exist, a mechanism by which selective
permeation allows speciﬁc solutes to cross the membrane must also exist. In contem-
porary cells, such processes are carried out by transmembrane proteins, which act as
channels and transporters. Examples include the proteins that facilitate the transport
of glucose and amino acids into the cell, channels that allow potassium and sodium
ions to permeate the membrane, and active transport of ions by enzymes that use
ATP as an energy source.
If we are to assemble an artiﬁcial cell, it will be necessary to overcome the mem-
brane permeability barrier. One possible way to accomplish this could be simple dif-
fusion across the bilayer. To give a perspective on permeability and transport rates
by di¤usion, we can compare the ﬂuxes of relatively permeable and relatively imper-
meable solutes across contemporary lipid bilayers. The measured permeability of lip-
id bilayers to small, uncharged molecules such as water, oxygen, and carbon dioxide
is greater than the permeability to ions by a factor of@10
9
. For instance, the perme-
ability coe‰cient of water is approximately 10
3
cm s
1
, and the permeability coe‰-
cient of potassium ions is 10
11
cm s
1
. By themselves, these values mean little, but
they make more sense in the context of time required for exchange across a bilayer.
Measurements show that half the water in a liposome exchanges in milliseconds,
whereas potassium ion half-times of exchange are measured in days.
We can now consider some typical nutrient solutes like amino acids and phos-
phates. Such molecules are ionized, which means that they would not readily cross
the permeability barrier of a lipid bilayer. Permeability coe‰cients of liposome
membranes to phosphate and amino acids have been determined (Chakrabarti and
Deamer, 1994) and were found to be in the range of 10
11
to 10
12
cm s
1
, similar
to ionic solutes such as sodium and chloride ions. From these ﬁgures one can esti-
mate that if an artiﬁcial cell depended on passive transport of phosphate across a
Experimental Approaches to Fabricating Artificial Cellular Life 25

lipid bilayer composed of a typical phospholipid, it would require several years to
accumulate phosphate su‰cient to double its DNA content, or pass through one
cell cycle. In contrast, a modern bacterial cell can reproduce in as short a time as 20
minutes.
Given that lipid bilayers are so impermeable to typical polar and ionic solutes,
how can we design artiﬁcial cells so that they will have access to essential nutrients?
One clue may be that highly evolved modern lipids are products of several billion
years of evolution, and typically contain hydrocarbon chains 16 to 18 carbons in
length. These chains provide an interior ‘‘oily’’ portion of the lipid bilayer that repre-
sents a nearly impermeable barrier to the free di¤usion of ions such as sodium and
potassium. The reason is related to the common observation that ‘‘oil and water
don’t mix.’’ That is, ion permeation of the hydrophobic portion of a lipid bilayer
faces a very high energy barrier called Born energy, which is associated with the dif-
ference in energy for an ion in a high dielectric medium (water with a dielectric con-
stant of 80) compared to the same ion in a low dielectric medium (hydrocarbon with
a dielectric constant of 2). This energy barrier is immense, up to 40 kcal mole
1
(Par-
segian, 1969).
However, recent studies have shown that permeability is strongly dependent on
chain length (Paula et al., 1996). For instance, shortening phospholipid chains from
18 to 14 carbons increases permeability to ions by a thousandfold (ﬁgure 2.4). The
reason is that thinner membranes have increasing numbers of transient defects that
open and close on nanosecond time scales, so that ionic solutes can get from one
side of the membrane to the other without dissolving in the oily interior phase of
the bilayer. Ionic solutes even as large as ATP can di¤use cross a bilayer composed
of dimyristoylphosphatidylcholine, a 14-carbon phospholipid (Monnard and
Deamer, 2001). We conclude that initial approaches to fabricating an artiﬁcial cell
do not necessarily depend on peptide channels to provide nutrient transport. It may
be su‰cient simply to prepare the membranes with a lipid composition that permits
relatively fast di¤usion of small substrate molecules, yet can maintain macromolecu-
lar components in the internal volume.
2.5 Growth Processes in Artificial Cells
Earlier reports (Walde, Goto, et al., 1994; Walde, Wick, et al., 1994) showed that
vesicles composed of oleic acid can grow and ‘‘reproduce’’ as oleoyl anhydride spon-
taneously hydrolyzed in the reaction mixture, thereby adding amphiphilic compo-
nents (oleic acid) to the vesicle membranes. This approach has recently been
extended by Hanczyc and coworkers (Hanczyc, Fujikawa, and Szostak, 2003; Hanc-
zyc and Szostak, 2004; see also chapter 5), who prepared myristoleic acid membranes
under deﬁned conditions of pH, temperature, and ionic strength. The process by
26 David Deamer

which the vesicles formed from micellar solutions required several hours, apparently
with a rate-limiting step related to the assembly of ‘‘nuclei’’ of bilayer structures.
However, if a mineral surface in the form of clay particles was present, the surface
in some way catalyzed vesicle formation, reducing the time required from hours to a
few minutes. The clay particles were spontaneously encapsulated in the vesicles. The
authors further found that RNA bound to the clay was encapsulated as well, and
remained within the vesicles for extended periods of time.
In a second series of experiments, Hanczyc, Fujikawa, and Szostak (2003; see also
chapter 5) found that the myristoleic acid vesicles could be induced to grow by add-
ing fatty acid to the medium, presumably by incorporating fatty acid molecules into
the membrane rather than by fusion of vesicles. If the resulting suspension of large
vesicles was then ﬁltered through a polycarbonate ﬁlter with pores 0.2mm in diame-
ter, the larger vesicles underwent a kind of shear-induced division to produce smaller
Figure 2.4
Stability and permeability of self assembled amphiphilic structures. Amphiphilic molecules such as fatty
acids having carbon chain lengths of 9 or more carbons form bilayer membranes when su‰ciently concen
trated. (A) Pure bilayers of ionized fatty acid are relatively unstable, but become markedly more stable
in long chain alcohols are added. (B) Dimensions of the amphiphile also play a role. Shorter chain
amphiphiles (9 10 carbons) are less able to form bilayers, while those of intermediate chain length (12 14
carbons) produce stable bilayers that also are permeable to ionic and polar solutes. Longer chain lengths
(16 18 carbons) produce bilayers that are increasingly less permeable to solutes (Paula et al., 1996).
Experimental Approaches to Fabricating Artificial Cellular Life 27

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CHAP. X.
VISIT TO THE NINGPO GREEN TEA DISTRICT.—MOUNTAIN
TRAVELLING CHAIR.—THE BUDDHIST TEMPLE OF TEIN-TUNG.—
SCENERY AROUND THE TEMPLE.—TRADITIONAL HISTORY
COMMUNICATED BY THE PRIEST.—THE TEMPLE AND ITS IDOLS.—
INVITATION TO DINNER WITH THE PRIESTS.—FIRST TRIAL WITH
CHOPSTICKS.—POLITENESS! OF THE CHINESE.—USUAL DINNER
COMPANY.—MY BED.—DEVOTIONS OF THE PRIESTS.—WILD BOAR
HUNT.—NARROW ESCAPE.—MODE OF FRIGHTENING THE ANIMALS
FROM THE BAMBOO PLANTATIONS.—MOUNTAIN SCENERY.—
BUDDHIST TEMPLE OF AH-YU-WANG.-POO-TO-SAN, OR THE
WORSHIPPING ISLAND.—ITS TEMPLES AND IDOLS.—BRONZE
GODS.—TREES AND SHRUBS.—GARDENS AND PET PLANTS OF THE
PRIESTS.—SALE OF GODS OR JOSSES.—OFFERINGS TO THE GODS
IN SHANGHAE AND NINGPO.—PROCESSIONS IN HONOUR OF THE
GODS.—CHRISTIAN MISSIONS.—MEDICAL MISSIONARY SOCIETY.—
ROMAN CATHOLICS.
Having despatched my collections to England by three different
vessels from Hong-kong, I sailed again, at the end of March 1844,

for the northern provinces. During the summer of this year, and in
that of 1845, I was able to visit several parts of the country, which
were formerly scaled to Europeans, and which contained subjects of
much interest.
About the beginning of May I set out upon an excursion with Mr.
Thom, the British consul, and two other gentlemen, to visit the
green tea district near Ningpo. We were informed that there was a
large and celebrated temple, named Tein-tung, in the centre of the
tea district, and above twenty miles distant, where we could lodge
during our stay in this part of the country. Twelve or fourteen miles
of our journey was performed by water, but the canal ending at the
foot of the hills we were obliged to walk, or take chairs for the
remainder of the way. The mountain travelling chair of China is a
very simple contrivance. It consists merely of two long bamboo
poles, with a board placed between them for a seat, and two other
cross pieces, one for the back and the other for the feet: a large
Chinese umbrella is held over the head to afford protection from the
sun and rain.
Mountain Chair.

The Chinese are quite philosophers after their own fashion. On our
way to the temple, when tired with sitting so long in our boat, we
several times got out and walked along the path on the sides of the
canal. A great number of passage-boats going in the same direction
with ourselves, and crowded with passengers, kept very near us for
a considerable portion of the way, in order to satisfy their curiosity. A
Chinaman never walks when he can possibly find any other mode of
conveyance, and these persons were consequently much surprised
to see us apparently enjoying our walk.
"Is it not strange," said one, "that these people prefer walking when
they have a boat as well as ourselves?" A discussion now took place
amongst them as to the reason of this apparently strange
propensity, when one, more wise than his companions, settled the
matter by the pithy observation, "It is their nature to do so;" which
was apparently satisfactory to all parties.
It was nearly dark when we reached the temple, and as the rain had
fallen in torrents during the greater part of the day, we were
drenched to the skin, and in rather a pitiable condition. The priests
seemed much surprised at our appearance, but at once evinced the
greatest hospitality and kindness, and we soon found ourselves quite
at home amongst them. They brought us fire to dry our clothes, got
ready our dinner, and set apart a certain number of their best rooms
for us to sleep in. We were evidently subjects of great curiosity to
most of them who had never seen an Englishman before. Our
clothes, features, mode of eating, and manners were all subjects of
wonder to these simple people, who passed off many a good
humoured joke at our expence.
Glad to get off our clothes, which were still damp, we retired early to
rest. When we arose in the morning, the view which met our eyes
far surpassed in beauty any scenery which I had ever witnessed
before in China. The temple stands at the head of a fertile valley in
the bosom of the hills. This valley is well watered by clear streams,
which flow from the mountains, and produces most excellent crops
of rice. The tea shrubs, with their dark green leaves, are seen dotted

on the lower sides of all the more fertile hills. The temple itself is
approached by a long avenue of Chinese pine trees. This avenue is
at first straight, but near the temple it winds in a most picturesque
manner round the edges of two artificial lakes, and then ends in a
flight of stone steps, which lead up to the principal entrance. Behind,
and on each side, the mountains rise, in irregular ridges, from one to
two thousand feet above the level of the sea. These are not like the
barren southern mountains, but are clothed nearly to their summits
with a dense tropical looking mass of brushwood, shrubs, and trees.
Some of the finest bamboos of China are grown in the ravines, and
the sombre coloured pine attains to a large size on the sides of the
hills. Here, too, I observed some very beautiful specimens of the
new fir (Cryptomeria japonica), and obtained some plants and seeds
of it, which may now be seen growing in the Horticultural Gardens at
Chiswick.
After we had breakfasted, one of the head priests came and gave us
a very pressing invitation to dine with him about mid-day; and in the
meantime he accompanied us over the monastery, of which he gave
the following history:—"Many hundred years ago a pious old man
retired from the world, and came to dwell in these mountains, giving
himself up entirely to the performance of religious duties. So earnest
was he in his devotions that he neglected everything relating to his
temporal wants, even to his daily food. Providence, however, would
not suffer so good a man to starve. Some boys were sent in a
miraculous manner, who daily supplied him with food. In the course
of time the fame of the sage extended all over the adjacent country,
and disciples flocked to him from all quarters. A small range of
temples was built, and thus commenced the extensive buildings
which now bear the name of "Tein-tung," or the "Temple of the
Heavenly Boys;" Tein signifying heaven, and tung a boy. At last the
old man died, but his disciples supplied his place. The fame of the
temple spread far and wide, and votaries came from the most
distant parts of the empire—one of the Chinese kings being amongst
the number—to worship and leave their offerings at its altars. Larger
temples were built in front of the original ones, and these again in

their turn gave way to those spacious buildings which form the
principal part of the structure of the present day.
All the temples are crowded with idols, or images of their favourite
gods, such as the "three precious Buddhas," "the Queen of
Heaven"—represented as sitting on the celebrated lotus or
nelumbium—"the God of War," and many other deified kings and
great men of former days. Many of these images are from thirty to
forty feet in height, and have a very striking appearance when seen
arranged in these spacious and lofty halls. The priests themselves
reside in a range of low buildings, erected at right angles with the
different temples and courts which divide them. Each has a little
temple in his own house—a family altar crowded with small images,
where he is often engaged in private devotion.
After inspecting the various temples and the belfry, which contains a
noble bronze bell of large dimensions, our host conducted us back to
his house, where the dinner was already on the table. The priests of
the Buddhist religion are not allowed to eat animal food at any of
their meals. Our dinner therefore consisted entirely of vegetables,
served up in the usual Chinese style, in a number of small round
basins, the contents of each—soups excepted—being cut up into
small square bits, to be eaten with chopsticks. The Buddhist priests
contrive to procure a number of vegetables of different kinds, which,
by a peculiar mode of preparation, are rendered very palatable. In
fact, so nearly do they resemble animal food in taste and in
appearance, that at first we were deceived, imagining that the little
bits we were able to get hold of with our chopsticks were really
pieces of fowl or beef. Such, however, was not the case, as our good
host was consistent on this day at least, and had nothing but
vegetable productions at his table. Several other priests sat with us
at table, and a large number of others of inferior rank with servants,
crowded around the doors and windows outside. The whole
assemblage must have been much surprised at the awkward way in
which some of us handled our chopsticks, and, with all their
politeness, I observed they could not refrain from laughing when,
after repeated attempts, some little dainty morsel would still slip

back again into the dish. I know few things more annoying, and yet
laughable too, than attempting to eat with the Chinese chopsticks
for the first time, more particularly if the operator has been
wandering on the hills all the morning, and is ravenously hungry.
The instruments should first of all be balanced between the thumb
and forefinger of the right hand; the points are next to be brought
carefully together, just leaving as much room as will allow the
coveted morsel to go in between them; the little bit is then to be
neatly seized; but alas! in the act of lifting the hand, one point of the
chopstick too often slips past the other, and the object of all our
hopes drops back again into the dish, or perhaps even into another
dish on the table. Again and again the same operation is tried, until
the poor novice loses all patience, throws down the chopsticks in
despair, and seizes a porcelain spoon, with which he is more
successful. In cases like these the Chinese themselves are very
obliging, although scarcely in a way agreeable to an Englishman's
taste. Your Chinese friend, out of kindness and politeness, when he
sees the dilemma in which you are, reaches across the table and
seizes, with his own chopsticks, which have just come out of his
mouth, the wished-for morsel, and with them lays it on the plate
before you. In common politeness you must express your gratitude
and swallow the offering.
During dinner our host informed us that there were about a hundred
priests connected with the monastery, but that many were always
absent on missions to various parts of the country. On questioning
him as to the mode by which the establishment was supported, he
informed us that a considerable portion of land in the vicinity
belonged to the temple, and that large sums were yearly raised from
the sale of bamboos, which are here very excellent, and of the
branches of trees and brushwood, which are made up in bundles for
fire-wood. A number of tea and rice farms also belong to the priests,
which they themselves cultivate. Besides the sums raised by the sale
of these productions, a considerable revenue must be derived from
the contributions of the devotees who resort to the temple for
religious purposes, as well as from the sums collected by those of

the order who are out on begging excursions at stated seasons of
the year. The priests are of course of all grades, some of them being
merely the servants of the others, both in the house and in the
fields. They seem a harmless and simple race, but are dreadfully
ignorant and superstitious. The typhoon of the previous year, or
rather the rain which had accompanied it, had occasioned a large
slip of earth on one of the hill-sides near the temple, and completely
buried ten or twelve acres of excellent paddy land. On our remarking
this, the priests told us with great earnestness that every one said it
was a bad omen for the temple; but one of them with true Chinese
politeness remarked that he had no doubt any evil influence would
now be counteracted, since the temple had been honoured with a
visit from us.
After inspecting the tea farms and the mode of manufacturing it, Mr.
Thom, Mr. Morrison, a son of the late Dr. Morrison, and Mr. Sinclair,
returned to Ningpo, leaving me to prosecute my research in natural
history in this part of the country. I was generally absent from the
temple the whole day, returning at dark with the collections of plants
and birds which I had been lucky enough to meet with in my
peregrinations. The friends of the priests came from all quarters of
the adjacent country to see the foreigner; and, as in the case of a
wild animal, my feeding time seemed to be the most interesting
moment to them. My dinner was placed on a round table in the
centre of the room, and although rather curiously concocted, being
half Chinese and half English, the exercise and fresh air of the
mountains gave me a keen appetite. The difficulties of the
chopsticks were soon got over, and I was able to manage them
nearly as well as the Chinese themselves. The priests and their
friends tilled the chairs, which are always placed down the sides of a
Chinese hall, each man with his pipe in his mouth and his cup of tea
by his side. With all deference to my host and his friends, I was
obliged to request the smoking to be stopped, as it was disagreeable
to me while at dinner; in other respects, I believe I was "polite"
enough. I shall never forget how inexpressibly lonely I felt the first
night after the departure of my friends. The Chinese one by one

dropt off to their homes or to bed, and at last my host himself gave
several unequivocal yawns, which reminded me that it was time to
retire for the night. My bed-room was upstairs, and to get to it I had
to pass through a small temple, such as I have already noticed,
dedicated to Tein-how, or the "Queen of Heaven," and crowded with
other idols. Incense was burning on the altar in front of the idols; a
solitary lamp shed a dim light over the objects in the room, and a
kind of solemn stillness seemed to pervade the whole place. In the
room below, and also in one in an adjoining house, I could hear the
priests engaged in their devotional exercises, in that singing tone
which is peculiar to them. Then the sounds of the gong fell upon my
ears; and, at intervals, a single solemn toll of the large bronze bell in
the belfry; all which showed that the priests were engaged in public
as well as private devotion. Amidst scenes of this kind, in a strange
country, far from friends and home, impressions are apt to be made
upon the mind, which remain vivid through life; and I feel convinced
I shall never forget the strange mixture of feelings which filled my
mind during the first night of my stay with the priests in the temple
of Tein-tung. I have visited the place often since, passed through the
same little temple, slept in the same bed, and heard the same
solemn sounds throughout the silent watches of the night, and yet
the first impressions remain in my mind distinct and single.
The priests, from the highest to the lowest, always showed me the
most marked attention and kindness. As many of them as I wished
cheerfully followed me in my excursions in the vicinity of the temple;
one carrying my specimen paper, another my plants, and a third my
birds, and so on. The gun seemed an object of great interest to
them, being so different from their own clumsy matchlocks; and
percussion caps were looked upon as most magical little objects. But
they were great cowards, and always kept at a most respectful
distance when I was shooting.
One evening a deputation, headed by the high priest, came and
informed me that the wild boars had come down from the mountains
at night, and were destroying the young shoots of the bamboo,
which were then just coming through the ground, and were in the

state in which they are highly prized as a vegetable for the table.
"Well," said I, "what do you want me to do?"
"Will you be good enough to lend us the gun?"
"Yes; there it stands in the corner of the room."
"Oh, but you must load it for us."
"Very well, I will;" and I immediately loaded the gun with ball.
"There, but take care and don't shoot yourselves." There was now a
long pause; none had sufficient courage to take the gun, and a long
consultation was held between them. At length the spokesman came
forward, with great gravity, and told me they were afraid to fire it
off, but that if I would go with them, and shoot the boar, I should
have it to eat. This was certainly no great sacrifice on the part of the
Budhist priesthood, who do not, or at least should not, eat animal
food. We now sallied forth in a body to fight the wild boars; but the
night was so dark that we could see nothing in the bamboo ravines,
and, perhaps, the noise made by about thirty priests and servants
warned the animals to retire to the brushwood higher up the hills.
Be that as it may, we could neither see nor hear any thing of them,
and I confess I was rather glad, than otherwise, as I thought there
was a considerable chance of my shooting, by mistake, a priest
instead of a wild boar.
The priests have two modes of protecting their property from the
ravages of these animals. Deep pits are dug on the hill sides, and, as
there are springs in almost all these places, the pits are scarcely
finished before they are half full of water. The mouth of each pit is
then covered over with a quantity of sticks, rubbish, and grass, to
attract the animal, and no sooner does he begin to bore into it with
his snout, than the whole gives way, and he is plunged, head
foremost, into the pit, from which it is quite impossible for him to
extricate himself, and he is either drowned or becomes an easy prey
to the Chinese. These pits are most dangerous traps to persons
unacquainted with the localities in which they are placed. I had
several narrow escapes, and once in particular, when coming out of

a dense mass of brushwood, I stept unawares on the treacherous
mouth of one of them, and felt the ground under my feet actually
giving way; but managing to throw my arms forward I caught hold
of a small twig which was growing near, and by this means
supported myself until I was able to scramble on to firmer ground.
On turning back to examine the place, I found that the loose rubbish
had sunk in, and a deep pit, half full of water, was exposed to my
view. The pit was made narrow at the mouth and widening inside
like a great China vase, being constructed in this manner to prevent
the boar from scrambling out when once fairly in it. Had I fallen in, it
would have been next to impossible to have extricated myself
without assistance, and as the pits are generally dug in the most
retired and wild part of the mountains, my chance would have been
a bad one. The fate of my predecessor, Mr. Douglas, who perished in
a pit of this kind on the Sandwich Islands, must still be fresh in the
recollection of many of my readers, and his melancholy end naturally
coming to my mind at the time, made me doubly thankful for my
escape.
The other method of protecting the young bamboos from the
ravages of the wild boar, is an ingenious one. A piece of bamboo
wood, about eight or ten feet long, and rather thicker than a man's
arm, is split up the middle to within a fourth of its length. This is
made fast to a tree in the bamboo thicket, and at an angle of about
forty-five degrees, the split part being left loose, a cord, also made
of bamboo, is fastened to it by one end, and the other is led to some
convenient place out of the thicket, where a man is stationed. When
the boars come down in the dead of night to attack the young
shoots, the man pulls the rope backwards and forwards, and clank,
clank, clank goes the bamboo, producing a loud and hollow sound,
which on a quiet evening may be heard at a great distance. The
animals are frightened and make off to their dens on the hills. The
first time I heard these things beating at night, all over the country, I
imagined that some religious ceremony was going on, the hollow
sounds of the bamboo being not unlike those produced by an
instrument used in the Budhist worship in all Chinese temples.

There are a large number of Budhist temples scattered over all this
part of the country. One, named Ah-yu-wang, which I also visited, is,
like Tein-tung, of great extent, and seemingly well supported. They
both own large tracts of land in the vicinity of the monasteries, and
have numerous small temples in different parts of the district which
are under their control. All the temples, both large and small, are
built in the most romantic and beautiful situations amongst the hills,
and the neighbouring woods are always preserved and encouraged.
What would indicate the residence of a country gentleman in
England, is in China the sign of a Budhist temple, and this holds
good over all the country. When the weary traveller, therefore, who
has been exposed for hours to the fierce rays of an eastern sun,
sees a large clean looking house showing itself amongst trees on the
distant hill-side, he may be almost certain that it is one of Budha's
temples, where the priests will treat him not only with courtesy, but
with kindness.
Poo-to, or the Worshipping Island, as it is commonly called by
foreigners, is one of the eastern islands in the Chusan Archipelago,
and seems to be the capital or stronghold of Budhism in this part of
China. This island is not more than five or six miles in circumference,
and, although hilly, its sides and small ravines are pretty well
wooded, particularly in the vicinity of the numerous temples. As it is
only a few hours' sail from Chusan, it had been visited at different
times by a number of our officers during the war, all of whom spoke
highly of its natural beauties and richness of vegetation. I was also
informed that the resident priests were fond of collecting plants,
particularly Orchidaceæ, and that their collections were much
increased by the itinerant habits of the begging priests, who visit the
most distant provinces of the empire, as well as by the donations of
the lay devotees, who come to Poo-to at stated seasons of the year,
to worship and leave their offerings in the temples. I therefore
determined to visit the place in order to judge for myself, and
accordingly set out in July, 1844, accompanied by my friend, Dr.
Maxwell, of the Madras army.

Leaving Chusan at night, with the tide in our favour, we reached the
island at sun-rise on the following morning. We landed, and pursued
our way over a hill and down on the other side by a road which led
us into a beautiful and romantic glen. It is here that the principal
group of temples is built, and when we first caught a glimpse of
them, as we wended our way down the hill, they seemed like a town
of considerable size. As we approached nearer, the view became
highly interesting. In front there was a large artificial pond, filled
with the broad green leaves and noble red and white flowers of the
Nelumbium speciosum,—a plant in high favour with the Chinese.
Every body who went to Poo-to admired these beautiful water-lilies.
In order to reach the monastery we crossed a very ornamental
bridge built over this pond, which, when viewed in a line with an old
tower close by, has a pretty and striking appearance.
The temples or halls which contain the idols are extremely spacious,
and resemble those which I have already described at Tein-tung and
Ah-yu-Wang. These idols, many of which are thirty or forty feet in
height, are generally made of wood or clay, and then richly gilt.
There is one small temple, however, of a very unassuming
appearance, where we met with some exquisite bronze statues,
which would be considered of great value in England. These, of
course, were much smaller than the others, but, viewed as works of
art, they were by far the finest which I saw during my travels in
China.
Having examined these temples, we pursued our way towards
another assemblage of them, about two miles to the eastward and
close on the sea-shore. We entered the courts through a kind of
triumphal arch, which looks out upon the sea, and found that these
temples were constructed upon the same plan as all the others. As
we had determined to make this part of the island our home during
our stay, we fixed upon the cleanest looking temple, and asked the
High Priest to allow us, without farther delay, to put our beds and
travelling baggage into it.

On the following day we inspected various parts of the island.
Besides the large temples just noticed, there are about sixty or
seventy smaller ones, built on all the hill sides, each of which
contains three or four priests, who are all under the superior, or
abbot, who resides near one of the large temples. Even on the top
of the highest hill, probably 1500 or 1800 feet above the level of the
sea, we found a temple of considerable size and in excellent repair.
There are winding stone steps from the sea beach all the way up to
this temple, and a small resting-place about half way up the hill,
where the weary devotee may rest and drink of the refreshing
stream which flows down the sides of the mountain, and in the little
temple close at hand, which is also crowded with idols, he can
supplicate Budha for strength to enable him to reach the end of his
journey. We were surprised to find a Budhist temple in such
excellent order as the one on the summit of the hill proved to be in.
It is a striking fact, that almost all these places are crumbling fast
into ruins. There are a few exceptions, in cases where they happen
to get a good name amongst the people from the supposed kindness
of the gods; but the great mass are in a state of decay.
From the upper temple on Poo-to-san the view is strikingly grand.
Hugged mountains are seen rising one above another and capped
with clouds. Hundreds of islands, some fertile, others rocky and
barren, lay scattered over the sea. When we looked in one direction
amongst the islands, the water was yellow and muddy; but, to the
eastward, the deep blue ocean had resumed its usual colour, and the
line between the yellow waters and the blue was distinctly and
curiously marked.
The wood on the island is preserved in the same manner as it is
around all the other Budhist temples. The principal species of trees
and shrubs met with are Pinus sinensis, Cunningham lanceolata,
yews, cypresses, the camphor tree, tallow tree, oaks, and bamboos.
The Camellia japonica grows spontaneously in the woods, where we
met with many specimens from twenty to thirty feet in height and
with stems thick in proportion. The variety, however, was only the

well-known single red. In other respects the flora of Poo-to is nearly
the same as on the island of Chusan.
A few pet plants were cultivated by all the priests who were
fortunate enough to have private residences at the little temples on
the sides of the hills. We were much pleased with the interest these
poor people took in their favourite flowers, but were disappointed in
the number and variety of plants, which, from the reports of others,
we expected to have found. Almost the only orchidaceous plant
which they had, proved to be the common and well-known
Cymbidium sinense. Daphne odorata, two or three species of
Gardenia, several varieties of Rose, the common Balsam, and the
favourite Nelumbium were nearly all the plants met with in the
gardens of the priests.
The island of Poo-to is set apart entirely as a residence for the
priests of the Budhist religion. Few other persons are allowed to live
there, and these are either servants or in some way connected with
the priests. No women are permitted to reside on the island, it being
against the principles of the Budhists to allow their priests to marry.
The number of priests are estimated at 2000, but many of them are
constantly absent on begging expeditions for the support of their
religion. This establishment, like Tein-tung, has also a portion of land
allotted to it for its support, and the remainder of the funds are
made up by the subscriptions of the devotees. On certain high days,
at different periods of the year, many thousands of both sexes, but
particularly females, resort to these temples, clad in their best attire,
to pay their vows and engage in the other exercises of heathen
worship. Little stalls are then seen in the temples or at the doorways
for the sale of incense, candles, paper made up in the form of the
ingots of Sycee-silver, and other holy things which are considered
acceptable offerings to the gods, and are either consumed in the
temples or carried home to bring a blessing upon the houses and
families of those who purchase them. The profits of these sales, of
course, go to the support of the establishment. When we consider
that these poor deluded people sometimes travel a distance of
several hundred miles to worship in the temples on Poo-to-san and

other celebrated places, we cannot but admire their spirit of
devotion. I was once staying in the temple of Tein-tung when it was
visited for three days by devotees from all parts of the country. As
they lined the roads on their way to the temple, clad in the graceful
and flowing costume of the East, the mind was naturally led back to
those days of scripture history when Jerusalem was in its glory, and
the Jews, the chosen people of God, came from afar to worship in its
temple.
Although no Christian can look upon the priests and devotees of the
Budhist creed without an eye of pity, yet he must give them credit
for their conduct, since he has every reason to believe them sincere,
and I am inclined to believe that justice has not been done them in
this respect. Mr. Gutzlaff, in describing his visit to Poo-to, is of a
different opinion. He says, "We were present at the vespers of the
priests, which they chanted in the Pali language, not unlike the Latin
service of the Romish church. They held rosaries in their hands,
which rested folded upon their breasts. One of them had a small
bell, by the tinkling of which their service was regulated; and they
occasionally beat the drum and large bell to rouse Budha's attention
to their prayers. The same words were a hundred times repeated.
None of the officiating persons showed any interest in the ceremony,
for some were looking around laughing and joking, while others
muttered their prayers. The few people who were present, not to
attend the worship, but to gaze at us, did not seem, in the least
degree, to feel the solemnity of the service. "What Mr. Gutzlaff says
is doubtless true, but after residing for months in their temples, at
different times, and in different parts of the country, I have no
hesitation in saying that such conduct is very far from being general.
In certain instances I have seen it myself, but this levity and
apparent want of attention was exhibited by the servants and
lookers on, who were taking no part in the ceremony, and not by the
respectable portion of the priests. On the contrary, I have generally
been struck with the solemnity with which their devotional exercises
were conducted. I have often walked into Chinese temples when the
priests were engaged in prayer, and, although there would have

been some apology for them had their attention been diverted, they
went on in the most solemn manner until the conclusion of the
service, as if no foreigner were present. They then came politely up
to me, examining my dress and every thing about me with the most
earnest curiosity. Nor does this apply to priests only; the laity, and
particularly the female sex, seem equally sincere when they engage
in their public devotions. Whether they are what they appear to be,
or how often they are in this pious frame of mind, are questions
which I cannot answer. Before judging harshly of the Chinese let the
reader consider what effect would be produced upon the members
of a Christian church by the unexpected entrance of a small-footed
Chinese lady, or a Mandarin, with the gold button and peacock
feather mounted on his hat, and his long tail dangling over his
shoulders. I am far from being an admirer of the Budhist priesthood;
they are generally an imbecile race, and shamefully ignorant of every
thing but the simple forms of their religion, but nevertheless there
are many traits in their character not unworthy of imitation.
There are two other sects in China, namely, the followers of Kong-
foo-tze or Confucius, and the sect of Taou or Reason. Although these
three sects form the principal part of the population, it is well known
that there are a great number of Mohommedans in every part of the
empire, who are not only tolerated, but admitted to offices under
government in the same manner as the members of the three
established sects. Jews also are found in several districts, but more
particularly at a place called Kae-foong-foo, in the province of
Honan.
The various religious ceremonies which the Chinese are continually
performing prove at least that they are very superstitions. In all the
southern towns every house has its temple or altar both inside and
outside. The altar in the inside is generally placed at the end of the
principal hall or shop, as the case may be, raised a few feet from the
ground, and having some kind of representation of the family deity
placed upon it. This is surrounded with gaudy tinsel paper, and on
the first of the Chinese month or other high days candles and
incense are burned on the table which is placed in front of it. The

altar on the outside of the door resembles a little furnace, in which
the same ceremonies are regularly performed. In the vicinity of small
villages, and sometimes in the most retired situations, the stranger
meets with little joss-houses or temples, gaudily decorated with
paintings and tinsel paper, and stuck round about with the remains
of candles and sticks of incense. In almost all Chinese towns there
are shops for the sale of idols of all kinds and sizes, varying in price
from a few "cash" to a very large sum. Many of those exposed for
sale are of great age, and have evidently changed hands several
times. I am inclined to believe that the Chinese exchange those gods
which do not please them for others of higher character, and which
they suppose are more likely to grant an answer to their prayers, or
bring prosperity to their homes or their villages.
The periodical offerings to the gods are very striking exhibitions to
the stranger who looks upon them for the first time. When staying at
Shanghae, in November, 1844, I witnessed a most curious spectacle
in the house where I was residing. It was a family offering to the
gods. Early in the morning the principal hall in the house was set in
order, a large table was placed in the centre, and shortly afterwards
covered with small dishes filled with the various articles commonly
used as food by the Chinese. All these were of the very best
description which could be procured. After a certain time had
elapsed a number of candles were lighted, and columns of smoke
and fragrant odours began to rise from the incense which was
burning on the table. All the inmates of the house and their friends
were clad in their best attire, and in turn came to Ko-tou, or bow
lowly and repeatedly in front of the table and the altar. The scene,
although it was an idolatrous one, seemed to me to have something
very impressive about it, and whilst I pitied the delusion of our host
and his friends, I could not but admire their devotion. In a short
time after this ceremony was completed a large quantity of tinsel
paper, made up in the form and shape of the ingots of Sycee silver
common in China, was heaped on the floor in front of the tables, the
burning incense was then taken from the table and placed in the
midst of it, and the whole consumed together. By and by, when the

gods were supposed to have finished their repast, all the articles of
food were removed from the tables, cut up, and consumed by
people connected with the family.
On another occasion, when at Ning-po, having been out some
distance in the country, it was night and dark before I reached the
cast gate of the city, near which I was lodged in the house of a
Chinese merchant. The city gates were closed, but two or three loud
knocks soon brought the warder, who instantly admitted me. I was
now in the widest and finest street in the city, which seemed in a
blaze of light and unusually lively for any part of a Chinese town
after nightfall. The sounds of music fell, upon my ear, the gong, the
drum, and the more plaintive and pleasing tones of several wind
instruments. I was soon near enough to observe what was going on,
and saw, at a glance, that it was a public offering to the gods, but
far grander and more striking than I had before witnessed. The table
was spread in the open street, and every thing was on a large and
expensive scale. Instead of small dishes, whole animals were
sacrificed on the occasion. A pig was placed on one side of the table,
and a sheep on the other, the former scraped clean, in the usual
way, and the latter skinned; the entrails of both were removed, and
on each were placed some flowers, an onion, and a knife. The other
parts of the table groaned with all the delicacies in common use
amongst the respectable portion of the Chinese, such as fowls,
ducks, numerous compound dishes, fruits, vegetables, and rice.
Chairs were placed at one end of the table on which the gods were
supposed to sit during the meal, and chopsticks were regularly laid
at the sides of the different dishes. A blaze of light illuminated the
whole place, and the smoke of the fragrant incense rose up into the
air in wreaths. At intervals the band struck up their favourite
plaintive national airs, and altogether the whole scene was one of
the strangest and most curious which it has ever been my lot to
witness.
There is another ceremony of a religious character which I
frequently observed in the northern cities—I allude to processions in
honour of the gods. I saw one of them at Shanghae, which must

have been at least a mile in length. The gods, or josses, were
dressed up in the finest silks, and carried about in splendid sedan-
chairs, preceded and succeeded by their numerous devotees,
superbly dressed for the occasion, and bearing the different badges
of office. The dresses of the officials were exactly the same as of
those who form the train of some of the high mandarins. Some had
a broad fan, made of peacock feathers, which they wore on the
sides of their hats, others were clad in glaring theatrical dresses,
with low caps, and two long black feathers stuck in them, and
hanging over their shoulders like two horns. Then there were the ill-
looking executioners with long, conical, black hats on their heads,
and whips in their hands for the punishment of the refractory. Bands
of music, placed in different parts of the procession, played at
intervals as it proceeded. Anxious to see the end of this curious
exhibition, I followed the procession until it arrived at a temple in
the suburbs, where it halted. The gods were taken out of the sedan-
chairs, and replaced with due honours, in the temple, from which
they had been taken in the morning. Here their numerous votaries
bent low before them, burned incense, and left their gifts upon the
altar. Numerous groups of well-dressed ladies and their children
were scattered over the ground in the vicinity of the temple, all
bending their knees and seemingly engaged in earnest devotion. A
large quantity of paper, in the form of the Sycee ingots, was heaped
up on the grass as it was brought by the different devotees, and,
when the ceremonies of the day were drawing to a close, the whole
was burned in honour of, or as an offering to, the gods. The sight
was interesting, but it was one which no Christian could look upon
without feelings of the deepest commiseration.
In the course of my travels in China I often met with Christian
missionaries, both Protestant and Roman Catholic, who have been
labouring amongst the Chinese for many years. Until very lately the
efforts of the Protestants had been chiefly confined to Macao and
Canton. Since the war, however, they have had an opportunity of
extending their operations, and some are now settled at all the new

ports which have been opened for foreign trade, as well as on our
Island of Hong-kong, which will now become their head-quarters.
The medical missionaries also act in conjunction with the others, and
are of great use in curing many of the diseases which prevail in the
country, while, at the same time, the truths of the Christian faith are
presented to the minds of their patients. Dr. Lockhart of the London
Missionary Society, who has established himself in the town of
Shanghae, had his hospital daily crowded with patients, many of
whom had come from very distant parts of the country. All were
attended to in the most skilful and careful manner, "without money
and without price." The Rev. Mr. Medhurst, who has laboured long
and zealously as a Christian missionary in the East, was also at
Shanghae. This gentleman is well known as an eminent Chinese
scholar, and, besides preaching to the people in their own tongue,
he has a printing establishment with Chinese type continually at
work, for the dissemination of the truths of the gospel. Several other
gentlemen and their families had arrived at the same port previous
to my departure, and were closely engaged in the study of the
language. Ning-po and Amoy were also occupied by missionaries,
both from England and America, and I suppose, ere this time, some
have also reached Foo-chow-foo on the River Min.
From my own experience of Chinese character, and from what I have
seen of the working of the Medical Missionary Society, I am
convinced that it must be a powerful auxiliary to the missionaries in
the conversion of the Chinese. I regret, however, to say, that up to
the present time little progress appears to have been made. One
portion of the people, and a large one, is entirely indifferent to
religion of any kind, and the rest are so bigoted and conceited, that
it will be a most difficult task to convince them that any religion is
better or purer than their own.
The Roman Catholic missionaries conduct their operations in a
manner somewhat different from the Protestants. They do not
restrict themselves to the out-ports of the empire, where foreigners
are permitted to trade, but penetrate into the interior, and distribute

themselves over all the country. One of their bishops, an Italian
nobleman, resides in the province of Keang-soo, a few miles from
Shanghae, where I have frequently met him. He dresses in the
costume of the country, and speaks the language with the most
perfect fluency. In the place where he lives he is surrounded by his
converts; in fact it is a little Christian village, where he is perfectly
safe, and I believe is seldom if ever annoyed in any way by the
Chinese authorities. When new Roman Catholic missionaries arrive,
they are met by some of their brethren or their converts at the port
nearest their destination, and secretly conveyed into the interior; the
Chinese dress is substituted for the European; their heads are
shaved, and in this state they are conducted to the scene of their
future labours, where they commence the study of the language, if
they have not learned it before, and in about two years are able to
speak it sufficiently well to enable them to instruct the people. These
poor men submit to many privations and dangers for the cause they
have espoused, and although I do not approve of the doctrines
which they teach, I must give them the highest praise for
enthusiasm and devotion to their faith. European customs, habits,
and luxuries are all abandoned from the moment they put their feet
on the shores of China; parents, friends, and home, in many
instances, are heard of no more; before them lies a heathen land of
strangers, cold and unconcerned about the religion for which they
themselves are sacrificing everything, and they know that their
graves will be far away from the land of their birth and the home of
their early years. They seem to have much of the spirit and
enthusiasm of the first preachers of the Christian religion, when they
were sent out into the world by their Divine Master to "preach the
gospel to every creature," and "to obey God rather than man."
According to the accounts of these missionaries, the number of
converts to their faith is very considerable; but I fear they, as well as
the Protestants, are often led away by false appearances and
assertions. Many of the Chinese are unprincipled and deceitful
enough to become Christians or in fact any thing else, in name, to
accomplish the object they may have in view, and they would

become Budhists the very next day should any inducement be
offered them to do so. Judging from appearances, the day must yet
be very distant when the Chinese, as a nation, will be converted to
the Christian faith. Could those individuals in our time, who predict
the near approach of the Millennium, see the length and breadth of
this vast country, with its three hundred millions of souls, they would
surely pause and reflect before they published their absurd and
foolish predictions.
CHAP. XI.
THE TEA-PLANT OF CHINA.—THE SPECIES FOUND IN THE GREEN
AND BLACK TEA DISTRICTS.—BEST SITUATION FOR TEA
PLANTATIONS.—REMARKS ON THEIR MANAGEMENT.—SEASONS,
AND METHODS, OF GATHERING THE LEAVES.—MANUFACTURE OF
TEA.—COTTAGES AMONGST THE TEA HILLS.—FURNACES AND
DRYING PANS.—FIRST APPLICATION OF HEAT.—ROLLING PROCESS.
—EXPOSURE OF THE LEAVES TO THE AIR.—SECOND HEATING.—
LENGTH OF TIME REQUIRED.—TWO KINDS OF TEA.—DIFFERENCE
IN THE MANUFACTURE OF EACH.—SELECTING AND PACKING TEAS.
—APPEARANCE AND COLOUR OF THE LEAF.—PECULIAR TASTE OF
FOREIGNERS FOR DYED TEAS.—GOOD SENSE OF THE CHINESE.—
TEA MERCHANTS.—THEIR VISITS TO THE TEA HILLS.—MODE OF
BUYING FROM THE SMALL GROWERS.—BLACK TEA DISTRICT IN
FOKIEN.—TEAS DIVIDED INTO TWO KINDS.—PECULIAR METHOD
OF PREPARING EACH.—CAUSE OF THEIR DIFFERENCE IN COLOUR.
—FLOWERS USED IN SCENTING THE FINER TEAS.—SIR JOHN
FRANCIS DAVIS'S REMARKS ON DIFFERENT KINDS OF TEAS SOLD
AT CANTON.
There are few subjects connected with the vegetable kingdom which
have attracted such a large share of public notice as the tea-plant of

China. Its cultivation on the Chinese hills, the particular species or
variety which produces the black and green teas of commerce, and
the method of preparing the leaves, have always been objects of
peculiar interest. The jealousy of the Chinese government, in former
times, prevented foreigners from visiting any of the districts where
tea is cultivated, and the information derived from the Chinese
merchants, even scanty as it was, was not to be depended upon.
And hence we find our English authors contradicting each other,
some asserting that the black and green teas are produced by the
same variety, and that the difference in colour is the result of a
different mode of preparation, while others say that the black teas
are produced from the plant called by botanists Thea Bohea, and the
green from Thea viridis, both of which we have had for many years
in our gardens in England.
During my travels in China since the last war, I have had frequent
opportunities of inspecting some extensive tea districts in the black
and green tea countries of Canton, Fokien, and Chekiang, and the
result of these observations is now laid before the reader. It will
prove that even those who have had the best means of judging have
been deceived, and that the greater part of the black and green teas
which are brought yearly from China to Europe and America are
obtained from the same species or variety, namely, from the Thea
viridis. Dried specimens of this plant were prepared in the districts I
have named by myself, and are now in the herbarium of the
Horticultural Society of London, so that there can be no longer any
doubt upon the subject.
In various parts of the Canton province, where I had an opportunity
of seeing tea cultivated, the species proved to be the Thea Bohea, or
what is commonly called the black tea plant. In the green tea
districts of the north—I allude more particularly to the province of
Chekiang—I never met with a single plant of this species, which is so
common in the fields and gardens near Canton. All the plants in the
green tea country near Ning-po, on the Islands of the Chusan
Archipelago, and in every part of the province which I had an
opportunity of visiting, proved, without exception, to be the Thea

viridis. Two hundred miles further to the north-west, in the province
of Kiang-nan, and only a short distance from the tea hills in that
quarter, I also found in gardens this same species of tea.
Thus far my actual observation exactly verified the opinions I had
formed on the subject before I left England, viz., that the black teas
were prepared from the Thea Bohea, and the green from Thea
viridis. When I left the north, on my way to the city of Foo-chow-foo,
on the River Min, in the province of Fokien, I had no doubt that I
should find the tea hills there covered with the other species, Thea
Bohea, from which we generally suppose the black teas are made;
and this was the more likely to be the case as this species actually
derives its specific name from the Bohee hills in this province. Great
was my surprise to find all the plants on the tea hills near Foo-chow
exactly the same as those in the green tea districts of the north.
Here were then green tea plantations on the black tea hills, and not
a single plant of the Thea Bohea to be seen. Moreover, at the time of
my visit, the natives were busily employed in the manufacture of
black teas. Although the specific differences of the tea-plants were
well known to me, I was so much surprised, and I may add amused,
at this discovery, that I procured a set of specimens for the
herbarium, and also dug up a living plant, which I took northward to
Chekiang. On comparing it with those which grow on the green tea
hills, no difference whatever was observed.
It appears, therefore, that the black and green teas of the northern
districts of China (those districts in which the greater part of the teas
for the foreign markets are made) are both produced from the same
variety, and that that variety is the Thea viridis, or, what is
commonly called the green tea plant. On the other hand, those black
and green teas which are manufactured in considerable quantities in
the vicinity of Canton are obtained from the Thea Bohea, or black
tea. And, really, when we give the subject our unprejudiced
consideration, there seems nothing surprising in this state of things.
Moreover, we must bear in mind that our former opinions were
formed upon statements made to us by the Chinese at Canton, who
will say any thing which suits their purpose, and rarely give

themselves any trouble to ascertain whether the information they
communicate be true or false.
The soil of the tea districts is, of course, much richer in the northern
provinces than it is in Quantung. In Fokien and Chekiang it is a rich
sandy loam, very different from the sample which will be found
noticed in the chapter on climate and soil. Tea shrubs will not
succeed well unless they have a rich soil to grow in. The continual
gathering of their leaves is very detrimental to their health, and, in
fact, ultimately kills them. Hence a principal object with the grower
is to keep his bushes in as robust health as possible; and this cannot
be done if the soil be poor.
The tea plantations in the north of China are always situated on the
lower and most fertile sides of the hills, and never on the low lands.
The shrubs are planted in rows about four feet apart and about the
same distance between each row, and look, at a distance, like little
shrubberies of evergreens.
The farms are small, each consisting of from one to four or five
acres; indeed, every cottager has his own little tea garden, the
produce of which supplies the wants of his family, and the surplus
brings him in a few dollars, which are spent on the other necessaries
of life. The same system is practised in every thing relating to
Chinese agriculture. The cotton, silk, and rice farms are generally all
small and managed upon the same plan. There are few sights more
pleasing than a Chinese family in the interior engaged in gathering
the tea leaves, or, indeed, in any of their other agricultural pursuits.
There is the old man, it may be the grand—father, or even the great-
grandfather, patriarch like, directing his descendants, many of whom
are in their youth and prime, while others are in their childhood, in
the labours of the field. He stands in the midst of them, bowed
down with age. But, to the honour of the Chinese as a nation, he is
always looked up to by all with pride and affection, and his old age
and grey hairs are honoured, revered, and loved. When, after the
labours of the day are over, they return to their humble and happy
homes, their fare consists chiefly of rice, fish, and vegetables, which

they enjoy with great zest, and are happy and contented. I really
believe that there is no country in the world where the agricultural
population are better off than they are in the north of China. Labour
with them is pleasure, for its fruits are eaten by themselves, and the
rod of the oppressor is unfelt and unknown.
In the green tea districts of Chekiang near Ning-po, the first crop of
leaves is generally gathered about the middle of April. This consists
of the young leaf-buds just as they begin to unfold, and forms a fine
and delicate kind of young hyson, which is held in high estimation by
the natives, and is generally sent about in small quantities as
presents to their friends. It is a scarce and expensive article, and the
picking of the leaves in such a young state does considerable injury
to the tea plantations. The summer rains, however, which fall
copiously about this season, moisten the earth and air, and if the
plants are young and vigorous they soon push out fresh leaves.
In a fortnight or three weeks from the time of the first picking, or
about the beginning of May, the shrubs are again covered with fresh
leaves, and are ready for the second gathering, which is, in fact, the
most important of the season. The third and last gathering, which
takes place as soon as new leaves are formed, produces a very
inferior kind of tea, which, I believe, is rarely sent out of the district.
The mode of gathering and preparing the leaves of the tea-plants is
extremely simple. We have been so long accustomed to magnify and
mystify every thing relating to the Chinese, that, in all their arts and
manufactures, we expect to find some peculiar and out of the way
practice, when the fact is, that many operations in China are more
simple in their character than in most other parts of the world. To
rightly understand the process of rolling and drying the leaves,
which I am about to describe, it must be borne in mind that the
grand object is to expel the moisture, and at the same time to
retain, as much as possible, of the aromatic and other desirable
secretions of the species. The system adopted to attain this end is as
simple as it is efficacious.

In the harvest seasons the natives are seen in little family groups on
the side of every hill, when the weather is dry, engaged in gathering
the tea leaves. They do not seem so particular, as I imagined they
would have been, in this operation, but strip the leaves off rapidly
and promiscuously, and throw them all into round baskets made for
the purpose out of split bamboo or rattan. In the beginning of May,
when the principal gathering takes place, the young seed vessels are
about as large as peas. These are also stripped off and dried with
the leaves; it is these seed-vessels, which we often see in our tea,
and which have some slight resemblance to young capers. When a
sufficient quantity of leaves are gathered, they are carried home to
the cottage or barn, where the operation of drying is performed.
The Chinese cottages, amongst the tea hills, are simple and rude in
their construction, and remind one of what we used to see in
Scotland in former years, when the cow and pig lived and fed in the
same house with the peasant. Scottish cottages, however, even in
these days, were always better furnished and more comfortable than
those of the Chinese are at the present time. Nevertheless, it is in
these poor cottages that a large proportion of the teas, with their
high-sounding names, are first prepared. Barns, sheds, and other
outhouses, are also frequently used for the same purpose,
particularly about the temples and monasteries.
The drying pans and furnaces in these places are very simply
constructed. The pans, which are of iron, and are made as thin as
possible, are round and shallow, and, in fact, are the same, or nearly
the same, which the natives have in general use for cooking their
rice. A row of these are built into brick work and chunam, having a
flue constructed below them, with the grating, or rather fire-place, at
one end, and the chimney, or, at least, some hole to allow the smoke
to escape, at the other. A chimney is a secondary consideration with
the Chinese, and in many instances which came under my
observation, the smoke, after passing below the drying-pans, was
allowed to escape, as it best could, through the doors and roofs of
the houses, which, indeed, in China, is no difficult matter.

When the pans are first fixed, the brick-work and chunam are
smoothed off very neatly round their edges and carried up a little
higher, particularly at the back of the pans, at the same time
widening gradually. When complete, the whole has the appearance
of a row of large high-backed basins, each being three or four times
larger than the shallow iron pan which is placed at its bottom,
immediately over the flue. When the fire is applied, the upper part of
these basins, which is formed of chunam, gets heated as well as the
iron pan, though in a less degree. The drying pans, thus formed,
being low in front, and rising very gradually at the sides and back,
the person, whose duty it is to attend to the drying of the leaves,
can readily manage them, and scatter them about over the back of
the basin. The accompanying sketch, which was made on the spot,
will render this description more clear.

Furnaces and Drying Pans.
The leaves having been brought in from the hills are placed in the cottage
or drying-house. It is now the duty of one individual to light the little fire
at the mouth of the flue, and to regulate it as nicely as possible. The
pans become hot very soon after the warm air has begun to circulate in
the flue beneath them. A quantity of leaves, from a sieve or basket, are
now thrown into the pans, and turned over, shaken up, and kept in
motion by men and women stationed there for this purpose. The leaves
are immediately affected by the heat. They begin to crack, and become
quite moist with the vapour or sap which they give out on the application
of the heat. This part of the process lasts about five minutes, in which
time the leaves lose their crispness, and become soft and pliable. They
are then taken out of the pans and thrown upon a table, the upper part
of which is made of split pieces of bamboo as represented in the annexed
sketch. Three or four persons now surround the table, and the heap of

tea leaves is divided into as many parcels, each individual taking as many
as he can hold in his hands, and the rolling process commences. I cannot
give a better idea of this operation than by comparing it to a baker
working and rolling his dough. Both hands are used in the very same
way; the object being to express the sap and moisture, and at the same
time to twist the leaves. Two or three times during the operation the little
bundles of rolled leaves are held up and shaken out upon the table, and
are then again taken up and pressed and rolled as before. This part of
the process also lasts about five minutes, during which time a large
portion of green juice has been expressed, and may be seen finding its
way down between the interstices of the bamboos. The leaves being now
pressed, twisted, and curled, do not occupy a quarter of the space which
they did before the operation.
The Rolling Process.
When the rolling process is completed the leaves are removed from the
table and shaken out for the last time, thinly, upon a large sort of screen,
also made out of split pieces of bamboo, and are exposed to the action of
the air. The best days for this purpose are those which are dry and
cloudy, with very little sun. The object being to expel the moisture in the
most gentle manner, and, at the same time, to allow the leaves to remain

as soft and pliable as possible. When the sun is clear and powerful the
moisture evaporates too rapidly, and the leaves are left crisp, coarse, and
not in a proper state to undergo the remaining part of the process. There
is no stated time for this exposure, as much depends upon the nature of
the weather and the convenience of the work-people; sometimes I have
seen them go on with the remaining part of the operation without at all
exposing the leaves to the air.
Having in this manner got rid of a certain part of the superfluous
moisture, the leaves, which are now soft and pliant, are again thrown
into the drying-pans, and the second heating commences. Again one
individual takes his post at the furnace, and keeps up a slow and steady
fire. Others resume their places at the different drying-pans—one at each
—and commence stirring and throwing up the leaves, so that they may all
have an equal share of the fire, and none get scorched or burned. The
process of drying thus goes on slowly and regularly. This part of the
operation soon becomes more easy, for the leaves, as they part with their
moisture, twist and curl, and consequently take up much less room than
they do at first, and mix together more readily. The tea leaves being now
rather too hot for the hand, a small and neat brush, made of bamboo, is
used instead of the fingers for stirring them up from the bottom of the
pan. By this means the leaves are scattered about on the smooth
chunam-work, which forms the back of the drying-pan, and, as they roll
down on this heated inclined plane they dry slowly, and twist at the same
time. During this operation the men and women who are employed never
leave their respective stations, one keeps slowly feeding the fire, and the
others continually stir the leaves. No very exact degree of temperature is
attempted to be kept up, for they do not use the thermometer, but a slow
and steady fire is quite sufficient; that is, the pan is made and kept so
hot, that I could not place my hand upon it for a second of time. In order
to get a correct idea of the time required to complete this second part of
the process, I referred to my watch on different occasions, and at
different tea farms, and always found that it occupied about an hour; that
is, from the time the leaves were put into the pan after exposure to the
air, until they were perfectly dry.
When the operation of drying is going on largely, some of the pans in the
range are used for finishing the process, while others, and the hottest
ones, are heating and moistening the leaves before they are squeezed

and rolled. Thus a considerable number of hands can be employed at
once, and the work goes on rapidly without loss of time or heat, the latter
of which is of some importance in a country so ill provided with fuel.
The tea prepared in the manner which I have just described is greenish
in colour, and of a most excellent quality. It is called by the Chinese in the
province of Chekiang, Tsaou-tsing, or the tea which is dried in the pan, to
distinguish it from the Hong-tsing, or that kind which is dried in flat
bamboo baskets over a slow fire of charcoal.
This latter kind—the Hong-tsing,—is prepared in the following manner:—
The first process, up to the period of rolling and exposure to the air, is
exactly the same as that which I have just described, but instead of being
put into the drying-pan for the second heating like the Tsaou-tsing, the
Hong-tsing is shaken out into flat baskets, which are placed over tubs
containing charcoal and ashes. The charcoal, when ignited, burns slowly
and sends out a mild and gentle heat. Indeed, the only difference
between the two teas consists in the mode of firing, the latter being dried
less and more slowly than the former. The Hong-tsing is not so green in
colour as the Tsaou-tsing, and I believe has rarely been exported.
After the drying is completed the tea is picked, sifted, divided into
different kinds and qualities, and prepared for packing. This is a part of
the operation which requires great care, more especially when the tea is
intended for the foreign market, as the value of the sample depends
much upon the "smallness and evenness" of the leaf, as well as upon its
other good qualities. In those districts where the teas are manufactured
solely for exportation, the natives are very particular in the rolling
process, and hence the teas from these districts are better divided and
more even—although I should doubt their being really better in quality—
than they are in the eastern parts of the province of Chekiang. When
they have been duly assorted, a man puts on a pair of clean cloth or
straw shoes, and treads the tea firmly into baskets or boxes, and the
operation is considered complete, in so far as the grower is concerned.
I have stated that the plants grown in the district of Chekiang produce
green teas, but it must not be supposed that they are the green teas
which are exported to England. The leaf has a much more natural colour,
and has little or none of what we call the "beautiful bloom" upon it, which
is so much admired in Europe and America. There is now no doubt that

all these "blooming" green teas, which are manufactured at Canton, are
dyed with prussian blue and gypsum, to suit the taste of the foreign
"barbarians:" indeed, the process may be seen any day, during the
season, by those who will give themselves the trouble to seek after it. It
is very likely that the same ingredients are also used in dying the
northern green teas for the foreign market; of this, however, I am not
quite certain. There is a vegetable dye obtained from Isatis indigotica
much used in the northern districts, and called Tein-ching, and it is not
unlikely that it may be the substance which is employed.
The Chinese never use these dyed teas themselves, and I certainly think
their taste in this respect is more correct than ours. It is not to be
supposed that the dye used can produce any very bad effects upon the
consumer, for, had this been the case, it would have been discovered
before now; but if entirely harmless or inert, its being so must be
ascribed to the very small quantity which is employed in the manufacture.
When the teas are ready for sale, the large tea merchants or their
servants come out from the principal towns of the district, and take up
their quarters in all the little inns or eating houses, which are very
numerous in every part of the country. They also bring coolies loaded
with the copper coin of the country, with which they pay for their
purchases. As soon as the merchants are known to have arrived in the
district, the tea growers bring their produce for inspection and sale.
These little farmers or their labourers may now be seen hastening along
the different roads, each with two baskets or chests slung across his
shoulder on his bamboo pole. When they arrive at the merchant's abiding
place the baskets are opened before him, and the quality of the tea
inspected. If he is pleased with its appearance and smell, and the parties
agree as to the price, the tea is weighed, the money paid down, and the
grower gets his strings of copper money slung over his shoulder, and
returns to his farm. But should the price offered appear too low, the
baskets are immediately shouldered with the greatest apparent
independence, and carried away to some opposition merchant. It,
however, sometimes happens that a merchant makes a contract with
some of the tea growers before the season commences, in which case
the price is arranged in the usual way, and generally a part paid in
advance. This, I understand, is frequently the case at Canton when a
foreign resident wishes to secure any particular kind of tea.

After the teas are bought up in the district where they are grown, they
are conveyed to the most convenient town, where they are assorted and
properly packed for the European and American markets. Such is the
system of green tea culture and manufacture which came under my own
observation in the province of Chekiang.
The black tea districts of Fokien, which I visited, are managed in the
same way as those of Chekiang.
I have already said that the species of plant which produces the black
teas near Foo-chow is the very same as that found in the green tea
districts of the north. Being further south, and of course in a hotter
climate, the tea plant of Fokien is generally grown at a high elevation
amongst the hills. At the risk of some little repetition I will insert an
account of my visit to the tea hills of Fokien.
Every cottager, or small farmer, has two or three patches of tea shrubs
growing on the hill sides, which are generally planted and kept in order
by the members of his own family. When the gathering season arrives,
the cottage doors are locked, and all proceed to the hills with their
baskets and commence plucking the leaves. This business, of course,
only goes on during fine days when the leaves are dry.
The first gathering takes place just when the leaf-buds begin to unfold
themselves in early spring. This tea is scarce and of a very superior
quality, being, in fact, the same, or nearly the same, as that which is
made from the young leaves in the green-tea district. The second
gathering produces the principal crop of the season; the third crop is
coarse and inferior.
When the leaves are brought home from the hills, they are first of all
emptied out into large flat bamboo sieves, and, providing the day is not
too bright, are exposed in the open air to dry off any superfluous
moisture. When this moisture has evaporated, convenient portions of the
leaves are brought in and thrown into a round flat iron pan, such as the
Chinese use for boiling their rice, and are exposed to the heat of a gentle
fire which is lighted below them. As soon as this heat reaches them, they
give out a large quantity of moisture with a crackling noise, and they
soon become soft and pliant. The person who attends to them stirs them
about with his hands, and in about five minutes takes them out and puts
in a fresh supply. The heated leaves are emptied out on a large round

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Protocells Bridging Nonliving And Living Matter Steen Rasmussen Mark A Bedau Liaohai Chen David C Krakauer David Deamer

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Protocells Bridging Nonliving And Living Matter Steen Rasmussen Mark A Bedau Liaohai Chen David C Krakauer David Deamer

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