4

Research Update

Acknowledgements

We thank Jane Burrell for producing

Figure 1, and Steve Swain and Greg
Symons for critical comments on the
manuscript. Our research is supported
by the Australian Research Council.
References
1 Beaudoin, N. et al. (2000) Interactions between
abscisic acid and ethylene signaling cascades.
Plant Cell 12, 11031115
2 Ghassemian, M. et al. (2000) Regulation of
abscisic acid signaling by the ethylene response
pathway in Arabidopsis. Plant Cell 12,
11171126
3 Alonso, J.M. et al. (1999) EIN2, a bifunctional
transducer of ethylene and stress responses in
Arabidopsis. Science 284, 21482152
4 Kepczynski, J. and Kepczynska, E. (1997)
Ethylene in seed dormancy and germination.
Physiol. Plant. 101, 720726
5 Debeaujon, I. and Koornneef, M. (2000)
Gibberellin requirement for Arabidopsis seed
germination is determined by testa
characteristics and embryonic abscisic acid. Plant
Physiol. 122, 415424
6 White, C.N. et al. (2000) Gibberellins and seed
development in maize. I. Evidence that
gibberellin/abscisic acid balance governs
germination versus maturation pathways. Plant
Physiol. 122, 10811088
7 Ross, J.J. et al. (2000) Evidence that auxin
promotes gibberellin A1 biosynthesis in pea. Plant

TRENDS in Plant Science Vol.6 No.1 January 2001

J. 21, 547552
8 Haga, K. and Iino, M. (1998) Auxin-growth
relationships in maize coleoptiles and pea
internodes and control by auxin of the tissue
sensitivity to auxin. Plant Physiol. 117,
14731486
9 Yang, T. et al. (1996) Genetic dissection of the
relative roles of auxin and gibberellin in the
regulation of stem elongation in intact lightgrown peas. Plant Physiol. 110, 10291034
10 Brian, P.W. and Hemming, H.G. (1955) The effect
of gibberellic acid on shoot growth of pea
seedlings. Physiol. Plant. 8, 669681
11 Davies, E. and Ozbay, O. (1975) Comparative
effects of indoleacetic acid and gibberellic acid on
growth of decapitated etiolated epicotyls of Pisum
sativum cv. Alaska. Physiol. Plant. 35, 279285
12 Sherriff, L.J. et al. (1994) Decapitation reduces
the metabolism of gibberellin A20 to gibberellin A1
in Pisum sativum L., decreasing the Le/le
difference. Plant Physiol. 104, 277280
13 Ross, J.J. (1998) Effects of auxin transport
inhibitors on gibberellins in pea. J. Plant Growth
Regul. 17, 141146
14 Lester, D.R. et al. (1997) Mendels stem length
gene (Le) encodes a gibberellin 3-hydroxylase.
Plant Cell 9, 14351443
15 Martin, D.N. et al. (1997) Mendels dwarfing gene:
cDNAs from the Le alleles and function of the
expressed proteins. Proc. Natl. Acad. Sci. U. S. A.
94, 89078911
16 Abel, S. and Theologis, A. (1996) Early genes and
auxin action. Plant Physiol. 111, 917
17 Guilfoyle, T.J. (1998) Aux/IAA proteins and auxin

signal transduction. Trends Plant Sci. 3, 205207

18 Lester, D.R. et al. (1999) Gibberellin 2-oxidation
and the SLN gene of Pisum sativum. Plant J. 19,
6573
19 Martin, D.N. et al. (1999) The SLENDER gene of
pea encodes a gibberellin 2-oxidase. Plant Physiol.
121, 775781
20 Reid, J.B. et al. (1983) Internode length in Pisum.
II. Additional information on the relationship and
action of loci Le, La, Cry, Na and Lm. J. Exp. Bot.
34, 349364
21 van Huizen, R. et al. (1997) Seed and hormonal
regulation of gibberellin 20-oxidase expression in
pea pericarp. Plant Physiol. 115, 123128
22 Barratt, N.M. and Davies, P.J. (1997)
Developmental changes in the gibberellininduced growth response in stem segments of
light-grown pea genotypes. Plant Growth Regul.
21, 127134
23 Law, D.M. and Davies, P.J. (1990) Comparative
indole-3-acetic acid levels in the slender pea and
other pea phenotypes. Plant Physiol. 93,
15391543
24 Coenen, C. and Lomax, T.L. (1997) Auxin-cytokinin
interactions in higher plants: old problems and new
tools. Trends Plant Sci. 2, 351356

John Ross*
Damian ONeill
School of Plant Science, University of
Tasmania, GPO Box 252-55, Hobart, Tasmania
7001, Australia.
*e-mail: john.ross@utas.edu.au

Molecular evidence for the evolution of photosynthesis

Robert E. Blankenship
Photosynthesis is a complex metabolic
process that originated on the early Earth.
Recently reported evidence based on the
molecular evolutionary analysis of the
chlorophyll biosynthetic pathway suggests
that the photosystem in the anoxygenic
purple photosynthetic bacteria is the most
ancient known, and that the photosynthetic
apparatus in the oxygen-evolving
cyanobacteria and relatives was the most
recent to appear. This and other evidence
indicate that photosynthesis has had a
complex and nonlinear evolutionary history
and that different parts of the photosynthetic
apparatus have distinct evolutionary origins.

The origin and early evolution of

photosynthesis has long been a subject for
analysis and speculation. In an earlier era,
the major method available was the
comparative biochemistry of existing
organisms, in which various characters such
as pigment contents or metabolic
capabilities were compared, and conclusions
were drawn concerning relationships. The

classic example of the power of this approach

was the recognition by Cornelis B. van Niel
that all (bacterio)chlorophyll-containing
photosynthetic organisms work at the most
basic level by light-induced redox chemistry.
This insight pointed the way to a unified
view of non-oxygen-evolving (anoxygenic)
and oxygen-evolving (oxygenic)
photosynthesis, and suggested that they
might have a common, ancient origin.
We can now employ many more tools to
address this question, including
biogeochemical evidence, microfossils,
molecular structural comparisons and
molecular sequence analysis. Many studies
have used these tools to address various
aspects of how photosynthesis began and
developed, and the results are at the same
time satisfying and frustrating. Satisfying,
in that some issues are now essentially
solved questions: for example, the
endosymbiotic origin of chloroplasts1.
Frustrating, in that our understanding of
other issues is not significantly further
advanced today than it was 25 years ago.

These issues include the nature of the

earliest photosystems and the origin and
development of the capability of oxygen
evolution by oxidation of water2. The
oxidation of water to form oxygen is one of a
handful of truly momentous developments
in the history of life, along with the origin of
life, the invention of information storage and
catalytic systems, and the advent of
eukaryotic cells. The accumulation of
molecular oxygen fundamentally changed
the redox balance on Earth and permitted
the development of aerobic metabolism and
advanced life forms. Biogeochemical
evidence3 indicates that oxygenic
photosynthesis was prevalent by at least
2 billion years ago and possibly much earlier.
Biomarkers indicate that organisms similar
to cyanobacteria were present 2.7 billion
years ago, and 3.5 billion-year-old
microfossils and stromatolites are often
interpreted as evidence that oxygenevolving photosynthesis was already
present at this early stage in the Earths
history. The advanced form of

http://plants.trends.com 1360-1385/01/$ see front matter 2001 Elsevier Science Ltd. All rights reserved. PII: S1360-1385(00)01831-8

Research Update

photosynthesis represented by
cyanobacteria is therefore itself an ancient
evolutionary development, indicating that
the emergence of more primitive forms of
photosynthesis took place even earlier.
The nature of early phototrophs

It seems certain that the first photosynthetic

organisms were simple and anoxygenic, and
that the cyanobacteria, with their much
more sophisticated apparatus composed of
two linked photosystems, were a later
development. But the nature of the earliest
photosynthetic cells has long been a
mystery. How similar were their properties
to those found today in the cyanobacteria
and the four known groups of anoxygenic
photosynthetic bacteria: the purple bacteria,
the heliobacteria, and the green sulfur and
green nonsulfur bacteria?
Within the past few years, gene sequence
data has become available for the reaction
centers of all these groups of organisms.
Along with biophysical studies, analysis of
these sequences has established that all
known photosynthetic reaction centers fall
into two main groups: those that use
pheophytins and quinones as intermediate
and terminal electron acceptors (including
photosystem 2, and the purple bacterial and
the green nonsulfur bacterial photosystems),
and those that use ironsulfur centers as
terminal acceptors (including photosystem 1
and the green sulfur and heliobacterial
photosystems)46. However, relating these
two types of reaction centers to each other
and establishing how they relate to possible
ancestral forms has proven elusive.
Sequence comparisons between the two
types of reaction centers reveal that, if they
are indeed evolutionarily related, then their
divergence occurred so long ago that they
have almost no residual sequence identity
(<10%). However, recent structural
comparisons have suggested that all
reaction centers are built on common
structural motifs, as would be expected if
they do indeed ultimately derive from a
common, distant ancestor7. Even within
each of the two classes of reaction centers,
precise relationships have been difficult to
establish in some cases. For example, the
number and timing of gene duplications in
the evolution of the pheophytinquinone
reaction centers is a subject of continuing
discussion5,8,9. It is likely that significantly
different functional constraints on the two
classes of photosystems have obscured the
phylogenetic signals that might reveal the
ancient relationships between them.
http://plants.trends.com

TRENDS in Plant Science Vol.6 No.1 January 2001

How to reveal ancient evolutionary events

Given that the divergences among the major

groups of photosynthetic reaction centers
are extremely ancient events, are molecular
evolution methods of no use in
understanding how photosynthesis
originated? How do all the classes of existing
photosynthetic organisms relate to each
other, especially given the many artifacts
that can influence the results10,11? Just
because the comparisons involving reaction
centers have proven unsatisfactory for
revealing ancient evolutionary events does
not necessarily mean that the task is
impossible. What is needed is a set of genes
that code for proteins that are found in and
have essentially the same function in all
types of photosynthetic organisms. It is also
important that these proteins be part of the
photosynthetic apparatus itself or
intimately involved in building or regulating
it. An increasing amount of evidence has
established that genomes are mosaics, made
up of parts from many sources12, so that if
one has any hope of understanding the
evolution of the process of photosynthesis,
then it is necessary to examine the genetic
information that is most intimately
concerned with photosynthesis.
Many parts of the photosynthetic
apparatus appear to be unsuitable for these
wide-scale comparisons. In addition to the
problems with reaction centers discussed
above, three apparently unrelated carbon
fixation pathways are found among the
various groups of photosynthetic organisms,
rendering their component enzymes (or the
genes encoding them) unsuitable for widerange comparisons. Many structurally and
functionally distinct classes of antenna
complexes are found in various
photosynthetic taxa, but there are no
antenna types that are universally
distributed. For these reasons, analyses of
carbon-fixation enzymes and antennas are
not useful for revealing deep relationships,
although they are important in establishing
more-detailed evolutionary relationships.
One set of genes that fits the abovementioned criterion and that has been
analyzed recently is the cytochrome bc
complexes13. However, the ideal subset of
the genetic information that codes for
photosynthetic complexes appears to be the
biosynthetic enzymes that make the
photosynthetic pigments such as
chlorophylls and carotenoids. Similar
pigments are found in all photosynthetic
organisms, and the differences that are
present almost certainly come at the end of

the process, so that most of the enzymes that

are involved in pigment biosynthesis have
precisely the same function in all cases.
New molecular data available

A recent paper by Carl Bauer and

colleagues14 has focused on sequencing and
molecular evolution analysis of many of
these pigment biosynthesis enzymes. They
have compared a series of chlorophyll
biosynthetic enzymes found in all five groups
of photosynthetic prokaryotes, plus several
algal and plant species. Their results
strongly support the assertion given above
that the anoxygenic form of photosynthesis
is evolutionarily more ancient than the
oxygenic form. The earliest branching taxa
in all cases were the purple photosynthetic
bacteria, with all the oxygenic taxa clustered
as the most recently derived. Their results
effectively disprove a strict interpretation of
the Granick hypothesis15, which states that
the pigment biosynthetic pathway
recapitulates the evolutionary appearance
of the pigment itself. Bacteriochlorophyll
biosynthesis goes through a chlorophyll-like
intermediate step16, and certain anoxygenic
photosynthetic bacteria contain traces of
chlorophyll-like pigments. These facts have
long been used to argue that chlorophyllcontaining organisms pre-date
bacteriochlorophyll-containing anoxygenic
photosynthetic organisms4, although the
results of Bauers group argue that
bacteriochlorophyll is indeed the more
ancient pigment. An ambiguity in this
conclusion is that it is not possible to tell
from this analysis whether the ancient
organisms contained chlorophyll or
bacteriochlorophyll, only that present-day
chlorophyll-containing organisms appear to
be more derived forms than the
bacteriochlorophyll-containing organisms.
Analysis of additional genes might provide
new insights into this issue.
The new analysis14 produced some
surprises. The heliobacteria consistently
grouped nearest to the oxygenic taxa, in
spite of the fact that biochemical analyses
indicate that this group contains by far the
most primitive photosynthetic apparatus of
any known organism. It has no antenna
complexes, a homodimeric reaction center, a
simple pigment complement and no capacity
for photoautotrophic growth17. However,
this close relationship of heliobacteria and
oxygenic photosynthetic organisms is also
suggested by the analysis of the cytochrome b
protein sequences13. In addition, the green
sulfur and green nonsulfur bacteria

Research Update

TRENDS in Plant Science Vol.6 No.1 January 2001

the Exobiology program of the US National

Aeronautics and Space Administration
(NASA) and the Arizona State University
NASA Astrobiology Institute.

?
?

Carbon fixation
pathway

Nonphotosynthetic
ancestor

Reaction
center

O2 evolution

References

Primitive anoxygenic
photosynthetic organism

Gene
recruitment

Cyanobacterium

Cytochrome f
?

TRENDS in Plant Science

Fig. 1. Illustration of the complex nature of the evolutionary history of the photosynthetic apparatus in
cyanobacteria, leading to a mosaic structure. Various inputs of genetic information are indicated, including inflow by
lateral gene transfer and gene recruitment from another metabolic pathway within an organism. Some possible
candidates for lateral transfer are indicated.

consistently grouped together in the

analysis of pigment biosynthetic enzymes.
These organisms contain a similar antenna
complex, the chlorosome, but are otherwise
distantly related organisms, both in
photosynthetic activity and nearly all other
aspects of cellular metabolism. Perhaps this
suggests that the pigment biosynthesis
genes and chlorosome structural proteins
have been laterally transferred between
these organisms.
An outgroup is often used to resolve the
ambiguity of where to root an evolutionary
tree. An outgroup is a sequence that is
homologous to the sequences under
consideration but is found only in distantly
related organisms or has diverged to a
different function in the same organism.
Bauers group14 used several different
sequences as outgroups, which themselves
reveal some interesting relationships
relevant to the evolution of photosynthesis.
For the light-independent
protochlorophyllide reductase complex they
used the nitrogenase complex as an
outgroup, which has been recognized for
some time as a related, and probably more
ancient, metabolic activity18. For the Mg
chelatase enzyme complex they used the
cobalt and nickel chelatase enzymes from
nonphotosynthetic organisms. Both of these
enzymes were almost certainly recruited
from other metabolic pathways, were
modified in function and now catalyze
biosynthetic steps that are unique to
photosynthetic systems.
The picture that is slowly emerging from
this and many other studies is that the
evolutionary development of photosynthesis
http://plants.trends.com

is a complex process that cannot be

described by a simple, linear, branching
evolutionary diagram. Rather,
photosynthesis emerged by recruiting and
modifying genes encoding components of
several other pre-existing metabolic
pathways, along with a few key innovations
and probably several lateral gene-transfer
events. The resulting view is that, like many
metabolic pathways19, photosynthesis is a
mosaic process that has no single well
defined evolutionary origin20.
Photosynthesis in different classes of
organisms or even different portions of the
photosynthetic apparatus in a single
organism might have significantly different
evolutionary histories (Fig. 1).
Bauers group14 has provided a quantum
leap forward in our knowledge of the
evolution of photosynthesis, in particular,
suggesting how the pigment biosynthesis
pathway might have developed. However,
their results cannot necessarily be
extrapolated to other major unsolved
questions in the field, such as the nature of
the earliest form of photosynthesis, the
origin of linked photosystems and the
advent of the oxygen evolution capability.
Information from a wide range of sources
and disciplines, including molecular
evolution studies of complete genome
sequences, biochemistry and geology, needs
to be assembled and integrated to provide a
deep understanding of the evolution of
photosynthesis.
Acknowledgements

Research in my laboratory on the origin and

evolution of photosynthesis is supported by

1 Margulis, L. (1993) Symbiosis in Cell Evolution:

Microbial Communities in the Archean and
Proterozoic Eons, W.H. Freeman
2 Blankenship, R.E. and Hartman, H. (1998) The
origin and evolution of oxygenic photosynthesis.
Trends Biochem. Sci. 23, 9497
3 Des Marais, D.J. (2000) When did photosynthesis
emerge on Earth? Science 289, 17031705
4 Olson, J.M. and Pierson, B.K. (1987) Evolution of
reaction centers in photosynthetic prokaryotes.
Int. Rev. Cytol. 108, 209248
5 Blankenship, R.E. (1992) Origin and early evolution
of photosynthesis. Photosynth. Res. 33, 91111
6 Nitschke, W. et al. (1998) Evolution. In
Photosynthesis: A Comprehensive Treatise
(Raghavendra, A.S., ed.) Cambridge University
Press, pp. 285304
7 Schubert, W.D. et al. (1998) A common ancestor for
oxygenic and anoxygenic photosynthetic systems: a
comparison based on the structural model of
photosystem I. J. Mol. Biol., 280, 297314
8 Blankenship, R.E. (1994) Protein structure,
electron transfer and evolution of prokaryotic
photosynthetic reaction centers. Antonie van
Leeuwenhoek J. Microbiol. 65, 311329
9 Lockhart, P.J. et al. (1996) Gene duplication and
the evolution of photosynthetic reaction center
proteins. FEBS Lett. 385, 193196
10 Yang, Z. (1996) Among-site rate variation and its
impact on phylogenetic analysis. Trends
Ecol.Evol. 11, 367372
11 Philippe, H. and Forterre, P. (1999) The rooting of
the universal tree of life is not reliable. J. Mol.
Evol. 49, 509523
12 Doolittle, W.F. (1999) Phylogenetic classification
and the universal tree. Science 284, 21242128
13 Schtz, M. et al. (2000) Early evolution of
cytochrome bc complexes. J. Mol. Biol. 300, 663675
14 Xiong, J. et al. (2000) Molecular evidence for the
early evolution of photosynthesis. Science 289,
17241730
15 Granick, S. (1965) Evolution of heme and
chlorophyll. In Evolving Genes and Proteins
(Bryson, V. and Vogel, H.J., eds), pp. 6788,
Academic Press
16 Beale, S.I. (1999) Enzymes of chlorophyll
biosynthesis. Photosynth. Res. 60, 4373
17 Amesz, J. (1995) The antenna-reaction center
complex of heliobacteria. In Anoxygenic
Photosynthetic Bacteria (Blankenship, R.E et al.,
eds), pp. 687697, Kluwer Academic
18 Burke, D.H. et al. (1993) Early evolution of
photosynthesis clues from nitrogenase and
chlorophyll iron proteins. Proc. Natl. Acad. Sci.
U. S. A. 90, 71347138
19 Lazcano, A. and Miller, S.L. (1999) On the origin
of metabolic pathways. J. Mol. Evol. 49, 424431
20 Blankenship, R.E. Molecular Mechanisms of
Photosynthesis, Blackwell Science (in press)

Robert E. Blankenship
Dept of Chemistry and Biochemistry, Center
for the Study of Early Events in
Photosynthesis, Arizona State University,
Tempe, AZ 85287-1604, USA.
e-mail: blankenship@asu.edu