i.:" ?

rtA-za

,.8

I TheAbsorptionof Light in Phoosynthesis

The first steps in photosynthesisare the absorption of Light
by o pigment molecule and the delivery of the absorbedenergy
ta other mo\eculescapable of entering into chemical'reclctions
by Govindiee and Rajni Govindjee

to
of the system by which the energy of one that has received them is said
photosynin
Thus
reduced.
been
have
sunlight is made available to the potothesiswater is oxidized and carbon diox,yrrtli"ti" machineryof the plant.
ide is reduced'
of
photoThe maio, org*i" prodicts
Ordinarily, of, course, water does not
sr-rbrnces,
r"
synthesis
and it is not oxi"uiUofryrates,
such as sugarsand sarches,whose com- reduce carbon dioxide,
to proceed
reaction
the
For
n'
b.T
dized
oi the empiri-b""u.rr"
position is some multiple
must^be
energy
cell,
the
plant
inside
carbhyi"t for*rrla tCgrOl.
of an
requirement
energy
The
supplied.
be
to
drates appear ,,rp"r6"i"tty
"ois
commonreaction
porrndr oi catbo' and water, it was oxidation-reduction
and-the electrori'l
iho.rgtt fo, *"nf-years that photo,syn- ly measured in volts,
photosynthesis I
in
involved
transport
thesii consistedi Jplitting o"rto' dioxpotential of ,
ide (CO2),which\'torrtd atl"owthe oxygen Poceegs-against an energy
volts.
1.2
about
fiee
(O2)
and
gas
a diatomic
to
An electrochemical gradient -of L'2
carbonto combinewith water.
tlre"r""p-'as
barrier,
It is now known that this scheme is volts representsa rather l,arge
that in
believe
to
reason
is
there
and
wawrong. Neither carbon oxide nor
overis
not
it
organisms
photosynthetic
or
split
be
ter cin properly be said to
apit
light;
of
quantum
single
a
by
net
o*"
TLe
de.o*p&e irr'photosynthesis.
reare
quanta
effect f tft" pro"r, is instead to transfer Pears_instead that two
be identiffed.
electron'
each
of
,
transport
for
the
water to carbon quired
In this article we are concerned main- hydrogen atims from
nllerby
is
supported
hypothesis
ihlt
comes from
ly with the first steps in photosynthesis: dioxid!; the oxygen evolved
the proc- ousrecent experiments;moreover' it acBecause
thJCO2.
not
H2O,
the
molea
speciffc
by
light
t'he absorption of
important observation
esstakes place in water solution it i-snot cords well with an
cule and the transfer of that energy from
by the- late
20
years-ago
almost
complet"
a
move
T".d"
one molecule to another, as in a bucket necessaryto actually
of llliUniversity
of
the
is t^rans- Robert Emerson
brigade, until it is eventually conveyed hydrogen atom; if r, electron
there
that
evidence
found
Emerson
or proton, nois.
to those few molecules that participate ferred, a hydrogen nucleus,
that
plants
in
t{tl:Tt
two
are
the alueous
Pig*"lt
in chemical reactiors. These initial Proc- can be drawn ater from
difof
slightly
tight
absorb
atom. Chmical Prefeteniiaily
essesare'called the "primary events" of medium ro complete the
(or
implying
wavelength
ierent
electrons
which
in
tdot)'
photosynthesis.They are physical.in na- processesof thiJkind,
molecule to an- that electron transport takes place in two
orr"
fro*
transferred
are
before
completed
be
must
they
rre,
i
re- main stages and involves two photo"rrd
the chemical activities of photosynthesis other, are called oxidation-reduction
"The Role of chloacons.The molecurethat has lost elec- chemicarvents [see
can begin
Eugene I'
Photosynthesis"'by
in
rophyil
trons is said to have been oxidized; the

IV
fhen the sun shineson e leaves
of a green Plant, somg of the
\[u/
Y V radiant energlr is utilized to
promote chemical reactions, with the uliimate result that water and carbon dioxide are converted into oxygen and organic compounds. Photosynthesis has
b""o tn*marized in is way since the
end of the lSth century. The summary is
essentially correct, but it describes the
in terms of what flows nto
only
Drocess
'the
'and
plant. Today a more deout of
precise explanation is
more
tailed and
know what haPPens
to
want
we
sought;
insie the illurriinated leaf. It is not suficient to say at light "promotes chemical reactions." Rather, the molecular
mechanisrn by which light is absorbed
and by which its energy is utilized must

Elecron Flow in PhotosYnthesis

: Investigations of the primary events
hav'e been hindered by the speed with
which the events take place and by their
complexity and inaccessibility;many can
be otservLd only in the living cell' Most
of the experiments intended to explore
their seqencehave by necessitybeen
indirect. Many of them have been quite
ingenious, however, and they have revealed several important characteristics

68

with
is stimulated by illumination
of the plant pigment chlorophyll
FLUORESCENCE
the
of
bottom
the
at
seen
be
can
'by
blue light. The natural green color or the chlorophyll
visible
made
light,
of
beam
The
page.
opposite
tbe
on
container iu the pUo,orl"nh
the top and ie absorbed by the pigment'
emoke particles auspended in the uir, t"rs from
of the absorbed light is dissipated as
energy
the
which is in an ether solution. Some of
in
part of the spectrum' chlorophyll
red
the
in
fluorescence
as
is
reradiated
heat; the rest
that
in
energy
the
of
moet
weakly;
only
the leavee of a living plant alao fluoresces, but
photosynthesis in the plant'
of
work
the
to
applied
is
light
red
eolution ie reemitred as
T. Lamme at Columlia University'
Tho photograph was made in the luboiutory of Alfred
K. Trench of Yale university'
Robert
ulaaby
seaweed
the
with chlorophyll extracred from

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in sltor{tr ir t'ross st'cti+rin ar clet'tron

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l
r
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t\j

is visilrlr., in urr tltjctror nri'

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r.r'grirr[ *r*{r: Iy (1. J. Arltztl. alstt
,I.lrr:
r . { r t t t r l ' , , . s , ; trl' l r l o r l r r l a s t
s r r . r " i l i l o r \ \ . l l l . r r ( ) r a I r : r ll r 1 ' f r l l , r z i r r g
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' f I l o : I r a { . t u r . ( l s r r r { a r : r : .. t t l o w r " r J r : f t i " i l ! 4 f
y i { u r l t l r l ' l a k g i r l r r l r r a r r : sl x 1 ' 1 ! r : g t r l u l l r - ' t la l a r l l o r t g i i u t i i r r a l l y '
graills lllat rtlstlllrl l:lll"
rr:r.,tulilrg inlrtrior surflcr,s crr,rtrcl rr'itlr
s
i
z
c
s illtl eal size ntuy lle as'
v

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i
r
.

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s
g
r
a
i

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s
9
f
Irlr:st{}rr:s.',fIern,:rrc
ol
s r r : i a t t : r lr r i t l l l l i l l r : r r l l r : l i 9 t l i t r t h t r t , r o ( : o s r " i r l r o t o s y l l l l r r s i s '

sl.ltUC.f.IrltE

Rabinowitch and Govindjee; Scrnrvrrrrc

HH

Aurncx, July, 1965].

Not all the moleculesassociatedwith
,\)(/,
the two pigment systems have been
HC
n."7\
identified, nor &re all their relations yet
4c-cn,
clear. Nevertheless,it is possibleto daw
Hsc
")"-^
at least a tentative map of the electronMg
transport pathway lsee illustration on
\*n"'''\"page 741. The scheme is based on a
"-\
model proposedby Robert Hill and Fay H 3 C - - 6
/"
Bendall of the University of Cambridge
2c_cH3
c
"t \qttt.lx.
[see "The N{echanismof Photosynthe"-"\
I
sis,"by R. P. Levine; ScrcNrr.r" *r*rHc
Ccex, December,1969].
" ,
-o
\
In the modified Hill-Bendall model,
\
light striking the aggregation of mole"
cules designatedpigment system II re*-/"-"
CHS
sults in the transfer of an electron from
\
a donor called Z to an acceptor called
T
,_/"-'
Q. Recentwork on the kinetics of oxygen
evoh*tion by Pierre Joliot of the Institut
\
CH
de Biologie Physico-Chimique in Paris
"
^,"-("
and BesselKok of the ResearchInstitute
for Advanced Studies in Baltimore has
shown that a molecule of oxygen il}
"r"\
evolved after Z has given up {our elec\
trons, and thus aecumulated four posi-i
tive charges;Z is eventually restored to o
\-"
F
n,"-("-"
neutrality by scavenging four electrons
L
from two water molecules. The oxygen
",(-""'
atoms from the water form an 02 mole)"-^
H-C
./"^'
cule and e four protons enter the solu-
*,"\--"
tion aspositive ions.
\
From reduced Q the electrons are
transferred, with the electrochemical
",(_"".
gradient, to pigment system I. Several
,/c^,
kinds of molecule serve as intermedi".\--"
HH
aries, including at least one crtochrome
(one of the proteins that also serve as
"^'
,,{
electron-carriers in cellular respiration),
an unidentified compound B, a plastoquinone and a copper-containing pro- PIGMENT MOLECULES are ilistinguished by systems of conjugated, or alternating,
tein, plastocyanin.On absorbinga quan- single and double bonds. Ifhen the pigment absorbs light an electron circulating tbrough.
tum of light, pigment systemI promotes out the system of bonds enters an excited state. fn chlorophyll {lett) rhe conjugated bonds
an electron from a "reacon-center" (colored band.) are in a complex ring called porphyrin. Attached to the ring is a *tail" ol
chlorophyll to another acceptor mole- phytol, made up of carbon atoms joined mostly by single bonds. Shown is chlorophyll a;
cule, labeled X. The oxidized reaction other formg difrer from it only slightly. Lr the carorenoid pigments (righ) the conjugated
bonds are located in a straight chain of carbon atoms that has a cyclic ring at each end.
center is then reduced by an electron
that flows from reduced Q via the intermediatesmentioned above.The reduced
PROTEIN LAYER
X then donatesthe electron to the ironPORPHYRIN
and-sulfur-containing protein f erredoxin,
reducing it. The ferredoxin, with the
help of e enzyme ferredoxin-NADP+
reductase, ultimately reduces nicotinamide adenine dinucleotide phosphate
(NADP+), a moleculewith many rolesin
metabolism.(In its reduced form it is abbreviated NADPH.) This sequence of
events is repeated for each of the four
electroys donated by Z, so that eight
electfbn transfers, and eight quanta of
ARRANGEMENT
OF PIGMENTS in tho lamellar membrane may be governed by tho
light, are required for each molecule of physical characteristics of the molecules. In this speculative model,
the phytol tail, which
orygen evolved.
is hydrophobic, or repellent to water, projects into the hydrophobic lipoid layer. Tho
Energetically the trahsfef of electrons porphyrin ring, a hydropbilic, or water-loving, group, associates with the hydrophilic
from reduced Q to the oxidized reaction- protein layer. Carotenoids are hydrophobic and are probably found in the lipoid portion.

ll

/il

^/Ir, T
l_o I
ll

._?"-'

T"'
tl

.f-"^"

,r1"

^-1"-"

*"-(-i{:-7').\'

7l

center molecule is a downhill process

and is coupled with the production of
adenosine triphosphate (ATP), the ubiquitous energy-carrying molecttle, fl'om
adenosine diphosphate (ADP) and inorganic phosphate. The existence of a cyclic flow of electrons around system l
has also been documented. The electrons
from rec{uced X, instead of reducing
NADP+, are cycled back to the reaction
center; when this cyclic flow goes via
plastoquinone it is coupled with the production of ATP. (Other sites have been
implicated in ATP production, but their
role is not yet confirmed.) The importance of cyclic electron flow varies from
plant to plant, but in most cases its contribution to photosynthesis is rather
small. It is ATP and NADPH that effect
the reduction of carbon dioxide and mediate its introduction into a carbohydrate
cycle fsee "The Path of Carbon in Photosynthesis," by J.A. Bassham; ScreNrrnc
AnrnncN, June, f962].
Pigment Molecules
The photosynthetic apparatus of the
higher plants is organized inside chloroplasts, the cellular organeiles that give

An aggregate of this size, they proposed,

should be considered the ultimate photosynthetic unit. We now know that eight
photochemically driven electron trans*
fr:rs are required to generate one molecule of oxygen, so that a smaller unit is
plausible, one containing 300 pigment
molecuies. This is the modern photosynindependent of the cell nucleus.
thetic unit, the smallest unit capable of
an
elaborateis
Inside the chloroplast
photochemical action.
ty folded network of membranes termed
Embedded in the thylakoid memlamellae. In chloroplasts from most
branes are the pigment molecules that
plants the lamellae form structures called
initiate the process of photosynthesis.
grana, which are separated by a material
Pigments are substances that by definiknown as stroma. The grana appear to be
which
tion strongly absorb visible light. Most
sacs,
dense stacks of rnembranous
absorb only in certain regions of the
are the basic units of the lamellae' These
spectrum and transmit light of all other
sacs, called thylakoids, contain lipids,
wavelengths; as a consequence they approteins and pigments. It is on and perpear colored. For example, chlorophyll,
haps between the thylakoids that the
the most important plant pgment, abbusiness of photosynthesis is transacted.
sorbs both the longer and the shorter
organizastmctural
further
level
of
A
waves in the visible spectrum: red and
tion can be inferred in the chloroplast,
orange and blue and violet. The transeven if it cannot be resolved with cermitted wavelengths, chiefly the yellow
tainty in the electron microscope. Experand green in the middle of the visible
iments performed more than 40 years
spectrum, combine to yield the green of
Arnold,
A.
William
ago by Emerson and
grass and trees lsee illustratian belouf .
who is now at the Oak Ridge National
The majority of organic molecules abLaboratory, suggested that a minimum
sorb most strongly in the ultraviolet; the
of 2,400 clorophyll molecules are revarious pigments of the chloroplast atrquired to evolve one molecule of oxygen.

plants their characteristic green color.

The chloroplasts are complex structures,
separated from the cytoplasm of the cell
by a membrane and apparently having
some autonomy: each has a bit of the genetic material DNA and is able to syntltcsize somc of the protcins it rcquircs

I
I

AI
I
I

a
ffL
4

a
co

l
t1
I
I

GREEN
W A V E L E N G T H( N A N O M E T E R S )
CHLOROPHYLLa
CHLOROPHYLLb
CAROTENOIDS
PHYCOERYTHRIN
PHYCOCYANIN
SOLAR SPECTRUM

72
,l
t

,-,-t-ti..

ir:. ":

i:i:11!fl

YELLOW

700
BED

the amount of light

ABSORPTION SPECTRUM of the phorosynrhetic pigments measures
bands, one in
absorption
two
have
chlorophylls
The
wavelengths.
absorb
at
various
they
intermediate wave'
the blue and one in the red. They are green because they transmit the
lengths. The carotenoids absorb shorter wavelengths and appear yellow or red' Phycoery'
absorbing long
th"irr, which absorbs blue through yellow light, is red and phycocyanin,
solar spectrum.
the
in
light
of
the
most
absorb
pigments
the
waves, appears blue. Together

sorb at the longer wavelengths of visible 670, 680, 685, 690 and 695 to720 nanolight becausothey havo chains or rings ,meters.The variations are probably proof carbon atoms connectedby "conjugat- duced by chlorophyll molecules in difed," or alternating, single and double ferent environments;they may be aggrebonds. In chlorophyll the system of con- gated differently with other chlorophyll
jugated bonds is located in a ring, and it
molecules, or they may be associated
encompassesnitrogen atoms as well as with proteins.
carbon atoms lsee top illustration on
Although the chlorophylls are unpage 711.The ring structureis one of the doubtedly the most important plant pigclass of compounds called porphyrins, ments,they are not the only onespresent
which are found widely in bo plants and they are not the only ones that parand animals. In chlorophyll the central ticipate in light absorption. The carotcavity of the ring is occuped by a mag- enoids, found in bacteria, algae and
nesium atom; e function of e magne- higher plants, absorbmainly blue wavesium is not yet fully understood. Porphy- lengths and are yellow, orange or red in
rin rings in the blood protein hemoglobin color. (Their name derives from the
and in the cytochromescontain an atom same root as "carrot," and they give the
of iron instead of magnesium.
carrot its orange color.) They consist of
Attached to the poqphyrin ti.rg in long chains of carbon atoms linked by
clorophyll is a long hydrocarbon "tail,"
conjugatedsingle and double bonds and
the phol chain. It consists of carbon bearing a six-carbon ring at each end
atoms linked together, but only one of
lsee top illustration on page 7Il. The
the bonds is double; for this reason the carotenoidsare divided into two classes:
phytol chain does not appear to play an the caotenes, which are hydrocarbons
important part in determining the chlo- and therefore consist of hydrogen and
rophyll absorption spectrum. Its function
carbon only, and the carotenols,which
may bo to anchor the molecule in the are alcoholsand ketonesand containoxythylakoid membrane. The phol chain gen aswell as carbon and hydrogen. The
is hydrophobic, that is, it repels water final category of pigments is the phycobut has an affinity fo oils and fats; the bilins, named for their resemblanceto
porphyrin ring, on the other hand, is hy- pigments in the bile of animals.They indrophilic, being drawn to water. It has ; clude the red phycoerythrins and the
therefore been proposed at the phytol : blue phycocyaninsand are found only in
portion of the moleculemay project into the blue-greenand red algae.The molethe lipid layer of the thylakoid mem- culesof both types have an open ring rebrane, securing the porphyrin at a par- lated to the porphyrin structure of the
ticular location in the protein part of the chlorophylls, but in the phvcobilins the
membrane lsee boltom illustration on ring is bound to a pro.teincomponent.
.-.1
page711.
Three major varieties of ctorophyll
Light Absorption
are distinguished by small difierences in
struchrre in one region of the porphyrin
When a photon is absorbed by an
ring. Chlorophyll c, the most abundant atom or molecule, its effect is to change
form, found in all higher plants and in the conffgurationof the electroniccharge
all algae,has a methyl group (-CH.) at associatedwith the valence, or outer,
at position. In chlorophyll b, which is electrons surrounding the atomic nufound only in the higher plants and in
cleus (or nuclei). Becausethe new conthe green algae, the methyl group is re- ffguration has more energy than the
"ground," or lowest, state, the atom or
placed by an aldehyde group (-CHO).
Finally, in bacteriochlorophyll, the form
moleculeis said to be in an excited state.
of the pigment found in photosynthetic The transition from the ground state to
bacteria, a single bond is ubstituted for
an excited state can take place only unone of the double bonds in e cojugat- der certain conditions prescribedby the
ed ring, and two hydrogen atoms are laws of quantum mechanics.The elecadded.
trons can occupy only specified,distinct
In addition to these structurally dis- energy states;regionsbetween statesare
tinct molecules, various spectral forms forbidden to them. Moreover,the energy
of chlorophyll have been detectedin livof the absorbed photon must exactly
ing cells: they ean be distinguished only match the energy of e transition. Since
by differences in their absoqption spec- the energy of a photon is inversely protra. C. Stacy French of the Carnegie portional to its wavelength, only certain
Institution of Washington's Department wavelengthscan be absorbed by a parof Plant Biology has detected forms of .ticular atom or molecule fsee top illuschlorophyll whose maxirnum absorp- tration on page 751.
tion of red light is at wavelengths of 660,
In atoms these restrictions are quite

PHOTONS

ANTENNA
PIGMENTS
(300oHLoRoPHYLLMOLECULES)
ANDOTHERPIGMENTS

tU
lrCJ)

z
cc
F

o
tJ

z
tu
I
I
I

I
I
I

I
I
I

R E A C T I O NC E N T E R
DONOR
PHOTOSYNTHETIC
UNIT ia thought to
consist of about 300 chlorophyll molecules;
in the higher plants and algae that seems to
be the minimum number necessary to bring
about the transport of a single electron. The
unit includes the bulk pigments, which
serve as an energy-gathering toantennarttand
a reaction center, where energy is trapped
and utilized to promote chemical reactions.

73

confining, since the permitted- energy

f"* and transitions between
levels
"tu
them are large' As a corlsequence the
absorption spctraof atomsusually consist oi a relatively few narrow lines' In
molecules,however, md particularly in
pigment molecules,a number of factors
i"ia to broaden the lines into bands' For
one thing, the conjugated bonds-in pigments biing about a great prolife-raon
of allowed states because each double
bond adds a pair of electrons shared by
thy' conjugatJd system as a whole' The

Two other factors also increase the

efiect of a large system of conjugated
quantum states in
bonds is thereforeto decreasethe transi- number of possible
and the rotavibrational
the
molecuies:
tion gap between the ground state and
These
molecules.
the
of
energy
tional
to
the fi-rs[,or lowest, excited state and
endiscrete
to
confined
also
are
motions
just
states
creaternany additional excited
more
rnuch
are
the levels
above the first. It is this effect that is re- ergy levels,but
so that the ground state
spaced,
cloiely
absoption
the
sponsible for shifting
excited statesbroaden
the
of
siectrum of pigment moleculesfrom the at d ""h
of
substates.In approxi- r l
manifold
a
into
ultraviolet region of the spectrum into
between the
transition
the
terms
mate
the visible t"glott; becauseof the smaller
state in
excited
first
the
and
state
ground
pigstate
transition to the ffrst excited
differenergy
an
a represents
ments are able to absorb photons of low- hlorophyll
electron
two
and
one
er energy and hence greaterwavelength' "rr"" oJ tetween

.T

I
t
d-r.o

-,4
J

'tr
z

l
F

- . ^

o
o.

PIGMENTSYSTEM I

z
I
F

3
o
o
tu
fr
I

z
o

*.e
E
o
xo

ANTENNA
PIGMENTSYSTEM II

+.4

ANTENNA

proceeds
oF ELECTRONS from water to carbon ilioxiile
requires t\Mo
volts-and
l'2
of
gradient
an electrochemicai
one at
urrrical evente. Four electrors must be transferred'
of
molecule
reduce-a
to lerate molecule of oxyged"and
absorption of a
the
with
begins
procesB
The
carbohydrate.
II' The energy of excita'
pbioo by the antenna of pigment system
in the reacrion center
moleculo
to a chlorophyll
iioo i"
"oo""y"il
ofthephotosyntheticunit;themoleculeistlesignatedP6B0be.
8pectrum is at 680 nano'
cause one of the banils in its absorption

FLO\tr
against
pf,oto.t
a time.
-a;;;

meterE.TheexciteilP6s0translersanelectrontotheacceptorQ,
the donor Z' Aftet Z
and subsequently recovers an electron from
oxidizing two mole'
by
them
regains
it
electrone
bas given op foo"
pasaed through a series of
is
elecrron
tlr"
From
water.
of
Q
culei
not been iilntified, plas'
carrier moleculee, including B, which has
(Crt), to plastocyanin (PC).
(pe)
cyrochrome
anil
|
ioqoiooo"
I' The reac'
Pltetocyanin injects the elecFon into pigment system

74

system I' designatecl P?00' is

tion-center chlorophyll of pigment
and promotee an elec'
pigments,
antenna
o*lr
its
excited through
trontotheacceptorX.Finally,theelectronispassedthroughferre.
(FRS), ferredoxin (FD) anil tho enzymo
doxin reducing subsrance
(R) to nicotinamide adenine dinu'
reductase
ferredoxin-NADPT
(NADP+),
which is thereby reduced to
cletide phosphate
product of these reactions' In
primary
is
the
NADPH
NADPH.
addition,duringtwooftheelectrontransfergadenoginetriphos.
diphosphate (ADP) and
phate (ATP) is generated from adenosine
is between plae'
generation
ATP
irorgu.ri" phosphate. One site for
with syetem II
associated
is
other
tle
toquinone and cytochrome /;
drive tho
ATP
the
and
but has not been located' The NADPH
processbywhichCOrisincorporatedintocarbohydrates(multiplee
of photosyntheei'
of ,fr" unit CII'O). fh"r* is also a cyclic system
through var'
return
then
X
and
to
P?00
in which electrons pass from
ATP is produced'
only
system
this
in
P700;
to
."rriJr.
ious electrot

i
I
i

-l

volts. (An electron volt is the energy acquired by an electron when it is accelerated through a potential of one volt.) The
vibrational substates are separated by
about .l electron volt and the rotational
substates by about .01 electron volt.
When a photon of appropriate energy
strikes a molecule of chlorophyll a, the
pigment enters an excited state almost
instantaneously,
within
10'15 sesond.
This is an extremely brief interval; in a
vacuum light travels only about three
ten-thousandths of a millimeter in l0'r5
second.
There are two main excited states
available in chlorophy o. The absorption of red light (wi a wavelength of
about 680 nanometers) raises the molecule to the lowest and most important of
the levels, called the first excited singlet
state. Blue light (with a wavelength of
about 440 nanometers) promotes the
molecule to e thfud excited singlet
state. (Transition to the second excited
singlet state, between e first and the
third, is weak.) The upward transition
may begin from any of the various vibrational and otational substates of e
ground state and end on any of the substates of the excited state.
Having reached an excited state, the
ctrlorophyll molecule cannot iinmediately and directly apply its energy to the
tasks of biochemistry, nor is useful work
the only possible outcome of the absorption of a photon. The excited molecule
can give up its energy in any of a number of ways, all of which conform to the
same laws of quantum mechanics that
govern light_ absolption.

THIRD
EXCITED
SINGLET
STATE

SECOND
EXCITED
SINGLET
STATE

J
INTERSYSTEM
CROSSING
FIRST
TRIPLET
STATE

Energy

GROUND
STATE

pHospHoREScENCE

EXCITATION
OF AN ELECTRON in a pigment molecule promotes the electron ro one
of a few discrete energy states; intermediate levels are forbidden. Superimposed on the
permitted energy states are substates representing the vibrational and rotational energy of
the molecule; the substates also are discrete, but they are closely spaced. When an electron
absorbs a photon i is first elevated to an energy level called an excited singlet state. De.
pending on the energy of the photon, the electron may reach the first or some higher excited singlet 6tate; it quickly subsides to the first, however, by dissipating part of its
energy as heat in a process callecl internal conversion. In the first excited singlet Btate the
molecule can utilize its energy in a chemical reaction, lose energy by internal conversion,
reradiate it as fluorescence or enter another excited state, the triplet state. Because the
triplet state is longer lived than the excited singlet states, light is emitted by molecules in
the triplet state only after a delay and is called phosphorescence instead of fluorescence.

onlv

molecules,

to the kinetic

motion

of the

not to photochemistrv.

Because of internal

For the purposes of photochemistry

the most important energy level in chlorophyll is the first excited singlet state.
N{olecules elevated to higher excited
states return to the ffrst excited singlet
state rapidly (in from 10'1a to 10-13 second), so that they reside in the higher
state too briefly to enter into any competing processes. The return to e first
excited state is achieved through the
small transitions separating the vibrational and rotational sublevels. Bach of
these transitions is so small that the
wavelengths associated with them are
not perceived as radiation, only as heat.
By the same mechanism a molecule that
has been raised to one of the higher sublevels of the first excited state wi quickly decay to one of the lower substates.
This process is called internal conversion; since it results in the degradation of
the received energy to heat, it contrib-

-----;

-/FLUORESCENCE
EXCITATION-

utes

The Fate of Excitation

conversion

a high-

energy photon of blue light is of no more

use to a plant than a reiatively low-energy red one. The squandering of the energy of short-wavelengthlight, however,

is less extravagant
seem.

Although

short

wavelength

than

it may

individual
are

the

at first

photons
most

of

ener-

getic, most of the energy of the solar flux

is distributed in the yellow and orange
and beyond. There are simply greater
numbers of photons at longer wave-

EXCITED
STATE

SINGLET

E X C I T E DS I N G L E T

DOUBLET

TRIPLET

SINGLET AND TRIPLET STATES are defined by the "spin" of the outer electrong in an
atom or molecule. In this diagram the arrows represent the spin axis. If the spins are antiparallel, the molecule is in a singlet state; if they are parallel it is in a triplet state. The
doublet state requires an unpaired electron. "Singletr' "doublet" and "triplet" refer to tho
number of ways the electrons can orient themselves with respect to a magnetic field.

---T--I

!--r--rl
l

HIGHER
EXCITED
SINGLET
STATES

'r
F
(5

I
F
CI

ul

J
tJ

ii r
j'i

>
3
(,

za

FIRST
EXCITED
SINGLET
STATE

'

TU

cr
o

z
I

tIJ

cc
o
z
I

ABSORPTION

3
o
=

II

;l

GROUND
STATE

ut
J
u

-->
ABSORPTTOT

associated. with light absorption and

ENERGY TRANSITIONS
are constrained by tbe requirement
fluorescence by chlorophyll
that the molecule occupy only diecrete energy states. OnIy photons
whoso energy correeponds to the energy difrerenco between the
ground state and an excited-stata can be absorbed; since the energy
ol a photon ie inversely related to its wavelength, the selective abeorption of photons ia refleeted in the absorption spectrum of the

FLUORESCENCE

FLUORESCENCEINTENSIW->

pigment (diagram and gratph at lelt). Even though chlorophyll absorbs light in several parta of the spectrumn it fluoresces only in the
red (diagram and graph at rght). The fluorescence is from the
first excited singlet state; molecules in higher stateE decay to this
ono by internal conversion. Fluorescence is at longer wavelengths
than the lowest-energy absorption band because molecules relax to
a low substate of the first excited singlet state before fluoreseing.

HIGHEREXCITED
SNGLETSTATES

INTERSYSTEMCROSSING

I
I
I
t
I
I

-i-I TNTERNAL I
I
lcoNvenspH
I'ATES Of' EXCITATION ENERGY are of varying probability
and of varying importanee to the living plant. The pathway indi'
eated in color ia the only one known to contribute to photosyn.

thesis. Transitions shown in eolid black neg take place in the plant
but do not result in useful chemical reaetione. Transitione indicated by broken Iines have not been observed in living cella.

t
lengths, and their aggregate energy is
much greater.
Once an excited clorophyll molecule
has subsided to one of the lower subIevels of e first excited singlet state,
several pathways are open to it. Through
further internal conversion it can ssipate its remaining energy of excitation as
heat; it can enter another excited state,
of a somewhat di-fferent character, called
the triplet state; it can return to the
ground state by emitting a quantum of
Iight as fluorescence; it can transfer its
energy to an appropriate neighboring
molecule, or it can enter into a chemical
reaction, such as those characteristic of
pigment systems I and II. The last process is obviously the most important, but
it is by no means the only one that takes
place.
Singlet and triplet states are distinguished by a quantum number called the
total spin number, which has to do with
the directions in rvhich the electrons in a
molecule "spin." In singlet states the
spins of a pair of electons are antiparallel, or opposte; in triplet states the spins
are parallel. f)oublet states, which are
also found in some photosynthetic pigments, occur when there are unpaired
electrons. The triplet state is so named
because electron pairs spinning in parallel can align themselves in three ways
with an external magnetic field; moreover, ey superimpose three additional
energy sublevels on the quantum state of
the molecule. In the singlet state all orientations wi respect to an external field
are equivalent, and no adtional'sublevels are intoduced [see bottont. illustration'on page 751.
The triplet state is metastable, that is,
it is stable for a comparatively long time.
Events that can reverse the spin of an
electron are relatively uncommon, and
transitions between the ground state and
the triplet state are therefore rare. The
triplet state can be readily entered, however, from an excited singlet state. There
is considerable evidence that molecules
in-the triplet state are present in e chloroplast. An analysis made in our laboratory at the University of Illinois suggested that they are present, but in low eoncentration; our data suggest at at any
given moment less than one chlorophyll
molecule in 10 million is in the triplet
state. J. S. Leigh, Jr., and L. P. Dutton of
t}re University of Pennsylvania School of
Medicine have found evidence for the
in
triplet state of bacteriochlorophyll
photosynthetic bacteria, but only at low
temperature and only when the photochemical oxidation-reduction processes
driven by the pigments were blocked.

I
I

6
z

T'J
F

z
ut
o
z
tJ

o
o
UJ

cc

f,
J
TL

600

6s0

70a
WAVELENGTH(NANOMETERS)

750

800

FLUORESCEICE
SPECTRA can discriminate
EMISSION
between pigment moleculeg
eerving fferent functione. fntact chloroplaste at room temperature (colored lne) have a
strong fluorescence peak at about 685 nanometere and a much smaller peak at about ?35
nanometers. At low temperature a more complex pattern emerges. Pigment system I (solid
blnck lne) fluoresceg most atrongly at ?35 nanometere, with minor bands at 684 anil 695. In
pigment Eystem lI (broken black lne) peaks at 685 and 695 nanometers are more important.

Even

if the triplet

illuminated

plants,

state is present
there

in

is no evidence

that it forms a part of the main photosynthetic pathway; there are many other
ways in which its energy could be dissipated, such as internal conversion and a
return directly to the ground state with
the emission of light. The latter effect is
termed phosphorescence:
it is character-

ized by light

emitted

at a considerably

longer

after

a delay

waveleng

and
than

the light absorbed. A molecule in the

triplet state can also return to the ffrst
excited singlet state if it absorbsa small
quantum of energlr; it then has before it
all the possible fates of other molecules
at that energy level.
Light emitted during a transitionfrom

E N E B G YL O S S

PHYCOERYTHRIN

TRAISFER OF ENERGY from one kind of pigment to another follows an established

Bequence. The carotenoids, which absorb blue light, pasa their energy of excitation on to
phycoerythrin. Energy received in this way, as well as the energy of greeu ght abeorbed
directly by phycoerhrin,
ie transferred to phycocyanin. Phycocyanin abgorbe orango
light, and pagges the accumulated energy to chlorophyll c. In each of theee traneitione
energy is lost by internal conversion, and in eome of tho transfers energy ie also dieeipated
as fluoreseenee. Chlorophyll b contrilutes its energy directly to chlorophyll a without loes.

PHOTON

,f

a singlet state directly to the grourld

state is terined ffuorescence instead of
phosphorescence.Ordinarily there is little delay between absorption and reemi.ssion,arrclthe wavelength of thr: fluorescent light is only slightly longer than
that of the light absorbed. The small incl'casc is causod l-ry thc loss of cnorgy
through internal conversion.

i;;i
..r,..

i',.r ll::.r
li': : ,ri:ir:,r:
iii: s
t ':,:'ii:t:;i)z,
a i.:i::,i

'i

:'

lrt*t3

I'-luorescerrce
Iiluorescence has provided sonie of
the most valuable techniques available
for thc investigation of photosynthesis,
but for the plant it is an entirely wasteful phenomenon. Ordinarily, holvever,
the rnagnitude of the e{Iect is rather'
snall. For every 100 photons absorbed,
only from three to six are recmittetl. For
this reason fluorescence in livrg plants
is faint and can rarely be detected by the
unaided eye. The fluoresceuce of chloroPHOTOI',1
i,,
ph;rll in solution, on the other hand, can
\
be rliiitc bright, since thc proccss tllrt
ii
would normally drain away most of the
E
energy of exciLation-photosynthesis-is
.:..
disrupted.
i.
Chlorophyll fluorescence is invariably
ii'i
tr
red, even if the exciting light is blue or
green or yellow. The fluorescent light is
PHOTONi::
E
ii.
emitted in a transition from the first excited singlet state, and molecules promoted to higher states by more energetic
merely dissipate part of their enphotons
PHOTON
,
ergy as heat before ffuoresc ng lsee tot
llustration on page 761. Lt r<om temw
perature the fluorescence spectrurn consists of a major band at 685 nanometers
il iii,iii{ii,:ii,ltltEt#. tffftfffffffff$f+8f#
f|t1li1!.!,:,
fi:,;liliir,i:il$ljll]',1,i'
f; lri'li'=r A ii1:..rj;,r..f
Wv.##FtyA
#ltffiffiRffiffiffieqffiffi$ffiffiWffi$ffi4ffiF,
*ii.'i.$[ii.\'l\ii$iii$,r"1g.<$,ri$r:r,,iii$tixr-q$til.Iii
$i,r,Nr,$N
and a minor band at about 740 nanomeffiiiffiffixffiffiffiffifffffwtfwwtj|.Fnv';r.'i;1;,f
F,$i):fg$irt$llliNt$ii:i"\-\:$;l*\\llii)ll\,\s\r.l,i$Hi.s
$iiii$f,.ii,i.li,I
ters. When the spectrum is measured at
W:?i'lit:r'it:t#'*il"*ij.ffi":lkffir;&tf*#fa1'#ltill?3"',?,'jbf'ii:{#.rW,Y,#WffiWWffiWffiffffiffiWffiffiWffiVYffiW}trffi:fi:3
ffiWffij::i:'l' low temperature, bands aPpear at 685,
Si$Wffi.W*Wffi-ffiHWffiffi$.$ffiffiffiffiffim'ffi,ffi
K\:$S N"\.\]$:$.Rs.\tliN{S.s.ffi*t$.
$\,$:li\\r,r[\\
S;lrffi'riiiri
K$*NNW&Xffiffiffiffi$#ffiffiffiWffiWl.sB
$l+irsi$N-ti$ ffiN &*ilti\l\\iil$]$ *.*i:,\iri$l*\il.S
1.:.{ 695 and 720 nanometers. Pigment sysF,#"qlit,?t,
F,##:l1,
i:ffi $$w tu\}i${t-\i}
:ffi
$LRffi$*i$il,$il$
fti$il,$il$s-.rffiffik-\ffiffiffiffiffiffiffiffiffi.s#ffiffifrl
s-rrffiffik-\ffiffiffiffiffiffiffiffiffi.g#ffiffifr#
*.ris.riir{i$
-{iiii\,fis{iir{i$
{:i'";E;
Nl}]$ Nt$|i$$
Nii;i,*NN $$l\\gi\SS.\N
Nji;i,*NN
Ntli"i-\$
Ri.lW$Tffi-*'$*ffi$$SffiS;9gggffitir.ili$
Ri.lW$Tffi;*'$*ffi$$SffiS;9gggffitir.ili$
$$l\\$i\,SS.\N
iili:i.N':li"s
N$i,iii.\\':li"
tem Itr is prirnarily responsible for the
{iii:!ii',::,,,
$;'ii:]$
ffi'$$t$ffiffi&.$ffiir,W$$*#,#e1ff\'.f$#t,:t;;,
li::iir$i,'jj
$;li$:{# ffi:tiljis#:iri
'fi.if,$S}i-iliii.?i$
':'::.'tl brndsat 685 ancl 695 nanomr:ters,ancl
{i.if,$ d;irlij
dlirliS trr:ills
tr:ills &d-i,fr$r,19$t:t.-ffiii:r"B$i;.f.'$*i-r,4t:}i!,nffi|,:,i4';::
&$-i,f$r,19$t:t.$fii:r"B$i;{.'s*i-r,4'{:};i!,n'{;i|ii'i,t;:i:'
$irli:irti
$$SXiiiI
$rli:irti
Ktr$'$SX*i1
Kti$
$ii

ii:ri:ir iii\iiiri::rl:
: : . :i :
&gi
l+i:
,!'4
i:,1.
. rl
, r'
: : : I \ \ - : r 1: : : : : N
: r . . i : : :

'll . :r"i

r,1

.:::

tr
ii:

:t

.: r':

Ii:ili; iii:iil

::r:lr:ii

;iii:ii

l : l l : i : 1 . : l l: l l : i
:,.!,i...i
*.l
:. .-i
$ :{
i d

|:a:

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fs\l
;:i i:i .'::
r:,:i
\::i ,:::r
)l:
1l, r
.i:I
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.'
.:..
lt

!,

iii:..:ii::\1

FJ-j!l;a!

..i

:!

i:l

i::i.

li:

j,i

L.;,r
iJ.i*ii..:{*5*,i$lNiE'iii;iti;ii;;ii:i,,E;',li:.1'n.;
N*$$ $S $l:ls'$ ii:j il"ii.,ii".i
i.:i::t;rt Nj'.ii$
'${iiiLii',$
r:,,;l:
$:r,i-,i' $r:*ld$ f,$. ,
#ffi jirilirfi$iil:fl$if tsi:tT*iij;iirr:,;,jlr]riii.il..*
H.\11i$
:'I...'.,
...:ji*SNir"\i\S\iii\inl$ SS Sirr,,::::li:: ll:$F.$i.ll::iri'ii:r:.irtiliN$$iilst:i.;,i...:
,'rir
I ;i ,l,i:,
Siii$ *!rls $";\'i:iii'.r,,.:a..,iftlrj
i\\:.SiSli"."*,i;ji)Sii:rri:r,ilii,tir;tl;.",rll
il"ii$iN $\ii$
, ,, ,, .i,i:j *..ili.+:j:i.' !,:;ii:tll,::i;.lji;j;,il;i.ir., ,. i r,,
$r,-r:.ri-ii::
l$ nijSs
r:fol:i::lj
:ir
: r r Jf. +
s . $ :i ]
i r ii .
: ! .:r ] .. .f: ..: :li ts
\ \ \.
r .$
Le$
s ! : i:
: : rjril si: il: $ $

] - i :i r ]i i.' i ::\ i:: ... . i- :.. . . . . ' . : : '. .: .... :.. r '- :.d ] a.,.: . :.: -.' .:. i .J l.: i :.: 1,: .: \ .lr $
.i:+ ,.: ii : ;., *'; .i 1l 3i i ' . : . ] i i " . i i ] . i . , l l i i ] i i i t l [ , :
'--ii,i, ''i i .i iii' ,
,,,,,
R.\i+i $.$uiii.:,:.i"r\;if';ii, ,,,,,,:i
lij,ilSl*
i-,Nl* $jlil.X ,$\iir.E*$'irli;iirrirsr::rl,"i,ii..,iirr:l$
ti:ti$i.\i:*tiiiiirli:;iidi:,ilitil*i.:,::i.;:,:L.ir.il:lt:.ii,i:[$i*';ul$$:lS.Tt*:iifij:r:,,itr.,ii..ir ..i I
i:rtil-.S
i:* i',,ii# i:ii i "i i": .,,f 'r'
ii,:i ij,ri:,i : :l ii:,:":ri rr,\$;i:ii:i ll$ i.l,,i$ irS-"ii{riliii -ir":i
i$ iiii i:ii ri i.: Liiiiii .:ii:r"
ij. ,,',.$
ii]:ji]ii$.Ii,iii1ilri1.:i^iiitt].i!lilii:,]i!.:iif$i.:::ij:t|i:ili|iJll}'sl.ji.:ii.r':.i.;iI;lj'{ii$iHtisIi"lci:$'
Hii:iri.jilj
iiilii lril:liiiLlti,irilit4!,,iii'$$p,r,;;r,i,iliJlr*,Sr.ji:ii.rtf,;r.I;rj

FLOW OF ENERGY throughchlorophyll

molecules in the thylakoitl mernbraneis
thought to be rapid and efficient,but undi'
rected.

In

the diagrams

at left

*,,,.]; r e p r e s e r l t s a p i g r n c n t m o l e c u l e ;
nSS*:iid**SSiii*$$.#H l;" i.i',.;i,,J'a,,.'.,;
tiiis\rS*Si\\:,s;r:i:Srir;;..,rir;*s:i:$
[r,,iiSri'";Ss
$j|+jriii'i...$$..l,$i:ii:\)lI:,.|'ii.it'i$$l.T{..--$!....r$s**ili.$iiiiijrYjlHsll$'t]ii$*"?s:*ff$s$r$$',T;f$$:t,f.tf.1f'.l];3
$|ljiiii'i...$N,$i:ii:\)lI:,.|'ii.it'i$$l.T{..--$!....r$s**ili.$iiiiijrYjlHsl'u't]ii$*"?s"*ff$s$r$$',T;f$$:t'f.1f.1f'.l];sf'i:i,f
reaction center (lIactrt squ.ures)

each square
there is one
for approxi'

$tlil1|lr:*ii"ti.l#
$i,ii.:;\ii,,iJiN"iri,i:iii:iili\$ $:SiR"$ $,-\ {iiiir:liiii:ii$Fi:r,l,,i*,.iljllitji,
mately 300 antenrla molecules" In the "lake
"t$Kjtt$i{i*\; f,ii:#l liilf.'i ir;i,;##
Ri:iiiiiilii,\l Nii\,$ **i);':t ,R. il$ -$,ili$ t1::ii ;jiiiil$$*Il;riri$,$tls\\i*\{$S$i:i3;iiii$i-t
model" (top\, energy absorbed by arry mole'
$i..jiii'$H:iiiritiii$$1i$'ii$.s"
$t$:iriiiiiii$iiiilT*$;}ll$.irt
lr\'f,jir$;ii
PHOTONitiiii,i$:N $+*.t$
"g,,iiiltr{l$
$ii\llri$i$iii{i;llfi',i$iiiii+.i1-Liliiiii
i$S:$iNrN.*lS iiliii\..\iii.lij$
$ltii:rtiriii: $ili$isi.ii:i,:"\:,r\,,,.
cule can wander through the entire mass of
iij,iit$r$l##i:l;ilid trii:+ii
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$i,tr,:l.i$l
il iS S$ii*tii;l-siii,,li'.Sr$ii'sj'.
iliiii#,:* s,i.iri:$#
pigments until it is trapped in a reaction
ii"li;i+;i+i{
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$tiili**tit:,iE*Y##i.l:ilr--\il$t$tf.l;it;ll;iflt#
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center or until it is reemirted. In the "iso'
i.r.l'.,.if;:'.i'*tj.ii;jitiii.lii#Il]i$i;tii..:]#*lli$ltIiliiiij'ji$iii:il.}x$.#*iii.i;il.$f,i.',t'jf
a single re'
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i:l:.'.'3.:#ii$$iiiiiii{Iiijli:i\$$...l.ii.i+1:i}iiiiiili$$$..iit:"i:i.!]$ili$$$i!i$'sil+j$]ii};iilii:-iijiii+#;.iiiii'|:i''ij..
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within ttrre confines of tlie 30O'rnolecule unit.

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of energy transfer, rvhich
Tbe mechanism
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7B

L,

if

pigment systemI for e 72O-nanometer

band.
In living cells the intensity of chlorophyll fluorescencechanges as a function of time. This efiect, discoveredby
the late Hans Kautsky of the University
of Heidetberg, consistsof a fast change,
completed about two secondsafter the
plant is illuminated, and a slower fluctuation that achieves a steady state only
after severalmimtes.Both temporal patterns could reflect e shifting relation
between pigment systemsI and II. The
fast change is sensitive to the rate of
electron flow through the transport system and might be explained as incating that pigment system II briefly gets
ahead of system I, and wastes the surplus energyin fluorescence.
The slow change is thought to be related to temporary alterations in the
thylakoid membrane. One hypothesis
suggeststhat if the two pigment systems
are spatially close together when the
light is first turned on, energy from pigment system II, which is highly fluorescent, might "spill over" into the weakly
fluorescent system I, thereby diminishing the overall light emission.If the two
systems were then separated, the spillover would be eliminated and the fluorescencewould increase.The conformaonal change in e thylakoid membrane
required by this theory might be caused
'by the movement of ions. An alternave
explanation postulates changes in the
density or spacing of ctorophyll molecules in pigment system II. Separating
the moleculesfrom one another could be
erpected to decreaseinternal conversion
and ths to increasefluorescence.
The lifetime of an excited state in a
fluorescent molecule is an important
parameter in photosynthesis, since it
gives an indication, albeit an indirect
one, of how much time is available for
e energy of excitation to reach a chemically active molecule. The lifetime is defined as the time required for fluorescence to decay ta L/e of its maximum intensity (a is approximately 2.7). As we
have mentioned, the lifetime of e higher singlet states is extremely brief, from
10-1{to 10-1ssecond;thesestates,however, do not enter into photochemistry.
The first excited. singlet state is much
longer lived. Its lifeme was first measured by Seymour S. Brody at the University of Iinois and by Aleksander N.
Terenin and his colleaguesat Leningrad
State University. New measurementsin
our laboratory of the lifetime of chlorophyll a in living cells, made with photosynthesisstopped by inhibitors or by low
temperature, or made in light so bright

A
e-'
t\-/

PHOTON

DONOR
PIGMENT

t\-/
o-'
ACCEPTOR

ea
^4.
( e-)

\_-/

,r ,\

DONOR

PIGMENT

ra
\-/
ACCEPTOR

l--l
DONOR

\\A

t/
PIGMENT

t?f
\--l

'ACCEPTOR

*uphill,"
TRANSPORT
against an electrochemical gradiento in the re.
OF ELECTRONS
action center of the chloroplast takes place in three stages. On abeorbing a photon (a), an
electron in a pigment molecule is promoted to an excited state. The electron can then be
transferred to the acceptor molecule (b); because the pigment is in an excited state this
process is 'odownhill,' that is, it is favored by the electrochemical grailient. fn the final
step (c)o the pigment regains an electron from a donor molecule, another downhill process.

it saturates the photochemical system,

yield a lifetime of about 2 x 10-e second.
Michael Seibert of the General Telephone and Electronics Research Laboratories and R. R. Alfano of the Cty College of the City University of New York
have recently employed a laser and an
instrument capable of marking extremely brief intervals in measuring fluorescence lifetimes in chloroplasts. At the resulting lorv average intensities they discovered two fluorescing species, one
with a lifetime of less than 10 X 10-12
second, the other with a lifetime of about
300 X 10-12 second. From the emission
spectra they have suggested that the two
species are molecules of ctrlorophyll in

different environments,the more rapidly

decayingone making up part of pigrnent
system I, the other contributing to pigment systemII.
The Transfer of Energy
Most of the pigment moleculesin the
chloroplast do not take part directly in
the chemical processesof photosynthesis. Of the 300 molecules in the basic
photosynthetic unit, almost all are
thought to serve merely as "antenna"
molecules. (The metaphor is not farfetched; after all, the molecules are
tuned to receive signals of a particular
wavelength.) The antenna pigments

lrurrrr", the energy they absorb to a reaction cbnter, which might consist of a
single specially deployed molecule of
chlorophyll . This molecule then introduces the energy into the oxidation-reduction cycle.
If such a systemis to operateefficiently, some method of rapidly transferring
the energy of excitation from one molecule to another must be provided. That
energy is indeed transferredis suggested
simple observation: green leaves
by
"
chlorophyll b, which is fluorescontain
cent in isolation, yet in living cells only
the fluorescenceof chlorophyll a can be
detected. Evidently the accessorypigments transfer their energy to chlorophytl so efficiently that their own fluorescenceis quenched.The effect can be
demonstrateddirectly in monomolecular
layersof mixed chlorophyll and b; even
when the mixture is'illuminated at a
wavelength that will stimlate only chlorophyll b,only chlorophyll fluorescence
is observed.
Among the accessorypigments a sequence of energy-carriers has been
worked out. It begns with the carotenoids, then passesthrough phycoerytlrin, phycocyanin and allophycocyanin

(another phycobilin) to clorophyll. In

plants that have no phycobilins, energy
can be transferred directly from the
carotenoids to the chlorophylls, but
when the phycobilins are present they
are invariably utilized in sequence. Each
transfer from one kind of pigment to another involves a modest loss of energy as
fluorescence or heat fsee bottom illustration on page 771.
Once the excitation energy has been
passed to a chlorophyll molecule it can
migrate through the entire population of
chlorophyll a molecules without loss.
That the migration does take place was
ffrst shown by Arnold and E. S. Meek at
Oak Ridge in 1956. Arnold and Meek illuminated chloroplasts with plane-polarized light and observed the resulting fluorescence. If the exciting energy had
been retained by each molecule during
its fluorescence lifetime, the polarization
would have been maintained in the fluorescent light; it was in fact destroyed
entirely, indicating extensive energy migration.
One possible explanation of the
mechanism underlying this migration is
ealled the Frster resonance theory, formulated in 1948 by the late Theodor

Frster of the Technical University in

Stuttgart. The theory holds that a molecule must first subside to the lowest vibrational sublevel of the ffrst excited singlet state before its energy can be transferred; once this requirement is met the
molecule is coupled to its neighbors by a
resonance fundamentally dependent on
their distance and orientation and on the
overlap between the fluorescence spectrum of the donor and the absorption
spectrum of the acceptor.
Faster mechanisms, in which the
molecule need not first relax to its lowest
vibrational state, have been proposed.
They cannot yet be excluded, but calculations based on measurements of fluorescence lifetime support lrrster's theory. An upper limit to the time required
for energy transfer has been provided by
an experiment devised by Peter M.
Rentzepis of Bell Laboratories. Working
with bacterial photosynthdtic systems
that had been stripped of all antenna
pigments, so that only the reaction centers remained, he observed an energy exchange in 6 r- 2 x l0-rz second.
The path taken by the migrating excitation energy in the antenna pigments
is thought to resemble a "random walk"

{
I

I
i

+I

PHOTON

PHOTON
J
r,",-

PIGMENT SYSTEM II

PIGMENTSYSTEMI

cARoTENoLS

cARoTENES

/
I

cHLoRoPHYLLb
\
a 660
.-C_HLoRoPHYLL

\
CHLOROPHYLLb

CHLOROPHYLLa 670

CHLOROPHYLLa 660

EXCHANGEOF EXCITATIONENERGY

cHLoRoPHvutaorc
f

>\

CHLOROPHYLLa670

CHLOR}

px-ir-i'"lst

CHLORO-

\envur

CHLOROPHYLL a 678

aaoo
f

705-720

CHLORO.
P H Y L La ,

\oasl

REACTIONCENTE

REACTIONCE
P700

e-

/--\
OF PIGMENTS in the higher planrs is difierent
DISTRIBUTION
in the two pigment systems.'Among the csrotenoids, carotenes
dominate in system I and carotenols in system II. Chlorophyll is
more abundant in pigment Bystem II. Chlorophyll a is the most im.

P080
e-

----\
portant pigment in both systems, but each has a characteristic assortment of forms of this pigment, distinguished by their absorp'
tion spectra $lach) and florescence spectra (color). The reaction
center pigments of the two systems also exhibit different spectra.

that continues until the energy qttrrtrtrttlt

happens to encounter an availrlrlel-eaction ccntcr, or until it is rentcliatcd by
fluorescetce.The reactiotl cclltcrs tllcllselves might be dispersed randomly in a
monolithic bed of bulk pigment, so that
any quantum of euergy could '"vandcr
throughout the mass and in principle
could eventually rerch any of the reaction centers. Alternatively, each reaction
center rnight be surrounded bY, and
served exclusively by, a fixed aggregate
of pigment molecules. These trvo hypotheses are called the "lake model" atr<l
lhe "isolated puddle model." The actual
situation apPears to be a hybrid of the
two, calling for a "conuected prrddle
model" lsee illustrations on page 7Bl'
The Reaction Center

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The elaborate system of pigmentation

in plant cells is intended solely to convey the er)ergy of light qttatrta to rcaction centers where the centrai events of
photosynthesis take place. Learning the
nature and the exact business of the reaction center has been an intractable
problem in the study of plant metaboIism, but some of the molecules active
there have now been at least labeled
spectrally if not identified chemically.
Louis N. M. Duysens of the University
of Leiden discovered the first of these
molecules when he observed a light-induced change in the absorption sPectrum of photosynthetic bacteria at 890
nanometers; the change was later sholvn
to be produced by bacteriochloropliyll in
the reaction center. Kok identified a similar change in the absorption of greerr
plants at 700 nanotneters; this light-indrrced absorption change is norv knorvn
to be caused by the reaction-center molecule of pigment system l, designated
P700. Exposure to light l-ras been demonstrated to oxidize P700. The analogous
molecule for pigment systern II was discovered by G. Doring and Ilorst T. Witt
of the Technical University in West lJerlin and is labeled P6B0 or l'690; whether
or not it is a direct party to au oxidationreduction process has not been definitely
determined.
These specialized pigmcnt rnolccules
appear to stand at the culmination o{ the
physical, energy:gathering stage of pliotosynthesis, and at the beginning of the
chemical, energy-storing phase. There
are a number of waYs of determining
whether or not they do mark that intersection. The chemical Processes they
participate in should be the first to take
plu"" after a brief flash of light, and they
shoulcl be in the main pathrvay of photo-

synthesis. F-rom experiments on the inhibition of photosynthesis by low temrcratrrrc we know that the reactions
slrould lrlsccontinuo cvcll at vcry low
ternperattrres (belorv 77 degrees Kelvin,
or 77 degrees Celsius above absolute
zcro). Thcse tests have been made and
the evidence that has accumulated is encouraging. P700 is in fact oxidized by
light at 77 degrees Kelvin (although the
pigment cannot be fully reduced at that
ternperature). The absorption change associated with P6B0 or P690 has been
observcd rtlow temperaturcs. A seriesof
calculations, beginning with early measurernents made by Kok, show that the
o.ridrtion of P700 is a highly efcient
reaction, and tlierefore it must be part of
the nrain pathway. (If it was not, it
'uvould compete successfully with the
nain course of events.)
Finally, if P6B0 and P700 ale the active molecules of the reaction center,
thcy rnrr.stbc capable of initiating the sequence of events that leads to the transfcr of electrons. They could do so, when
they are in the photoexcited state, by
donating one of their own electrons to
the acceptor molecule, then recovering
iL r,vhen in the ground state from the
clonor lsee illustration on page 79]. In
the case of pigment system II, for example, P680, after absorbing a Photon,
would transfer one of its valence electrons to the molecule labeled Q and in
the process would return to the ground
state. It could then accept an electron
frorn the donor molecule Z. This hypothesis irnplies that the reaction-center pigments bear a positive charge during the
electron transfer, and the presence of
chlorophyll ions in the chloroplast has
inclccd been demortstrated lly electronspin-resonance techniques that monitor
unpaired electrons.
If this sequence of events is correct,
sorne means must be provided to prevent
Z and Q from reacting with each other
to annul the charges developed on them.
A direct reaction between them is thernrodynatnically the most favorable of all
rcactions, rndit is known to take place,
although only to a minor extetrt' (It is responsible for the delayed light emission
dis"overed in l95l by Arnold and Bernard L. Strehler at Oak Ridge.) The
back-reaction betweenZ and Q must not
l>e allowed to dissipate a significant
unount of the al>sorbed erlcrgy, however, if photosynthesis is to achieve useful results. What mechanism intervenes
to scparate them is for llow entirely unknown. That is only one of several mysteries, however, that remain to be solved
by furtlrcr shrdy of photo.synthesis.