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Photosynthesis Chapter 8 2

Photosynthesis Overview Energy for all life on Earth ultimately comes from photosynthesis 6CO2 + 12H2O C6H12O6 + 6H2O + 6O2 Oxygenic photosynthesis is carried out by Cyanobacteria 7 groups of algae All land plants – chloroplasts

Chloroplast Thylakoid membrane – internal membrane Contains chlorophyll and other photosynthetic pigments Pigments clustered into photosystems Grana – stacks of flattened sacs of thylakoid membrane Stroma lamella – connect grana Stroma – semiliquid surrounding thylakoid membranes

Courtesy Dr. Kenneth Miller, Brown University Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Cuticle Epidermis Mesophyll Vascular bundle Stoma Vacuole Cell wall 1.58 mm Inner membrane Chloroplast Outer membrane Courtesy Dr. Kenneth Miller, Brown University

Stages Light-dependent reactions Require light Capture energy from sunlight Make ATP and reduce NADP+ to NADPH Carbon fixation reactions or light-independent reactions Does not require light Use ATP and NADPH to synthesize organic molecules from CO2

Light-Dependent Reactions Calvin Cycle Organic molecules Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Sunlight Photosystem Photosystem H2O O2 O2 Thylakoid Light-Dependent Reactions ADP + Pi ATP NADP+ NADPH Calvin Cycle Organic molecules CO2 Stroma

Discovery of Photosynthesis Jan Baptista van Helmont (1580–1644) Demonstrated that the substance of the plant was not produced only from the soil Joseph Priestly (1733–1804) Living vegetation adds something to the air Jan Ingenhousz (1730–1799) Proposed plants carry out a process that uses sunlight to split carbon dioxide into carbon and oxygen (O2 gas)

F.F. Blackman (1866–1947) Came to the startling conclusion that photosynthesis is in fact a multistage process, only one portion of which uses light directly Light versus dark reactions Enzymes involved Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Maximum rate Excess CO2; 35ºC Temperature limited Increased Rate of Photosynthesis Excess CO2; 20ºC Light limited CO2 limited Insufficient CO2 (0.01%); 20ºC 500 1000 1500 2000 2500 Light Intensity (foot-candles)

C. B. van Niel (1897–1985) Robin Hill (1899–1991) Found purple sulfur bacteria do not release O2 but accumulate sulfur Proposed general formula for photosynthesis CO2 + 2 H2A + light energy → (CH2O) + H2O + 2 A Later researchers found O2 produced comes from water Robin Hill (1899–1991) Demonstrated Niel was right that light energy could be harvested and used in a reduction reaction

Pigments Molecules that absorb light energy in the visible range Light is a form of energy Photon – particle of light Acts as a discrete bundle of energy Energy content of a photon is inversely proportional to the wavelength of the light Photoelectric effect – removal of an electron from a molecule by light

Increasing wavelength Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Increasing energy Increasing wavelength 0.001 nm 1 nm 10 nm 1000 nm 0.01 cm 1 cm 1 m 100 m UV light Gamma rays X-rays Infrared Radio waves Visible light 400 nm 430 nm 500 nm 560 nm 600 nm 650 nm 740 nm

Absorption spectrum When a photon strikes a molecule, its energy is either Lost as heat Absorbed by the electrons of the molecule Boosts electrons into higher energy level Absorption spectrum – range and efficiency of photons molecule is capable of absorbing

high Absorbtion Light low 400 450 500 550 600 650 700 Wavelength (nm) Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. high carotenoids chlorophyll a chlorophyll b Absorbtion Light low 400 450 500 550 600 650 700 Wavelength (nm)

Pigments in Photosynthesis Organisms have evolved a variety of different pigments Only two general types are used in green plant photosynthesis Chlorophylls Carotenoids In some organisms, other molecules also absorb light energy

Chlorophylls Chlorophyll a Chlorophyll b Main pigment in plants and cyanobacteria Only pigment that can act directly to convert light energy to chemical energy Absorbs violet-blue and red light Chlorophyll b Accessory pigment or secondary pigment absorbing light wavelengths that chlorophyll a does not absorb

Structure of chlorophyll porphyrin ring Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Structure of chlorophyll porphyrin ring Complex ring structure with alternating double and single bonds Magnesium ion at the center of the ring Photons excite electrons in the ring Electrons are shuttled away from the ring Chlorophyll a: R = CH3 H2C CH H R Chlorophyll b: R = CHO H3C CH2CH3 Porphyrin head N N H Mg H N N H3C CH3 H H H O CH2 CO2CH3 CH2 O C O CH2 CH CCH3 CH2 CH2 CH2 CHCH3 Hydrocarbon tail CH2 CH2 CH2 CHCH3 CH2 CH2 CH2 CHCH3 CH3

Copyright © The McGraw-Hill Companies, Inc Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. high Oxygen-seeking bacteria Absorbtion Light Filament of green algae low Action spectrum Relative effectiveness of different wavelengths of light in promoting photosynthesis Corresponds to the absorption spectrum for chlorophylls

© Eric Soder/pixsource.com Carotenoids Carbon rings linked to chains with alternating single and double bonds Can absorb photons with a wide range of energies Also scavenge free radicals – antioxidant Protective role Phycobiloproteins Important in low-light ocean areas Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Oak leaf in summer Oak leaf in autumn © Eric Soder/pixsource.com

Photosystem Organization Antenna complex Hundreds of accessory pigment molecules Gather photons and feed the captured light energy to the reaction center Reaction center 1 or more chlorophyll a molecules Passes excited electrons out of the photosystem

Antenna complex Also called light-harvesting complex Captures photons from sunlight and channels them to the reaction center chlorophylls In chloroplasts, light-harvesting complexes consist of a web of chlorophyll molecules linked together and held tightly in the thylakoid membrane by a matrix of proteins

Photosystem Electron acceptor Photon e– Electron donor e– Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Photosystem Electron acceptor Photon e– Electron donor e– Reaction center chlorophyll Chlorophyll molecule Thylakoid membrane Thylakoid membrane

Reaction center Transmembrane protein–pigment complex When a chlorophyll in the reaction center absorbs a photon of light, an electron is excited to a higher energy level Light-energized electron can be transferred to the primary electron acceptor, reducing it Oxidized chlorophyll then fills its electron “hole” by oxidizing a donor molecule

Excited chlorophyll molecule Chlorophyll reduced Chlorophyll oxidized Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Excited chlorophyll molecule Light Electron donor Electron acceptor e– e– e– e– Chlorophyll reduced Chlorophyll oxidized Donor oxidized Acceptor reduced + – + – e– e– e– e–

Light-Dependent Reactions Primary photoevent Photon of light is captured by a pigment molecule Charge separation Energy is transferred to the reaction center; an excited electron is transferred to an acceptor molecule Electron transport Electrons move through carriers to reduce NADP+ Chemiosmosis Produces ATP Capture of light energy

Cyclic photophosphorylation In sulfur bacteria, only one photosystem is used Generates ATP via electron transport Anoxygenic photosynthesis Excited electron passed to electron transport chain Generates a proton gradient for ATP synthesis

b-c1 complex Reaction center (P870) Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. High Excited reaction center e– Electron acceptor Energy of electrons b-c1 complex e– ATP Reaction center (P870) Low Photon Electron acceptor e– Photosystem

Chloroplasts have two connected photosystems Oxygenic photosynthesis Photosystem I (P700) Functions like sulfur bacteria Photosystem II (P680) Can generate an oxidation potential high enough to oxidize water Working together, the two photosystems carry out a noncyclic transfer of electrons that is used to generate both ATP and NADPH

Photosystem I transfers electrons ultimately to NADP+, producing NADPH Electrons lost from photosystem I are replaced by electrons from photosystem II Photosystem II oxidizes water to replace the electrons transferred to photosystem I 2 photosystems connected by cytochrome/ b6-f complex

Noncyclic photophosphorylation Plants use photosystems II and I in series to produce both ATP and NADPH Path of electrons not a circle Photosystems replenished with electrons obtained by splitting water Z diagram

Proton gradient formed Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Excited reaction center 2. The electrons pass through the b6-f complex, which uses the energy released to pump protons across the thylakoid membrane. The proton gradient is used to produce ATP by chemiosmosis. Ferredoxin 2 e– Fd Excited reaction center NADP reductase 2 Plastoquinone e– e– NADP+ + H+ NADPH PQ 2 b6-f complex Plastocyanin Reaction center Energy of electrons 2 e– PC Photon H+ Proton gradient formed for ATP synthesis Reaction center 3. A pair of chlorophylls in the reaction center absorb two photons. This excites two electrons that are passed to NADP+, reducing it to NADPH. Electron transport from photosystem II replaces these electrons. Photon 2 H2O e– 2H+ + 1/2O2 Photosystem I Photosystem II 1. A pair of chlorophylls in the reaction center absorb two photons of light. This excites two electrons that are transferred to plastoquinone (PQ). Loss of electrons from the reaction center produces an oxidation potential capable of oxidizing water.

Two photosystems high Rate of Photosynthesis low Far-red light on Off Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. high Rate of Photosynthesis low Far-red light on Off Red light on Off Both lights on Off Time

Photosystem II Resembles the reaction center of purple bacteria Core of 10 transmembrane protein subunits with electron transfer components and two P680 chlorophyll molecules Reaction center differs from purple bacteria in that it also contains four manganese atoms Essential for the oxidation of water b6-f complex Proton pump embedded in thylakoid membrane

Photosystem I Reaction center consists of a core transmembrane complex consisting of 12 to 14 protein subunits with two bound P700 chlorophyll molecules Photosystem I accepts an electron from plastocyanin into the “hole” created by the exit of a light-energized electron Passes electrons to NADP+ to form NADPH

Antenna complex Thylakoid membrane NADP reductase ATP synthase Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Photon Light-Dependent Reactions Photon H+ ATP ADP + Pi ATP NADP NADPH ADP NADPH Antenna complex Calvin H+ + NADP+ Cycle Thylakoid membrane Fd 2 2 e– PQ e– 2 2 e– Stroma 2 2 e– 2 2 PC H2O H+ Proton H+ Plastoquinone Plastocyanin Ferredoxin gradient H+ Water-splitting enzyme H+ Thylakoid space 1/2O2 2H+ NADP reductase ATP synthase Photosystem II b6-f complex Photosystem I 1. Photosystem II absorbs photons, exciting electrons that are passed to plastoquinone (PQ). Electrons lost from photosystem II are replaced by the oxidation of water, producing O2 2. The b6-f complex receives electrons from PQ and passes them to plastocyanin (PC). This provides energy for the b6-f complex to pump protons into the thylakoid. 3. Photosystem I absorbs photons, exciting electrons that are passed through a carrier to reduce NADP+ to NADPH. These electrons are replaced by electron transport from photosystem II. 4. ATP synthase uses the proton gradient to synthesize ATP from ADP and Pi enzyme acts as a channel for protons to diffuse back into the stroma using this energy to drive the synthesis of ATP.

Chemiosmosis Electrochemical gradient can be used to synthesize ATP Chloroplast has ATP synthase enzymes in the thylakoid membrane Allows protons back into stroma Stroma also contains enzymes that catalyze the reactions of carbon fixation – the Calvin cycle reactions

Production of additional ATP Noncyclic photophosphorylation generates NADPH ATP Building organic molecules takes more energy than that alone Cyclic photophosphorylation used to produce additional ATP Short-circuit photosystem I to make a larger proton gradient to make more ATP

Carbon Fixation – Calvin Cycle To build carbohydrates cells use Energy ATP from light-dependent reactions Cyclic and noncyclic photophosphorylation Drives endergonic reaction Reduction potential NADPH from photosystem I Source of protons and energetic electrons

Calvin cycle Named after Melvin Calvin (1911–1997) Also called C3 photosynthesis Key step is attachment of CO2 to RuBP to form PGA Uses enzyme ribulose bisphosphate carboxylase/oxygenase or rubisco

3 phases Carbon fixation Reduction Regeneration of RuBP RuBP + CO2 → PGA Reduction PGA is reduced to G3P Regeneration of RuBP PGA is used to regenerate RuBP 3 turns incorporate enough carbon to produce a new G3P 6 turns incorporate enough carbon for 1 glucose

3-phosphoglycerate (3C) (PGA) Ribulose 1,5-bisphosphate (5C) (RuBP) Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Stroma of chloroplast Light-Dependent Reactions 6 molecules of Carbon dioxide (CO2) ADP + Pi ATP NADP+ NADPH Calvin Cycle Rubisco 12 molecules of 6 molecules of 3-phosphoglycerate (3C) (PGA) Ribulose 1,5-bisphosphate (5C) (RuBP) 12 ATP PHASE 1 12 ADP 6 ADP Regeneration of RuBP 12 molecules of PHASE 3 Calvin Cycle 1,3-bisphosphoglycerate (3C) 6 ATP 12 NADPH PHASE 2 4 Pi Reduction 12 NADP+ 10 molecules of 12 Pi Glyceraldehyde 3-phosphate (3C) 12 molecules of Glyceraldehyde 3-phosphate (3C) (G3P) 2 molecules of Glyceraldehyde 3-phosphate (3C) (G3P) Glucose and other sugars

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Output of Calvin cycle Glucose is not a direct product of the Calvin cycle G3P is a 3 carbon sugar Used to form sucrose Major transport sugar in plants Disaccharide made of fructose and glucose Used to make starch Insoluble glucose polymer Stored for later use

Energy cycle Photosynthesis uses the products of respiration as starting substrates Respiration uses the products of photosynthesis as starting substrates Production of glucose from G3P even uses part of the ancient glycolytic pathway, run in reverse Principal proteins involved in electron transport and ATP production in plants are evolutionarily related to those in mitochondria

Heat Sunlight Photo- system II Photo- system I O2 Electron Transport Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Heat Sunlight Photo- system II Photo- system I O2 Electron Transport System ATP H2O ADP + Pi ADP + Pi ATP NADP+ NADPH NAD+ NADH Calvin Cycle CO2 Krebs Cycle ATP Glucose Pyruvate ADP + Pi ATP

Photorespiration Rubisco has 2 enzymatic activities Carboxylation Addition of CO2 to RuBP Favored under normal conditions Photorespiration Oxidation of RuBP by the addition of O2 Favored when stoma are closed in hot conditions Creates low-CO2 and high-O2 CO2 and O2 compete for the active site on RuBP

Under hot, arid conditions, leaves lose water by Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Leaf epidermis Heat H2O H2O O2 O2 CO2 CO2 Stomata Under hot, arid conditions, leaves lose water by evaporation through openings in the leaves called stomata. The stomata close to conserve water but as a result, O2 builds up inside the leaves, and CO2 cannot enter the leaves.

Types of photosynthesis C3 Plants that fix carbon using only C3 photosynthesis (the Calvin cycle) C4 and CAM Add CO2 to PEP to form 4 carbon molecule Use PEP carboxylase Greater affinity for CO2, no oxidase activity C4 – spatial solution CAM – temporal solution

a. C4 pathway b. C4 pathway Mesophyll cell Bundle-sheath cell CO2 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Mesophyll cell Bundle-sheath cell CO2 Mesophyll cell RuBP Calvin Cycle 3PG (C3) G3P Stoma Vein a. C4 pathway Mesophyll cell Bundle- sheath cell CO2 Mesophyll cell C4 Bundle- sheath cell CO2 Calvin Cycle G3P Stoma Vein b. C4 pathway a: © John Shaw/Photo Researchers, Inc. b: © Joseph Nettis/National Audubon Society Collection/Photo Researchers, Inc.

C4 plants Corn, sugarcane, sorghum, and a number of other grasses Initially fix carbon using PEP carboxylase in mesophyll cells Produces oxaloacetate, converted to malate, transported to bundle-sheath cells Within the bundle-sheath cells, malate is decarboxylated to produce pyruvate and CO2 Carbon fixation then by rubisco and the Calvin cycle

CO2 Mesophyll cell Phosphoenolpyruvate (PEP) Oxaloacetate AMP + PPi Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. CO2 Mesophyll cell Phosphoenolpyruvate (PEP) Oxaloacetate AMP + PPi ATP + Pi Pyruvate Malate Pyruvate Malate Bundle-sheath cell CO2 Calvin Cycle Glucose

C4 pathway, although it overcomes the problems of photorespiration, does have a cost To produce a single glucose requires 12 additional ATP compared with the Calvin cycle alone C4 photosynthesis is advantageous in hot dry climates where photorespiration would remove more than half of the carbon fixed by the usual C3 pathway alone

CAM plants Many succulent (water-storing) plants, such as cacti, pineapples, and some members of about two dozen other plant groups Stomata open during the night and close during the day Reverse of that in most plants Fix CO2 using PEP carboxylase during the night and store in vacuole

When stomata closed during the day, organic acids are decarboxylated to yield high levels of CO2 High levels of CO2 drive the Calvin cycle and minimize photorespiration

(inset): © 2011 Jessica Solomatenko/Getty Images RF Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. night CO2 C4 day CO2 Calvin Cycle G3P (inset): © 2011 Jessica Solomatenko/Getty Images RF

Compare C4 and CAM Both use both C3 and C4 pathways C4 – two pathways occur in different cells CAM – C4 pathway at night and the C3 pathway during the day