Critical Reviews in Microbiology, 19( 1):43-59 (1993)

The Prochlorophytes: Are They More Than

Just Chlorophyll a/b-Containing
Cyanobacteria?
George S. Bullerjahn*
Department of Biological Sciences, Center for Photochemical Sciences, Bowling Green State
University, Bowling Green, OH 43403
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13

Anton F. Post
Department of Microbial and Molecular Ecology, Life Sciences Institute, Hebrew University, Givat
Ram, Jerusalem 91904 Israel

*To whom all comspondcncc should be addressed.

ABSTRACT: The prochlorophytes are a diverse group of photosynthetic prokaryotes that fall within the cy-
anobacterial lineage, yet lack phycobilisomes as light harvesting structures. Instead, the prochlorophytes have
a light-harvesting apparatus composed of the higher plant pigments chlorophylls a and b. This review discusses
the evolutionary relationships among these bacteria, with focus on the structure and function of the photosynthetic
apparatus. This analysis yields a consensus from studies both on Prochloron sp. and Prochlororhrix hollandica
For personal use only.

as to how the thylakoid membrane is organized. Overall, we propose that the structure of the light-harvesting
complexes (LHC) from prochlorophytes is very different from those of chloroplast systems, and is evolutionarily
very ancient. The functional association of the light-hawesting apparatus with photosystem I (PSI) in both
Prochlorothrix and Prochloron, as well as a demonstrated capacity for PSI-dependent anoxygenic photosynthesis
in Prochlororhrir, may indicate that there is an increased dependence on cyclic photophosphorylation in these
organisms. Finally, the structure of the prochlorophyte thylakoid membrane is discussed with respect to the
forces that drive thylakoid membrane stacking in prochlorophytes and chloroplasts. We suggest that the light-
harvesting structures in prochlorophytes play little, if any, role in this process.

KEY WORDS: cyanobacteria, chlorophyll, photosynthesis, thylakoid, Prochlorothrir, Prochloron, Prochlo-

rococcus, photosynthetic antenna.

I. INTRODUCTION chlorophytes and even considered to fall within

Until 1975, it was common knowledge that
the combination of chlorophylls a and b (Chl a
and b) was unique to the (eukaryotic) green algae
and the higher plants. During the last 2 decades,
reports have appeared on a few photosynthetic
prokaryote species that carry Chl a and b. These
findings triggered intense discussion regarding
the&possible role in the evolution of green chlo-
roplasts. As a result, they were grouped as pro-
a separate taxonomic group. We use the term
‘‘prochlorophytes” here meaning prokaryotes
containing chlorophylls a and b with no reference
whatsoever to them being prokaryotic chloro-
phytes. This review focuses mainly on the current
knowledge of thylakoid structure, organization,
and functioning in prochlorophytes. This focus
is used for a critical review of molecular phy-
logeny and the position of prochlorophytes among
photosynthetic organisms; additionally, we ex-

1040-841)(/93/%.50
Q 1993 by CRC Press, Inc.

43
 amine the properties of the photosynthetic ap-
paratus, aiming to explain how such features sup-
port competitiveness in their natural environ-
ments.
II. INITIAL CHARACTERIZATION OF
PROCHLOROPHYTES
As described previously, the most obvious
feature of the prochlorophytes is the presence of
Chl b in addition to Chl a. While Chl a is an
essential component mediating both light-har-
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
vesting and electron transport functions in all ox-
ygenic phototrophs. Chl b is a pigment dedicated
to light-harvesting. For the sake of clarity, those
Chl alb pigment-protein complexes dedicated to
light-harvesting henceforth are referred to as an-
tenna complexes, while the core Chl a complexes
comprising photosystems I and I1 (PSI and PSII)
are termed reaction center complexes.
Prochlorophytes are found in different en-
vironments. Until now, three genera having one
species each have been described: Prochloron
For personal use only.
didemni, Prochlorothrix hollandica, and Pro-
chlorococcus marinus. Prochloron didemni is
found in a symbiotic relationship with marine
tunicates in (sub)tropicalcoastal waters. Upon its
discovery,' it was at first thought to be a marine
cyanobacterium containing two distinct chloro-
phyll types. Identification of these pigments as
Chl a and b and the apparent absence of cyano-
bacterial antenna phycobilins subsequently formed
the basis for the creation of a new phylum, the
Prochlorophyta.2 For a decade, Prochloron was
the only known photosynthetic prokaryote con-
taining Chl Q and b, and it was considered by
some a living fossil, proof for the theory of endo-
symbiosis..
The second prochlorophyte was discovered
much along the same lines that characterized the
finding of Prochloron. From a series of enrich-
ment experiments aimed at the isolation of char-
acteristic planktonic photosynthetic organisms in
a shallow lake in the Netherlands emerged a light
green subculture lacking phycobilins. Although
never published under that name, it was at first
identified as Oscillatoria limnetica. After estab-
lishing the prokaryotic nature of the organisms
together with the presence of Chl a and b, it
44
became evident that a second prochlorophyte was
found.3 It differs from Prochloron in that it is
filamentous, free-living, and thrives in a fresh-
water environment; thus it was described as the
new species Prochlorothrix hollandica in the new
family of the Prochlor~thricaceae.~ With the find-
ing of Prochlorothrix came the realization that
prochlorophytes may form a group of related Chl
alb-carrying prokaryotes, whose representatives
were hitherto overlooked due to a lack of criteria
for recognition and the apparent difficulties met
when trying to bring these organisms into culture.
The discovery of a third prochlorophyte in
part stemmed from work that had described for
over a decade the presence of a distinctly differ-
ent type of chlorophyll in marine waters at var-
ious locations in the Atlantic O ~ e a n .The
~ . ~oc-
currence of this chlorophyll species was corre-
lated with the presence of picophytoplankton. It
took the on-board deployment of a flow cyto-
meter capable of detecting fluorescence emission
by individual cells to detect the smallest prochlo-
rophyte so far.' They are found in large numbers
in marine surface waters as free-living photosyn-
thetic coccoid cells, and contain divinyl-Chl a
and b. After its isolation, this prochlorophyte was
named Prochlorococcus marinus.8The discovery
of this organism will most probably effect a shift
in the focus of prochlorophyte research away from
the evolutionary origin of the chloroplast toward
characterization of the photosynthetic apparatus
in an ecologically important group of organisms.
111. TAXONOMIC STUDIES AMONG THE
PROCHLOROPHYTES
As one reviews much of the literature on the
prochlorophytes, it becomes clear that taxonomic
classification and the inferring of relationships
between organisms can be a risky enterprise.
Some microbiologists have been eager to show
a direct relationship between prochlorophytes and
the chloroplast, whereas others were determined
to place them among the cyanobacteria. The un-
derlying problem in such exercises lies both in
the danger of overemphasizing one aspect while
belittling others and in the import of externally
imposed bias into the judgment of a certain prop-
 erty. Here we try to summarize the available in-
formation used for the determination of homol-
ogy and structural similarity among prochloro-
phytes and other groups of phototrophs.
A. Morphology
Studies of the prochlorophyte ultrastructure
using light and electron microscopy clearly es-
tablished the prokaryotic nature of the three
prochlorophytes (Figure 1 a to d). The various
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
For personal use only.
prochlorophytes differ considerably in cell size.
Pruchloron is found as coccoid cells with a di-
ameter of 9 to 30 The spherical cells of
Pruchlorucuccus are 1.2 to 1.6 pm in length and
0.6 to 0.8 p m in width.* The only filamentous
species, Pruchluruthrir, are cells of 3 to 10 pm
in length and 0.5 to 1.5 p m in diameter during
exponential g r o ~ t hThe
. ~ filaments or trichomes
consist of at least 5 cells, but may increase to up
to 100 cells or more in the absence of shearing
forces such as stirring or aeration. In all three
species, the thylakoid membranes lack the phy-

FIGURE 1. Electron micrographs of Prochloron (a); Prochlorococcus (b); Prochlorothrix (c and

d) cells. Note the presence of carboxysomes (arrows) and thylakoid membranes arranged in
parallel arrays. (Scale bars = 0.5 Km.) (Photos courtesy of Drs. Hewson Swift, Sallie Chisholm,
Ken Miller, and Karen Milbauer-Warner.)

45
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
For personal use only.
FIGURE 1 (continued)
cobilisomes characteristic for cyanobacteria, and
the membranes are found spread in the cyto-
plasm, often running parallel to one other and to
the cell wall. Modest membrane stacks are seen,
which appear as these parallel arrays of mem-
branes, but it is difficult to distinguish well-de-
fined stacked granal and unstacked stromal mem-
branes characteristic of higher plant chloro-
46
plasts. l o This difference in overall membrane or-
ganization is especially interesting in light of the
thylakoid organization as studied in Prochfo-
rothrix (see later discussion).
Further prokaryotic traits found in prochlo-
rophytes are the different inclusion bodies. Car-
boxysomes, which contain the enzymes essential
for efficient photosynthetic carbon fixation, have
 been shown to occur in Prochioron".'2 and in
Prochlorothrix.l 3 Gas vacuoles were found at both
sides of the cell wall separating the longitudinal
cells in the Prochlorothrix filaments. l 3 The cell
wall itself bears great similarity to that of Gram-
negative bacteria and cyanobacteria. Transmis-
sion electron micrographs clearly show a cyto-
plasmic membrane, a periplasmic space, an outer
membrane, and an outer sheath in all three spe-
c i e ~ . ~ - ~ In
. ~summary,
* ~ . ' ~ . these
~ ~ features show
that the prochlorophytes are morphologically more
similar to cyanobacteria than to chloroplasts. The
'only exception to this observation, thylakoid
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
stacking, is dealt with in some detail in Section
IV.
B. Cell Composition
The macromolecular building blocks of the
prochlorophyte cell bear great homology to those
of cyanobacteria. On the cell surface of Proch-
lorothrix, one finds an abundant S-layer protein,
which binds the carotenoid zeaxanthin. l 4 This
For personal use only.
protein has been observed only in Prochlorothrix
until now, although a similar carotenoid-protein
configuration may exist in Prochloron based on
the observation that Prochloron secretes zeax-
anthin into media and buffers used during phys-
iological determinations. Is The cell wall contains
a multilayered A 1-gamma-type peptidoglycan in
both Pr~chloron'~*'' and Prochlorothrix.l8 Such
an outer sheath is very similar to that found in
cyanobacteria. Prochlorophyte membranes con-
sist of four major lipid components, manogalac-
tosyl-diacylglycerol, digalactosyl-diacylgly-
cerol, sulfoquinovosyl-diacylglycerol,and phos-
phatidylglycerol, but not phosphatidylcholine,
which is specific for the green chloroplast. 19-22
Like cyanobacteria, prochlorophytes contain
monoglucosyl-diacylglycerol, their overall fatty
acid compositions are similar, i9-22 and so are the
buoyant densities of both the thylakoids and the
cytoplasqic membrane. I 5 Major fatty acids in
Prochloron and Prochlorothrix are 14:0,14: lw5,
16:0,- 16:lw7, and two novel fatty acids unique
to Prochlorothrix: 16: l w l 2 (hexadec-4-anoic
acid) and a 16:2 isomer.22The DNA is generally
found in the central regions of the cell surrounded
by the thylakoid rnembrane~.'.~.~~ The genome
size of Prochloron and Prochiorothrix has been
estimated at 3.5 to 4.0 X lo9 bp,23.'4 while the
GC composition of Prochloron and Prochloroth-
rix DNA is between 40 to 50%.4.24
C. Phylogenetic Studies
Study of gene homology is presently consid-
ered the most powerful tool for taxonomic and
phylogenetic purposes. An initial study of Proch-
loron 5s rRNA has already established a position
for Prochloron among the cyanobacteria, distant
from the eubacteria and the chloroplast.2s The
only sequences known for all three prochloro-
phytes are the genes encoding 16s rRNA and
DNA-dependent RNA polymerase, and these have
been used for phylogenetic Such work
places the prochlorophytes as divergent members
within the cyanobacterial radiation, and the re-
sults from these studies have led to the conclusion
that Chl b evolved more than
Of the three bacteria, the most sequences
available for phylogenetic comparisons to other
phototrophs have been determined from Proch-
iorothrix. The cloned genes analyzed mostly en-
code different components of the photosynthetic
apparatus. To date, the following gene sequences
are available for comparison: psbA, encoding the
D1 protein of PSII;29 psbB, encoding the PSII
Chl-protein CP47;30psbH, encoding the 10-kDa
PSII pho~phoprotein;~' petBD, encoding apocy-
tochrome b, and subunit 4 of the cyt bdf com-
p l e ~ petE,
; ~ encoding pla~tocyanin;~~ and rbcLS,
encoding the subunits of ribulose bisphosphate
carboxylase/oxygenase. 33*34 While many of these
sequence analysis studies support a close affinity
of Prochlorothrix to the cyanobacteria, analysis
of the PsbA gene product revealed that both the
Prochlorothrix and chloroplast homologs lack a
carboxyl-terminal seven amino acid domain seen
in all cyanobacterial sequences so far.29 It re-
mains to be seen what functional and evolution-
ary significance can be ascribed to these different
primary structures of the D1 protein, although it
has been noted that the appearance of Chl b is
associated with the truncated form of the poly-
~ e p t i d eAll
. ~ ~other gene sequences and their op-
eron structure reflect that Prochlorothrix falls
within the cyanobacteria; for example, compar-
47
 ison of plastocyanin sequences from Prochlo-
rothrix and other sources indicates that Proch-
forothrix is highly divergent from the chloroplast
lineage (Figure 2) .32*3s-40 Additionally, while
rbcLS are linked in cyanobacterial and Pro-
chlorothrix, in eukaryotes rbcL is a plastid gene
and the rubisco small subunit is encoded by a
nuclear multigene family .33*34 Chloroplasts also
have a conserved operon transcription unit of
psbBHpetBD , but in Prochlorothrix and cyano-
bacteria only petBD are linked in an o ~ e r o n . ~ '
However, the identification of similar putative
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
cis-acting regions upstream from the Prochlo-
rothrix petBD and psbH homologs may indicate
that these genes are coordinately expressed in this
ba~terium.~' Finally, recent sequence data avail-
r fort to identify characteristics that may distin-
P. hollandka
For personal use only.
Lr
Entemmorphaap.
Anabama sp.
Plartocyanin, prokaryotic and eukaryotic sources
FIGURE 2. Phylogenetic parsimony analysis based
on plastocyanin protein sequences from Prochlorothrix,
Anabaena, chlorophyte (Chlorella fusca and Entero-
morpha), and green plant (Spinacea oleracea, Hor-
deum vulgare, and Arabidopsis thaliana) sources.
Branch lengths reflect the number of character state
changes yielding that branch. Sequence data were re-
trieved from references cited in the
48
able on the atpBE operon, encoding the p and E
subunits of the thylakoid CF1 complex, also sup-
port the positioning of Prochloron within the
~yanobacteria.~'
All these data together indicate that the
prochlorophytes are outside of the chloroplast li-
neage, but more information on Prochlorococcus
and Prochloron is necessary to establish their
relationship to one another within the cyanobac-
terial radiation. While it has been stated that the
prochlorophytes are a diverged polyphyletic
group, some evidence has accumulated to suggest
that the antenna apoproteins of Prochloron and
Prochlorothrix are structurally similar,42and thus
the antennae share a common origin. A major
emphasis of this review is to provide insight into
the structure and physiological properties of the
prochlorophyte photosynthetic apparatus in an ef-
guish them from other phototrophs. Particular
emphasis is given to a detailed discussion of the
antennaheaction center interactions, which so far
appear to be unique among photosynthetic
systems.
IV. PHOTOSYNTHETIC APPARATUS OF
THE PROCHLOROPHYTES
The thylakoid complexes involved in elec-
tron transport have been characterized in some
detail in Prochloron, but the structure and dy-
namics of the photosynthetic mechanism are best
understood for Prochlorothrix. Such information
has been useful in providing a focus for similar
studies on Prochlorococcus, for which little is
currently known. In the following sections, the
relevant characteristics of the photosynthetic
complexes that identify these bacteria as being
functionally distinct from the green chloroplast
and phycobilisome-containing organisms are
described.
A. Composition of the Prochlorophyte
Photosynthetic Membrane
1, Photosynthetic pigments
. Presented in Table 1 is the chlorophyll and
carotenoid composition of Prochloron, Proch-
 TABLE 1
Pigment Composition of the Prochlorophytes

Organism Major carotenoid Chl a/b ratio (cells) Chl a/b ratio (antennae)

Prochloron @-Carotene 4-7 2.4

Zeaxanthin
Prochlorothrix p-Carotene 8-9 2.5
Zeaxanthin
Prochlorococcus a-Carotene 1' ?
Zeaxanthin
Chloroplasts Lutein 2-3 1.4
Violaxanthin
p-Carotene
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13

Pigments are the divinyl-chlorophyllsa and b.

Data compiled from references cited in t e ~ t . ~ - ~ ~ ' + ' ~ ~ ~ ~

lorothrix, and P ~ o c ~ ~ ~ ~ o c o c c uAss . 'from ~ ~ P* r~o c~h~l o'r ~~ n~The

mentioned earlier, Prochlorococcus is unique
among this group by the fact that it contains di-
vinyl chlorophylls. The characteristics of the di-
vinyl Chl a and b result in a slightly red-shifted
Soret absorption maximum red-shifted 8 to 10
nm; this absorbance change may be advantageous
when competing for light in the deep euphotic
For personal use only.
. ~~~~antibody
~ ~ ~ ~did
* not cross-
react with the chloroplast LHCII apoproteins, and
antibodies against LHCII failed to identify these
Prochloron and Prochlorothrix antenna apopro-
t e i n ~ . ~These
' . ~ were the first data to suggest that
the prochlorophyte antenna is structurally dissim-
ilar to that of the green chloroplast. Circular di-
chroism (CD) studies on the purified pigment-

zone of marine waters. All three prokaryotes have protein complexes also indicate that the organi-
a carotenoid composition distinct from chloro- zation of Chl b molecules is different from that
plasts of green algae or higher plants.43 in LHCII, as the negative CD band at 470 nm
ascribed to trimeric Chl b is absent in the Pro-
chlorothrix antenna.49 Moreover, unlike the sit-
2. Antenna Structures uation in higher plants, there is growing evidence
that the prochlorophyte antenna has a functional
The major Chl ulb antenna polypeptides of association with PSI; while one report has claimed
Prochloron and Prochlorothrix are hydrophobic that the antenna is largely bound to PSII parti-
intrinsic thylakoid proteins of 30 to 35 kDa.42.4547 c l e ~ a, number
~~ of papers from three separate
Pigment analysis of purified antenna complexes laboratories have reported that the bulk of the
yields a relatively high Chl a:b ratio in compar- Chl b both copurifies and is energy coupled to
ison to the major green chloroplast antenna, light- PSI in both Prochloron and Prochloro-
harvesting complex I1 (LHCII). The antenna t h r i ~ . ~Of~ course,
~ . ~ ' this does not mean that
complex of Prochloron has been shown to have these bacteria lack a PSII antenna, just that the
an a:b ratio of 2.4,46whereas a ratio of 2.5 has bulk antenna is arranged differently, allowing ef-
been reported for Prochlorothrix.48Overall, these ficient energy transfer to PSI in vivo. In Pro-
ratios yield the high a:b ratio of whole cells com- chlorothrix, the 35-kDa antenna apoprotein pro-
pared to green algae and higher plants (see Table tein is largely associated with PSI particles, while
1). The antenna proteins appear to form a small, there is some evidence that the 33-kDa antenna
homologous family of structurally related poly- protein may be associated with To sum
peptides as judged from immunological studies; up, while the precise partitioning of these antenna
an antibody prepared against the Prochlorothrix proteins in the photosynthetic membrane is cur-
30-kDa antenna apoprotein crossreacted with two rently not fully understood, it is clear that the
additional Prochlorothrix proteins of 33 and 35 overall arrangement of the Prochlorothrix an-
kDa, and to the major 34-kDa antenna apoprotein tenna clearly is quite different from that in green

49
 chloroplasts. How this organization might con-
tribute to the phototrophic metabolism of the
prochlorophytes is addressed later in this section.
Freeze-etch electron microscopy of Pro-
chlorothrix and Prochloron membranes demon-
strates that there is some degree of partitioning
between PSI and PSII centers in the thylakoid
m e m b ~ a n e . ~ This
’ . ~ ~ feature, termed lateral het-
erogeneity, is a widespread feature of chloroplast
systems in which the PSII centers are located in
the stacked, p a l membranes, while PSI is found
in the unstacked, stromal thylakoids. A notable
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
observation from these studies is that the particles
of the Prochloron and Prochlorothrix EF, frac-
ture face, which have been described as PSII plus
antenna particles, are -30% smaller than those
seen in green c h l o r o p l a s t ~ Transmission
.~~~~~
electron microscopy of Prochlorothrix and
Prochloron membranes revealed that while some
degree of membrane appression is evident, the
degree of stacking is less widespread than in
chloroplast t h y l a k o i d ~ . ~ * * , ~ ~ . ~
The antibody to the 30-kDa Prochlorothrix
antenna protein was used to retrieve immuno-
For personal use only.
positive lambda ZAP clones encoding fragments
of the antenna polypeptides .55 Sequence analysis
also indicated that the antenna proteins are struc-
turally dissimilar from the LHC proteins (Cab
polypeptides) of chloroplast systems; currently,
it is unclear whether the antenna proteins have
evolved independently from the LHC polypep-
tides encoded by the plant and algal Cub multi-
gene family.56This suggests once again that the
structure and overall organization of the light-
harvesting apparatus in both Prochforon and
Prochlorothrix is fundamentally different from
that in chloroplasts.
3. Reaction Centers and Electron
Carriers
PSII and PSI particles that exhibit photo-
chemical activity have been prepared from
Prochloron and Prochlorothrix; immunological
analysis of these particles and whole thylakoid
membranes demonstrates the presence of the ho-
mologous reaction center polypeptides also found
in cyanobacteria and chloroplasts. For example,
PSI particles from Prochloron and Prochloroth-
50
rix yield a polypeptide composition very similar
to PSI preparations from higher plants or cyano-
b a ~ t e r i a . ~The
~ . ~composition
’ of PSII particles is
somewhat less clear, but the major components
of the PSII complex have been identified. These
include the Chl a-binding polypeptides CP47 and
CP43;45 the reaction center proteins D1 and
D2;29,58the 10-kDa psbH prqtein;” and the 33-
kDa PsbO protein involved in stabilizing the do-
nor (water-splitting) side of PSII.59 In addition,
homologs of the small, functionally enigmatic
hydrophobic PSII proteins have been found in P .
hollundica; this includes the so-called PsbJ and
PsbK proteins, whose presence in both cyano-
bacteria and chloroplasts has been well
documented. 58.60
While no active preparations of the cyto-
chrome bdf complex from prochlorophytes have
been reported, analysis of whole membranes by
difference spectroscopy and tetramethylbenzi-
dine staining have identified both cytochromes f
and b, in both Prochloron and Prochf~rothrix.~~
Furthermore, the petBD genes encoding apocy-
tochrome b, and subunit 4 have been cloned and
~ e q u e n c e d .Finally,
~~ examination of soluble
fractions from Prochlorothrix has shown that the
primary electron donor to PSI is pla~tocyanin;~~
this is in contrast to an earlier paper that stated
that a cytochrome cSsJserves this function in
Prochlorothrix.6’In summary, virtually all of the
components known to be associated with whole
chain electron transport in oxygenic photosyn-
thesis have been identified in either Prochloron
or Prochlorothrix, thus it is reasonable to assume
that the overall protein composition of such com-
plexes is not substantially different from those
reported for cyanobacteria and chloroplast
systems.
4. Modes of Photosynthesis
With the presence of two photosystems and
all electron carriers known from oxygenic pho-
tosynthesis in cyanobacteria and chloroplasts, it
is hardly surprising that photosynthetic oxygen
evolution and carbon fixation do take place in
the prochlorophytes at rates capable of supporting
growth. However, the firm positioning of
prochlorophytes within the cyanobacterial radia-
 tion has prompted the search for anoxygenic pho-
tosynthesis as an alternative mode of photosyn-
thetic energy conservation in Prochlorothrix.
Anoxygenic photosynthesis is found among fila-
mentous ~yanobacteria~) and involves a dual role
for sulfide: as an inhibitor of oxygen evolution
at the donor side of PS I1 and as an electron donor
to the plastoquinone pool .62-67 Prochlorothrix was
shown to have a constitutive but low capacity of
anoxygenic photosynthesis at approximately 5%
of light-saturated oxygenic photosynthesis.68This
capacity was accompanied by a high resistance
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
of the oxygen-evolving complex of Prochforolh-
rix against sulfide stress. These data suggest that
Prochlorothrix may have evolved from a bacter-
ium adapted to reduced environments and regular
periods of anoxic conditions. Green chloroplasts
are in general very sensitive to sulfide inhibition,
and anoxygenic photosynthesis has not been found
in these systems.
B. Dynamics of the Photosynthetic
Apparatus
For personal use only.
1. Regulation of Energy Distribution by
Phosphorylation
While the structure and composition of the
Prochloron and Prochforothrix photosynthetic
apparatus are becoming well understood, more
recent studies have focused on the adaptive re-
sponses of the light reactions in response to
changes in environmental light conditions. In both
the chloroplast and cyanobacterial systems,
mechanisms exist to regulate energy imbalances
between PSII and PSI. This phenomenon, termed
the state 1>2 transition, acts to maximize elec-
tron transport should the wavelength of the am-
bient light favor absorption by one photosystem
over the other. In a general sense, this is achieved
by changes in antenna orientation such that en-
ergy transfer is preferentially redirected toward
the photosystem that less efficiently absorbs the
ambient light. For example, in low light and in
far-red light preferentially absorbed by PSI, en-
ergy transfer favors energy trapping by PSII. This
is termed state I . In a state 2 situation, in satu-
rating light or in red light preferentially absorbed
by PSII, energy is redistributed to PSI. Such a
transition is defined by changes in the PSII flu-
orescence yield; state 1 is the high fluorescence
condition, while adaptation to state 2 results in
a marked decrease in PSII fluorescence. This
phenomenon occurs in both green chloroplast and
cyanobacterial systems, but the mechanisms
driving this adaptation are different for each. In
green chloroplasts, the major antenna LHCII is
reversibly phosphorylated in a light-dependent
fashion by a thylakoid kinase that is activated by
sensing the redox state of electron transport in-
termediates between PSII and When PSII
is overexcited relative to PSI, the kinase is ac-
tivated, and it phosphorylates a subset of the an-
tenna. The phosphorylated antenna then decou-
ples from PSII to balance energy flow between
PSI and PSII.69*70 Accompanying these events is
the destacking of the thyIakoid granal membranes
as a consequence of the movement of phosphoryl-
ated LHCII away from PSII center^.^^,^^ In the
phycobilisome-containing organisms (cyanobac-
teria and red algae), state transitions do not occur
by this mechanism; instead reorganization of the
photosynthetic apparatus occurs such that energy
transfer is enabled by spillover from the Chl a
PSII core to PSI.74.75
More recently, emerging studies have doc-
umented the existence of state transitions in P.
' * ~ ~ an initial study could not
h o l l a n d i ~ a . ~While
correlate changes in redox state with antenna pro-
tein pho~phorylation,~~ additional work has shown
that the 35-kDa Chl ulb antenna protein is re-
versibly phosphorylated in a light-dependent
manner both in vivo and in vitro by a thylakoid-
bound protein k i n a ~ e . ~White* light at moderate
to saturating intensity and PSII light (650 nm)
yielded enhanced phosphorylation at the same
timescale (-30 min) as a state 1>2 transition,
as judged by the decrease in PSII fluorescence
~ i e l d . ~ Inhibitors
**~' of phosphatases also yielded
a highly phosphorylated antenna, and such cells
were locked in state 2 in v ~ v o . ~ '
Moreover, mon-
itoring PSI function in vivo by measuring non-
photochemical fluorescence quenching (an indi-
rect measure of cyclic electron flow) indicates
that a transition to state 2 is accompanied by an
increase in PSI a ~ t i v i t y .All
~ ' these data together
suggest the involvement of protein phosphoryla-
tion in regulating energy distribution between the
photosystems; thus this reversible light-depen-
51
 dent mechanism is one feature that demonstrates
some similarity to what has been observed in
chloroplast systems, and has probably been con-
served through the evolution of oxygenic
phototrophs .48*s'
Examining the antenna complexes of Pro-
chloron revealed that the major 34-kDa Chl alb
polypeptide is highly phosphorylated irrespective
of light treatment.47Additionally, state 1>2 tran-
sitions were not detected; this observation led to
the proposal that the cells were locked in a state
2 situation in response to the abundance of PSII
light in their natural symbiotic habitat.47Because
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
these experiments were performed in isolated
Prochloron membranes and cells, it is unknown
whether reversible phosphorylation can occur in
Prochloron while maintained in their native sym-
biotic lifestyle.
2. Antenna-Reaction Center Interactions
The observation that state 1>2 transitions
occur in Prochlorothrix led to additional studies
For personal use only.
aimed at understanding short-term changes in an-
tenna size and function in response to light. From
analysis of oxygen evolution rates in response to
changes in irradiance, the PSII antenna size was
calculated and shown to be -40% larger than in
cells adapted to high light and subsaturating light
intensities.s' Inasmuch as state transitions have
not been reported for Prochlorococcus and
Prochloron, it remains to be seen whether such
dynamic short-term responses occur in other
prochlorophytes. However, as described previ-
ously, there is a phosphorylation mechanism act-
ing on the Prochloron antenna47and thus it is
possible that state transitions play a role in an-
tenna function in hospite.
As mentioned briefly earlier, evidence has
accumulated indicating that the bulk of the an-
tenna is functionally bound to PSI.4648"' This
differs considerably from the situation in green
chloroplasts and cyanobacteria, where the anten-
nae are largely PSII-associated. The bases for this
evidence take into consideration both biophysical
and biochemical data. For example, low tem-
perature fluorescence emission studies of Pro-
chlorothrix cells indicate that PSI fluorescence
is enhanced when cells are excited by wave-
52
lengths absorbed by Chl b.sl This demonstrates
efficient coupling of a Chl b-containing antenna
to the PSI reaction center. Due to the bulk of the
Chl alb antenna associated with PSI, fluores-
cence induction measurements of Prochlorothrix
cells revealed efficient fluorescence quenching
by PSI at room tempe~ature.~' The functional
partitioning of the major Chl alb antenna protein
in PSI particle preparations also has been dem-
onstrated for both Prochlorothrix and Prochlo-
ron, and fluorescence excitation spectra of these
preparations show efficient energy transfer to the
PSI reaction enter.^^.^,^^.^ *
3. Ljgh &Shade Adaptation
Additional studies have examined the long-
term effects of different environmental light con-
ditions on Prochlorothrix and Prochloron. Com-
parison of Prochlorothrix and Prochloron cul-
tures grown at limiting vs. saturating intensities
for photosynthesis revealed that the low light-
grown cells had an increased Chl content of five-
and twofold, r e ~ p e c t i v e l y . ~ ~This
, ' ~ . ~increase
~
correlated with an increase in the photosynthetic
unit size of PSII; this value is calculated as the
number of Chl molecules per molecule of O2
evolved. In addition to changes in pigment con-
tent, long-term adaptation of Prochlorofhrir to
low light resulted in a 65% increase in the ratio
of PSI to PSII centers,62while in Prochloron,
the ratio of reaction centers remained constant.77
Changes in reaction center ratios are a common
adaptation in cyanobacteria to changes in photon
fluxes, while in chloroplasts, the PSVPSII ratio
is less variable in response to environmental
changes.79In both organisms, the rate of linear
electron transport appears to remain constant un-
der the different light regimes .62.78
Current work on Prochlorococcus has fo-
cused on the growth characteristics of the dif-
ferent marine isolates. Strain SS 120, originating
from deep water in the Sargasso Sea, yields an
optimal growth rate of 0.55 generations per day;
light saturating for photosynthesis is achieved at
40 kmol quantalm2/s.80While this strain is well
adapted for growth in very low light (3 pmol
quantalm*/s), supersaturating light yielding pho-
toinhibitory photodamage to PSII occurs at 70
 pmol quanta/m2/s.80By comparison, the strain
MED4, taken from near the surface in the Med-
iterranean, yields similar optimal growth rates
but was clearly better adapted to higher light en-
vironments; photosynthesis saturates at 60 kmol
quanta/m2/s, while the minimum light supporting
growth is approximately 16 pmol quanta/mz/s.80
MED4 is also comparatively resistant to photo-
inhibition. 8o Future work aimed at understanding
antenna function and dynamics in Prochlorococ-
cus should identify the physiological bases for
the adaptation to surface vs. deep water light
environments.
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
An additional response seen in Prochloroth-
rix during long-term adaptation to supersaturat-
ing light is the accumulation of zeaxanthin pig-
ments at the cell surface.14 This results in the
selective screening of wavelengths in the blue
region of the spectrum. The zeaxanthin is bound
to a protein complex that appears to coat the cell
surface, thus this complex has been proposed to
represent an S (surface)-layer that assembles un-
der light stress. Cultures shifted to high light
exhibit a 2.3-fold increase in the level of this
For personal use only.
protein within 24 h,I4 and RNA blot analysis
indicates that the mRNA for this complex is un-
der transcriptional control by light." Overall, the
accumulation of this carotenoid-protein layer may
account for much of the 40% decrease in the
quantum efficiency for O2evolution in high light-
adapted cells.62
4. Model for the Photosynthetic
Apparatus of Prochlorophytes
Studies in recent years have established that
the organization of the photosynthetic membrane
in both Prochloron and Prochlorothrix is some-
what different from that in cyanobacterial and
chloroplast systems. While many parallels can be
drawn, particularly with respect to the structure
and composition of PSI and PSII reaction centers,
the association of the reaction centers within the
antennae demands that a new model be proposed.
Taking into consideration that there is lateral het-
erogeneity of PSI and PSII centers in the mem-
brane, and that the antenna can be reversibly
phosphorylated and associated with PSI, we rec-
oncile these facts with the model proposed in
Figure 3. In state 1, both PSI and PSII share a
common light-harvesting antenna composed
largely of the 35-kDa Chl alb antenna protein in
its dephosphorylated state. This condition should
exist both in the dark and upon treatment with
fk-red (PSI) light. Transition to state 2 in both
high light and red light results in the phosphor-
ylation of the 35-kDa protein and the decoupling
of the bulk antenna from PSII. This results in the
exclusion of PSII centers from the major light-
harvesting apparatus in the cell. It is this situation
that, we argue, results in the lateral heterogeneity
seen in freeze-etch electron micrographs. As
mentioned previously, the PSII EF, particles seen
following freeze-etch are smaller than those in
chloroplasts, consistent with the model stating
that the PSII centers should lack a large antenna
complex. One possible flaw in the model is that
it does not take into account the fact that thyla-
koid membrane stacking occurs in both Pro-
chloron and Prochlorothrh. However, it is un-
clear what molecular mechanism(s) might drive
stacking in the first place, and what physiological
role stacking may play in the overall functioning
of the photosynthetic mechanism.*' While ini-
tially it was believed that stacking is mediated
largely by the chloroplast PSII antenna,73Aru-
bidopsis mutants lacking chloroplast LHCII are
state 1 (dark. far md:
energy to PSI
P and II)
state 2 (high light, red:
energy to PSI)
FIGURE 3. Model for state transitions in Prochlor-
othrix. In state 1, both PSI1 and PSI centers share a
common antenna, composed largely of the 35-kDa an-
tenna polypeptide. Transition to state 2 in red (PSII) or
in saturating light results in phosphorylation of the 35-
kDa protein and exclusion of PSI1 centers from the bulk
antenna. The 30- and 33-kDa proteins are Chl a/b an-
tenna apoproteins, which are thought to be associated
with PSI and PSII, respectively.
53
 capable of exhibiting thylakoid’stacking.82Thus,
it is more likely that the membrane stacks form
primarily from interaction of the PSII centers
themselves and not through the direct agency of
the LHCII antennae complexes. Nonetheless, it
is probable that chloroplast LHCII contributes to
the stabilization of thylakoid yielding
the extensive grana seen in green plants. From
the limited data available, we would propose that
the degree of stacking and lateral heterogeneity
in Prochloron and Prochlorothrix arises from in-
teractions among the PSII centers excluded from
the bulk antenndPS1 complexes, thus resulting
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
in increased stacking in a state 2 situation (see
Figure 3). This is opposite to what is known to
occur in chloroplasts, in which state 1 promotes
s t a ~ k i n g . ~The
~ . overall
~ ~ . ~ lower
~ degree of stack-
ing in prochlorophytes can be attributed both to
the lack of an LHCII complex to stabilize mem-
brane appression, and an overall high PSUPSII
ratio in comparison to ~ h l o r o p l a s t s .Of
~ ~course,
.~~
this model is speculative, but it is testable and
will help address directly the structural and me-
chanistic requirements necessary to drive thyla-
For personal use only.
koid stacking. This hypothesis also would predict
that the absence of stacking in cyanobacteria re-
sults from steric hindrance of membrane appres-
sion by the extrinsic phycobilisome complexes.
The positioning of the phycobilisome also would
act to interfere with any direct interactions among
PSII centers, which also may be involved in the
stacking process.
Finally, our model for antenna organization
should not be interpreted to mean that these or-
ganisms lack a PSII antenna, rather that the an-
tenna is organized such that energy transfer to
PSI is favored due to a physical association of
the antenna to PSI in state 2. A functional as-
sociation of the antenna to both PSI and PSII
occurs in state 1, resulting in an increased PSII
antenna size. Moreover, recent evidence suggests
that a 33-kDa antenna protein (see Figure 3) ex-
clusively copurifies with PSII.48.50 Thus, we rec-
ognize that a PSII antenna exists, but it is not
the extensive array of PSII pigment-proteins seen
in green chloroplasts and phycobilisome-contain-
ing organisms.
54
V. DISCUSSION
A. What Is the Prochlorophyte Antenna?
The discovery of Prochloron and Prochlo-
rothrix has led to several studies aimed at un-
derstanding the structure and dynamics of the
photosynthetic apparatus. Such work has pro-
vided a great deal of information on the function
of the antennae complexes and their relationship
to the reaction centers. One observation we have
noted from the accumulated data is the apparent
lack of a chloroplast LHCII-like complex in these
bacteria. In contrast, the major antenna polypep-
tides appear to have a strong functional coupling
to PSI, and the characterization of the isolated
pigment-proteins suggests profound differences
between LHC and the prochlorophyte antennae.
While one report has claimed that the antenna
can copurify with PSII,50there are no convincing
data supporting efficient energy transfer to PSII
centers, and studies of antenna function in vivo
demonstrate energy coupling to PSI centers. These
structural and functional differences are partic-
ularly noteworthy as several papers have inde-
pendently suggested that Chl b antennae evolved
more than In light of all these data, we
would propose that the term “LHC,” which has
been used indiscriminately to denote a Chl alb
antenna, be reserved for antennae types seen in
chloroplast systems only. To resolve beyond all
doubt whether the prochlorophyte antenna is ev-
olutionarily distinct or divergent from the LHCII
antennae will require more detailed sequencing
studies examining the primary structure of the
Prochloron, Prochlorothrix, and Prochlorococ-
cus antenna apoproteins.
B. Why An Antenna Having Efficient
Coupling to PSI?
The most unusual feature of Prochloron and
Prochlorothrix is the functional association of the
antenna to PSI. All other cyanobacteria and chlo-
roplast systems are arranged such that the major
antenna complex is tightly coupled to PSII. This
 arrangement evidently results from a strong de-
pendence on linear electron transport to contrib-
ute both to a protonmotive force and an accu-
mulation of reducing equivalents. Such a system
would thus be less dependent on cyclic photo-
phosphorylation around PSI, which generates
protonmotive force only. The antenna organi-
zation of Prochloron and Prochlorothrix appears
to yield a situation in which energy transfer to
PSI is favored, thus favoring cyclic electron flow
over linear. In order for this to make good bio-
logical sense, the demands for reducing equiv-
alents must be met by other means. Thus, we are
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
suggesting that Prochlorothrix and other pro-
chlorophytes may have evolved from a bacterium
exhibiting a mixotrophic lifestyle, in which ox-
idation of reduced substrates is coupled to
NAD(P)H accumulation. Consequently, ATP de-
mands could be met by PSI activity, and pools
of reducing power could be maintained by both
PSII and oxidative pathways. Inasmuch as
Prochlorothrix is found in shallow, eutrophic
lakes containing relatively high concentrations of
organic material, it also is possible that reduced
For personal use only.
carbon is capable of serving as an alternative
electron donor. Studying the interactions of
Prochlorothrix with reduced substrates should be
an important priority in determining whether there
is any dependence on mixotrophic or photohet-
erotrophic modes of growth. This is of particular
importance when considering that Prochlorothrix
can exhibit low rates of anoxygenic photosyn-
thesis in the presence of sulfide;68 perhaps this
antenna organization is better adapted for such
alternative modes of electron transport.
C. Evolution of Major Antenna Systems
A fundamental question that remains con-
cerns the relationship of the prochlorophyte an-
tenna to other antenna types seen in chloroplasts
and cyanobacteria. Current phylogenetic data
suggest that Chl b evolved more than once, but
whether it has multiple origins within the pro-
chlorophytes is a matter of debate. An argument
in favor of a monophyletic origin for the proch-
lorophyte antenna comes from work demonstrat-
ing immunological. relationships between the
Prochloron and Prochlorothrix antenna apopro-
t e i n ~ and
, ~ ~ their functional associations with
The presence of the prochlorophytes
within the cyanobacterial lineage also suggests
that both the ProchlorothrixlProchloron anten-
nae and phycobilisomes are antenna types that
could have arisen from a common ancestral pho-
totroph. Such an ancestor would have both a Chl
alb PSI antenna and a biliprotein PSII antenna.
While no such organism has been described so
far, it is consistent with the observation that both
these antennae have ancient origins within a pro-
karyotic lineage. Organisms having either only
phycobilisomes or only Chl alb antennae arose
by losing one of the two antenna types. The big-
gest problem with this proposal is that it is dif-
ficult to imagine what selective pressures could
result in the loss of the phycobilisome, an ex-
tremely efficient antenna and hence an ideal ad-
aptation for light-limited environments. 84 As al-
luded to previously, an antenna coupled to PSI
would be favored in an environment with reduced
substrates, thus favoring photoheterotrophy and
PSI-mediated cyclic electron flow. Under such
conditions, light-harvesting by PSII centers could
possibly become dispensable. However, PSII
centers are retained in the bulk chlorophyll an-
tenna, as autotrophy would be selected for in
environments where reduced substrates become
transiently scarce.
D. Prochlorophytes and Plastid
Evolution
To date, the sequence data have prompted
the theory that Chl b arose multiple times during
the evolution of phototrophs. Based on the lim-
ited sequence data and immunological studies on
the Chl alb antenna apoproteins, we believe that
the antenna of Prochlorothrix is structurally dif-
ferent from LHCII and has a separate evolution-
ary origin. However, although no data are avail-
able on the Prochlorococcus antenna, the Pro-
chloron and Prochlorothrix antennae appear to
share a common origin. Thus, it is quite certain
that the prochlorophytes are not the progenitors
of green plastids, and the true ancestor to these
chloroplasts, having an LHCII-like antenna, is
as yet unknown.
Evidence is accumulating to suggest that the
red algal and green chloroplasts have a polyphy-
55
 letic origin, while addibonal proposals have been
raised regarding horizontal gene transfer in play-
ing a role in the evolution of novel prokaryotic
species and chloroplast types.3335 Furthermore,
a problem remains in explaining the overall high
AT content of organellar DNA, while photosyn-
thetic prokaryotes are, by comparison, relatively
GC-rich. It has been argued that the AT bias arose
following endosymbiosis, but this debate is
unre~olved.~~
VI. CONCLUDING REMARKS
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
The question posed in the title of this review
takes into consideration the fact that the taxon-
omic studies tell us that the prochlorophytes are
cyanobacteria, but physiological studies indicate
that their photosynthetic apparatus is organized
in a unique way. The evolutionary origin(s) of
the prochlorophyte antennae are still unknown,
but the special features of the Prochforon and
Prochlorothrix photosynthetic mechanism may
prove useful in understanding general rules for
For personal use only.
how photosynthetic organisms function and re-
spond to changes in environmental light and nu-
trient conditions. Thus, we argue that under-
standing the photosynthetic features of these pro-
karyotes is necessary to understand fully both
their evolution as well as their ability to colonize
specialized marine and aquatic environments.
These organisms have several special properties
that demand further study.
Much work remains to be done regarding the
prochlorophytes. Our major interest has been to
examine the physiological properties of the pho-
tosynthetic apparatus in an effort to understand
how this different antenna system functions, and
thus how it contributes to the growth and metab-
olism of the cell. Two areas need,to be addressed
in the near future. First, the complete sequence
of the Prochlorothrix Chl alb-binding antenna
apoproteins should be obtained and analyzed for
any domains that may cooperate in Chl binding.
This study is essential for establishing the degree
of divergence from the antenna apoproteins of
green chloroplasts. Second, a detailed study on
the structure and function of the Prochforococcus
antenna is necessary to prove that it is homolo-
gous to the Prochforon and Prochlorothrix an-
56
tennae. Hampering such studies is the difficulty
in growing Prochforococcus to high density in
batch culture; future biochemical work will de-
pend on improving the culture conditions so this
can be achieved. Additional important studies
include examining the molecular basis for the
anoxygenic, sulfide-resistant photosynthesis;
perhaps such work can give clues toward under-
standing the functional significance of a PSUan-
tenna complex.
ACKNOWLEDGMENTS
The authors thank Drs. Karen Milbauer-
Warner, Kenneth Miller, Hewson Swift, and
Sallie Chisholm for the electron micrographs that
appear in this article. Dr. Susan Golden and Lisa
W.-J. Moore provided work prior to publication
and helpful discussions. The work compiled from
the authors’ laboratories was supported by grants
DCB-8903060 and MCB-9204672 from the Na-
tional Science Foundation (to G.S.B.) and award
88-00341 from the U.S./Israel Binational Sci-
ence Foundation (to A.F.P.).
REFERENCES
1. Lewin, R. A., A marine Synechocystis (Cyanophyta,
Chrwoccales) epizoic on ascidians, Phycologia, 261,
153, 1975.
2. Lewin, R. A., Prochlorophyta as a proposed new
division of algae, Nature, 261, 697, 1976.
3. Burger-Wiersma, T., Veenhuis, M., Korthals,
H. J., Van Der Wiel, C. C. M., and Mur, L. R.,
A new prokaryote containing chlorophylls a and b,
Nature, 320, 262, 1986.
4. Burger-Wiersma, T., Stal, L. J., and Mur, L. R.,
Prochlorothrix hollandica gen. nov., spec. nov.: a
filamentous oxygenic photoautotrophic prokaryote
containing chlorophylls a and b, Int. 1. Sysr. Bac-
teriol., 39, 250, 1989.
5. Gieskes, W. W. and Kraay, G. W., Unknown chlo-
rophyll u derivatives in the North Sea and the tropical
Atlantic Ocean revealed by HPLC analysis, Limnol.
Oceanogr., 28, 757, 1983.
6. Veldhuis, M. J. W. and Kraay, G. W., Vertical
distribution and pigment composition of a picoplank-
tonic prochlorophyte in the subtropical North Atlan-
tic: a combined study of HPLC-analyis of pigments
and flow cytometry, Mar. Ecol. Prog. Ser., 68, 121,
1990.
 7. Chisholm, S. W., Olson, R. J., Zettler, E. R.,
Goericke, R., Waterbury, J. B., and Welschmeyer,
N. A., A novel free-living prochlorophyte abundant
in the oceanic euphotic zone, Nature, 334,340,1988.
8. Chisholm, S. W., Frankel, S. L., Goericke, R.,
Olson, R. J., Palenik, B., Waterbury, J. B., West-
Johnsrud, L., and Zettler, E. R., Prochlorococcus
marinus nov. gen. nov. sp.: an oxyphototrophic ma-
rine prokaryote containing divinyl chlorophyll a and
b, Arch. Microbiol.. 157, 297, 1992.
9. Lewin, R. A. and Cheng, L., Collection and han-
dling of Prochloron and its ascidian hosts, in Proch-
loron, Microbial Enigma, Lewin, R. A. and Cheng,
L., Eds., Chapman and Hall, New York, 1989, 9.
10. Thorne, S.W., Newcomb, E. H.,and Osmond,
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
C. B., Identification of chlorophyll b in extracts of
prokayotic algae by fluorescence spectroscopy, Proc.
Natl. Acad. Sci. U.S.A., 74, 575, 1977.
11. Schulz-Baldes, M. and Lewin, R. A., Fine structure
of Synechocystis didemnii (Cyanophyta: Chroococ-
cales), Phycologia, 15, 1, 1976.
12. Swift, H. and Leser, G., Cytochemical studies on
prochlomphytes: localization of DNA and ribulose
1,s bisphosphate carboxylaseloxygenase, J . Phycol.,
25, 751, 1989.
13. Golecki, J. R. and Jiirgens, U. J., Ultrastructural
studies on the membrane systems and cell inclusions
of the filamentous prochlorophyte Prochlorothrix
For personal use only.
hollandica, Arch. Microbiol., 152, 77, 1989.
14. Engle, J. M., Burkhart, W., Sherman, D. M.,
and Bullerjahn, G. S., Purification and character-
ization of a surface-associated carotenoid-binding
complex from the photosynthetic prokaroyote,
Prochlorothrix hollandica, Arch. Microbiol., 155,
453, 1991.
15. Omata, T., Okada, M., and Murata, N., Sepa-
ration and partial characterization of membranes from
Prochloron sp., Plant Cell Physiol., 26, 579, 1983.
16. Moriarty, D. J. W., Muramic acid in the cell walls
of Prochloron, Arch. Microbiol., 120, 191, 1979.
17. Stackebrandt, E. and Kandler, O., The murein
type of Prochloron, Zentralbl. Bakteriol. Hyg.. I.
Abt. Orig., 3 , 354, 1982.
18. Jhrgens, U. J. and Burger-Wiersma, T., Peptid-
oglycan-polysaccharide complex in the cell wall of
the filamentous prochlorophyte Prochlorophyie hol-
landica. J. Bacteriol., 171, 498, 1989.
19. Perry, G. J., Gillan, F. T., and Johns, R. B.,
Lipid composition of a prochlorophyte, J. Phycol.,
14, 369, 1978.
20. Johns, R. B., Nichols, P. D.,Gillan, F. T., Perry,
G. J., and Volkman, J. K., Lipid composition of
a symbiotic prochlorophyte relation to its host, Camp.
Biochem. Physiol.. 69B, 843, 1981.
21. Murata, N. and Satoh, N., Analysis of lipids in
Prochloron sp.: occurrence of monoglucosyl diacyl-
glycerol, Plant Cell Physiol., 24, 133, 1983.
22. Volkman, J. K., Burger-Wiersma, T., Nichols,
P. D.,and Summons, R. E., Lipids and chemotax-
onomy of Prochlorothrix hollandica, a planktonic
prokaryote containing chlorophylls a and 6,J . Phy-
col., 24, 554, 1988.
23. Herdman, M., Deoxyribonucleic acid base com-
position and genome size of Prochloron, Arch. Mi-
crobial., 129, 314, 1981.
. 24. Engle, J. M. and Bullerjahn, G. S., unpublished
data, 1991.
25. McKay, R. M., Salgado, D., Bonen, I.,
Stackebrandt, E., and Doolittle, W. F., The 5s
ribosomal RNAs of Paracoccus denitrificans and
Prochloron, Nucleic Acids Res., 10, 2963, 1982.
26. Turner, S., Burger-Wiersma, T., Giovannoni,
S. J., Mur, L. R., and Pace, N. R., The relation-
ship of a prochlorophyte Prochlorothrix hollandica
to green chloroplasts, Narure, 337, 380, 1988.
27. Palenik, B. and Haselkom, R., Multiple evolu-
tionary origins of prochlorophytes, the chlorophyll 6
containing prokaryotes, Nature, 355, 265, 1992.
28. Urbach, E., Robertson, D. L., and Chisholm,
S. W., Multiple evolutionary origins of prochloro-
phytes within the cyanobacterial radiation, Nature,
335, 267, 1992.
29. Morden, C. W. and Golden, S. S., psbA genes
indicate common ancestry of prochlorophytes and
chloroplasts, Nature, 337, 382, 1988.
30. Greer, K. L. and Golden, S. S., Nucleotide se-
quence of psbB from Prochlorothrix hollandica, Plant
Mol. Biol., 17, 915, 1991.
31. Greer, K. L. and Golden, S. S., Conserved rela-
tionship between psbH and petBD genes: presence of
a shared upstream element in Prochlorothrix hoilan-
dica, Plant Mol. Biol., 19, 355, 1992.
32. Seeburg, D. and Bullerjahn, G. S., unpublished
data, 1992.
33. Morden, C. W. and Golden, S. S., Sequence anal-
ysis and phylogenetic reconstruction of the genes en-
coding the large and small subunits ribulose-I ,5-bis-
phosphate carboxylase/oxygenase from the chloro-
phyll alb-containing prokaryote, Prochlorothrix hol-
landica. J . Mol. Evol., 32, 379, 1991.
34. Golden, S. S., Morden, C. W., and Greer, K. L.,
Comparison of sequences and organization of pho-
tosynthesis genes among the prochlorophyte Pro-
chlorothrix hollandica, cyanobacteria and chloro-
plasts, in Symbiogenesis, Prochlorophytes and the
Origins of Plastids, Lewin, R. A., Ed., Chapman
and Hall, New York, 1992, in press.
35. Aitken, A., Prokaryote-eukaryote relationships and
the amino-acid sequence of plastocyanin from Ana-
baena variabilis, Biochem. J . , 149, 657, 1975.
36. Kelly, J. and Ambler, R. P., The amino acid se-
quence of plastocyanin from Chlorellafusca,Biochem.
J., 143, 681, 1974.
37. Simpson, R. J., Moritz, R. L., Nice, E. C., Grego,
B., Yoshizaki, F., Sugimura, Y., Freeman, H.C.,
and Murata, M.,Complete amino-acid sequence of
platocyanin from a green alga, Enteromorpha pro-
lifera. Eur. J . Biochem., 137, 497, 1986.
57
 38. Nielsen, P. S. and Gausing, K., The precursor of
barley plastocyanin: sequence of cDNA clones and
gene expression in different tissues, FEES Lett., 225,
159, 1987.
39. Rother, C., Jansen, T., Tyagi, A., Tittgen, J.,
and Herrmann, R. G., Plastocyanin is encoded by
an uninterrupted nuclear gene in spinach, Curr. Ge-
net.. 11, 171, 1986.
40. Vorst, O., Oosterhoff-Teertstra, R., Vankan, P.,
Smeekens, S., and Weisbeek, P., Plastocyanin of
Arabidopsis thaliana; isolation and characterization
of the gene and chloroplast import of the precursor
protein, Gene, 65, 59, 1988.
41. Lockhart, P. J., Beanland, T. J., Howe, C. J.,
and Larkum, A. W. D., Sequence of Prochloron
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
didemni arpBE and the influence of chloroplast origins,
Proc. Natl. Acad. Sci. U.S.A.. 89, 2742, 1992.
42. Bullerjahn, G. S., Jensen, T. C., Sherman, D. M.,
and Sherman, L. A., Immunological characteriza-
tion of the Prochlorothrix hollandica and Prochloron
sp. chlorophyll alb antenna proteins, FEMS Micro-
biol. Lert., 67, 99, 1990.
43. Powls, R. and Britton, G., A series of mutant strains
of Scenedesmus obliquus with abnormal carotenoid
compositions, Arch. Microbiol., 113, 275, 1977.
44. Withers, N., Alberte, R. S., Lewin, R. A.,
Thornber, J. P., Britton, G., and Goodwin, T. W.,
Photosynthetic unit size, carotenoids , and chloro-
For personal use only.
phyll-protein composition of Prochloron sp., a pro-
karyotic green alga, Proc. Narl. Acad. Sci. U.S.A..
75, 2301, 1978.
45 Bullerjahn, G. S., Matthijs, H. C. P., Mur,
I 58. Krugh, B. and Bullerjahn, G. S., unpublished data,
L. R.,and Sherman, L. A., Chlorophyll-protein
composition of the thylakoid membrane from Pro-
chlorothrix hollandica, a prokaryote containing chlo-
rophyll b, Eur. J. Biochem., 168, 295, 1987.
46. Hiller, R. G. and Larkum, A. D. W., The chlo-
rophyll-protein complexes of Prochloron sp. (Pro-
chlorophyta), Biochim. Biophys. Acta, 806, 107,
1985.
47, Schuster, G., Owens, G. C., Cohen, Y., and
Ohad, I., Thylakoid polypeptide compositions and
light independent phosphorylation of the chlorophyll
alb protein in Prochloron, a prokaryote exhibiting
oxygenic photosynthesis, Biochim. Biophys. Acta,
767, 596, 1984.
48, Post, A. F., Gal, A., Ohad, I., Milbauer, K. M.,
and Bullerjahn, G. S., Characterization of light-
activated reversible phosphorylation of a chlorophyll
alb antenna apoprotein in the photosynthetic pro-
karyote. Prochlororhrix hollandica, Biochim. Bio-
phys. Acta, 1100, 75, 1992.
49. Matthijs, H.C. P., van der Staay, G. W. M., van
Amerongen, H.,van Grondell, R., and Garab, G.,
- Structural organization of chlorophyll b in the pro-
chlorophyte Prochlorothrix hollandica, Biochim.
Biophys. Acta, 975, 185, 1989.
50. Van der Staay, G. W. M., Brouwer, A., Baard,
R. L., Van Mourik, F., and Matthijs, H. C. P.,
Separation of Photosystems I and I1 from the oxy-
58
chlorobacterium (prochlorophyte) Prochlorothrix
hollandica and association of chlorophyll b binding
antennae with Photosystem 11, Biochim. Biophys.
Acta, 1102, 220, 1992.
51 Post, A. F., Cohen, I., Milbauer-Warner, K. M.,
and Bullerjahn, G. S., Regulation of excitation en-
ergy distribution in Prochlorothrir hollandica: in-
volvement of a chlorophyll alb antenna associated
with PSI, Biochim. Biophys. Acra. in press.
52 Giddings, T. H.,Withers, N. W., and Staehelin,
L. A., Supramolecular structure of stacked and un-
stacked regions of the photosynthetic membranes of
Prochloron sp., a prokaryote, Proc. Natl. Acad. Sci.
U.S.A., 77, 352, 1980.
53 Miller, K. R., Jacob, J. S.,Berger-Wiersma, T.,
and Matthijs, H. C. P., Photosynthetic membrane
structure in Prochlbrothrir hollandica: a chlorophyll
b-containing prokaryote, J. Cell Sci., 91, 577, 1988.
54. Whatley, J.M., The tine structure of Prochloron,
New Phytol., 79, 309, 1977.
55. Bullerjahn, G. S., Krugh, B., and Evans, T. T.,
unpublished data, 1992.
56. Chitnis, P. R. and Thornber, J. R., The major
light-harvesting complex of photosystem 11: aspects
of its molecular and cell biology, Phorosynrh. Res.,
16, 41, 1988.
57. Schuster, G., Neuchushtai, R., Nelson, N., and
Ohad, I., Purification and composition of photosys-
tern I reaction center of Prochloron sp., an oxygen-
evolving prokaryote containing chlorophyll b, FEES
Lett., 191, 29, 1985.
1991.
59. Bullerjahn, G. S. and Sherman, L. A., unpub-
lished data, 1987.
a.Ikeuchi, M., Eggers, B., Shen, G., Webber, A.,
Yu,J., Hirano, A., Inoue, Y.,and Vermaas, W.,
Cloning of the psbK gene from Synechocystis sp.
PCC6803 and characterization of photosystem I1 in
mutants lacking PSII-K, J. Biol. Chem., 266, I 1 1 11,
1991.
61. Burger-Wiersma, T. and Matthijs, H. C. P., The
biology of the Prochlorales, in Autototrophic Micro-
biology and One-Carbon Metabolism, Codd, G.A.,
Ed., Kluwer, Dordrecht, 1990, I .
62. Burger-Wiersma, T. and Post, A. F.,A functional
characterization of the photosynthetic apparatus of
Prochlorothrix hollandica (Prochlorales), a chloro-
phyll b containing procaryote, Planr Physiol., 91,
770, 1989.
63. Garcia-Pichel, F. and Castenholz, R. W., Com-
parative anoxygenic photosynthetic capacity in seven
strains of a thermophilic cyanobacterium, Arch. Mi-
crobiol., 153, 344, 1990.
64.Cohen, Y., Jorgensen, B. B., Padan, E., and
Shilo, M., Sulfide dependent anoxygenic photosyn-
thesis in the cyanobacterium Oscillatoria limnetica.
Nature. 257, 489, 1975.
 65. Cohen, Y., Padan, E., and Shilo, M., Facultative
anoxygenic photosynthesis in the cyanobacterium Os-
cillatoria limnetica, J. Bacteriol.. 123, 855, 1975.
66. Oren, A., Padan, E., and Malkin, S., Sulfide in-
hibition of photosystem I1 in cyanobacteria (blue-
green algae) and tobacco chlorplasts, Biochim. Bio-
phys. Acta, 546, 270, 1979.
67. Arieli, B., Padan, E., and Shahak, Y., Sulfide-
induced sulfide-quinone reductase activity in thyla-
koids of Oscillatoria limnetica, J. Biol. Chem., 266,
104, 1991.
68. Post, A. F. and Arieli, B., Photosynthesis of Pro-
chlorothrix hollandica under sulfide-induced anoxic
conditions, J. Bacteriol., in press.
69. Kyle, D. J., Haworth, P., and Arntzen, C. J.,
Critical Reviews in Microbiology Downloaded from informahealthcare.com by University of Sydney on 10/08/13
Thylakoid membrane phosphorylation leads to a de-
crease in connectivity between PSI1 reaction centers,
Biochim. Biophys. Acta, 680, 336, 1982.
70. Kyle, D. J., Kuang, T.-Y., Watson, J. L., and
Arntzen, C. J., Movement of a subpopulation of
LHC-I1 from grana to stoma lamellae as a conse-
quence of its phosphorylation, Biochim. Biophys.
Acra, 765, 89, 1984.
71. Bennett, J., Shaw, E. K., and Michel, H., Cyto-
chrome b,lfcomplex is required for phosphorylation
of light-harvesting chlorophyll alb complex I1 in chlo-
roplast photosynthetic membranes, Eur. J. Biochem..
171, 95, 1988.
For personal use only.
72. Allen, J. F., How does protein phosphorylation reg-
ulate photosynthesis?, Trends Biochem. Sci., 17, 12,
1992.
73. Staehelin, L. A. and Amtzen, C. J., Regulation of
chloroplast membrane function: protein phosphoryla-
tion changes the spatial organization of membrane
components, J . Cell Biol., 97, 1327, 1983.
74. Biggins, J., Campbell, C. L., and Bruce, D.,
Mechanism of the light state transition in photosyn-
thesis. 11. Analysis of phosphorylated polypeptides
in the red alga Porphyridium cruentum, Biochim.
Biophys. Acta, 767, 1984.
75. Bruce, D., Biggins, J . , Steiner, T., and
Thewalt, M., Mechanism of the light state transition
in photosynthesis. IV. Picosecond fluorescence spec-
troscopy of Anacystis nidulans and Porphyridium
cruentum in state 1 and state 2 at 77K, Biochim.
Biophys. Acta, 806, 237, 1985.
76. Matthijs, H. C. P., van der S h y , G., and Mur,
L. R., Phosphorylation and dephosphorylation of
membrane proteins from the prochlorphyte Pro-
. chlororhrix hollandica in fixed redox states, Biochim.
Biophys. Acta, 975, 317, 1989.
77. Alberte, R. S., Cheng, L., and Lewin, R. A.,
Photosynthetic characteristics of Prochloronlascidian
symbiosis. I. Light and temperature responses of the
algal symbiont of Lissoclinum patella, Mar. Biol.,
90,575, 1986.
78. Alberte, R. S., Cheng, L., and Lewin, R. A.,
Characteristics of Prochloronlascidian symbiosis. 11.
Photosynthesis-irradiance relationships and carbon
balance of associations from Palau, Micronesia, Sym-
biosis, 4, 147, 1987.
79. Fujita, Y. and Murakami, A., Regulation of elec-
tron transport composition in the cyanobacterial pho-
tosynthetic system: stoichiometry among photosys-
tem I and 11 complexes and their light-harvesting
antennae and cytochrome bdf complex, Plant Cell
PhysioL, 28, 1547, 1987.
80. Moore, L. W.-J. and Chisholm, S., unpublished
data, 1992.
81. Miller, K. R. and Lyon, M. K., Do we really know
why chloroplast membranes stack?, Trends Biochem.
Sci., 10, 219, 1985.
82. Murray, D. L. and Kohorn, B. D., Chloroplasts
of Arabidopsis thaliana homozygous for the ch- 1 lo-
cus lack chlorophyll b, lack stable LHCPII and have
stacked thyfakoids, Plant Mol. Biol., 16, 71, 1991.
83. Chow, W. S., Miller, C., and Anderson, J. M.,
Surface charges, the heterogeneous distribution of the
two photosystems, and thylakoid stacking, Biochim.
Biophys. Acra, 1057, 69, 1991.
84. Glazer, A. N., Light guides: directional energy trans-
fer in a photosynthetic antenna, J. Biol. Chem., 264,
1, 1989.
85. Smith, J. M., Dawson, C. G., and Spratt, B. G.,
Localized sex in bacteria, Nurure, 349, 29, 1991.
59