ae Molecules C1.3 Photosynthesis ‘There are many different forms of energy, such as chemical ‘changed into chemicaleneray. The carbon compounds the energy that previously was light. 1. Converting light energy to chemical energy in photosynthesis energy, heat (tnermal energy) and light (a form ofradiant energy), Energy can be changed from one form into another. Lightenergy istransformediintorchemicalienerayin photosynthesis. This happens when light is absorbed by pigments (coloured substances such as chlorophyll} and is iat are produced iny photosynthesis contain the chemical 2. Carbon dioxide is converted to glucose in photosynthesis The ratio of atoms in a glucose molecule is carbon : 2 hydrogens : | oxygen. The ratio in carbon dioxide molecules s 1 carbon : 2 oxygens. To convert carbon dioxide into glucose, half of the oxygen must therefore be removed and hydragen must be added Hydrogen is ebtained by splitting water molecules into thydrogenandiaxygen. Tris can only happen when light is avaiable because eneray is required, Twelve water molecules are spit for each glucose molecule made: 12H, 24H +60, Halfof the hyclrogen atoms become part of a glucose molecule, The other half are used to remove one oxygen atom from carbon dioxide and convert ito water: 6CO, + 24H C,H,,0, + 6H,0 3. Equation for photosynthesis Synthesis of glucose by photosynthesis can be summed up with a simple equation: carbon dioxide + water + ight > glucose + oxygen Oxygen isa by-product of spliting water to release hydrogen, Three groups of organisms carry out photosynthesis: plantsyalgaeand:cyanobacteria. These organisms use some ofthe axygjen that they produce in aerobic cell respiration (for example the roots of plants) but most of itis excreted into the environment. In organisms that ve underwater, the oxygen produced during photosynthesis can sometimes be seen emerging as bubbles, which rise to the water surface. In terrestrial organisms the oxygen diffuses into the atmosphere unseen, 4. Chromatography Tear up a eafinte 2. Gina the leafpieces wth small pieces sharp sand and propanone to extractthe leaf pigments pestle F 5, Add afew drops of propanone te dissolve the pigments mortar transferring tiny drops of solution| Jom 8. Remove the srp from the tube 8 when the running solvent has nearly reached thetop. Drawa penciling to value. Ryis 6. Buld up a concentrated spot 10 mm from the end of a stip of chromatography paper or LC strip {thin ayer chromatography) by 3. Transfera sample of etractto awatch glass 4. Evaporate to ddpyness with hot air from ahairlyer = 7. Suspend the stip ina ‘ube with the base dipping into running solvent chromatography strip concentrated spot of pigment running solvent The pigmentin each spot ean be ilentfied omits colour and ts Ry distance maved by a sistance moved by spot '~ Gistance moved by solver! showhow fr stance moved byaspot spot, asa proportion ofthe distance moved by the salvent. Pigments move atdifferentrates, The rate depends carotene (0.9) — chlorophyll 9(0.65) chlerophyl 6 (0.6) $ cistance moved by solvent ‘on whether pigment is more atvracted 4 tothe hydrophobic running solvent orto xanthophyll zal the hydrophilic chromatographystrip. (0.30.5), Typical Rvalues are shown on the agrar right 32 distance:Molecules co (EJ Wavelengths of light absorbed in photosynthesis ‘The Sun emits @ wide spectrum of electromagnetic radiation, Particularly large amounts in the range of wavelengths from 400 nm to 700 nm reach the Earth's surface and this range is used both in photosynthesis and human vision. Violetlighthasithe:shortestwavelength and red has the longest. The shorter the wavelenatth, the more energy photon of light has Pigments such as chlorophyll have a structure that allows, an electron within the molecule to jump from one energy levelup toa higher level, using energy obtained by absorbingaphotonoflight. This is how solar energy is transformed into chemical energy in photosynthesis. The electron is said to be “excited. Rhotosyntheticpigments ‘can pass excited electrons on to other molecules. Much ofthe energy camed by the excited electrons ends up in glucose or other carbon compounds Forany photosynthetic pigment, only specific wavelengths of light have the amount of energy needed to raise an electron toa higher energy level and only these wavelengths are absorbed, Other wavelengths are reflected. A graph showing which wavelengths are absorbed is an absorption spectrum, Key —chlorophyila <== ehlorophyllb 0 4860 500 550 600 650 700 violet indigo blue green yellow orange red wavelength of light/ nm: Although chlorophylls are the main pigments in photosynthesis others are used, including xanthophylland carotene, | 6. Comparing absorption and action spectra ‘A.graph showing relative amounts of photosynthesis at different wavelengths of ightis an absorption spectrum, The date needed for its obtained experimentally ‘The wavelength of ightis the independent variable. It ‘can be varied using LEDs or colour filters. ‘The dependent variable isa measure of the rate ‘offphotosynthesis. This can be the rate of oxygen production or carbon dioxide consumption. Bubbles of ‘oxygen rising from 2 pondweed can be counted or the rise in pH due to CO, consumption can be measured. bubbles of oxygen rising to the water surface pondweed inverted and with a cut stem LEDs emitting a specific wavelength A 20 0 4860 500 550 600 650 700 violet indigo blue green yellow orange red wavelength of ight /nm %useo 338 2 Molecules 7. Limiting factors on the rate of photosynthesis Processes such as photosynthesis are affected by various factors, but usually just one of these factors is actually limiting the rate ata particular time. Thisis the factor that is nearest to its minimum and is called the limiting factor. ‘The three possible limiting factors for photosynthesis sociumhydrogencatbonate solution dea tovarythe carbon dioxide concentration \ \- 7] bubbles Gf stones | | pendent | tht Source that tanbemovedt | dlsancns ovary veterats theight otenaty tempersre (—themosately conte at [inating factor | Method of varying the factor Temperature | Place pondweed nwaterina thermostatically controlled water bath oron ahot plate to vary the | suggested range S°Cto.45°CinS oF 10°Cintervals ‘These principles should be remembered when designing an experiment to investigate the effect of a Imiting factor on photosynthesis 1. Only one limiting factor should be investigated at a time—thisis the independent variable 2. Asuitable range for the independent variable should bbe chosen, from the lowest possible level, to alevelaat which the factors no longer limiting 3. An accurate method should be chosen for measuring the rate of photosynthes's. This is the dependent variable and is usvally a measure of oxygen production pper unit time. 4, So the independent variable is the only factor affecting the rate of photosynthesis, all other factors must be kept constant. These are the control variables. Of the three factors temperature, ight intensity and CO, concentration, ane will be the independent variable and the other two will be controlvariables. Controlling the factor Set the thermostat at 25°C and kept there throughout the experiment temperature Lightintensty | Movelight source to aiferent clstances and measure lant intenstywith alc meter | tiobt intensity = Vatstance’) Start wth oiled, cooled water {00 then add measured antes of NBHCO, to increase the CO, concenation Carbon dioxide concentration 4,5,7,10and 14 em and no ight gives a good range o intensities Oto 50 mmol dry? in YO mmolam” intervals Keep the light source ata constant istance, such as Som ‘Add enough NaHCO, to.gvea high CO, concentration (60 mmolam) Hypotheses in biological research Hypotheses are provisional explanations. They require repeated testing. Hypotheses about the effects of three limiting factors on rates of photosynthesis are shown as annotations on the graphs below. You can test them using techniques in the table above, See 34 ce vous8. Carbon dioxide enrichment experiments ‘The carbon dioxide concentvalion of the atmosphere was 270 ppm untilthe late 18th century. Human activities have since increased it’oy more than 50%. The CO, concentration is forecast during the 21st century to rise tomote then double the presndustral level. This massive increase will have many effects on the Earth. There is already evidence of faster rates of photosynthesis, because CO, concentration commonly isthe limiting factor. There is also evidence of increases in growth and. accumulation of plant biomass, locking up carbon and helping to lessen the rise in atmospheric CO,, Future rates of photosynthesis and plant growth can be predicted from results of experiments in which CO, concentration is artificially increased, with other factors unchanged. Experiments in greenhouses have been followed up by fee air cartoon dioxide enrichment (FACE) experiments where the plants are notin an enclosed space. Inthe fist series of FACE experiments, plant growth in agricutual crops and young tree plantations was investigated. The second series is being conducted in natural or semi-natural forests around the world. Circles ‘of towers are built from which carbon dioxide can be released. CO, concentration is monitored inside the circles and whenitdops below 550 pom, CO, isteleased on theupnind side. Control plots are included, with air rather than carbon dioxide released from the circles oftoners, ‘Many variables are moritored inthe crcularareas from ‘hich the consequences ofa global ise to 550 ppm CO, canbe predicted for plants, other organisms and wnole ‘ecosystems n particular, the hypothes's that increased photosynthesis and plant growth will moderate risesin CO, concentration can be tested AHL 9. Photosystems Photosystems are molecular arrays of chlorophyll and other accessory pigments with special chlorophylls as the reaction centre, from which pairs of excited electrons are emitted. Photosystems are located in thylakoid membranes. A thylakoid is a saclke vesicle. In photosynthetic eukaryotes the thylakoids are flattened and arranged in stacks inside | chloroplasts. They have two types of photosystem (PS land PS IN with dlfferent structures and functions, tines Reson, PSI swvorna PS Lis positioned where the thylakoid membrane | Js exposed to the surrounding stroma, whereas | PS Il does not need to be in contact with the stroma. Cyanobacteria also have thylakoids with PS land I, but they are not located in chloroplasts and the | pigments are arranged differently, Molecules co 10. Pigment molecules in a photosystem Each photosystem has two types of functional unit: aveaction centre that emits excited electrons and antenna complexes that harvest ght energy and funnel itto the action centre. Antenna complexes have many pigment molecules, of multe types, na precise arrangement. These are the advantages + Awiderarray witha liger number ofpigrnent molecules | intercepts more photons oflightand therefore supplies energy to thereaction centre atafaster ate + Each type of pigment absorbs a narrow range of wavelengths. Awider range of wavelengths can be absorbed and a greater proportion of the energy in sunlight s used because photosystems contain different ypes of pigment rather than just one + Eneray/s transferred from pigment to pigment by excitation energy transfer. In general, X* + Y > X+Y*, where X and Y are adjacent pigments and the asterisked pigment has exctation energy. The energy passes because Vis at lower eneray level when excited than X. The precisely structured array cof pigments inan antenna complex ensures that energy is funneled tothe reaction centre ofthe photosystem, Part ofthe array in PS lis shown, Feastian centre 11. Photolysis of water in PSII Absorption of ight allows a special chlorophyll (P680)in | the reaction centre of PS IIto emitan excited electron. | ‘Asa consequence, °680 is reduced and remains so Untiit regains an electron, released when wateris soi. This happens in the oxygen-evolving complex (OFC) ‘of PSilon the surface of the photosystem facing the | space inside the thylakoid. The OEC binds a total of wo water molecules and splits them into four electrons, four protons and one molecule of oxygen, | 2H,O-90, + 4H" + 4e~ The electrons pass to the P680. The protons are released into the thylakoid space, contributing to a proton gradient across the thylakoid membrane. The ‘oxygen molecules are a waste product and diftuse ‘outof the thylakoids. This spliting of water is called photolysis because it only happens inthe light when the P68O chlorophyll has lost electrons, Evolution of photolysis, billions of years ago, initiated the accumulation of oxygen in the atmosphere, allowing | organisms to respite aerobically and causing oxidation of dissolved iron and other minerals in the oceans, 35ae Molecules 12. ATP production by chemiosmosis in thylakoids [ATP one ofthe two useful products of reactions inthe thylakoids —the ightdepenclent reactions. Asin mitochondria, ATP is produced by chemiosmosis (described in Section C1.2.15). | ATP synthase is located in thylakoid membranes. A proton gradient is generated by chains of electron carriers in | thylakoid membranes, Excited electrons pass along these chains, releasing energy which is used to pump protons | from the stroma to the thylakoid! space. The electrons can be supplied in two ways: 1. Cyclic photophosphoryiation Pairs ofexcited electrons are = emitted by PS | and after passing ‘along the chain of carriers they retumto PSI [ae 2. Non-cyclic photophespherylation —> Pairs of excited electrons are emitted by PS land 13. NADPis reduced by PSI Reduced NADP is one of the two useful products of the ight-dependent reactions in thylakoids. Itis produced | Using excited electrons emitted by photosystem |. Each | | electron emitted is accepted by an electron carrier | (ferredoxin) on the surface of the thylakoid membrane, The reduced electron carrier binds to an enzyme (NADP reductase) also on the surface of the thylakoid membrane and transfers the electron itis carrying to the ‘enzyme. Two electrons are needed to reduce NADP. | NADP + 2e —>reduced NADP NADP is dissolved in the stroma. Ibis reduced when active site. Electrons emitted by PS | are replaced | byelectrons from PS Il and passed along the chain of electron caries. Production of reduced NADP (NADPH is therefore accompanied by non-cyclc photophosphorylation. NADP reductase Gorn) PSI thylakoid svora membrane 36 {after passing along the chain of carriers they flow to PS the random molecular motion brings tte the enzyme’s. “14. Thylal Thylakoids ate systems because they consist of interacting and interclependent components. ‘The thylakoid membrane is the key component: + Impermeable to protons so a proton gradient can develop + Enclosesa small volume of fluid so a gradient develops rapily + Made of phospholipids so can hold photosysterns composed of hydrophobic pigment molecules + Holds other components in the correct relative positions—AlP synthase, election carriers and NADP reductase. Each thylakoid component depends on other ‘components: + PS!lsupplies excited electrons to the electron cartier chain + The electron carrier chain supplies energy to proton pumps + Proton pumps create a proton gradient used iy ATP synthase + PS supplies excited electrons to NADP reductase + PSilresupplies PS Iwith electrons ‘+ Waterin the thylakoid space supplies electrons toPSILx 15. Rubisco: © Carbon dioxide diffuses into the stroma ofthe chloroplast and is then fixed by being converted into, 2 more complex carbon compound. This happens ina carboxylation reaction catalysed by the enzyme Rubisco. Carbon dioxide is combined with ribulose bisphosphate (RuBP), which isa five-carbon sugar. The product of the reaction is an unstable sic-carbon compound, which immediately spits to form two molecules of glycerate 3-phosphate. RuBP + CO, RUDISCO > 9 atycerate 3-phosphate 16. Synthesis of triose phosphate Molecules co Light energy is not used directly in carbon fixation, soitis alight‘ndependent reaction of photosynthesis, High concentrations of Rubisco are needed in the stroma of chloroplasts, because it works relatively slowly and is not effective in low carbon dioxide concentrations. Because of this and because of the abundance of photosynthesizing organisms, Rubisco is the most abundant enzymne on Earth, Giycerate 3-phosphate, formed by carbon fixation, is an organicacid. is converted into triose phosphate (a three- carbon sugat) by 2 reduction reaction in the stroma, The hydrogen needed for ths s supplied by reduced NADP. Energy s also needed and is supplied by ATP. glycerate reduced 3ephosphate "NADP + ATP tiiose phosphate * NADP + ADP This reaction is part of the Calvin cycle. all reactions in the Calvin cycle are lightindependent because they can happen in the dark, es long es ATP end reduced NAD? are available, 17. The Calvin cycle For the Calvin cycle to continue, every RuBP consumed rust be replaced. Triose phosphate is used to regenerate RUBP. Five molecules oftriose phosphate are converted bya series ofreactions into three molecules of RUBP. This process requires the use of eneray in the form of ATP. The. reactions are surnmarized in the diagram below. For every sixmolecules of tiose phosphate made by the lightsindependent reactions, five are used to regenerate RUBP and one exits the cycle. alycerate 3-phosphate CO, \ Calvin ribulose evel tose bisphosphate phosphate carbohydrates _ 18. Using the products of the Calvin cycle All ofthe carbon in compounds in photosynthesizing organisms s fixed in the Calvin cycle. Glucose is produced by inking together two triose phosphates, Itcan be converted into other carbohydrates such as sucrose for transport, or starch for storage. Pathways for making fatty acids and amino acids start with glycerete 3-phosphate or triose phosphate from the Calin cycle, or with intermediates from pathways of aerobic respiration. ‘Mineral nutrients supply allelements other than carbon, hydrogen and oxygen in compounds made by photosynthesizing organisms, + Nitrogen for making amino acids is supplied by nitrate or ammonium ions. + Sulfurforamino acids s supplied as sulfate. + Phosphorus for making phospholipids, ATP and nucleotides for DNA and RNA is supplied 2s phosphate 19, Interdependence of photosynthesis reactions The Iightsndependent reactions cannot happen without supply of ATP and reduced NAD? from the light-dependent reactions. Sim aby te light- | dependent reactions require ADP ancé NADP, which the light-ndependent reactions produce 37