T'S NOT love that makes the world go round, it's photosynthesis .

Virtually all plants and animals depend on it . Photosynthesis is the process by which plants trap sunlight and turn it into chemical energy. Today, most plants take in carbon dioxide from the air and combine it with water using light energy, to produce sugars and oxygen . It is a complicated chemical process that took a long time to evolve . Even now the process has its shortcomings . The most obvious one is that it does not make the most of the light available. Plants look green because they contain a pigment, chlorophyll, which absorbs light at the red and blue ends of the spectrum and uses it for photosynthesis . The plant reflects the wavelengths in between, principally those of green light; this is what we see . Ideally, a photosynthetic pigment should be black, that is, it should absorb light at all visible wavelengths so that it wastes none of its energy . So why did nature choose green? To find an answer, we first have to go to a salt lake and then back in time, almost to the beginning of life . The salt lake probably contains an unusual photosynthetic bacterium, called Ilalobaclerium holohium . This species bears all the hallmarks of a living fossil . Halobaclerium lives in salt lakes where the concentration of salt may be as high as 20 per cent (seawater is only 4 per cent). Almost all other organisms would die in salt at this concentration, Halobacterium dies if the salt concentration falls to levels at which other organisms can grow . The bacterium is therefore isolated in an environment free of competition from other organisms. Because of this, much of evolution has passed it by ; it still

The countryside is green. In an ideal world it would be black . A new theory explains why Andrew Goldsworthy
has many of the attributes of early forms of life . One of Halohacterium's ancient characteristics is that it has unusual membranes. These contain branched terpenoids in place of the unbranched fatty acid molecules that modern organisms have in their membranes. Perhaps terpenoids predate fatty acids as components of the cell's membranes. The way lIalobaclerium photosynthesises is also extremely primitive. The bacterium has no chlorophyll. Instead, it contains a purple pigment, bacteriorhodopsin, so called because it is similar to rhodopsin, the light-sensing pigment in the eyes of animals. lIalobacterium's photosynthetic pigment is based on a terpenoid molecule of the same size as those in its membrane and might have evolved from them . It is difficult to say exactly how old Halobacterium is because organisms as simple and as small as this leave no fossils-at least not that we would recognise easily. However, Melvin Calvin, who discovered a vital biochemical pathway in photosynthesis (the Calvin cycle) found terpenoids resembling those in Halobacterium's membranes in rocks about 3000 million years old. If these compounds are relics of organisms similar to Ilalobacterium, they must have been widespread by this time . They, might have originated much earlierperhaps close to the dawn of life, which was probably between 3000 and 4000 million years ago. We know a good deal about the photosynthesis and metabolism of Ilalobacterium, mainly as a result of research in the 1970s by Walther Stoeckenius of the University of California at San Francisco. Unlike organisms that contain chlorophyll, Halobacterium cannot take carbon dioxide and water and

Why trees are green

New Scientist 10 December 1987

49

convert them to sugars by photosynthesis . It needs a supply of organic substances to grow . Instead, the bacterium uses light energy to swim, to absorb nutrients and for making adenosine triphosphate (ATP), which, in turn, makes energy for other metabolic reactions (see Box 1) . Such an organism would be well suited to the conditions in the so-called "primeval broth" of complex organic materials when life was just beginning . The chemicals in the "broth" had gradually accumulated in the seas over millions of vears as a result of the action on relatively simple chemicals of ultraviolet light and electrical discharges in the atmosphere . The purple colour of bacteriorhodopsin probably resulted from natural selection for a photosynthetic pigment that absorbed a broad band of wavelengths in the middle of the visible spectrum, Only wavelengths in or near the visible region of the spectrum have the right amount of energy to be useful in photosynthesis . Furthermore, water strongly absorbs light outside this region, so the light would not have been available to the aquatic organisms in which photosynthesis first evolved . Bacteriorhodopsin lies in the bacterium's external membrane ; it needs light energy to pump positively charged hydrogen ions (protons) out of the cell . As a result, a concen-

tration gradient builds up across the membrane and the bacterial cell becomes negatively charged relative to its surroundings . The cell then releases the energy stored in this gradient when it allows protons back into the cell . This process drives the active uptake of nutrients, provides power for the whiplike flagella that propel the bacteria and generates ATP . Organisms like Halobacterium would have had the advantage over earlier forms of life that depended for their energy on the anaerobic fermentation of nonrenewable resources in the primeval broth . The advent of these simple photosynthetic bacteria provided an almost unlimited supply of metabolic energy . The bacteria could even recycle the spent products of earlier fermentations, using solar energy . The biosphere gained a new lease of life . These simple photosynthetic organisms must have been extremely successful compared with their nonphotosynthetic ancestors and they would probably have become the dominant form of life . In those days, the seas might well have been purple . Successful as they might have been, purple bacteria such as Halobacterium had a serious failing that ultimately led to their downfall . they could not fix carbon dioxide . The fermentation reactions of all the nonphotosynthetic

ORGANISMS similar to Halobacterium ~,J were probably among the first to use light energy for photosynthesis . With it, they could absorb food more efficiently and make virtually unlimited amounts of ATP to provide energy for metabolism . These organisms probably pioneered the chemiosmotic production of ATP which is now almost universal . As in the rest of evolution, Halobacterium's type of photosynthesis and its coupling to transport and the synthesis of ATP must have arisen in small manageable stages, with each step having a selective advantage over the one before, We will probably never know exactly how it happened, but we can make some educated guesses . The first step was probably the evolution of selective systems of passive transport, followed closely by the development of active transport and the production of ATf, The earliest organisms had very simple metabolism and obtained most of their metabolites ready-made from the primeval broth . Their cell membranes . like those of organisms today . had to control the passage of materials to allow the entrv of nutrients but prevent the loss of metabolites . To do this, most of the membrane is made of oily materials. Although the membrane is only two molecules thick, the water-soluble contents of the cell cannot dissolve in it and remain trapped within the cell . To allow the entry of nutrients that are soluble in water, there are "carrier" proteins "floating" in the membrane . Their job is to recognise the nutrient molecules and let them in . They do this by combining reversibly with specific nutrients. The resulting compound is free to change its shape so that the part attached to the nutrient can pass freely back and forth through the membrane . These simple systems are selective . but allow passage only down a diffusion gradient . Sometimes, organisms need to transport materials actively against a diffiision

l : From purple pigment to modern methods of making energy

gradient from a dilute solution into a expelled protons yielded an added bonus ; it concentrated one . This needs a pumping provided the basis for the so-called comechanism driven by energy . Probably one transport mechanisms that Halobacteriumn of the first systems of active transport to and virtually all modern plants and bacteevolve was a pump for the excretion of ria use to take up nutrients . To do this, all hydrogen ions (protons) . The cell generates that was needed was a slight modification protons in the course of metabolism . It of the carrier molecules to give them a must remove the protons, usually against a binding site for protons next to that for the diffusion gradient, to keep the pH of the nutrient . Movement was permitted only cell at the right value for its enzymes to when both sites were full or both were function properly . This may require a empty, so that protons and nutrients considerable input of energy because the always crossed the membrane together . By concentration of hydrogen ions outside a linking the passage of the two, the steep cell may be more than 100 times greater diffusion gradient for protons trying to rethan that within . enter the cell could overcome a weaker Modern organisms contain three main diffusion gradient for nutrients trying to types of proton pump depending on their leave . Thus, cells could absorb nutrients source of energy . Some are driven by the from solutions that were more dilute oxidation-reduction reactions of respirathan its own contents . Mechanisms such as tion, some by photosynthesis and others by this enabled organisms to continue to ATP . Those driven by ATP probably absorb nutrients as the primeval broth ran evolved first because the primitive atmo- out of carbon . sphere did not contain oxygen and photoBacteriorhodopsin probably brought an synthesis has to stop at night . But ATP even more important bonus . The very steep generated by anaerobic fermentations proton gradient created by its photocould provide the steady source of energy chemical reactions could drive the ATPneeded to keep the cell's pH constant day linked proton pumps backwards . Protons and night . pumped out of the cell by the light could Unfortunately, anaerobic fermentations now flow back in, generating ATP as they are very inefficient at producing ATP, and did so . Because the force that drives this with the depletion of the primeval broth, reaction is similar to osmosis, the process is the necessary substrate was running called chemiosmosis. This may have been out . The first proton pump driven by how the chemiosmotic production of ATP light, perhaps based on bacteriorhodopsin, in virtually all modern organisms first must have given its owner a considerable evolved . 11 advantage . Bacteriorhodopsin consists of a single protein molecule attached to an unsaturated terpenoid (retinal) which gives it its colour . It lies in the external membrane and . when it absorbs light, it changes its shape and thrusts a proton out of the cell . The evolution of a system that actively Halobacterium halobium, theJirst photosynthesiser?

so

New Scientist 10 December 1981

The mast advanced plants have the least <fcient system for absorbing light. :Mosses, ferns and higher plants are "backtivard"in their photosynthesis . Brown algae (right) have extra pigments to make the most oJ'the light that filters through the murky ivater

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Accessory pigments absorb more light.

organisms in the broth generated a great deal of carbon dioxide as a waste product . In the absence of any effective means to refix the carbon dioxide into organic chemicals, this must have led to a steady loss of carbon from the biosphere . If the broth ran out of carbon, life would almost certainly have become extinct . Fortunately, an organism with the potential to fix carbon dioxide was waiting in the wings. This organism was probably a bacterium that lived deep underwater on the surface of sediments . The organism had a different photosynthetic pigment, probably similar to chlorophyll a, the green pigment that modern plants contain . Like bacteriorhodopsin, chlorophyll lies in the cell membrane . But unlike bacteriorhodopsin, which functions on its own, chlorophyll works in conjunction with a number ofother molecules in the membrane, not only to pump protons out of the cell but also to pump electrons in . This capacity to pump electrons enabled it to reduce carbon dioxide to sugars {see Box 2}, In simple chemical terms, chlorophyll uses light energy to drive a system of redox reactions (reduction and oxidation), to reduce carbon dioxide using what, at first sight, would

seem an impossibly weak reducing agent. Bacteria with chlorophyll probably evolved as a result of natural selection as the supply of organic materials in the primeval broth dwindled and carbon dioxide was the only major source of carbon left. Life still would not have been easy for these bacteria. They needed a supply ofsuitable compounds in their environment to provide them with electrons for their type of photosynthesis . As with modern photosynthetic bacteria, these organisms probably used compounds from decaying organic matter in the sediments on which they lived . They might have made use of a variety of electron donors, but the most important were probably sulphur compounds such as hydrogen sulphide . The primeval broth had an ample supply of sulphur compounds because the nonphotosynthetic organisms in the sediment produced them as waste products during anaerobic fermentation . As the sediments on which the new type of photosynthesisers lived were deep underwater, the bacteria must have been short of light. What little light they had was at the red and blue ends of the spectrum-all that remained after

centre) absorb red and orange light. Red algae (top) are almost the ideal colour and absorb even green light

1he primitive cyanobacteria (above and

New Scientist 70 Qecerriber 7987

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using electrons taken From a donor molecule . The transfer of' electrons between molecules is called a redox reaction: one substance is oxidised when it donates electrons to another, and the second compound is reduced in the process . For electrons to flow between molecules in this way, the receiving molecule must have a greater attraction for the electrons than the donor. This difference can be measured as a voltage, or redox potential . Most coinmonly available compounds in the environment do not release their electrons with enough potential to reduce carbon dioxide. Photosynthesis based on chlorophyll solved this problem. Unlike bacteriorhodopsin, which uses light energy to transfer protons across the cell membrane . chlorophyll uses light to transfer electrons . Chlorophyll takes these electrons from relatively weak donor molecules outside the cell . When light excites the chlorophyll, the pigment molecule ejects these electrons into the interior with enough energy to reduce carbon dioxide. The chlorophyll does not reduce the carbon dioxide directly . Instead . the electrons it ejects feed into a chain of electron "carrier" molecules which pass them on from one to another until they finally enter a biochemical pathway called the Calvin cycle. 'I he reactions in the Calvin cycle reduce carbon dioxide to sugar. The cycle needs ATP as well as electrons. Photosynthesis with chlorophyll provides the ATP . Some of the electron carrier molecules are arranged in the membrane so that as they transport the "high-energy" electrons coming from chlorophyll, they combine with protons on the inside of the cell and release them on the outside. In this way. some nf' the energy of the ciectrom helps to generate a proton gradient across

P HOTOSYNTIIESIS dioxide chlorophyll with reduces carbon to sugars

Z : Photosynthesis with green pigments

the membrane for the manufacture of ATP. If the cell needs extra ATP for any reason . some of the electrons, instead of being passed to carbon dioxide, are returned to the starting point via a chain of electron carriers and go through the photochemical system again . Each time the v electrons pass through the circuit they add to the proton gradient and d generate ATP. In modern photosynthetic bacteria, chlorophyll is concenCareen plants keep their chloruphy11 in chloroplasts trated in folds in the outer membrane called chromalophores . energy from the other chlorophyll moleThis enhances the effectiveness of' the cules and use it to transfer electrons across proton transport system . After crossing the the membrane . membrane, the protons remain confined In bacteria, the photosynthetic memwithin the chromatophore [which, tech- branes are simply infoldings of the cell's nically, is outside the cell] and build up a outer membrane . In higher plants, the steeper gradient . The steeper the gradient, photosnthetic membranes lie within the more efficient the production of A 1'P special' organelles called chloropiasls . when the protons return to the cell . Chloroplasts bear a remarkable resemThe system of electron carriers and blance to photosynthetic bacteria, but they enzymes that services the chlorophyll are surrounded by an extra membrane . molecule is complex and bulky . To avoid Biologists think that chloroplasts originated the need for such a system for each chloro- when a primitive, nonphotosynthetic phyll molecule, photosynthetic organisms organism engulfed one of the types of evolved a phenomenon called resonance photosynthetic bacteria that produce transfer, which allows many pigment oxygen . Once inside a vacuole (now the molecules to share the same enzymes. outer membrane of the chloroplast), the Resonance transfer enables one pigment bacterium lived in a svitthiotic relationship molecule to transfer to another the energy with the organism that had engulfed it . The it absorbs from light. Chlorophyll mole- inner membrane of the chloroplast . which cules are arranged in groups of between corresponds to the outer membrane of the about 5(1 and 3U(1, called photosynthetic original bacterium, is thrown into a units. At the hub of each unit . at the so- complex pattern of folds, the thylakoids, called reaction centre, are specialised which contain chlorophyll and correspond chlorophyll molecules that collect the to the bacterial chromatophores . 0 As the biosphere ran out of organic compounds, the ability to fix carbon dioxide became more important. The green bacteria began to have the advantage as the purple bacteria starved to death . Life had won a reprieve when organisms began to fix carbon dioxide using electrons from sulphur compounds. But the relative scarcity of electron donors limited the amount of carbon dioxide these organisms could recapture from the atmosphere . Salvation came in the form of photosynthesis that produced oxygen . The principle of this process is simple . Instead of taking electrons from relatively scarce sulphur compounds, organisms took them from water. of which there is an almost unlimited sit pplv . The removal of electrons from water to reduce carbon dioxide results in the decomposition of water molecules with the production of oxygen . This apparently simple act, however, involved a huge change in biochemistry . The voltage that a single lightenergised chlorophyll molecule can generate by its electronpumping action is not enough to strip electrons from water and give them to carbon dioxide. To accomplish this, nature used the simple expedient of taking two such electron pumps and arranging them in series, rather like putting two cells into a flashlight to obtain a higher voltage. A second photochemical electron pump evolved alongside the first. A chain of electron carriers connected the two electrically_ so that electrons had to pass through each pump in turn on their way from water to carbon dioxide and so received a double dose

the purple bacteria swimming above them had taken their share. It is not surprising, then, that they evolved a photosynthetic pigment that absorbed specifically at these wavelengths. The red and blue absorption peaks of chlorophyll a complement almost exactly the green absorption peak of bacteriorhodopsin ( see Diagram below) . Modern photosynthetic bacteria that live in sediments are similar in this respect . However, because modern bacteria must cope not with purple bacteria swimming above them, but with organisms containing chlorophyll, they contain special chlorophylls, such as bacteriochlorophyli, in which the main absorption peaks are pushed even further outwards into the infrared . The absorption spectrum rf chlorophyll a superimposed on the absorption spectrum of, membranes containing hacteriorhndop5in (shadedj . Chlorophyll's absorption peaks ft neat[ oneither side of bacteriorhudnpsin's

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350 400 450 500 550 600 650 700 Wavelength (nm)

The absorption spectra of the main accessory pigments from the red algae. Light energy passes from left to right along this chain of overlapping pigments and then to the red absorption peak of chlorophyll a

chlorophyll, which absorbs red light

Why leaves are green the cells within a leaj'are packed with

of energy . The old electron pump (now called photosystem 1) delivered electrons to the systems that fixed carbon dioxide, and the new one (photosystem 2) removed the electrons from water and delivered them to photosystem 1 . The evolution of photosystem 2 was perhaps one of the most important steps in evolution . For the first time, fixation of carbon dioxide was independent of the amount of decaying organic material and sulphur compounds in the environment. As a result, life began to flourish . The amount of carbon in the biosphere grew steadily until the carbon dioxide had all but disappeared from the atmosphere and the massive amounts of oxygen produced allowed the evolution of the animal kingdom . By the time this new form of photosynthesis had evolved, most of the purple bacteria had probably disappeared . This meant that much more light in the centre of the spectrum became available and natural selection sought ways to use it . Yet black plants-which would absorb all this light--did not evolve . Because photosynthesis is such a complex process, it would have been difficult to exchange chlorophyll for another, fundamentally different, pigment . Instead, a process evolved in which a number of "accessory pigments" absorbed light in the hitherto unused centre of the spectrum and passed their energy on to chlorophyll by a simple physical process called resonance transfer . Different plants have evolved with accessory pigments absorbing at different wavelengths . Often they have several such pigments with overlapping absorption bands so that they make the most of a large part of the spectrum (see diagram above) . Perhaps the simplest organisms to make extensive use of accessory pigments were the cyanobacteria . Their pigments, phycocyanin and allophycocyanin, absorb in the orange region, but even here they d not cover the whole spectrum . An improvement came with the evolution of red algae . Red algae have not only the orange-absorbing pigments of the cyanobacteria but also contain large amounts of a red pigment, phycoerythrin, which absorbs in the green . Between them, these pigments cover a much larger part of the spectrum . Despite their name the red algae tend to be very dark, approaching black-the ideal colour. Similarly, the dark colour of brown algae, the familiar seaweeds, is . due to a combination of chlorophyll and the accessory pigment Andrew Goldsworthy lectures in plant physiology and biochemistry fucoxanthol . Both red and brown algae usually grow in relaat Imperial College, London . tively deep water where the light is dim . The pressure on

plants to absorb light more efficiently has produced accessory pigments that cover a good range of the spectrum . Where selection pressure has been weaker, plants have not developed such a range of pigments. Green algae, which live near the surface of water, and the land plants that evolved from them, are not so short of light and depend almost entirely on chlorophyll . Green plants have two kinds of chlorophyll . Chlorophyll a is the main photosynthetic pigment ; chlorophyll b, which has two peaks of absorption near the centre of the spectrum, functions only as an accessory pigment . Although chlorophyll b absorbs some of the light missed by chlorophyll a there is still a large gap between its twin absorption peaks where light escapes unabsorbed . Carotenoids partly filled this gap, absorbing light towards the blue end of the spectrum . But these orange and yellow pigments do not pass their energy efficiently to chlorophyll and no pigment absorbs significantly ~in the green, Consequently, the plant wastes a considerable amount of energy in the middle of the spectrum and reflects green light . Paradoxically, the most advanced plants have the least advanced system for absorbing light-because they grow in adequate light and do not have to extract as much energy as possible from light . This "backwardness" does have a bonus . Few of us would like to see our green countryside turn black in the interests of photosynthetic efficiency . Instead, we may owe our thanks for pleasant green surroundings to the sea of purple bacteria countless millions of years ago . These tiny purple organisms probably provided the selection pressure that made modern vegetation green . Not only that, their photosynthetic pigment gave rise to our visual pigment and so gave us the eyes to see it . 11 7 _r