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PHOTOSYNTHESIS IN ACTION
PHOTOSYNTHESIS
IN ACTION
Harvesting Light, Generating
Electrons, Fixing Carbon

Edited by

ALEXANDER RUBAN
School of Biological and Behavioural Sciences, Queen Mary University of London, United Kingdom

CHRISTINE H. FOYER
School of Biosciences, College of Life and Environmental Sciences, University of Birmingham,
Edgbaston, United Kingdom

ERIK H. MURCHIE
Division of Plant and Crop Sciences, School of Biosciences, University of Nottingham, Sutton Bonington,
Loughborough, United Kingdom
Academic Press is an imprint of Elsevier
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 Contributors

Martin Battle School of Life Sciences, University of Jeremy Harbinson Laboratory of Biophysics, Helix
Essex, Colchester, Essex, United Kingdom Building, Wageningen University, Wageningen,
Roel Brienen School of Geography, Faculty of Earth The Netherlands
and Environment, University of Leeds, Leeds, Tanja A. Hofmann Suffolk One Sixth Form College,
United Kingdom Ipswich, Suffolk, United Kingdom
Alexandra J. Burgess School of Biosciences, Univer- Elias Kaiser Horticulture and Product Physiology
sity of Nottingham, Sutton Bonington, Loughbor- Group, Radix Building, Wageningen University,
ough, United Kingdom Wageningen, The Netherlands
Amanda P. Cavanagh School of Life Sciences, Uni- Diana Kirilovsky Institute for Integrative Biology
versity of Essex, Colchester, Essex, United of the Cell (I2BC), CEA, CNRS, Paris-Saclay Uni-
Kingdom versity, Gif-sur-Yvette Cedex, France
Luca Dall’Osto Department of Biotechnology, Uni- Anja Krieger-Liszkay Institute for Integrative Biol-
versity of Verona, Verona, Italy ogy of the Cell (I2BC), CEA, CNRS, Paris-Saclay
Robyn Emmerson School of Life Sciences, Univer- University, Gif-sur-Yvette Cedex, France
sity of Essex, Colchester, Essex, United Kingdom Tracy Lawson School of Life Sciences, University of
Christine H. Foyer School of Biosciences, College of Essex, Colchester, Essex, United Kingdom
Life and Environmental Sciences, University of Bir- Lorna McAusland School of Biosciences, University
mingham, Edgbaston, United Kingdom of Nottingham, Sutton Bonington, Loughborough,
David Galbraith School of Geography, Faculty of United Kingdom
Earth and Environment, University of Leeds, Jun Minagawa National Institute for Basic Biology,
Leeds, United Kingdom Division of Environmental Photobiology, Okazaki,
Emanuel Gloor School of Geography, Faculty of Japan
Earth and Environment, University of Leeds, Alejandro Sierra Morales Center for Crop Systems
Leeds, United Kingdom Analysis, Radix Building, Wageningen University,
Rodrigo L. Gomez Department of Biotechnology, Wageningen, The Netherlands
University of Verona, Verona, Italy Conrad W. Mullineaux School of Biological and
Zeno Guardini Department of Biotechnology, Uni- Behavioural Sciences, Queen Mary University of
versity of Verona, Verona, Italy London, London, United Kingdom
Guy Hanke School of Biological and Chemical Sci- Erik H. Murchie School of Biosciences, University
ences, Queen Mary University of London, London, of Nottingham, Sutton Bonington, Loughborough,
United Kingdom United Kingdom

ix
x Contributors

Jacob Pullin School of Life Sciences, University of Herbert van Amerongen Laboratory of Biophysics,
Essex, Colchester, Essex, United Kingdom Wageningen University, Wageningen, The
Christine A. Raines School of Life Sciences, Netherlands
University of Essex, Colchester, Essex, United Shellie Wall School of Life Sciences, University of
Kingdom Essex, Colchester, Essex, United Kingdom
Andrew J. Simkin School of Biosciences, University Emilie Wientjes Laboratory of Biophysics, Wagen-
of Kent, Canterbury, Kent, United Kingdom ingen University, Wageningen, The Netherlands
 Foreword

In the rapidly changing world, profound into the energy and matter of life. That is why
knowledge of the processes that enable its exis- photosynthetic organisms are called autotrophs;
tence is crucial for humankind. Our planet is they are feeding themselves. They do so on a diet
entering a new geological era—the Anthropo- of carbon dioxide, light, and water—a tough
cene. Throughout the history of humanity, we diet, but it has been working for several billions
have changed Earth’s biosphere: first, its of years without any detrimental impact on the
nature—plants, animals, microbes—then our biosphere. In reality, almost all energy we use
nature with advances in medicine, and finally, today came and keeps coming via photosyn-
its climate. We began shaping the world armed thetic organisms, including fossil fuels.
with science and technology, and driven by the So, what is the purpose of this book, and why
ambition of economic growth. As a result, we does it claim to be different from the numerous
came to gradually realise that our engagement photosynthesis books published in the past? The
in the evolution (or deterioration) of the bio- following are the objectives of the book: (i) to
sphere is here to stay. The recent violent mani- reveal to the scholars and readers the fundamen-
festations in climate patterns, such as heat tal knowledge of the stages (and the integration)
waves, floods, forest fires, and hurricanes, of energy and matter transformation in photo-
prompt conscientious people to think about synthesis, that supports all life; (ii) to show
how they can protect and preserve life on our how flexible and adaptable to the environment
planet. these stages have evolved to be; and (iii) to
The driving force behind life on Earth is the explain how we can alter these stages, their prin-
Sun that gives us energy in the form of light. This ciples, mechanisms, and efficiency in order to
energy feeds the biosphere’s ‘wheel of life’ con- improve the photosynthesis for our own benefits
sisting of the two opposing chemical processes: and the benefit of the biosphere. Increasing pho-
the oxidation and reduction of oxygen. The for- tosynthetic adaptability, productivity, and sur-
mer is photosynthesis, the process that gives us vival in the changing environment is necessary
molecular oxygen to breathe, food to eat, and to meet the food demands of the Anthropocene.
many materials that we rely upon. The latter This is photosynthesis in action as we currently
process is respiration. Most lifeforms, even those know it. This is up-to-date knowledge, and it is
that possess no lungs or gills, need to respire—to our duty to address the demands of this new
use oxygen for slowly and intelligently ‘burn- geological era. The book is written by a group
ing’ organic matter in order to build the struc- of excellent experts working in the all-
tures of life. Photosynthesis, however, starts it encompassing spectrum of the topics of photo-
all. It captures the Sun’s energy and converts it synthesis research. It took me little effort to

xi
xii Foreword

convince them to commit to this project, since the book has the potential to teach a broad
all of them believe in the beauty and strength audience of students, researchers, engineers,
of the photosynthesis research for the sake of and policymakers.
humankind. Have we succeeded? The answer,
I assume, is here for the reader to learn, judge, Alexander Ruban
and, hopefully, act. I, therefore, believe that
 Preface

Food production is facing one of the most agriculture. The information provided in this
critical challenges of our times. As argued in book will enable and support readers in
the recently approved Green Deal, “our food (1) understanding the mechanisms of photosyn-
system is under threat and must become more thesis at a functional level, (2) quantitatively
sustainable and resilient”—not only at Euro- determining the relationships between each
pean level but also on a global scale (European individual mechanism with the context of the
Commission, 2020). It comes as no surprise that operation of photosynthesis as a whole, and
the 2020 Nobel Prize for peace was awarded to (3) highlighting possible interventions where
the ‘World Food Program’, which has addressed the improvement of efficiency and function
the economic, social, and ecological emergencies might be achieved. The 12 chapters that com-
that threaten food security worldwide. The main prise this book are from outstanding scientists,
objective of the food security program is to who are internationally recognised experts in
increase the production yield of cereals around their respective fields. Each author has not only
the globe and thereby ensure the incremental provided essential information but also
demand for food, animal feed, and biofuels included personal perspectives and insights that
(Miraglia et al., 2019; Prosekov and Ivanova, allow a deeper understanding of the topic and
2018). The Green Deal rules out the extensive its importance.
use of fertilisers due to their high pollution Chapters 1, 4, and 7 introduce the readers to
potential. Genetic improvement is an essential the principles of light harvesting (1) adaptations
part of the strategy, as is the increasing the effi- of the processes of light capture to abiotic stress
ciency of plant use of resources (Bailey-Serres (4) and possible ways to manipulate the process
et al., 2019). Photosynthesis is the major driver in order to improve the photosynthetic effi-
of carbon gain, biomass production, and crop ciency and protection against environmental
yield. It is hence a central target for the improve- factors (7).
ment and sustainable production of crops in a Chapters 2, 5, and 8 provide a comprehensive
changing climate. Climate change is considered introduction to the photosynthetic electron
the most important factor limiting the produc- transport system in plants (2), the differences
tion and yield of cereals. Understanding the fun- between the photosynthetic electron transport
damental mechanisms of photosynthesis, its systems in chloroplasts and in cyanobacteria
functional operation, and how the photosyn- (8), and the systems that limit the production
thetic processes are regulated in response to of reactive oxygen species (ROS) and hence chlo-
changing environmental inputs and signals, as roplast signalling.
well as the central role of photosynthesis in plant Chapters 3, 6, and 9 introduce the mecha-
growth, development, and survival, is a key nisms and limitations of CO2 uptake and assim-
driver of strategies for environmental-friendly ilation in leaves and chloroplasts, the barriers to

xiii
xiv Preface

diffusion and the metabolic processes underly- carbon gain and crop yield, as well as its
ing conversion of this gas into carbohydrates. responses to a changing climate. The content
Environmental ‘stress’ places restrictions on of the book will also be of interest to specialists
photosynthesis in leaves that can commonly who are working and studying various aspects
restrict productivity and yield (6). Improving of photosynthesis and its dynamic regulation,
carbon fixation in crops to underpin yield as well as nonspecialists, who are simply
improvement will be a substantial contribution interested in knowing more about this
to global food security, and routes for achieving environment-friendly and evolving process that
this via the Calvin Benson pathway are sustains all life on Earth.
described (9).
Chapters 10 and 11 provide an integration of A.V. Ruban, E. Murchie, and C.H. Foyer
the multiple scales on which photosynthesis
operates. The role of photosynthesis in shaping References
Earth’s climate and biosphere is described. Reg-
Bailey-Serres, J., Parker, J.E., Ainsworth, E.A., Oldroyd, G.E.
ulation of photosynthesis can be perceived at the D., Schroeder, J.I., 2019. Genetic strategies for improving
molecular, cellular canopy, and ecosystem level, crop yields. Nature 575, 109–118.
and this is integrated using tools such as math- European Commission, 2020. A Farm to Fork strategy for a
ematical modelling. fair, healthy and environmentally-friendly food system.
Miraglia, M., Marvin, H.J.P., Kleter, G.A., et al., 2019. Climate
We recommend this book to undergraduate
change and food safety: an emerging issue with special
students, postgraduate students, and resear- focus on Europe. Food Chem. Toxicol. 47 (5), 1009–1021.
chers who are interested in gaining a deeper Prosekov, A.Y., Ivanova, S.A., 2018. Food security: the chal-
knowledge of photosynthesis and its role in lenge of the present. Geoforum 91, 73–77.
 C H A P T E R

1
Harvesting light
Herbert van Amerongen and Emilie Wientjes
Laboratory of Biophysics, Wageningen University, Wageningen, The Netherlands

1 Introduction, what is light harvesting The RCs will be discussed in more detail in
and why is it needed? Chapter 2; here, we summarise a few essential
aspects. In organisms performing oxygenic photo-
As already mentioned in the foreword of this synthesis, two types of RCs are present, type I and
book, the driving force behind life on earth is the II. The type II RC complex, for instance, is a large
sun that provides energy in the form of light. pigment–protein complex consisting of several
Photosynthesis is the process that captures this closely associated proteins with a number of
energy and converts it into the energy of life. cofactors, including several chlorophyll a (Chl a)
In this chapter, we address how the energy is pigments. If one of them, the primary electron
captured and delivered to the reaction centres donor, is in the (first) excited state, it can donate
(RCs), entities that are located in the thylakoid an electron to a neighbouring cofactor, which
membrane and where the conversion process forms the start of a cascade of electron transfer
starts by a charge separation. The combination steps, ultimately leading to the generation of a
of capturing and delivery is usually called light chemical potential that is fuelling ATP formation
harvesting and is performed by the light- and the generation of reductive power.
harvesting complexes, also called antennae. The pigment–protein ratio in the RCs is rather
low and a type II RC contains only 6 Chls a (less
Electron transfer or charge separation: The than 6 kDa), whereas the total molecular mass is
transfer of an electron from a donor molecule in hundreds of kDa. One Chl a in full sunlight can
an excited state to a neighbouring cofactor. In typically absorb
10 photons per second and in
photosynthesis this process occurs in a isolation the RC would be inactive most of the
photosynthetic RC. time. To use their ‘expensive’ RCs more effi-
Excitation-energy transfer: The nonradiative ciently, photosynthetic organisms usually have
transfer of excitation energy from an excited pig- ‘cheap’ light-harvesting complexes (LHCs) that
ment, the donor, to a neighbouring pigment, the surround the RCs to provide them with excita-
acceptor. Upon transfer the donor falls back to its tions. In that case, the excited-state energy is
ground state and the acceptor goes to the excited transferred from pigment to pigment towards
state. This process allows to deliver the excitations the RC where it can be used for charge separa-
to the RCs. tion. The highly abundant light-harvesting

Photosynthesis in Action 3 Copyright # 2022 Elsevier Inc. All rights reserved.

https://doi.org/10.1016/B978-0-12-823781-6.00007-1
4 1. Harvesting light

FIG. 1 (A) PSI and PSII are embedded in the thylakoid membrane. Here, the PSI core (Caspy et al., 2020, pdb 6YAC) and the
PSII core (Su et al., 2017, pdf 5XNL) from higher plants are shown. The polypeptides are displayed as transparent coloured
cartoons, the chlorophylls in green, the RC Chl in red, pheophytin (PSII) in orange, the manganese cluster in purple, and the iron
sulphur clusters (PSI) in yellow/orange. (B) Top view of PSII and PSI core.

complex II (LHCII) in plants, for instance, has a generation is not high enough, this causes a sub-
far higher pigment density (15 kDa of pigments stantial loss of energy because of charge recom-
per 25 kDa of protein) than the RC, like all mem- bination processes. A light-harvesting system is
bers of the LHC family. The antenna systems therefore a cheap way to increase the turn-over
usually contain hundreds of pigments per RC. rate of the RC by typically two orders of magni-
The combination of a type II RC in organisms tude, at the same time facilitating multielectron
performing oxygenic photosynthesis and its redox processes.
associated LHCs is called photosystem II (PSII)
whereas photosystem I (PSI) contains a type
I RC. Fig. 1 shows the PSI and PSII core com- 2 The concepts of light-harvesting
plexes, which are composed of the RC and capacity and efficiency
tightly associated LHCs. This minimal antenna
system is usually enlarged by peripheral LHCs. The absorption capacity or absorption cross-
The high number of antenna pigments in PSII is section σ abs of an antenna system associated
also required because the conversion process with an RC can be increased by augmenting
initiated by the RCs involves multielectron the number of LHCs. This may be desirable in
redox processes. If the rate of electron low-light conditions when the number of

I. Principles
 2 The concepts of light-harvesting capacity and efficiency 5

FIG. 2 PSII supercomplex with different antenna sizes: C2S2M2, C2S2, and C2S supercomplexes. PSII core proteins yellow,
LHCII S-trimer blue, LHCII M-trimer red, minor antenna (CP24, CP26, CP29) orange, chlorophylls green sticks, carotenoids
orange sticks, Mn cluster purple spheres, pheophytin red spheres, and RC Chls 405–406 orange spheres. The bottom schematic
diagram reflects the fact that a photosystem with a small antenna has a relatively low σ abs and high ϕCS, whereas a larger
antenna corresponds to a higher value of σ abs and a lower one of ϕCS.

excitations delivered to the RC is below the max- These counteracting effects lead to an estimated
imum photosynthetic capacity. The term σ abs is maximum value of the light-harvesting capacity
used here in an intuitive way to denote the for PSII when the antenna contains 200–250 Chls
capacity to absorb available photons; a doubling a (Wientjes et al., 2013). For PSI, this number is
of σ abs implies that the number of absorbed pho- significantly higher (Croce and Van
tons doubles when the available light intensity Amerongen, 2020). The maximum capacity can
remains the same. However, an increase of σ abs be useful in low-light conditions, but when the
does not necessarily mean that also the light- organisms are exposed to high-light intensities,
harvesting capacity increases. The latter also saturation of the photosynthesis process may
depends on the quantum efficiency of charge occur, the surplus of excitations may lead to
separation ϕCS after absorption. The value of photodamage and the light-harvesting capacity
ϕCS is simply the fraction of absorbed photons should decrease rapidly to minimise deleterious
that leads to charge separation in the RC. The effects. This can be done by a decrease of σ abs
total light-harvesting capacity scales with the and/or ϕCS, and both processes occur in nature
product σ abs ϕCS. Whereas the value of σ abs goes as a quick response to increasing light intensi-
up when the number of LHCs increases, the ties. σ abs can be lowered on a short time scale
value of ϕCS typically goes down because on by detaching part of the antenna system,
average excitations need to travel longer to reach whereas ϕCS can be decreased by reducing the
the RC and the probability to get lost via compet- excited-state lifetime of the photosystem by
ing deexcitation processes increases (Fig. 2). introducing excitation quenchers. When

I. Principles
6 1. Harvesting light

organisms grow in high-light conditions, the systems with (exceptionally) high extinction
maximum light-harvesting capacity is usually coefficients of the order of
1  105 M1 cm1
not needed and the antenna size is smaller. (Fig. 3).
Below we present the solar spectrum and Together, Chl a and β-carotene are responsi-
introduce various pigments, their structures, ble for all the absorption of solar photons in
and absorption spectra and illustrate that differ- the cores of PSI and PSII. The carotene molecules
ent pigments are used to deal with the available transfer their excitation energy within 1 ps to a
light and optimise the absorption cross-section. neighbouring Chl a molecule, after which excita-
Various examples will be given of light- tion energy transfer (EET) proceeds between Chl
harvesting complexes occurring in nature. After a molecules until one of the primary donors
that, we discuss the properties and mechanisms becomes excited and charge separation can take
that underlie the quantum efficiency ϕCS and place.
discuss the variability of the photosystems and Additional light-harvesting complexes, also
some regulatory aspects. called outer antenna complexes, can increase
the absorption cross-section of the photosys-
tems. In plants and green algae, additional major
(LHCII) and minor complexes are present that
3 Solar spectrum and its coverage by show a large homology to each other and they
photosynthetic pigments will be discussed in more detail in the next par-
agraph. In addition to Chl a, they also contain
In Fig. 3A, the solar irradiance spectrum at the Chl b (Fig. 3) and a number of different xantho-
surface of the earth is given. Also, the absorption phylls (oxygenated carotenoids) like lutein,
spectrum of Chl a is shown, the most important neoxanthin, and violaxanthin, together broaden-
pigment in oxygenic photosynthesis, which ing/increasing the absorption cross-section
shows prominent peaks around 670 and (Fig. 4).
430 nm. Both fall within the visible region rang- Another class of outer antenna complexes can
ing from 400 to 700 nm, often called photosyn- be found in diatoms, which form a very diverse
thetically active radiation (PAR). Photons phytoplanktonic group that is responsible for
absorbed in this wavelength region can drive 20% of the global primary productivity. Besides
oxygenic photosynthesis, although also photons Chl a, they contain Chl c molecules (Fig. 3) as
that are slightly outside this region can contrib- pigments together with a number of caroten-
ute to some extent (Fig. 3A). There are some bac- oids. These Chl a/c complexes show a large
teria that can use light between 700 and 1000 nm homology with the Chl a/b complexes of plants
for anoxygenic photosynthesis, but here, we will and green algae. However, the absorption spec-
focus on the oxygenic process (Fig. 3A). trum of Chl c shows more absorption in the blue
The primary electron donors in PSI and PSII part of the spectrum, which is useful in the
are both Chls a and they are often indicated as aquatic environments in which diatoms live
P700 and P680, referring to the wavelengths of and where substantially less red light is present.
their respective absorption maxima. In Fig. 3A, These Chls are structurally peculiar because
the absorption spectrum of β-carotene is also they do not contain a long hydrophobic phytol
given, which is partly complementary to that chain that is common to the other Chls and
of Chl a. Both pigments can absorb visible light BChls, leaving space in the light-harvesting
due to their extended conjugated π-electron complexes for additional carotenoids, which

I. Principles
 3 Solar spectrum and its coverage by photosynthetic pigments 7

FIG. 3 (A) Spectrum of photosynthetic pigments and sunlight. The sun spectrum is based on the American Society of Test-
ing and Materials G-173 spectrum converted to photons m2 s1 scale (U.S. Department of Energy (DOE)/NREL/ALLI-
ANCE). The spectra of the pigments are on a molecular extinction coefficient scale, the pigment spectra are reported in
80% acetone, only that of bacteriochlorophyll a (BChl a) is in toluene. BChl a is only found in a number of organisms that
perform nonoxygenic photosynthesis. (B) Structures of the pigments of (A). Note that Chl c lacks the phytol chain, which
is present in all other chlorophylls.

I. Principles
8 1. Harvesting light

FIG. 4 (A) Structure of LHCII monomer from plants (Su et al., 2017). The protein is shown in grey, Chl a in light green, Chl b in
dark green, violaxantin in red, neoxanthin in yellow, and lutein in orange. For clarity, the phytol chains of the chlorophylls are not
shown. (B) Phycobilisome of the red alga Porphyridium purpureum (Ma et al., 2020). Polypeptides are displayed as cartoons:
phycocyanins are displayed in orange, allophycocyanins in red, and all others in grey. Pigments are displayed as spheres: phy-
courobilin in cyan (abs max 498 nm), phycoerythrobilin in green (abs max 540 and 565 nm), and phycocyanobilins in orange (abs
max 620 nm in phycocyanin and 650 nm in allophycocyanins). For comparison, the PSII core and LHCII monomer are dis-
played on the same scale. Polypeptides in grey cartoons, pigments in spheres: chlorophylls in green, carotenoids in orange,
and the manganese cluster in purple. (C) Bottom view of PBS displayed in B showing that the pigments that absorb energy
at lower energy are located at the bottom where PBS interacts with the photosystem.

can further increase the absorption cross-section number of bilins, attached to phycobiliproteins
in the blue/green. that absorb above 700 nm.
Cyanobacteria form another important class
of photosynthetic organisms. Most of them live
in a water environment, and they are the only 4 Light-harvesting complexes, a few
bacteria performing oxygenic photosynthesis. examples
They exist nearly everywhere on our planet, as
long as there is some light available. The outer The most abundant light-harvesting complex
antenna complexes of cyanobacteria and some on earth is LHCII (Fig. 4A) that usually occurs as
red algae, called phycobilisomes, are completely a trimer in plants and green algae. It is mainly a
different from those of plants, algae, and dia- light-harvesting complex for PSII, but a variable
toms. They are huge, consisting of many fraction (dependent on light conditions) can also
water-soluble pigment–protein complexes, and bind to PSI. It is composed of pigments, which
they are appressed to the membrane-embedded are coordinated by a protein scaffold. The pro-
photosystems (Fig. 4). Their pigments are bilins, tein is folded in three transmembrane helices
open-chain tetrapyrroles that are covalently plus some parts that are located outside the
bound to the proteins. They typically absorb membrane. One monomer typically harbours
between 550 and 650 with an absorption maxi- eight Chls a and six Chls b together with four
mum that is strongly dependent on the protein carotenoids: two luteins that form a central
environment and pigment configuration. cross, one neoxanthin, and one violaxanthin
Finally, while most cyanobacteria contain Chl (Fig. 4A). A crucial aspect of this structure,
a as their only type of chlorophyll, there are a which is determined by the protein, is the dense
few species that contain Chl d or Chl f (Fig. 3), pigment packing that guarantees short distances
which allows them to absorb light in the near- between pigments, which is advantageous for
infrared. These cyanobacteria also contain a efficient EET. The dense packing also minimises

I. Principles
 5 Pigment properties in more detail: Absorption shifts and broadening 9
protein synthesis, thereby saving energy. The protein to pigment weight ratio can vary from
presence of some protein seems to be essential 7.5 to 16. It also implies that EET is less efficient
in oxygenic photosynthesis because similar because of the larger pigment-to-pigment dis-
Chl a concentrations in solution lead to efficient tances, leading to slower EET steps. However,
excitation quenching (converting the excited- these antenna systems have an efficient funnel-
state energy into heat), also called concentration like organisation, meaning that there is a gradi-
quenching, which would be detrimental for effi- ent of high-energy pigments to low-energy
cient photosynthesis. Other important aspects to pigments going from the outside of the radial
notice are the close contacts between the carot- rods to the central allophycocyanin rods, with
enoids and chlorophylls and the short distances some red-shifted bilins, which are in contact
between Chls b and Chls a, which will be with the photosystems. The advantage of such
addressed in the next paragraph. a funnel is that the excitation energy is directed
LHCII is, in fact, a combination of three pro- towards the RC. This is not the case for the
teins in different compositions: lhcb1, lhcb2, and antenna systems of higher plants and algae in
lhcb3, which are slightly different. In addition, which the peripheral antenna are almost isoe-
most plants also have three additional minor nergetic with the RC.
light-harvesting complexes, lhcb4, lhcb5, and
lhcb6 (see Fig. 2) as part of PSII, also called
CP29, CP26, and CP24, respectively. These occur 5 Pigment properties in more detail:
as monomers only and share a large sequence Absorption shifts and broadening
homology with LHCII. On the other hand, lhca1,
lhca2, lhca3, and lhca4 bind to PSI but also show In Fig. 5, an energy level diagram is given for
a large sequence homology. Noticeably, green Chl a, Chl b, and a typical carotenoid.
algae do not contain lhcb4, whereas they have A transition from the ground state to the Qy state
in addition to lhca1–4 also lhca5–9. All these of either Chl a or Chl b corresponds to the
complexes contain Chl a and b and a variety of lowest-energy absorption band in the red part
different carotenoids. of the solar spectrum. The transitions to the Soret
The outer light-harvesting complexes of dia- bands of these Chls correspond to absorption of
toms are called fucoxanthin-chlorophyll pro- blue light. The transition from the ground state
teins or FCPs, and they are also members of to the triplet state T of Chl is forbidden and will
the large LHC family, but instead of Chl b they not be observed in the absorption spectrum. For
bind Chl c. They show substantial sequence carotenoids, the transition from the ground state
homology with LHCII, and the pigment organi- to the T state and the S1 state are both forbidden,
sation has many common motifs. It is of interest whereas absorption to S2 is allowed.
to mention that again all carotenoids are in van Although the energy level diagram suggests
der Waals contact with Chl molecules and that that the optical transitions correspond to sharp
each Chl c molecule is in the close vicinity of a absorption lines, it is obvious from the absorption
Chl a molecule. spectra above that this is not the case. In fact, there
In Fig. 4, the structure of a phycobilisome is are many ways in which the (protein) environ-
also shown. A striking difference, as compared ment of the pigments can shift the absorption lines
to the LHC family, is the far lower pigment/pro- of individual pigments. In a dynamic and some-
tein ratio in phycobilisomes for these evolution- what heterogeneous protein environment, this
ary old light-harvesting complexes, which is leads to inhomogeneous broadening of the
costly from a protein-synthesis point of view. absorption spectrum. Moreover, the electronic
Depending on the type of subcomplex, the transitions couple to vibrations of the pigments

I. Principles
10 1. Harvesting light

FIG. 5 (A) Absorption spectra and energy level diagram of ground and singlet excited states of Chl a, Chl b, and carotenoids.
The optical forbidden S1 state of Cars is indicated as a dashed line. Absorption spectra are of pigments in 80% acetone. (B) Triplet
excited state energy levels of Chl and Car relative to the singlet excited state levels. Absorption (A), fluorescence (F), internal
conversion (IC), intersystem crossing (ISC), and triplet–triplet energy transfer (T-T ET) are indicated in the figure.

themselves and of the protein environment (pho- Z  

nons), which leads to additional homogeneous ! !
μ¼ Ψ∗f e r Ψi dxdydz
broadening of the absorption spectrum. The elec-
trostatic environment of the pigments is a well-
where Ψf and Ψi are the excited-state and
known reason for absorption shifts to occur, but
ground-state
  wavefunctions of the pigment
also H-bonding and puckering of the Chl rings !
contribute. Whereas the structural deformation and e r represents the 3-D dipole moment
of Chls and, thus, the shifts in absorption are !
operator. μ has a vectorial character, and the
somewhat limited, the effects can be larger for transition probability is proportional to
carotenoids and, in particular, for bilin molecules, ! 2
leading to substantial shifts in absorption. Also, !
E  μ ¼ E2 μ2 cos 2 θ
the occurrence of excitonic states and charge-
transfer states can substantially contribute, but Here, E2 is a measure of the incident light inten-
before addressing those phenomena, we need to sity, μ2 is also called the dipole strength of a par-
introduce the transition dipole moment. ticular transition, which scales with the molar
extinction coefficient εA for absorption (see
equation below),
!
and θ is the angle between
5.1 Transition dipole moment the vectors E and μ .
!

The optical transition from the ground state of The relation between εA and μ2 is given by
a pigment to one of its excited states is the result  0


 8π 3 N ν 

of the interaction between the electric field com- εA ν ¼ μ2 ν
! 3 ð ln ð10Þhcn Þ
ponent of the incident light E and the transition
! !

dipole moment μ of the pigment. μ is a quantum where ν is the wavenumber in cm1, N0 is Avo-
mechanical parameter defined as gadro’s constant divided by 1000, h is Planck’s

I. Principles
 5 Pigment properties in more detail: Absorption shifts and broadening 11
constant, c is the speed of light in vacuum in cm/ dipole moments of molecule 1 and 2, R12 is the
s, and n is the refractive index of the solvent/ centre-to-centre distance between the pigments,
environment. Integration over the entire absorp- and Rc12 is the corresponding normalised vector.
tion band leads to the well-known relation The parameter κ is given by
Z   
 



μ2 ¼ 9:186  103 n ε ν =ν dν κ ¼ μb1  μb2  3 Rc
12  μb1 Rc
12  μb2

When two Chls are located close together in which μb1 and μb2 are the normalised transition
(
1 nm), they will usually strongly interact with dipole moment vectors.
a coupling strength V12. This interaction leads to In case of two isoenergetic pigments (having
two excited states (exciton states) at different equal site energies), the two exciton states are sep-
energies that are shared between both pigments; arated by 2V12 as shown in Fig. 6. In addition to the
each exciton state is, in fact, a linear combination exciton splitting by 2V12, a shift of the average
of the excited states of the individual pigments. energy level, denoted as displacement energy,
One could say that both molecules form a new may also occur. The dipole strength of the absorp-
supermolecule with delocalised excitations. tion transitions to each exciton level depends on
The coupling strength is often approximated the relative position and orientation of both pig-
by the dipole–dipole interaction leading to the ments and two extreme examples are given in
following expression for V12: Fig. 6. In the head-to-tail orientation of the transi-
tion dipole, the transition to the lowest exciton
0! !   1
 μ  μ  3 Rc 
!
μ Rc 
!
μ
level is strongly allowed and the dipole strength
1 @
1 2 12 1 12 2
A is approximately equal to twice the dipole
V12 ¼ 3
4πεε0 R12 strength for a monomer. In the parallel staggered
 
1 ! !  κ configuration, the situation is reversed.
¼ μ1 μ2 The splitting in energy between the two exciton
4πεε0 R312
states is 2j V12 j, when both monomers are
isoenergetic, i.e., their site energies are equal.
in which ε is the dielectric constant, ε0 is the vac- When the site energies of both monomers differ,
! !
uum permittivity, μ 1 and μ 2 are the transition the excitonic coupling increases the separation

FIG. 6 Excitonically coupled dimer. If two monomeric pigments of similar energy have a strong excitonic coupling (V1,2),
they form an excitonically coupled dimer. The average energy level is lowered by the displacement energy (D), and the energy
levels are split by 2V12. On the right side, the allowed electronic transitions for two specific orientations of the transition dipole
moments (represented by the arrows in the plane of the Chls are shown).

I. Principles
12 1. Harvesting light

between the energy levels. This effect becomes within several hundreds of femtoseconds.
smaller the larger the difference in site energies Whereas a Car stays in this state for typically
is, and for large differences in site energies, the 10 ps, the excited-state lifetime of an individual
excitonic effect becomes negligible. A direct con- Chl is several ns. The Chl molecule can sponta-
sequence is that only a slight broadening of the neously fall back from the excited state to the
absorption bands will occur and no visible split- ground state by emitting a photon, a process
ting if the excitonic coupling is small compared called fluorescence. The corresponding decay
to the absorption bandwidth of the monomers. rate (radiative rate or rate of fluorescence kF) is
In case there are more than two interacting pig- directly proportional to μ2, which is the dipole
ments, the number of exciton states will go up pro- strength that also corresponds to absorption
portionally although this may not be noticeable in from the ground state to the first excited state.
the absorption spectrum because of overlap due to The relation between this rate and the dipole
broadening. strength is given by the following equations:
Depending on the protein environment Chl a
 
 3 
can also function as an electron donor or an elec- 64π 4 n ν
tron acceptor, which is what is happening in the 
 

A ν ¼ μ2 ν
RCs where Chl a molecules function both as pri- 3h
mary electron donor and acceptor. But there Z
may also be partial electron transfer from one 


A ν d ν ¼ kF
Chl to another in the excited state, leading to a
so-called charge-transfer (CT) state. CT states
are characterised by increased spectral broaden- For Chl a, a typical radiative rate is
0.05 ns1,
ing and bathochromic shifts to lower energies. but the exact value depends on the environ-
Transitions to CT states are usually only weakly ment/solvent. It can also fall back via internal
allowed, but if these states couple to excitonic conversion in which case the excited-state
states, they may borrow dipole strength and energy is transformed into heat with a typical
become visible in absorption. An example of this rate constant kIC ¼ 0.02 ns1 for Chl a in a solvent.
situation are the red-forms in the antenna of PSI. In the thylakoid membranes, this rate may be at
The red-forms correspond to Chl a dimers that least a factor 10 larger. In addition, the excited
due to excitonic interaction and mixing with a state may relax to a lower-lying triplet state,
CT state absorb around 710 nm, which is more a process called intersystem crossing (see
than 30 nm red-shifted from the absorption of Fig. 5B), and for isolated Chls, this is the most
the bulk Chl a molecules. This red-shifted likely decay pathway with a typical rate con-
absorption is thought to be important under can- stant of kISC ¼ 0.13 ns1. In the absence of excita-
opies, where the blue and red light is absorbed tion energy transfer or electron transfer, these
by upper leaves and the available light is three processes determine the excited-state life-
strongly enriched in the far-red region. time τ ¼ (kd)1 ¼ (kF + kIC + kISC)1 of individual
Chls, which is around 4–5 ns in solution,
whereas it is presumably around 2 ns for Chl a
6 Pigment properties in more detail: pigments in the antenna of PSII. The processes
Decay of the excited states of excitation energy transfer and electron trans-
fer are crucial in photosynthesis, they substan-
After an individual Chl or Car has reached tially lower the excited-state lifetime of
one of its excited states by light absorption, it individual Chls. It is important to mention
falls back to the first excited electronic state that triplet formation on Chls is potentially

I. Principles
 7 Excitation energy transfer, the F€orster equation 13
dangerous for the organism because Chl triplets is defined above, see exciton coupling) and the
can react with oxygen to produce singlet oxy- refractive index n refers to the (protein) medium
gen, which is very reactive and damages lipids, between the two pigments. Combining this
amino acids, and nucleic acids. To prevent this, expression with the expressions for the (fluores-
every Chl a in the antenna is in van der Waals cence) dipole strength for the donor and the
contact with a carotenoid molecule and Chl trip- (absorption) dipole strength of the acceptor
lets are efficiently transferred to these Cars. leads to the F€
orster equation
Because Car triplets are substantially lower in 
energy, they do not lead to singlet oxygen for- R0 6
kDA ¼ kF
mation and Cars can even scavenge singlet oxy- RDA
gen in case it is produced. Here, kF is the radiative rate or rate of fluores-
cence that was introduced above and the F€ orster
radius R0 is given below:
7 Excitation energy transfer, the F€
orster Z 
 

9 lnð10Þ 2 fD ν εA ν 4

equation R0 ≡
6
κ
dν
128π 5 n4 N 0 ν
The Chl excitations created in the antenna It is important to note that the overlap integral
need to be transported to the primary donor of should be determined from the normalised fluo-
one of the RCs, and to obtain a high quantum rescence spectrum fD of the donor (the area of
efficiency, this process of excitation-energy which should be normalised to 1 in this equation)
transfer (EET) should be much faster than the and the absorption spectrum εA of the acceptor,
competing decay processes introduced above. both in situ. Although the appearance of the
How is this achieved? Excitation energy transfer F€orster equation might suggest that the donor
is based on Coulombic interactions between the molecules emit a fluorescence photon that is
pigments and this leads to radiationless EET absorbed by the acceptor molecule, this is not a
between the various pigments. In first approxi- correct view. As already stated earlier, the trans-
mation, many of the transfer steps can be fer is a radiationless process, and the size of the
described by the F€ orster equation. The latter is overlap integral reflects how well the energy
an expression for the rate of EET from molecule levels of donor and acceptor match with each
1 to molecule 2 and it is a manifestation of other. The orientation factor κ2 and the distance
Fermi’s Golden rule and scales with V212, where RDA can be obtained from structural data about
the coupling strength V12 between molecules 1 the pigment–protein complexes, if available.
and 2 is approximated by the dipole–dipole cou- Strictly speaking, the F€
orster equation gives
pling, that was introduced above. This is repre- only a good approximation if the coupling
sented by the following equation: between the pigments is relatively weak, mean-
Z ing that the coupling strength is substantially
4π 2 κ2 
 


kDA ¼ 2 4 6 μ2D ν μ2A ν d ν smaller than the absorption and fluorescence
h cn RDA bandwidths of the pigments involved. More-
in which the subscripts 1 and 2 have been over, the pigments should be far enough apart
replaced by D (donor) and A (acceptor). The fact for the dipole–dipole approximation to be cor-
that the coupling strength is squared leads to the rect. In practice, this requirement is satisfied
R6
DA dependence of the transfer rate kDA. κ is an
2 when the centre-to-centre distances of the pig-
orientational factor depending on the relative ments are 1.5 nm or more, but even with a dis-
positioning and orientation of both pigment (κ tance of around 1.0 nm, the approximation still

I. Principles
14 1. Harvesting light

works rather well (Van Amerongen et al., 2000; hierarchical equations of motion (HEOM) the-
Croce and Van Amerongen, 2020). However, for ory. However, those theories go beyond the
a better description, more sophisticated theories level of the current chapter and more details
are required (see later). can be found in the following reviews
The spectral overlap integral is larger for (Valkunas et al., 2018; Mirkovic et al., 2017;
downhill EET, when the acceptor molecule has Novoderezhkin and Van Grondelle, 2018).
a lower excited-state energy than the donor than
for uphill EET, but the reverse process can also
take place. The ratio of downhill and uphill 9 Overall trapping in photosynthetic units
transfer rates is ruled by Boltzmann statistics
and described by the detailed balance equation The average time between the absorption of a
k12/k21 ¼ eΔE/kT. Here, k12 is the rate of EET from photon somewhere in a photosystem and charge
molecule 1 to 2 and k21 is the rate for reverse EET, separation in the RC is called the trapping time
ΔE is the excited-state energy difference of both τtrap, and the corresponding rate kphot ¼ τ1 trap is
pigments, k is the Boltzmann constant, and T is sometimes called the rate of photosynthesis,
the absolute temperature. which is a bit of a misnomer. The trapping time
τtrap can be considered as the sum of two contri-
butions, the migration time τmig and the overall
8 Excitation energy transfer, beyond the charge separation time τcs, i.e., τtrap ¼ τmig + τcs.
F€
orster equation The migration time τmig, also called first passage
time, reflects the average time for an excitation
As was pointed out earlier, the F€ orster equa- to reach the primary electron donor. The overall
tion is strictly speaking only valid if the coupling rate of charge separation (kcs) is the inverse of the
between the pigments is weak and the dipole– overall charge separation time τcs, which is
dipole approximation is sufficiently accurate. determined both by the intrinsic rate of charge
When, on the other hand, the coupling is very separation kics between primary donor and
strong (larger than the bandwidth), exciton acceptor and the probability ppd that the excita-
splitting will take place and in that case relaxa- tion resides on the primary donor according to
tion between delocalised exciton levels will the Boltzmann distribution, i.e., kcs ¼ (τcs)1 ¼
occur that can be described by Redfield theory. kics/ppd. If kics would, for instance, be 1 ps1
Excitations are differently distributed over the and the entire photosystem would contain 100
pigments contributing to the different exciton isoenergetic Chls, τcs would be 100 ps. More gen-
states and the energy corresponding to the dif- erally, ppd is given by
ference in energy levels is either dissipated as
thermal energy to the direct environment of eEpd =kT
the pigments or reversely taken up from this X
eEi =kT
environment. However, such strong coupling
i
does typically not occur in photosynthetic sys-
tems, and we end up with an intermediate situ- where Epd is the excited-state energy of the pri-
ation in which coupling energies are of the same mary donor and Ei reflects the excited-state
order of magnitude as the bandwidths and nei- energy of all the pigments connected to the pri-
ther the F€ orster equation nor the Redfield theory mary donor, including the primary donor itself.
can be applied and more sophisticated theories T is the absolute temperature, and k is the Boltz-
are needed, like generalised F€ orster theory, mann constant.
modified Redfield theory, combined Redfield– When the migration is infinitely fast and τmig
F€orster approach, Lindblad theory, and can be ignored, the overall trapping is called

I. Principles
 10 Summary: The ideal antenna system—The role of the protein 15
trap-limited, whereas in the opposite case with 250 Chl a molecules, but its excited-state lifetime
infinitely fast charge separation, the process is is as short as
50 ps, corresponding to a value of
termed migration-limited. In reality, the truth ϕphot ¼ 0.97–0.98 for the quantum efficiency,
lies probably somewhere in between, but in PSII, using a value of 0.5 ns1 for kd. In contrast, the
there is a substantial contribution of the migra- largest PSII-LHCII complexes of plants can con-
tion time to the overall trapping time (Croce tain around 130 Chl a molecules per RC, corre-
and Van Amerongen, 2020). Whatever the dom- sponding to a lifetime of
300 ps and a value
inating term in the expression for the overall of ϕphot ¼ 0.85.
trapping time, it is imperative that τtrap is sub- As was already mentioned earlier, the photo-
stantially smaller than the normal average systems can adapt to changing light conditions,
excited-state lifetime of the pigments involved which can either be an increase/decrease of light
(i.e., the lifetime τ in the absence of trapping) intensity or a change in spectral composition.
to avoid that too many excitations get lost. For The response can be a change of antenna size
this reason, antenna systems cannot exist as of PSII and/or PSI but also the reversible induc-
1-D systems, because the excitations would ‘dif- tion of excitation quencher. This will be further
fuse’ far too slowly. However, as we know, the discussed in Chapter 4, ‘Abiotic stress and adap-
photosystems are organised in 2-D thylakoid tation in light harvesting’, by Jun Minagawa.
membranes and the pigment–protein complexes
even have some 3-D characteristics with pig-
ments on the stromal and luminal sides. The 10 Summary: The ideal antenna system—
relation between trapping, excited-state lifetime, The role of the protein
and quantum efficiency of excitation trapping
ϕphot can be expressed by the following equa- An ideal light-harvesting antenna has to fulfil
tions: ϕphot ¼ kphot/(kphot + kd). In the case of PSII, a list of requirements. First, it needs to be able to
this is equal to 1 τopen/τclosed ¼ 1  F0/Fm. Here, strongly absorb the light spectrum that is avail-
τopen and τclosed are the excited-state lifetimes able. This is achieved by selecting the right pig-
with the RC in the open and closed state, respec- ments, but also by the protein that tunes the
tively, and F0 and Fm are the corresponding absorption wavelengths of the pigments. The
steady-state fluorescence levels in the presence absorbed light energy needs to be transferred
of open and closed RCs, respectively. When to the RC and trapped by the RC before the exci-
applying the above equations, it is implicitly tation energy is lost by other decay processes.
assumed that we are dealing with separate pho- This means that the excited state lifetime of the
tosynthetic units, consisting of a reaction centre antenna complexes in the absence of photosyn-
and accompanying antenna system. In reality, thesis should be long, e.g., in ns range. The other
different photosystems can be connected to each side of the coin is that the excitation energy trans-
other, and if one RC is closed, the excitation fer must be fast. As such, the protein has to keep
might migrate to another RC. Moreover, there the pigments at a proper orientation and distance
may be some heterogeneity in the composition from each other that allows for efficient energy
of the various photosystems and some care transfer. Furthermore, the Chls a should be in
should be taken while interpreting the data that close proximity to Cars to avoid the production
are usually collected with time-resolved or of reactive singlet oxygen by the reaction of Chls
steady-state fluorescence measurements. a in the triplet state with molecular oxygen.
In general, the quantum efficiency ϕphot of PSI Finally, the antenna should be adaptive. In low
is very high, even when the antenna size is very light, its absorption cross section should be large,
large. For instance, the PSI-LHCI complex of the and its decay rate should be low. On the other
green alga Chlamydomonas reinhardtii has around hand, when the light intensity is high, and more

I. Principles
16 1. Harvesting light

energy is absorbed than can be used for photo- Novoderezhkin, V.I., Van Grondelle, R., 2018. Modeling of
synthesis, the antenna system should become energy transfer in photosynthetic light harvesting. In:
Croce, R., van Grondelle, R., van Amerongen, H., van
smaller, and/or the decay rate of the antenna Stokkum, I. (Eds.), Light Harvesting in Photosynthesis.
should increase. The on and off switch of the CRC Press, pp. 269–303. Chapter 13.
antenna is regulated at the protein level. Su, X., Ma, J., Wei, X., et al., 2017. Structure and assembly
mechanism of plant C2S2M2-type PSII-LHCII supercom-
plex. Science 357, 815–820.
References Valkunas, L., Chmeliov, J., Van Amerongen, H., 2018. The
exciton concept. In: Croce, R., van Grondelle, R., van
Caspy, I., Borovikova-Sheinker, A., Klaiman, D., et al., 2020. Amerongen, H., van Stokkum, I. (Eds.), Light Harvesting
The structure of a triple complex of plant photosystem in Photosynthesis. CRC Press, pp. 249–268. Chapter 12.
I with ferredoxin and plastocyanin. Nat. Plants 6, Van Amerongen, H., Valkunas, L., Van Grondelle, R., 2000.
1300–1305. Photosynthetic Excitons. World Scientific, Singapore.
Croce, R., Van Amerongen, H., 2020. Light harvesting in oxy- Wientjes, E., Van Amerongen, H., Croce, R., 2013. Quantum
genic photosynthesis: structural biology meets spectros- yield of charge separation in photosystem II: functional
copy. Science 369, 6505eaay2058. effect of changes in the antenna size upon light acclima-
Ma, J., You, X., Sun, S., et al., 2020. Structural basis of energy tion. J. Phys. Chem. B 117, 11200–11208.
transfer in Porphyridium purpureum phycobilisome.
Nature 579, 146–151.
Mirkovic, T., Ostroumov, E.R., Anna, J.M., Van Grondelle, R., Further reading
Scholes, G.D., 2017. Light absorption and energy transfer
in the antenna complexes of photosynthetic organisms. Blankenship, R.E., 2014. Molecular Mechanisms of Photosyn-
Chem. Rev. 117, 249–293. thesis, second ed. Wiley Blackwell, Hoboken, NJ.
 C H A P T E R

2
Transport of electrons
Anja Krieger-Liszkay and Diana Kirilovsky
Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Paris-Saclay University, Gif-sur-Yvette
Cedex, France

1 General principles of photosynthetic by emitting a photon, fluorescence emission; (3)

conversion of light energy the exciton can be transferred to other pigment
(chlorophyll or carotenoid); and finally (4) it
Chlorophyll (Chl) is the major light-absorbing can be used for photochemistry. In the last case,
pigment in most photosynthetic organisms. In the excited Chl (Chl*), which is a strong reducing
cyanobacteria, in addition to chlorophyll, the agent, transfers an electron to an acceptor mole-
phycobilins, open tetrapyrroles that are cova- cule, which will be reduced whilst the Chl is oxi-
lently bound to phycobiliproteins, forming the dised, creating a hole-charge pair (Chl+/
phycobilisome, the cyanobacterial extramem- Acceptor
). This is possible since the electron
brane antenna, are very important light harvest- in its excited state is on a higher energy level
ing pigments. Each time a photon is absorbed by and is easier to be transferred to another molecule
a Chl molecule, an electron in the ground state is than in its ground state. The cationic Chl+ will
excited and it populates one unoccupied excited come back to its initial state by oxidising another
state. In the chlorophyll absorbance spectrum, molecule or by charge recombination (Chl+ +
two principal bands are observed, one in the Acceptor
¼ Chl + Acceptor) since the couple
red and the other one in the blue region of the Chl+/Acceptor
is an unstable system. In addi-
spectra. The first one corresponds to the transi- tion to these four ways of energy dissipation at
tion to the first excited state (S1) and the second the level of the S1 state, there exists also the pos-
one to the second excited state (S2). This state sibility for the forbidden transition to a triplet
decays very rapidly by internal conversion to state of excited Chl. This triplet state can either
the more stable S1 state. From the S1 state, the be directly quenched by carotenoids in its neigh-
excited Chl molecule can dissipate the excitation bourhood or react with oxygen, 3O2, a triplet in its
energy in three different ways: (1) internal con- fundamental state, to the highly reactive and oxi-
version where the excitation energy is converted dising singlet oxygen, 1O2.
into heat whilst the electron returns to the The Chls in the antenna complexes, which
ground state; this is the most rapid process; (2) play a light-absorption role, principally transfer
the excited electron returns to the ground state the energy to an adjacent Chl molecule. The

Photosynthesis in Action 17 Copyright # 2022 Elsevier Inc. All rights reserved.

https://doi.org/10.1016/B978-0-12-823781-6.00003-4
18 2. Transport of electrons

energy can be transferred to a large number of 2 Linear electron transport

molecules (in the same or different proteins)
during the lifetime of the excited state. When In linear photosynthetic electron transport
the excitation energy arrives at a particular Chl chain, the two photosystems are arranged one
a or a pair of Chl a molecules (primary electron after each other with H2O as substrate for photo-
donor) in the reaction centre, charge separation system II (PSII) at the beginning of the chain and
takes place and the electron is transferred to the with photosystem I (PSI) and the final acceptor
primary electron acceptor in a few picoseconds. NADP+ at the end. The order of events was dis-
This is the reaction in which light energy is con- covered by Emerson in 1957. When chloroplasts
verted into chemical energy and it occurs in all were illuminated with light at wavelengths
reaction centres: Type I reaction centres includ- >690 nm, the relatively constant quantum yield
ing those of green sulphur bacteria and reaction of O2 evolution dropped dramatically, whilst it
centre I (RCI) (cyanobacteria, algae, and plants) reached back its maximum by illuminating at
and Type II reactions centres including those of 650 nm and 700 nm. The far-red light absorbed
purple bacteria, Chloroflexi, and reaction centre by PSI can only be efficiently used for O2 evolu-
II (RCII) (cyanobacteria, algae, and plants). tion when light of a shorter wavelength is pre-
The nature of the primary acceptors and the sent, which is absorbed by PSII. The Emerson
characteristics of the special Chl pair (primary effect can be used to determine the action spec-
donor), at the start of the electron transport pro- tra of the two photosystems. Compared to linear
cess, are different in each reaction centre (see the electron transport, cyclic electron flow (see
sections on PSII and PSI). Upon charge separa- below) does not show an Emerson effect.
tion, the positive charge can be shared by the Fig. 1 shows the ‘Z-scheme’ of the electron
two Chl of the dimer as it is the case in RCI or transport system first proposed by Hill and
be centred on one Chl (like in photosystem II Bendall in 1960. In PSII, charge separation takes
(PSII)). In each reaction centre, the primary place after excitation of a chlorophyll a molecule,
donor is designated P (for pigment) followed P680, the primary acceptor is a pheophytin (a
by a number that indicates its wavelength of chlorophyll molecule without the Mg2+ ion),
absorbance (bleaching when the system absorbs and then the electron is transferred to the pri-
light): P680 in PSII, P700 in PSI. The primary mary plastoquinone acceptor QA, a one electron
acceptor in PSII is a pheophytin (Pheo) mole- acceptor, and from there to the secondary qui-
cule, whilst in PSI, it is a Chl molecule (Ao). none acceptor QB, a two electron acceptor. After
As mentioned earlier, the couple P+/ a second charge separation event and electron
Acceptor
is unstable since the small distance transport to QB
, it becomes double reduced
between them promotes a rapid charge recombi- and protonated. The plastoquinol (QH2) leaves
nation: the acceptor molecule gives back the elec- its binding pocket (QB-site) and diffuses to the
tron to the donor. In the recombination reaction, plastoquinone pool. A plastoquinone binds to
all the energy is lost (mostly as heat) and cannot the QB-site and becomes QB. At the donor side
be stored. To avoid this, fast reactions (much fas- of PSII, the water-splitting complex, a Mn4O5Ca
ter than recombination reactions) occur involv- cluster, is bound. After four excitations of PSII,
ing more spaced secondary acceptors and the Mn4O5Ca cluster accumulates four positive
donors that will separate (in distance) and stabi- charges, 4 electrons are taken from 2 H2O mole-
lise the charges in both sides of the membrane. cules, and 1 molecule O2 is generated.
The charges are stabilised not only by increases After excitation of the chlorophyll a dimer,
in distance but also by energy losses in each reac- P700, in the reaction centre of PSI, an electron is
tion step, leading to a further stabilisation. transferred to the acceptor A0, a monomeric

I. Principles
 2 Linear electron transport 19
A
PSII PSI Fd

PQ Cytb6f

Pc
2H2O O2 + 4H+

B *P700

-1,0 -1,0
redox potential (Volt)

*P680
-0,5 Fd -0,5
Fd
NADP+
0,0 PQ/PQH2 0,0
Cyt b6f Pc
0,5 P700+ 0,5

1,0 H2O/O2 1,0

PSI
P680+

PSII
FIG. 1 Photosynthetic electron transport.(A) Simplified scheme of linear photosynthetic electron transport. Yellow flashes,
excitation by light; red flashes, charge separation, and electron transport within the reaction centres; blue flashes, electron trans-
port from water to ferredoxin (Fd), including the Q-cycle; Cyt b6f, cytochrome b6f complex; Pc, plastocyanin; PQ, plastoqui-
none; PS, photosystem. (B) Z-scheme of the photosynthetic electron transport. The red arrows indicate the potential difference
between the excited and the ground states of the central chlorophylls of the reaction centres, P680 in photosystem II (PSII) and
P700 in photosystem I (PSI). The linear electron transport is shown by dark blue flashes, cyclic electron flow by light blue, and the
Q-cycle by orange flashes. OEC, oxygen evolving complex; TyrZ, redox-active tyrosin residue of the D1 protein in PSII; Pheo,
pheophytin; QA, QB plastoquinone acceptors in PSII; PQ, mobile plastoquinone; cyt bl and cyt bh, hemes of the cytochrome b6
with a low redox potential (bl) or a high redox potential (bh); 2Fe–2S, iron–sulphur cluster of the Rieske protein; cyt f, cyto-
chrome f; PC, plastocyanin; A0, A1, electron acceptors in PSI; Fx, FA, FB, 4Fe–4S clusters in PSI; Fd, ferredoxin. This scheme
shows the functional relationship and not the distribution of the complexes within the thylakoid membrane. Credit: Made
by authors.

chlorophyll a, then to the acceptor A1 (phylloqui- PQH2 molecule, one electron is donated from the
none) and afterwards to three 4Fe–4S clusters (see cyt bl (low potential) to the cyt bh (high potential)
below for more details on PSI). and the second one reduces the 2Fe–2S cluster of
An intermediate redox chain connects the two the Rieske protein, which reduces cyt f. Plastocy-
photosystems. QB delivers electrons to the anin, a copper-containing soluble protein in the
mobile plastoquinone pool that diffuses within thylakoid lumen, is reduced by cyt f and delivers
the membrane. The cytochrome b6f complex oxi- an electron to P700+.
dises plastoquinol by reduction of its b-type cyto- At the acceptor side of PSI (stromal side), the
chromes cyt bl and cyt bh. Upon the oxidation of a terminal acceptor FAFB reduces the soluble

I. Principles
20 2. Transport of electrons

ferredoxin (Fd) that contains a 2Fe–2S cluster. the lumen whilst oxidation of a PQH2 in the
The flavin-containing ferredoxin-NADP+-oxido- Qi site releases 2 protons to the stroma.
reductase (FNR) finally transfers the electrons to 3. The reduction of NADP+ at the stoma side
the acceptor NADP+. consuming one H+ per e
.
Experimental evidence for the Z-scheme was
As a consequence, per e
, which are moved
provided by the spectra of the oxidation or reduc-
from H2O to NADP+, two H+ are transferred
tion of the cofactors. Measurements of the differ-
to the thylakoid lumen: one at the PSII, one at
ence spectra of cyt f in intact chloroplasts, for
the cyt b6f, and one H+ is consumed in the stroma
example, have shown that preferential excitation
upon reduction of NADP+.
of PSI induces an oxidation of this redox compo-
During electron transport, in parallel with the
nent whilst excitation of PSII leads to a reduction
ΔpH, an electrochemical gradient ΔΨ is gener-
of cyt f. Similar results have been found for plas-
ated across the thylakoid membrane. The ΔpH
toquinone and P700, to name just a few of the com-
together with ΔΨ forms the proton motive force
ponents of the linear electron transport chain.
(pmf ), which is the energetic basis for the pho-
Another approach to identify the components
tosynthetic generation of ATP by the ATP
of the electron transport chain is the use of spe-
synthase of the thylakoid membrane.
cific inhibitors. A large number of substances is
To achieve oxidation of water, positive poten-
available that interrupt electron transport at
tials are required (+0.82 V) whilst the negative
well-defined sites (see Fig. 2).
midpoint potential of ferredoxin (
430 mV) per-
The electron transport from H2O to NADP+ is
mits the reduction of NADP+ to NADPH (Em
accompanied with the generation of a proton gra-

320 mV). The energy for these chemically
dient (ΔpH) across the thylakoid membrane.
highly demanding reactions comes from the
Three reactions contribute to the formation of
absorption of sunlight (680nm in the case of PSII
the ΔpH:
and 700 nm in the case of PSI), followed by charge
1. At the level of PSII, four H+ are taken from the separation. The oxidation potential of P680 + is
stroma upon reduction of 2 PQ to PQH2 and estimated to be +1.2 to +1.3 V, the highest known
four protons are released in the lumen upon in biology. The midpoint potentials of the elec-
oxidation of two water molecules to form one tron carriers are shown in the Z-scheme (Fig. 1).
oxygen molecule. The electron transfer reactions following the
2. At the level of cyt b6f complex, oxidation of charge separation events are exergonic, leading
two PQH2 in the Q0 site releases 4 protons to to a stabilisation of the radical pairs.

DCMU DBMIB Ferricyanide

Formate
Bromoxynil DNP-INT
MV O2
H2O
OEC PSII QA QB PQ Cytb6f PC PSI Fd

½ O2 DPC
SiMo DCBQ DCPIP DCPIP/ascorbate
PBQ
TMPD
bypass

FIG. 2 Action of inhibitors (red), artificial electron donors (orange), and acceptors on photosynthetic electron transport.
DBMIB, 2,5-dibromo-6-isopropyl-3-methyl-1,4-benzoquinone; DCBQ, 2,6-dichloro-1,4-benzoquinone; DCMU, 3-(3,4-dichlor-
ophenyl)-1,1-dimethylurea; DCPIP, dichlorphenopindophenol; DNP-INT, 2,4-dinitrophenylether of iodonitrothymol; DPC,
diphenylcarbazide; MV, methylviologen or paraquat; PBQ, phenyl-p-benzoquinone; SiMo, silicomolybdate; TEMPD, N,N,
N0 ,N0 -tetramethyl-p-phenylenediamine. Credit: Made by authors.

I. Principles
 4 The protein complexes involved in electron transport and ATP synthesis 21

3 Cyclic electron transport is located in the grana region of the thylakoid

membrane. In the membranes, the PSII is a
Cyclic electron transport (CET) involves the dimer formed by two identical complexes that
transfer of electrons from the PSI acceptor side function independently. Each complex, contain-
via ferredoxin to the PQ pool and their subse- ing 36 chlorophyll and 9 carotenoids and around
quent return to PSI via the cyt b6f complex and 20 subunits, is largely conserved in all oxygenic
plastocyanin. CET results in no net formation photosynthetic organisms (Fig. 3). Whilst the
of NADPH, but it contributes protons to the proteins involved in charge separation and
ΔpH and leads thereby to the production of extra charge stabilisation are almost identical, differ-
ATP. It is important for well regulating the ATP/ ences appear in secondary proteins, especially
NADPH ratio needed for the variable demands in those involved in the stabilisation of the com-
of the metabolism. Together with other regula- plex. The photochemical reaction centre is
tory mechanisms, CET plays an important role formed by two homologue proteins, D1
in protecting the photosynthetic apparatus (encoded by the psbA gene) and D2 (encoded
against photo-oxidative damage when more by the psbD gene), forming a symmetric hetero-
light is absorbed than needed to fulfil the meta- dimer that binds all the cofactors needed for
bolic demands. Two Fd-PQ dependent CET charge separation and stabilisation. The pres-
pathways do exist: (1) the NDH-dependent path- ence of the cytochrome b559 (formed by two sub-
way involving the NDH complex, a homologue units PsbE and PsbF) is also essential for PSII
of mitochondrial complex I that uses Fd as a sub- activity since the minimum PSII complex able
strate and couples electron transfer to PQ with to perform charge separation is composed of
proton pumping to the lumen and (2) the D1/D2/cyt b559. D1 and D2 comprise five
PGR5-PGRL1-dependent antimycin-A sensitive α-helices exposing the N-termini to the stromal
pathway. The exact pathway of this second type side of the thylakoid membrane. An inorganic
of CET is still unclear. PGR5-PGRL1 may act complex, Mn4CaO5, called oxygen evolving
directly as a Fd-PQ reductase or alternatively complex (OEC), involved in the oxidation of
somehow regulate PQ reduction at the Qi site water and production of oxygen, is attached to
of cyt b6f by Fd. Heme ci (see below) of cyt b6f the luminal side of D1. Specific luminal extra-
may play a role as a redox module facilitating membrane proteins (OE33 (or PsbO), OE23
the reduction of PQ by cyt bh at the Qi site. Cyclic (PsbP), and OE16 (PsbQ) in plants and PsbO,
electron transport around PSI may involve mem- PsbV (cyt c550), and PsbU in cyanobacteria) sta-
brane reorganisation and supercomplex forma- bilise the binding and activity of the OEC com-
tion between PSI and cyt b6f as has been plex. In cyanobacteria, there are several
evidenced in the green alga Chlamydomonas isoforms of D1 with slight differences rendering
reinhardtii. the protein more or less resistant to high light
stress. As a consequence, these isoforms are dif-
ferentially expressed depending on the light
4 The protein complexes involved in conditions. D1 is the first component of PSII that
electron transport and ATP synthesis is damaged under high light intensities and
must be replaced very often. To increase the
light absorption, two antenna subunits are
4.1 Photosystem II
located surrounding the D1/D2 core. CP43
In the chloroplast of plants and green algae, (CP for Chl binding protein) is composed by five
photosystem II (PSII) with its light harvesting α helices and contains 14 chlorophylls whereas
complex II (LHCII), the external antenna of PSII, CP47 contains 16 chlorophylls and is composed

I. Principles
22 2. Transport of electrons

B C
D1 D1 D2
D2
CP43 Fe
CP47 QB QA

Pheo Pheo
CAR Chl
Cyt b559
Chl
CAR

P680

OEC

FIG. 3 Photosystem II structure from the thermophilic cyanobacterium Thermosynechococcus elongatus. (A) Photosystem II
dimer. The PSII is shown from the side, with the luminal side of the membrane at the bottom and the stromal side at the top of
the figure. (B) The reaction centre II (D1/D2/cyt b559) and the inner antennae (CP43 and CP47). (C) The cofactors and pigments
attached to the heterodimer D1/D2. Chl, chlorophyll; CAR, carotenoid; Pheo, pheophytin; QA and QB, primary and secondary
quinone acceptors; OEC, oxygen evolving complex. Credit: Figure generated from PDB file 2axt using Pymol.

by six α helices and three β-sheets. The other structure was first based on the knowledge from
subunits are small, and they are generally com- the purple bacteria reaction centre. Neverthe-
posed by only one transmembrane α helix. They less, comparative studies led to the discovery
stabilise the dimer D1/D2 structure and of several differences between PSII and the bac-
improve electron transport activity. In plants terial reaction centre. Here is described what we
and green algae, trimeric and monomeric LHCII knows nowadays about electron transport
proteins are attached to the dimeric PSII com- in PSII.
plex forming supercomplexes containing differ- The heterodimer D1/D2 contains all the
ent quantities of these LHC proteins. In cofactors involved in electron transport in PSII
cyanobacteria and red alga, these membrane (Fig. 3C): 4 Chls (PD1, PD2, ChlD1, ChlD2), 2
LHCII complexes do not exist, and the principal pheophytins (PhD1 and PhD2; only PhD1 is
antenna of the PSII is a huge extramembrane involved), two quinones (QA covalently bound
complex, named phycobilisome. to D2 and QB attached to D1), an Fe atom, two
As mentioned earlier, the PSII belongs to the redox-active tyrosine residues (TyrZ and TyrD),
type II reaction centres. The first and best char- and finally the manganese cluster (Mn4CaO5),
acterised type II reaction centre is that of purple formed by 4 Mn, 5 O, and 1 Ca, principally
bacteria. Our understanding of photosystem II attached to D1 (Fig. 4).

I. Principles
 4 The protein complexes involved in electron transport and ATP synthesis 23
FIG. 4 Structure of the water oxidising Mn4O5Ca cluster in PS II
with four Mn ions (Mn1 to Mn4, purple) and one Ca (yellow),
bridged by oxygen ligands (red). Three Mn ions (1 to 3) and the
Ca form a distorted cube bridged by oxygen ligands, the fourth
Mn (Mn4) is dangling. Mn4 and the Ca carry two water molecules
each (W1 to W4, orange). The coordination of the metal ions by
amino acid ligands from D1 and CP43 is also shown. Credit: From
Lubitz, W., Chrysina, M., Cox, N., 2019. Water oxidation in photosystem
II. Photosynth Res. 142, 105–125. https://doi.org/10.1007/s11120-019-
00648.

In the purple bacteria reaction centre when the charges at the Mn4O5Ca cluster and to oxidise
energy arrives to the special Chl pair PLPM, which two water molecules. At the acceptor side, QA

are strongly coupled, the Chl gives an electron to gives the electron to QB, forming QB
in
the Chl BChlL1 (with BChl being bacteriochloro- 0.2–0.4 ms. QB
has a strong affinity for its binding
phyll that absorbs at much longer wavelengths pocket in D1 and waits for a second electron. Once
than Chl a) forming the redox pair P+ BChlL
. This double reduced and taking two protons (QBH2), it
pair that is formed just three picoseconds after the leaves its binding pocket at D1 and it is replaced by
formation of excited state P* is stabilised by pass- a new oxidised quinone molecule from the plasto-
ing the electron to the bacterial pheophytin (BPh) quinone pool of the membrane. In darkness,
in 1 ps generating P+ BPh
BChlL. depending on the redox state of the plastoquinone
In PSII, the special pair is less strongly coupled, (PQ) pool, not only QB but QB
can also be present
and the energy arriving at the reaction centre is in the QB pocket. In this case, QA
gives the elec-
shared by the four Chls (PD1PD2ChlD1ChlD2). tron to QB
in 0.8 ms forming directly QH2. The
The energy is not localised on a specific Chl, lead- plastoquinol molecule is reoxidized by the cyto-
ing to different charge separation reactions chrome b6f complex (see the next section).
between the Chls. A few tens of picoseconds after Nowadays, the structure of the oxygen-
the first charge separation, the redox pair evolving complex (Mn4CaO5) and the mecha-
PD1+ PhD1
(equivalent to P+ BPh
ChlL) is formed nism of charge accumulation and water
(Fig. 5). The positive charge is localised at PD1. oxidation are the most active research subjects
Then, the electron is transferred from Pheo to on PSII, the catalytic mechanism is still a matter
QA in 400 ps with loss of energy forming the more of debate, and the picture is changing very often.
stabilised pair PD1+ QA
. PD1+ is a strong oxidant Joliot and Kok were the first who observed that
(1.2–1.4 V) and takes an electron from the Tyr160 the quantity of oxygen formed by each flash in a
of D1 (TyrZ) in several tens of nanoseconds (50 series of flashes varies. When a dark adapted
to 250 ns) depending on the oxidation state of sample is illuminated by a series of flashes, no
the Mn cluster (see Fig. 6). Finally, the neutral tyr- or very little O2 is formed in the first two flashes;
osyl radical TyrZ. oxidises the Mn cluster in then a maximum of O2 is produced in the third
around 55μs (in S1) to 1 ms (in S3; for an explana- flash and finally less in the fourth flash. Thus,
tion of the S-states, see Fig. 6). Four charges sepa- the production of oxygen oscillates with a
rations are needed to accumulate 4 positive period of four (Fig. 6A). This is due to the fact

I. Principles
24 2. Transport of electrons

FB
FA

FX

QB QA B A
Fe
quinoneA1
PhD1
ChlA0
PhD2
Chl

ChlD2 ChlD1
PD2 PD1
P680 P700
Reaction Center II Reaction Center I

FIG. 5 Electron transport in photosystem II and photosystem I reaction centres (RC). In photosystem II, 4 chlorophyll a
molecules (PD1, PD2, ChlD1, ChlD2), 2 pheophytins (PhD1 and PhD2; only PhD1 is involved in electron transport), two plasto-
quinones (primary quinone QA covalently bound to D2 and secondary quinone QB attached to D1), and a Fe atom with bicar-
bonate as ligand between QA and QB are shown. The energy arriving at RCII is shared by the 4 Chls; charge separation reaction
occurs, and finally, the redox pair P+D1 Ph

D1 is formed in a few tens of ps. Then, the electron is transferred to QA and then to QB.
A second photon and charge separation is needed to double reduce QB and form PQH2 that leaves the RCII. In reaction centre I,
the two symmetric branches from P700 to Fx are active. P700 donates very fast an electron to the chl A+A forming the P+700A
0A
(A
+
0B) redox couple. Then, the electron is transferred to A1A (A1B) (a phylloquinone) creating the more stable P700A1A (A1B)

redox couple and finally to FX. Then, the electron is transferred to FA or FB Fe–S clusters in PsaC.

that the Mn cluster must accumulate 4 positives (unstacked thylakoids) in the chloroplasts. Plant
charges in order to oxidise two water molecules PSI is monomeric whereas cyanobacterial PSI
and to produce one oxygen molecule. Four can be monomeric or trimeric depending on the
charge separation reactions are needed to accu- growth conditions. Most of the time it is isolated
mulate these 4 positives charges in the as a trimer. In some cyanobacteria strains, even
(Mn4CaO5) complex, which goes through 5 dif- the presence of tetrameric PSI was observed.
ferent redox states in which the oxidation of The PSI monomer is formed by 12 subunits, in-
Mn increases: S0, S1, S2, S3, and S4 (Fig. 6B). cluding the heterodimer PsaA–PsaB (82–83kDa)
In darkness, most of the centres are in S1. This containing all the cofactors needed for charge sep-
explains why the maximum of oxygen is formed aration and stabilisation: the special chlorophyll
in the third flash. Each flash advances the com- pair (P700), four additional chlorophylls (pair
plex to the next S state (Fig. 6A). A and pair A0), two phylloquinones molecules,
and an iron–sulphur (4Fe–4S) cluster (Fx)
(Fig. 7). Three extrinsic proteins at the stroma side
4.2 Photosystem I of the thylakoid membrane, PsaC, PsaD, and PsaE,
Photosystem I (PSI), the other transmembrane are also essential for PSI activity. PsaC binds
pigment-protein complex of the photosynthetic two other 4Fe–4S clusters (FA and FB) that are
apparatus in which light-induced charge separa- located next to FX at the stromal side. The Fe–S
tion occurs, is present in the stroma lamellae clusters are ligated by cysteine residues. PsaF, a

I. Principles
 4 The protein complexes involved in electron transport and ATP synthesis 25
FIG. 6 (A): Release pattern of O2 measured polarographi-
cally after illumination with successive light flashes of spinach
thylakoids at 4°C. O2 release follows a 4-flash pattern (the start-
ing dark stable state is S1). The original experiment was per-
formed by Pierre Joliot as early as 1969 . (B) Water oxidation
cycle (Kok cycle) detailing the five basic S states (S0 to S4),
the light-induced 1e
oxidation steps and the proton release
pattern, the uptake of the two substrate waters, and the Mn oxi-
dation states. The reaction times for the single electron oxida-
tion steps are also indicated. Note that here the ‘S’ stands for
‘state’. Credit: From Lubitz, W., Chrysina, M., Cox, N., 2019. Water
oxidation in photosystem II. Photosynth Res. 142, 105–125. https://
doi.org/10.1007/s11120-019-00648.

transmembrane protein subunit, forms the dock- (PsaA) and AB (PsaB)). Although it was gener-
ing site of plastocyanin at the donor side of PSI. ally assumed that P700 gives the electron to A0
In addition to the cofactors needed for electron (another Chl) to form the first pair radical, more
transport, the PsaA–PsaB dimer attaches around recently it was proposed that AA (or AB) gives
79 chlorophyll molecules that serve as internal the electron to A0 and then very fast P700 donates
antenna. Other 11 chlorophylls and 22 carotenoids an electron to A+A forming the P+700A


0A (A0B)
are coordinated to the small subunits PsaJ, PsaK, redox couple in around 3.7 ps. Then the electron
PsaL, PsaM, and PsaX. In plants, LHCI antenna is transferred in 20–30 ps to A1A (A1B) (a phyllo-
complexes are attached to the monomeric PSI. quinone) creating the more stable P+700A


1A (A1B)
In PSI, there are two symmetric branches for redox couple and finally to FX in about 200 ns.
electron transport from P700 to Fx(Fig. 5). Both Then, the electron is transferred to FA or FB
branches are active. The PSI photoactive core 4Fe–4S clusters in PsaC. Together, PsaC, PsaD,
is formed by 4 excitonically coupled chloro- and PsaE form the docking site of ferredoxin,
phylls: Chl a (PsaB) and Chla’ (PsaA) forming the soluble electron acceptor containing a 2
P700 and two accessory chlorophylls (AA Fe–2S cluster. Ferredoxin has a very negative

I. Principles
26 2. Transport of electrons

FIG. 7 Photosystem I structure from the thermophilic cyanobacterium Thermosynechococcus elongatus. (A) Photosystem
I monomer. The PSI is shown from the side, with the luminal side of the membrane at the bottom and the stromal side at
the top of the figure. The following subunits are visible in this figure: PsaA (green), PsaB (cyan), PcaC (magenta), PsaD (lemon),
psaD (pink), psaJ (orange), psaX (yellow), and psaF (grey). Subunits psaI, psaK, psaL, and psaM are not visible. PsaC, PsaD, and
PsaE are extra membrane proteins. (B) The three proteins carrying all the pigments and cofactors are as follows: PsaA, PsaB,
and PsaC. All the chlorophylls (green) and carotenoids (orange) are shown in addition to the cofactors involved in electron
transport. (C) The cofactors involved in electron transport. Credit: Figure generated from PDB file 1jb0.

reduction potential (
430 mV), and it is easily the plastocyanin dissociates from the cytochrome
reduced by FA or FB clusters (
550 mV). It and diffuses in the lumen till it reaches PSI. The
accepts only one electron, which is shared by binding to PSI is facilitated by its PsaF subunit.
the 2 Fe molecules. In cyanobacteria and green algae, in addition
The final step of linear electron transport is the to, or instead of plastocyanin, cytochrome c6, a
reduction of the NADP+ by Fd via the ferredoxin- c-type cytochrome, is present. Under low concen-
NADP reductase (FNR), a 35–45kDa soluble pro- trations of copper in the medium, the amount of
tein. FNR accepts two electrons, one at a time, cytochrome c6 increases.
from Fd. The electrons are stored at the Flavin
adenine dinucleotide (FAD) cofactor, which can
exist in three forms: fully oxidised, semireduced, 4.3 Cytochrome b6f
and completely reduced going from FAD to The cytochrome b6f complex (cyt b6f ) is the
FADH to FADH2. Then, it realises the two elec- third essential membrane complex involved in
tron reduction of NADP+ into NADPH. photosynthetic electron transport and is equally
P700+ is re-reduced by the luminal soluble pro- distributed between stacked and unstacked
tein plastocyanin that binds a Cu atom. The plas- regions of the thylakoid membrane. It is the link
tocyanin attaches to the cytochrome b6f and between PSII and PSI. Cyt b6f oxidises the mem-
receives an electron from the cytochrome f. Then, brane diffusing plastoquinol molecule formed in

I. Principles
 4 The protein complexes involved in electron transport and ATP synthesis 27

A cyt b6 B
Car
Heme bh
subunit 4
Heme ci Chl

Heme bl
ISP (Rieske)
cyt f FeS

Heme

FIG. 8 Structure of the cytochrome b6f complex from the cyanobacterium Nostoc PCC 7120. (A) Cytochrome b6f monomer.
The cyt b6f is shown from the side, with the luminal side of the membrane at the bottom and the stromal side at the top of the
figure. In the figure, the following subunits are visible: cyt b6 (green), cyt f (magenta), Rieske (ISP) (yellow), subunit IV (cyan),
PetG (subunit 5, violet), and PetN (subunit 8, orange). PetL (subunit 6) and PetM (subunit 7) are not visible. (B) The cofactors of
the cyt b6f monomer. Credit: Figure generated from PDB file 2zt9.

PSII and reduces the luminal plastocyanin (or IV and cyt b6. In each monomer, there are two
cytochrome c6) that reduce P700+ in PSI. Plastocy- quinone binding pockets: one is at the luminal
anin diffusion is a crucial regulatory element of side (Q0), binds a plastoquinol molecule, and
plant photosynthetic electron transport. The cyt is involved in its oxidation, and the other one
b6f, like PSII, is a dimer. Each monomer contains is in the stromal side (Qi) binding an oxidised
four large polypeptide subunits, including two plastoquinone.
cytochromes (cytochrome b6 and cytochrome The electron transfer within the complex is
f ), a 2Fe–2S protein (the Rieske protein) and pro- known as the ‘Q cycle’. One plastoquinol
tein IV, and four small subunits consisting of one (PQH2) binds to the Q0 site at cytochrome b6,
transmembrane α-helix (PetG, PetL, PetM, and its oxidation leads first to the transfer of one elec-
PetN) with a structural role (Fig. 8). The large tron to the Rieske Fe–S centre and the release of
subunits contain the redox cofactors. The cyt b6 one proton to the lumen. The Rieske protein
possesses four transmembrane helices and coor- moves away (around 20 Å) from the cyt b6
dinates two heme groups (bL and bH) and, an towards the cyt f. Once the cyt f is reduced, the
additional heme group, ci that is electronically Rieske protein comes back to its original posi-
coupled with bH. Cytochrome f is formed by a tion and the oxidised PQ molecule dissociates
single transmembrane helix and a large soluble from the Q0 site. The cyt f gives the electron to
luminal domain, which binds a heme group. The the plastocyanin (or cytochrome c6). The second
Rieske Fe–S protein consists also in a single electron is transferred to the low redox potential
transmembrane helix and a soluble luminal heme (bL) and then to the high redox potential
domain, which coordinates a 2Fe–2S cofactor heme (bH), both in the cyt b6, and a second pro-
(instead of a heme). A chlorophyll molecule ton is released to the lumen. A plastoquinone
and a carotenoid molecule of unknown roles molecule attached to the stromal Qi site is semi-
are also bound to the complex between subunit reduced by the electron coming from bH.

I. Principles
28 2. Transport of electrons

A proton is taken up from the stroma. The semi- is activated by the cyt b6f when the PQ pool is
quinol remains in the site till a second electron largely reduced.
coming from the reoxidation of a second PQH2
in the Q0 site. The full Q cycle drives the oxida-
tion of one PQH2, the reduction of two plastocy-
4.4 ATP synthesis
anins and the accumulation of two extra protons The formation of an anhydride binding
in the lumen. between adenosine diphosphate, ADP, and
The cyt b6f is the point of control and regula- phosphate, Pi, generates adenosine triphos-
tion of the photosynthetic electron transport phate, ATP. ATP is often referred to as the
chain. In most of environmental conditions, ‘molecular unit of currency’ of the cell. This gen-
the rate of the electron transport depends on eration of ATP requires energy that is provided
the rate of reoxidation of the PQ pool by the by the proton motive force (pmf ), with
cyt b6f. In addition, cyt b6f is inhibited when pmf 
60 mv ∙ΔpH + Δ Ψ at room temperature.
the lumen pH is very low, a situation occurring The most widely accepted values is 4 for the
at high light intensities and other stress condi- H +/ATP ratio in plant ATP synthase. Plants
tions. Finally, cyt b6f is an essential element of produce ATP by the chloroplast cFocF1-ATP
a photoprotective mechanism called ‘States tran- synthase, which is very similar to the ATP
sitions’, which regulates the relative activities of synthase in the mitochondria. It consists of a
the photosystems by partial migration of LHCII hydrophobic, membrane-spanning part (Fo)
complexes from PSII to PSI and vice versa. This and a hydrophilic, catalytically active head
mechanism involves a LHCII specific kinase that (F1) (Fig. 9). The total molecular weight of the

FIG. 9 Cryo-electron microscopy structure of chloroplast ATP synthase from

spinach (PDB ID 6fkh). The differently coloured subunits are labelled in Greek
(CF1) and Latin letters (CFo). The redox domain in the γ-subunit, exclusively
found in chloroplasts of the green lineage, is coloured in light green. Credit: From
Buchert, F., 2020. Chloroplast ATP synthase from green microalgae. Adv. Bot. Res. 96,
75–118. https://doi.org/10.1016/bs.abr.2020.07.001 (Elsevier).

I. Principles
 4 The protein complexes involved in electron transport and ATP synthesis 29
ATP synthase is about 500 kDa. The ATP and δ act as a stator to prevent unproductive
synthase consists of 26 protein subunits, 17 of rotation of cF1 with cFo. The head is composed
them wholly or partly membrane-embedded. of 3 αβ-subunits, the nucleotide, and phosphate
ATP synthesis in the hydrophilic α3β3 head binding sites, which are connected to the F0 via
(cF1) is powered by the cFo rotary motor in the subunits δ, ε, and γ. The activity of the
the membrane. cFo contains a rotor ring of enzyme depends on the pmf and additionally
14 c subunits, each with a conserved protonata- on the thiol-modification of the regulatory cyste-
ble glutamate. Subunit a conducts the protons to ine residues of the γ-subunit. Reduction of the
and from the c-ring protonation sites. The cen- disulphide bridge by the thioredoxin system
tral stalk of subunits γ and ε transmits the torque activates the ATP synthase. This regulation is
from the Fo motor to the catalytic cF1 head, important to limit the reverse reaction, i.e.,
resulting in the synthesis of three ATP per com- ATP-hydrolysis, in the dark.
plete turn. The peripheral stalk subunits b, b0 ,

I. Principles
Another random document with
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similar in form to other species of the genus. We represent the
development of this larva in Fig. 237. We may call attention to the fact
that this figure illustrates the large size of the paunch, which is so
extraordinary in some of the states of the Termitidae.

It will be recollected that the genus Calotermes is destitute of workers.

There is another genus, Anoplotermes, in which the reverse condition
prevails, and the soldier is absent; this is the only case yet known in
which such a state of affairs exists. The species is called A. pacificus by
Fritz Müller; it differs from other Termitidae in possessing a proventriculus
destitute of triturating ridges. The nests of this species are utilised by a
little Eutermes (E. inquilinus Müller) for its own advantage; whether by
first destroying the Anoplotermes or whether by merely taking possession
of the nests abandoned by their owners is not known. It is a most
remarkable fact that the Eutermes resembles the Anoplotermes so
extremely that the two can scarcely be distinguished, though anatomically
they are quite different. The resemblance is indeed so great that it
deceived Von Jhering into supposing that the two genera were alternate
generations of a single species, one generation possessing soldiers, the
other being without them. Subsequently, by anatomical investigation, he
recognised[299] the error into which he had fallen—an error that, under
such peculiar circumstances, was quite pardonable.

Fig. 237.—Changes in external form of the young larva of Calotermes

rugosus. A, Newly hatched with nine joints in antennae, × 8; B, older
larva with ten joints, × 8; C, next stage with eleven joints, × 8; D, larva
with twelve joints; the position of the parts of the alimentary canal are
shown—v, crop; m, stomach; b, paunch; e, intestine; r, dorsal vessel, ×
16⁄3 (After Fritz Müller.)
Hagen has suggested[300] that Hodotermes japonicus never produces
winged forms. Very little, however, is actually known as to this species.

Marching and Harvesting Termites.—Smeathman alluded to a

remarkable Termes seen by him in Africa, giving it the name of T. viarum.
Nothing further is known of this Insect, which, according to Smeathman's
account, may possibly be the most remarkable of the family. T. viarum is
said to be larger than T. bellicosus, and was discovered issuing in large
numbers from a hole in the ground and marching in columns consisting of
workers directed by soldiers of enormous size, some of whom climbed up
plants and gave audible signals to the army, which immediately
responded with a hissing noise and by increasing their pace with the
utmost hurry; they continued marching by the spot where Smeathman
observed them for upwards of an hour. He was not able to find their
nests, and no specimens have been preserved; both soldiers and workers
possessed eyes. Marching in this way by daylight is contrary to the nature
of ordinary Termites, and some doubt has existed as to the correctness of
Smeathman's observation, which has in fact remained for upwards of a
century without confirmation.

Fig. 238.—Eyed, grass-cutting Termite, Hodotermes havilandi, A, soldier; B,

worker. South Africa. In life the head is carried horizontally, so the
piece of grass sticks up like a flag-pole.

Mr. G. D. Haviland has, however, this year discovered in Natal a Termite

which shows that there are species in Africa of the kind described by
Smeathman, the workers and soldiers being possessed of facetted eyes.
Mr. Haviland states that the workers of this species issue from holes in
the ground during the heat of the day and cut grass both dead and green.
They carry it, in lengths of about two inches, to the mouths of the holes,
often leaving it there and going at once to fetch more. Under acacia
bushes they carry acacia leaflets as well as grass. In the middle of the
day more grass accumulates at the entrance to the holes than can be
taken in, but as the heat of the day diminishes the workers cease to
forage and take in the accumulation. When the grass is all in they
sometimes close the mouth of the hole with moistened pellets of earth
brought in their mouths. The soldiers remain in the holes; when disturbed
they jerk themselves like soldiers of other species to frighten away the
intruder; when they bite, their grip is very tenacious. The holes are about
⅓ of an inch in diameter, and there are usually several of them a few
yards apart; around each of them is a patch over which the grass has
been cut quite short. Mr. Haviland followed these holes by digging for a
distance of 20 feet and to a depth of 5½ feet; they remain uniform in size
except that near the entrance there may be one or two chambers in which
the grass is temporarily stored, but these do not hold more than would be
collected in an hour or two. As the burrow descends it is occasionally
joined by another, and at the point of junction there is usually a
considerable widening. Sometimes they run straight for 6 or 7 feet,
sometimes they curve abruptly, sometimes they are nearly horizontal, but
near the mouth may be almost vertical in direction. These Termites are
very local, but the specimens are numerous when found. Mr. Haviland
dug for these Insects at two places on the Tugela river, one of them being
at Colenso. It is much to be regretted that he was unable to reach the
nest. We figure a soldier selected from specimens sent by Mr. Haviland to
the Cambridge University Museum. This Insect is apparently much
smaller than Smeathman's T. viarum. Other species of Termitidae have
been described[301] as forming underground tunnels in Africa, but none of
the species have yet been satisfactorily identified.

It was stated by Smeathman that some species of Termites had

chambers in their habitations in which grew a kind of fungus used by the
Insects for food; Mr. Haviland is able to confirm Smeathman in this
particular; he having found fungus-chambers in the nests of more than
one species both in Singapore and South Africa (Fig. 240).

Habitations.—In nothing do Termites differ more than in the habitations

they form. Sometimes, as we have mentioned in the case of Calotermes,
there is no real structure formed; only a few barriers being erected in
burrows or natural hollows in wood. In other cases very extensive
structures are formed, so that the work of the Termites becomes a
conspicuous feature in the landscape. This is of course only the case in
regions that are not much interfered with by man; the great dwellings
spoken of by Smeathman and others soon disappear from the
neighbourhood of settlements, but in parts of Africa and in Australia large
dwellings are still formed by these creatures. In the latter part of the world
there exists a very remarkable one, formed by an undetermined species
called by the officers and crew of her Majesty's ship Penguin the
"compass ant." The outline of one of the structures formed by this Termite
we represent in Fig. 239. Mr. J. J. Walker, to whom we are indebted for
the sketch from which this figure is taken, has also favoured us with the
following extract from his diary, of date 4th August 1890: "The most
interesting feature in the scenery (about forty miles inland from Port
Darwin) was the constant succession of huge mounds raised by the
Termites, of which I had seen some comparatively small examples in my
rambles near Port Darwin; but these exceeded in dimensions all I had
ever seen. The most frequent as well as the largest kind was usually of a
reddish or ferruginous colour outside, and generally almost cylindrical in
shape with obtusely-pointed top, but nearly always more or less weather-
worn, with great irregular buttresses and deep ruts down the sides; many
of them look like ruined towers in miniature. Their usual height was from 8
to 10 feet, but many were much higher, and some attained an (estimated)
elevation of at least 20 feet. Another kind, seen only in one or two places
along the line, was of a much more singular character; they averaged
only 4 to 5 feet high, were built of a dark-gray mud, and in shape were
like thin flat wedges set upright (see Fig. 239), reminding one of
tombstones in a churchyard. But the most remarkable feature about these
mounds was that they had all the same orientation, viz. with the long
faces of the wedge pointing nearly north and south. Why this is so I am
quite at a loss to imagine, and I much regret that I had no opportunity of
closely examining these most singular structures. A third kind of mound,
usually not exceeding 2 feet in height, was of a simple, acute, conical
figure, and generally of a gray colour somewhat paler than the last."

Fig. 239.—Termitarium of compass or meridian Termite of North Australia.

A, face extending south and north; B, cross-section.
The material used for the construction of the dwellings is either earth,
wood, or the excrement of the Termites. The huge edifices mentioned by
Smeathman are composed of earth cemented together so as to look like
stone or brick, and the buildings appear to be almost as strong as if they
were actually constructed with these materials. In many cases the
substance used is comminuted wood that has passed one or more times
through the alimentary canal of the Insects, and may therefore be called
excrement. Whether the stone-like material is made from earth that has
passed through the alimentary canal or from grains gathered for the
purpose has not been well ascertained. In any case the material is
cemented together by means of the secretions of glands. Dudley and
Beaumont have described the process of construction, in a species
observed by them, saying that earth is brought and placed in position by
the mandibles, and cemented by liquid from the abdomen.[302] Von
Jhering says[303] that some species form the exterior walls of their
dwellings of stone-like material, but make use of woody matter for the
construction of the interior. Smeathman has described the nest of Termes
bellicosus. The whole of the very strong external wall consists of clay-like
material, cemented by the secretions of the Termites to a very firm
consistence. The royal cell is built of the same material as the framework
of the nest; whilst the nurseries in which the young are chiefly found are
built of woody material, and are always covered with a kind of mould—the
mycelium of a fungus—and plentifully sprinkled with small white bodies,
which, under the microscope, are found to be filled with a number of
oblong, spore-like cells.

Fig. 240.—Fragment of Termitarium of Termes angustatus, S. Africa,

showing fungus chambers and orifices of communication.

These nurseries rest on the clay-like framework of the nest, but are not
attached thereto; they in no way support it, or one another, indeed they
have the appearance of being constantly added to on their upper margins
and constantly eaten away on their under parts. Fig. 240 represents the
appearance of the upper boundary of a nursery taken from a nest of
Termes angustatus. The small white bodies, mentioned above, have
disappeared: the mycelium of the fungus, though not shown in the figure,
is still visible on the specimen from which it was drawn, and gives rise to a
whitish, glaucous appearance.

In various parts of the world nests formed on trees by Termites are to be

seen; these tree nests are, it would appear, in some cases only parts of a
community, and are connected with the main body by galleries. In other
cases nests are formed in various positions of advantage; Messrs.
Hubbard and Hagen have given us an account[304] of some of these—
probably the work of Eutermes ripperti—as seen in Jamaica. They
describe the nests as spherical or conical masses, looking externally as if
composed of loamy earth; they are placed on trees, fences, or walls; they
vary in size from that of a man's fist to that of a hogshead; they appear to
be composed of finely comminuted wood fastened together by saliva.
These nests are formed on the same principle as those of the wasps that
make nests hanging to trees and bushes, as they consist of an external
protecting envelope covering a comb-like mass in the interior. At the
bottom of the nest there is a covered gallery leading to the earth, where
the main nest appears to be situate; galleries also are constructed so as
to lead to the tops of trees and other places, in such a manner that the
Termite can still keep up its peculiarity of working and travelling in tunnels
and yet roam over a large area; the activity of these Termites continues
day and night. In each nest there is a queen, who lays eggs that are
removed by the worker Termites to the bottom of the nest. The young are
fed on a prepared food, consisting apparently of comminuted vegetable
matter, of which considerable masses are laid in store. Some of the nests
are rich in containing many pounds' weight of this material, while others
are apparently quite destitute of it. There is a soldier form and at least two
kinds of workers. Some species of true ant frequently shares the nest of
these white ants, but on what terms the two kinds of Insects live together
is not stated.

Termite Ravages.—In countries whose climate is favourable to their

constitutions certain kinds of Termites become of great importance to our
own species. Owing to their taste for woody matter and to their habit of
working in concealment, it is no uncommon thing for it to be discovered
that Termites have obtained access to a building and have practically
destroyed the wooden materials used in its construction; all the interior of
the wood being eaten away and only a thin outer shell left intact. A
Termite, T. tenuis, was introduced—in what manner is not certainly
known[305]—to the Island of St. Helena, and committed such extensive
ravages there that Jamestown, the capital, was practically destroyed and
new buildings had to be erected. Other such cases are on record.
Destructive species can sometimes be destroyed by placing in the nests
a portion of arsenicated food. This is eaten by some individuals, who
perish in consequence; and their dead bodies being consumed by their
comrades, the colony becomes checked if not exterminated.

The number of described species of Termitidae does not much exceed

100, but this is certainly only a small portion of those existing, the total of
which may probably reach 1000 species.

Termitidae are classed by some naturalists with the Orthoptera, and they
have a great deal in common with some of the cursorial division of that
Order, more particularly Forficulidae and Blattidae; but they differ from
Orthoptera in the nature and form of the wings. They are also classed by
some, with a few other forms, as a separate Order of Pseudo-Neuroptera
called Corrodentia, but this is not a very satisfactory course, as the
Termitidae do not agree closely with the forms associated with them,
while the aggregate so formed is far from being very distinct from other
forms of Neuroptera. On the whole the best plan appears to be to treat
the Termitidae as forming a distinct family of the Order Neuroptera, or to
make it a distinct Order, as proposed by Grassi. Packard now associates
Termites in an Order with the biting-lice, and calls it Platyptera.

Fossil Termites.—Termitidae were very abundant in Tertiary times, and

the genera appear to have been then much the same as at present. In
Mesozoic strata the remains of true Termitidae apparently exist in the Lias
in Europe, but farther back than this the family has not been satisfactorily
traced. It was formerly supposed that Termitidae existed in the
Carboniferous strata, but this appears to be very doubtful; and the fossil
remains of that epoch, which were presumed to be those of Termites, are
now referred by Scudder and others to the Neuropteroid division of the
Order Palaeodictyoptera, an Order which is formed entirely of Palaeozoic
fossil remains.
 CHAPTER XVII

NEUROPTERA CONTINUED—PSOCIDAE (BOOK-LICE AND DEATH-WATCHES)—

THE FIRST FAMILY OF AMPHIBIOUS NEUROPTERA (PERLIDAE, STONE-FLIES).

Fam. IV. Psocidae—Book-Lice, Death-Watches.

Minute Insects with slender, thread-like, or hair-like antennae; four

delicate membranous wings, the front pair of which are the larger;
their neuration is not abundant and is irregular, so that the cells are
also irregularly arranged; the transverse nervules are only one or two
in number.[306] Prothorax very small, in the winged forms quite
concealed between the head and the large mesothorax; this latter
closely connected with, or fused with, the metathorax. Species quite
wingless, or with wings unfitted for flight, exist; in them the prothorax
is not so extremely small, while the mesothorax is smaller than in the
winged forms. Tarsi of two or three segments. Metamorphosis slight,
marked chiefly by the development of wings and ocelli.

Fig. 241.—Psocus fasciatus, England. (After M‘Lachlan.)

The Psocidae are without exception small and soft-bodied Insects, and
are only known to those who are not entomologists by the wingless forms
that run about in uninhabited or quiet apartments, and are called dust-lice
or book-lice. They are perhaps more similar to Termitidae than to any
other Insects, but the two families differ much in the structure of their
wings, and are totally dissimilar in the nature of their lives.

Fig. 242.—Transverse horizontal section of head of Psocus: f, fork or pick; t,

lingua; mx, left maxilla; c, cardo; p, stipes; m.m, muscles; m.s, socket
 of mandible.

Fig. 243.—A, Front of head of Psocus heteromorphus; cl, post-clypeus; g,

epicranium: B, transverse horizontal section of post-clypeus of Psocus:
cl, post-clypeus; c.m, clypeal muscles; g, epicranium; t, tendons; l.m,
labial muscle in section; oe, oesophagus; oe.b, oesophageal bone.
(After Burgess and Bertkau.)

The antennae consist of eleven to twenty-five joints, or even more, about

thirteen being the usual number; the basal two are thicker than the others,
and are destitute of setae or pubescence such as the others possess.
The maxillae and labium are remarkable. The former possesses a
peculiar hard pick or elongate rod; this is considered by many naturalists
to be the inner lobe, but Burgess thinks it more probably an independent
organ,[307] as it has no articulation of any kind with the outer lobe. The
latter is remarkably thick and fleshy; the palpus is 5-jointed. Other
authorities consider the pick to be certainly the inner lobe; if it be not, the
latter is quite wanting. Hagen agrees with Burgess in stating that the pick
slides in the outer lobe as in a sheath. The labium has a large mentum
and a ligula divided anteriorly into two lobes; at each outer angle in front
there is a globular projection, which is doubtless the labial palpus;
reposing on the labium there is a large free lingua. The labrum is large,
attached to a distinct clypeus, behind which there is a remarkable post-
clypeus, which is usually prominent as if inflated; to its inner face are
attached several muscles which converge to be inserted on a plate
placed below the anterior part of the oesophagus, and called by Burgess
the oesophageal bone; under or within the lingua there is a pair of lingual
glands. Judging from Grosse's study of the mouth of Mallophaga, we may
conclude that the oesophageal bone will prove to be a sclerite of the
hypopharynx. The eyes of the winged forms are frequently remarkably
convex, and there are also three ocelli, triangularly placed on the vertex.
The head is free and very mobile. The coxae are rather small, exserted,
contiguous; the sterna small. The abdomen has usually ten segments,
though sometimes only nine can be detected.

The thorax in Psocidae usually looks as if it consisted of only two

segments. This is due to two opposite conditions: (1) that in the winged
forms the prothorax is reduced to a plate concealed in the fissure
between the head and the mesothorax bearing the first pair of wings; (2)
that in the wingless forms (Fig. 247), though the prothorax is distinct, the
meso- and metathorax are fused into one segment.

Fig. 244.—Reproductive organs of Clothilla pulsatoria. A, Male; a, vesiculae

seminales; b, testes; c, vasa deferentia; d, ejaculatory duct. B, Female;
a, b, egg-tubes; c, oviduct; d, uterus, containing egg; e, accessory
gland (the enveloping sac in section); f, its duct. (After Nitzsch.)

The internal anatomy is only very incompletely known. Nitzsch[308] has,

however, described the alimentary canal and the reproductive organs of
Clothilla pulsatoria. The former is remarkably simple: no proventriculus or
crop was found; the stomach is very elongate, and consists of a sac-like
anterior portion and an elongate, tubular posterior part. There are four
Malpighian tubes. The posterior part of the canal is remarkably short, the
small intestine being scarcely as long as the rectum. The ovaries (Fig.
244, B) consist of five egg-tubes on each side; connected with the oviduct
there is a peculiar accessory gland consisting of a sac containing other
small sacs each with an elongate efferent duct; the number of the
secondary sacs varies from one to four according to the individual. The
testis (Fig. 244, A, b) is a simple capsule; connected with the base of the
ejaculatory duct there is a pair of elongate accessory glands or vesiculae
seminales.

The life-history has never been satisfactorily sketched. The young greatly
resemble the old, but have no ocelli or wings, and sometimes the tarsi are
of two joints, while in the adult they have three. The antennae have also
in these cases a less number of joints in the young stage. The food is
animal or vegetable refuse substances; many live on fungoid matter of
various kinds, mouldy chaff being, it is said, a favourite pabulum; the
mould on palings is a source of food to many; others live on the rust-fungi
of leaves, and many frequent the bark of trees. They are able to spin
webs, probably by the aid of the lingual glands; the eggs are deposited, in
some cases, on leaves and covered with a web. Hagen says that a
peculiar organ, possibly a gland—he calls it a hose[309]—exists at the
base of the tarsal claws. In our climate most of the species pass the
winter in the egg-state. There may be two generations in a year, perhaps
more.

The nomenclature of the wing-veins of Psocidae has given rise to much

discussion.[310] The system shown in the accompanying figure is
probably the most convenient; the subcostal vein (2) is always obscure,
and sometimes can only be detected by very minute examination. Some
interesting information as to the minute structure and mode of formation
of the wings and their nervures has been given by Hagen.[311]

Fig. 245.—Anterior wing of Elipsocus brevistylus. (After Reuter.) 1, Costal

vein; 2, subcostal; 3, radial; 4, cubitus; 4a, branches of cubitus; 5,
sector of the radius; 5a, forks thereof.

In the young the wings first appear as buds, or outgrowths of the sides of
the meso- and meta-thorax; afterwards the prothorax decreases, while
the other two thoracic segments and the wing-rudiments attached to them
increase. The wings from their very origin appear to be different from
those of the Orthoptera, and the changes that take place in the thoracic
segments in the course of the development, differ from those that occur in
Orthoptera.
 Fig. 246.—Micropterous form of Mesopsocus unipunctatus. a, a, Wings.
(After Bertkau.)

There are several peculiarities connected with the wings. Frequently they
exist, though of no use for flight; some Psocidae that have perfectly-
formed wings are so reluctant to use them that, M‘Lachlan says, they will
allow themselves to be crushed without seeking to escape by flight. At
certain periods, however, some Psocidae float on the wing in
considerable numbers, especially in a moist still atmosphere, and then
drift about like the winged Aphididae, which are frequently found with
them. There is evidence that individuals, or generations, of some of the
winged species occur with only rudimentary wings; although this has
been denied by Kolbe, there can be no doubt about it. The form figured
above (Fig. 246) was described by Bertkau[312] as a distinct genus, but
was afterwards recognised by him[313] to be a short-winged form of
Mesopsocus unipunctatus. It is probable that the adult female of this
species has the wings always micropterous, while the male has these
organs of the full size. In other species the condition of the rudimentary
wings seems to be quite constant. The facts concerning the wings of
Psocidae are so peculiar that Kolbe came to the conclusion that the
organs exist not because they are of use for flight, so much as because it
is the nature of an Insect to develop wings.[314]

Some of the species of Psocidae have never any trace of wings. These
apterous forms are mostly included in the division Atropinae, and are
usually very minute; it has been again and again erroneously stated that
they are the young state of winged forms. Hagen kept a large colony of
Atropos divinatoria for some years in confinement, so that he saw
numerous generations as well as many specimens. He found the
apterous condition quite constant.

The association of ocelli with wings is nearly constant in Psocidae. The

genus Clothilla—allied to Atropos—possesses very rudimentary wings but
no ocelli. Hagen, however, found[315] that in a certain locality no less than
12 per cent of the individuals of this species were provided with ocelli,—a
most extraordinary variation.
In some of these apterous forms there is found on each side of the
prothorax a tubercular prominence which, according to Hagen, can be
considered only as the rudiment of a wing that never develops. Though
no existing Insect is known to possess rudimentary wings on the
prothorax, we have previously mentioned (p. 344) that in the
Carboniferous epoch appendages of the nature alluded to were not very
rare.

A genus of living forms—Hyperetes—in which the three thoracic

segments are well developed, but in which there are no alar appendages
or rudiments, is considered by Hagen to be more primitive than the
Psocidae found in amber to which we shall subsequently allude.

The number of described species of Psocidae does not reach two

hundred; we have, however, thirty species or more in Britain.[316] Nietner
observed about the same number in the immediate vicinity of his house in
Ceylon. The isolated and remote Hawaiian group of islands is remarkably
rich in Psocidae. Two thousand is a moderate estimate of the number of
existing species. The largest forms yet discovered belong to the Brazilian
genus Thyrsophorus; they attain, however, a breadth of only about one
inch with the wings fully expanded. The Cuban genus Embidopsocus is
said to be of great interest from its approximation to Embiidae. It is at
present very inadequately known.

One (or more) very minute Insects of this family—Clothilla pulsatoria

according to Hagen, Atropos[317] divinatoria according to some other
authors—is widely known under the name of the death-watch, owing to its
being believed to make a peculiar ticking noise, supposed to be prophetic
of the decease of some individual—a human being we fancy, not a death-
watch. It is difficult to believe that so minute and soft an Insect can
produce a sound audible to human ears, and many entomologists are of
opinion that the sound in question is really produced by a beetle—of the
genus Anobium—which lives in wood, and that as the beetle may be
concealed in a hole, while the Clothilla is seen running about, the sound
is naturally, though erroneously, attributed to the latter. But the rapping of
the Anobium is well known, is produced while the Insect is at large, and is
said to be a different noise from that of the Psocid; evidence too has been
given as to the production of the sound in a workbox when the Psocid
was certainly present, and the most careful search failed to reveal any
beetle.

Fig. 247.—A, Atropos divinatoria; B, Clothilla pulsatoria. (After M‘Lachlan.)

The Rev. W. Derham, who two hundred years ago was Rector of
Upminster, in Essex, and was well known as a distinguished writer and
philosopher, gave an account of the ticking of death-watches to the Royal
Society.[318] This gentleman was a most accurate and minute observer;
he was well acquainted with the ticking of the greater death-watch—
Anobium—which he describes very accurately, as well as the acts
accompanying it, the details he mentions being exactly such as occur at
the present time. He not only heard the ticking of the Psocid or lesser
death-watch, but repeatedly witnessed it. He says: "I am now so used to,
and skilful in the matter as to be able to see, and show them, beating
almost when I please, by having a paper with some of them in it
conveniently placed and imitating their pulsation, which they will readily
answer." He also states that he could only hear them beating when it was
done on paper, and that this death-watch will tick for some hours together
without intermission, with intervals between each beat, so that it much
resembles the ticking of a watch. The act of ticking was accompanied by
rapping the front of the head on the paper, but Mr. Derham could not be
sure that the sound was produced in that manner, because each stroke
was also accompanied by a peculiar shudder, or recoil. After a prolonged
ticking he observed that another individual of the other sex made its
appearance. The species figured by Mr. Derham more resembles a
Hyperetes than it does either of our two known book-lice, Atropos and
Clothilla.
 Fig. 248.—The lesser death-watch of Upminster. (After Derham.) A,
magnified; B, natural size.

Fig. 249.—Sphaeropsocus kunowii. From amber. × 30. (After Hagen.)

Numerous species of Psocidae are preserved in amber; Hagen[319] has

made a careful study, based on a considerable number of specimens, of
about thirteen such species. They belong to no less than nine genera and
five sub-families. Sphaeropsocus is the most remarkable; this Insect has
a well-developed prothorax, as is the case in the wingless Psocids, and a
pair of large wings or tegmina meeting by a straight suture along the
back, as is usual in beetles, though quite unknown in existing Psocidae.
Another species, Amphientomum paradoxum, has the body and
appendages covered with scales like a butterfly or moth; other species,
found in gum-copal or still living, have scales on various parts of the body,
but not to so great an extent as this amber species. The genus
Amphientomum is still represented in Ceylon and elsewhere by living
forms; Packard has figured some of the scales;[320] they appear to be
extremely similar to those of Lepidoptera or Thysanura. The facts
connected with this fauna of amber Psocidae would seem to show that
the family was formerly more extensive and important than it is at present;
we should therefore expect to find numerous fossil forms in strata of date
anterior to that of the amber; but this is not the case, all that is known as
to fossil Psocidae being that Scudder has recently ascribed traces of an
Insect found in the Tertiary rocks of Utah to this family as a distinct genus.

Fam. V. Perlidae.
 Insects of moderate or large size, furnished with four membranous
wings; these are usually complexly reticulate; the hind pair are much
the larger, and have a large anal area of more simple venation, which
becomes plicate when folded. The coxae are small, the legs widely
separated. The larvae are aquatic in habits; the metamorphosis is
slight.

Fig. 250.—Pteronarcys frigida, male. (After Gerstaecker.)

The Perlidae form a small family of Insects unattractive in their general

appearance. The life-history of each individual consists of two abruptly
contrasted portions; the earlier stage being entirely aquatic, the later
aerial. Hence the Perlidae come into the amphibious division of
Neuroptera. The definition we have given above would, except as regards
the texture of the front wings and the aquatic habits of the larvae, apply to
many Insects of the Order Orthoptera. The Phryganeidae, another family
of Neuroptera, have aquatic larvae and wings somewhat similar in form to
those of the Perlidae, but the members of the two families cannot be
confounded, as the Phryganeidae have hairy front wings and large and
contiguous coxae.

The antennae of the Perlidae are long, very flexible, and composed of a
very large number of joints. The parts of the mouth vary a good deal. The
mandibles and maxillae are usually rather small, and all the parts of the
mouth are of feeble consistence or even membranous; the maxillary palpi
are, however, well developed and exserted from the mouth, five-jointed.
The labium is short and but little conspicuous. The mandibles in some
forms are almost membranous, but in other genera they are firmer and
are toothed. The labium is composed of a very large mentum, beyond
which is a large piece, usually undivided, bearing the four terminal lobes;
the three-jointed palpus is seated on the side of the large middle sclerite,
which is no doubt of composite nature. Considerable variety as to the
lower lip prevails. The head is broad and flat; there is an indistinctly-
indicated clypeus, three—more rarely two—ocelli, and on each side an
eye neither very large nor perfect. The prothorax is free, and has a flat,
margined notum. The meso- and the meta-thorax are large, equal
segments. The pro-, meso-, and meta-sternum are large pieces; between
the first and second, and between the second and third there is an
intervening membrane. The metasternum is much prolonged backwards,
and has on each side a peculiar slit; similar orifices exist on the other
sterna (Fig. 254, o). Newport, who has examined them in Pteronarcys,
says that they are blind invaginations of the integument; he calls them the
sternal or furcal orifices.[321] According to this naturalist these very
peculiar openings pass into the body "as strong bone-like tubes, diverging
from the axis to the periphery of the body in the immediate vicinity of
some of the principal tracheae, but that they do not in any way
communicate with them, as they terminate abruptly as caecal structures."
He thinks them analogous with the endo-skeleton of other Insects; a view
which cannot be considered sufficiently established. Laboulbène
states[322] that when Perla parisina is seized and placed on its back, it
does not move, but emits a liquid at the base of the articulation of the
legs. This suggests that it may come from these sternal orifices. The
abdomen consists of ten dorsal plates, the first being short, and of nine
ventral; the dorsal plates are much more ample transversely than the
ventral. Frequently the hind body is terminated by two long, many-jointed
cerci, looking like antennae. The coxae are small, not prominent, and are
directed outwards. The legs are slender, the tibiae often grooved. The
tarsi are three-jointed, terminating in two claws and a more or less distinct
pad. In the genus Isopteryx an auditory organ has been described as
existing in the legs, in a position similar to that of the analogous structures
in Termitidae and Blattidae. The wings when closed repose flat on the
back, and fold and overlap so that only one is seen (Fig. 251); in this state
the costal portion of each front wing is turned downwards, so as to protect
to some extent, the sides of the body.
 Fig. 251.—Perla maxima. (After Pictet.)

Fig. 252.—Perla sp., nymph, showing tracheal gills. Pyrénées orientales.

The early stages are known, but have not been described minutely, and
there appears to be very little information as to the youngest life. All the
species are, when immature, aquatic in their habits; the larvae greatly
resemble the perfect Insects in form, though differing in not possessing
wings and in the ocelli being merely opaque spaces. They have rather
large compound eyes; the future wings are represented by lobe-like
prolongations—varying in length according to age—of the meso- and
meta-notum. In the Nemourae the cerci are absent in the imago though
present in the young. The larvae of Perlidae are carnivorous and are able
to swim well, the legs being provided with abundant swimming hairs; they,
however, as a rule, prefer to walk at the bottom of the pool, or on rocks or
boulders in the water they live in.

One of the most peculiar features of the Perlidae is their respiratory

system. Unfortunately the greatest differences of opinion have prevailed
on various matters in connexion with this subject, and there are several
points about which it is not possible at present to express a decided
opinion.
 Fig. 253.—Tracheal gill and portion of a trachea of Pteronarcys. (After
Newport.)

The larvae have no stigmata; it appears to be generally agreed that there

is in them no means of admitting air to the tracheal system by means of
orifices. Some breathe entirely through the integument, the process being
aided by the accumulation of tracheae at the spots where the breathing
orifices should be, and where the integument is more delicate. Others,
however, possess gills in the form of protruded bunches of filaments,
connected with tracheae in the manner shown in Fig. 253. These
filamentous branchiae occur in numerous species of the family, and are
situate on various parts of the body, but many species are destitute of
them in genera, other members of which possess the filaments. In some
Nemourae instead of bunches of filaments there are tubular projections
on the prothoracic segment; and in Dictyopteryx signata similar structures
occur even in the cephalic region, Hagen stating[323] that there exists a
pair on the submentum and another on the membrane between the head
and the thorax. In the imago state, stigmata are present in the normal
fashion, there being two thoracic and six abdominal pairs. In several
species the filaments persist in the imago, so that in these cases we meet
with the curious condition of the coexistence of branchiae with a well-
developed and functionally active system of spiracles; this is the more
curious because the creatures usually have then nothing to do with the
water, it having been ascertained that in these cases the species live out
of the water as other terrestrial and aerial Insects do. These instances of
persistence of branchiae during the aerial life have been the source of
some perplexity; the condition was shown to exist in Pteronarcys by
Newport, and has since been demonstrated in various other forms.
Newport believed that the imago of Pteronarcys breathes by means of the
gills, although it lives out of the water and possesses spiracles; and he
informs us that Mr. Barnston observed the Insect when on the wing
"constantly dipping on the surface of the water." Hence Newport
concluded that Pteronarcys in the winged state is "an amphibious animal."
That a winged Insect should live in the air and yet breathe by means of
gills would be truly extraordinary, and there can be little doubt that
Newport's idea was erroneous. Hagen[324] was able to examine living
imagos of the species in question. He found that they avoided the water,
and though he placed some individuals therein, yet they did not use the
gills. He also informs us that the branchiae have, during life, a shrivelled
appearance, indicating that they are not functionally active, but are merely
useless organs carried over to the imago from the previous instar, in
which they were truly the means of obtaining air. Hagen also ascertained
that the spiracles of the imago are in a normal state, being adapted for
breathing, even as far back as the seventh abdominal segment.

Fig. 254.—Under side of body of Pteronarcys regalis, imago. (After

Newport.) g, Tracheal gills; o, sternal orifices.

Great difference of opinion has prevailed as to the relations of the

branchiae to the stigmata, it having been contended that the falling off of
some of the branchiae left the stigmatic orifices. The facts appear to be
only consistent with the conclusion that the two are totally independent
organs. This subject has been investigated by Palmén,[325] who finds that
in Perlidae—contrary to what occurs in may-flies—the species are either
entirely destitute of gills, or these organs are persistent throughout life. It
is not to be inferred from this that the gills in the perennibranchiate
Perlidae are as conspicuous as they are in the exceptional Pteronarcys:
for it appears that at the final moult the gills usually become very much
contracted and concealed by the new integument; in some cases they
merely appear as slight prominences in the neighbourhood of the
stigmata.

Pictet, Dufour, Newport, and Imhof[326] have studied the internal anatomy.
The alimentary canal is remarkable for the enormous oesophagus; there
is no distinction between this and the crop. A proventriculus is quite