Plant defense responses Hypersensitive response Prepare a 10’ talk for Friday March 3 on plant defense responses or describe interactions between plants& pathogens, pests or symbionts Plant defense responses Hypersensitive response Systemic acquired resistance Innate immunity Phytoalexin synthesis Defensins and other proteins Oxidative burst Some possible pests Nematodes Rootworms Aphids Thrips Gypsy moths hemlock woolly adelgid Some possible pathogens Agrobacterium tumefaciens Agrobacterium rhizogenes Pseudomonas syringeae Pseudomonas aeruginosa Viroids DNA viruses RNA viruses Fungi Oomycetes Some possible symbionts N-fixing bacteria N-fixing cyanobacteria Endomycorrhizae Ectomycorrhizae

Converts light to chemical energy Photosynthesis Converts light to chemical energy 6 CO2 + 6 H2O + light energy <=> C6H12O6 + 6 O2

Photosynthesis 2 sets of rxns in separate parts of chloroplast

Photosynthesis 1) Light rxns use light to pump H+ use ∆ pH to make ATP by chemiosmosis

Photosynthesis 1) Light rxns use light to pump H+ use ∆ pH to make ATP by chemiosmosis 2) Light-independent (dark) rxns use ATP & NADPH from light rxns to make organics

Photosynthesis 1) Light rxns use light to pump H+ use ∆ pH to make ATP by chemiosmosis 2) Light-independent (dark) rxns use ATP & NADPH from light rxns to make organics only link: each provides substrates needed by the other

Light Rxns 3 stages 1) Catching a photon (primary photoevent)

Light Rxns 3 stages 1) Catching a photon (primary photoevent) 2) ETS

Light Rxns 3 stages 1) Catching a photon (primary photoevent) 2) ETS 3) ATP synthesis by chemiosmosis

Catching photons photons: particles of energy that travel as waves Energy inversely proportional to wavelength () visible light ranges from 400 -700 nm

Catching photons Photons: particles of energy that travel as waves caught by pigments: molecules that absorb light

Pigments Can only absorb certain photons

Pigments Can only absorb certain photons Photon has exact energy to push an e- to an outer orbital

Pigments Can only absorb certain photons Photon has exact energy to push an e- to an outer orbital from ground to excited state

Pigments Photon has exact energy to push an e- to an outer orbital from ground to excited state each pigment has an absorption spectrum: l it can absorb

Pigments Chlorophyll a is most abundant pigment chlorophyll a looks green -> absorbs all l but green Reflects green

Accessory Pigments absorb l which chlorophyll a misses chlorophyll b is an important accessory pigment

Accessory Pigments absorb l which chlorophyll a misses chlorophyll b is an important accessory pigment others include xanthophylls & carotenoids

Accessory Pigments action spectrum shows use of accessory pigments l used for photosynthesis

Accessory Pigments action spectrum shows use of accessory pigments l used for photosynthesis plants use entire visible spectrum l absorbed by chlorophyll work best

Light Reactions 1) Primary photoevent: pigment absorbs a photon

Light Reactions 1) Primary photoevent: pigment absorbs a photon e- is excited -> moves to outer orbital

Light Reactions 4 fates for excited e-: 1) returns to ground state emitting heat & longer  light = fluorescence

Light Reactions 4 fates for excited e-: 1) fluorescence 2) transfer to another molecule

Light Reactions 4 fates for excited e-: 1) fluorescence 2) transfer to another molecule 3) Returns to ground state dumping energy as heat

4 fates for excited e-: 1) fluorescence 2) transfer to another molecule 3) Returns to ground state dumping energy as heat 4) energy is transferred by inductive resonance excited e- vibrates and induces adjacent e- to vibrate at same frequency

4 fates for excited e-: 4) energy is transferred by inductive resonance excited e- vibrates and induces adjacent e- to vibrate at same frequency Only energy is transferred

4 fates for excited e-: 4) energy is transferred by inductive resonance excited e- vibrates and induces adjacent e- to vibrate at same frequency Only energy is transferred e- returns to ground state

Photosystems Pigments are bound to proteins arranged in thylakoids in photosystems arrays that channel energy absorbed by any pigment to rxn center chlorophylls

Photosystems Pigments are bound to proteins arranged in thylakoids in photosystems arrays that channel energy absorbed by any pigment to rxn center chls Need 2500 chlorophyll to make 1 O2

Photosystems Arrays that channel energy absorbed by any pigment to rxn center chls 2 photosystems : PSI & PSII PSI rxn center chl a dimer absorbs 700 nm = P700

Photosystems Arrays that channel energy absorbed by any pigment to rxn center chls 2 photosystems : PSI & PSII PSI rxn center chl a dimer absorbs 700 nm = P700 PSII rxn center chl a dimer absorbs 680 nm = P680

Photosystems Each may have associated LHC (light harvesting complex) (LHC can diffuse within membrane) PSI has LHCI: ~100 chl a, a few chl b & carotenoids

Photosystems Each may have associated LHC (light harvesting complex) (LHC can diffuse within membrane) PSI has LHCI: ~100 chl a, a few chl b & carotenoids PSII has LHCII: ~250 chl a, many chl b & carotenoids Proteins of LHCI & LHCII also differ

Photosystems PSI performs cyclic photophosphorylation Absorbs photon & transfers energy to P700

cyclic photophosphorylation Absorbs photon & transfers energy to P700 transfers excited e- from P700 to fd

cyclic photophosphorylation Absorbs photon & transfers energy to P700 transfers excited e- from P700 to fd fd returns e- to P700 via PQ, cyt b6/f & PC

cyclic photophosphorylation Absorbs photon & transfers energy to P700 transfers excited e- from P700 to fd fd returns e- to P700 via PQ, cyt b6/f & PC Cyt b6/f pumps H+

Cyclic Photophosphorylation Transfers excited e- from P700 to fd Fd returns e- to P700 via cyt b6-f & PC Cyt b6-f pumps H+ Use PMF to make ATP

Cyclic photophosphorylation first step is from P700 to A0 (another chlorophyll a) charge separation prevents e- from returning to ground state = true photoreaction

Cyclic photophosphorylation first step is from P700 to A0 (another chlorophyll a) next transfer e- to A1 (a phylloquinone) next = 3 Fe/S proteins

Cyclic photophosphorylation first step is from P700 to A0 (another chlorophyll a) next transfer e- to A1 (a phylloquinone) next = 3 Fe/S proteins finally ferredoxin

Cyclic photophosphorylation Ferredoxin = branchpoint: in cyclic PS FD reduces PQ

Cyclic photophosphorylation Ferredoxin reduces PQ PQH2 diffuses to cyt b6/f 2) PQH2 reduces cyt b6 and Fe/S, releases H+ in lumen since H+ came from stroma, transports 2 H+ across membrane (Q cycle)

Cyclic photophosphorylation 3) Fe/S reduces plastocyanin via cyt f cyt b6 reduces PQ to form PQ-

Cyclic photophosphorylation 4) repeat process, Fe/S reduces plastocyanin via cyt f cyt b6 reduces PQ- to form PQH2

Cyclic photophosphorylation 4) repeat process, Fe/S reduces plastocyanin via cyt f cyt b6 reduces PQ- to form PQH2 Pump 4H+ from stroma to lumen at each cycle (per net PQH2)

Cyclic photophosphorylation 5) PC (Cu+) diffuses to PSI, where it reduces an oxidized P700

Cyclic photophosphorylation energetics: light adds its energy to e- -> excited state Eo' P700 = +0.48 V Eo' P700* = -1.3 V

Cyclic photophosphorylation energetics: light adds its energy to e- -> excited state Eo' P700 = +0.48 V Eo' P700* = -1.3 V Eo' fd = - 0.42 V

Cyclic photophosphorylation energetics: light adds its energy to e- -> excited state Eo' P700 = +0.48 V Eo' P700* = -1.3 V Eo' fd = - 0.42 V Eo' cyt b6/f = +0.3V

Cyclic photophosphorylation energetics: light adds its energy to e- -> excited state Eo' P700 = +0.48 V Eo' P700* = -1.3 V Eo' fd = - 0.42 V Eo' cyt b6/f = +0.3V Eo' PC = +0.36V

Cyclic photophosphorylation energetics: light adds its energy to e- -> excited state Eo' P700 = +0.48 V Eo' P700* = -1.3 V Eo' fd = - 0.42 V Eo' cyt b6/f = +0.3V Eo' PC = +0.36V e- left in excited state returns in ground state

Cyclic photophosphorylation e- left in excited state returns in ground state Energy pumped H+

Cyclic photophosphorylation Limitations Only makes ATP

Cyclic photophosphorylation Limitations Only makes ATP Does not supply electrons for biosynthesis = no reducing power

Photosystems PSI performs cyclic photophosphorylation Makes ATP but not NADPH: exact mech for PQ reduction unclear, but PQ pumps H+

Photosystem II Evolved to provide reducing power -> added to PSI

Photosystem II Evolved to provide reducing power Added to PSI rxn center absorbs 680 nm (cf 700 nm)

Photosystem II rxn center absorbs 680 nm (cf 700 nm) can oxidize H2O redox potential of P680+ is + 1.1 V (cf + 0.82 V for H2O)

Photosystem II rxn center absorbs 680 nm (cf 700 nm) can oxidize H2O redox potential of P680+ is + 1.1 V (cf + 0.82 V for H2O) Use e- from H2O to reduce NADP+ (the e- carrier used for catabolic reactions)

Photosystem II rxn center absorbs 680 nm (cf 700 nm) can oxidize H2O redox potential of P680+ is + 1.1 V (cf + 0.82 V for H2O) Use e- from H2O to reduce NADP+ (the e- carrier used for catabolic reactions) use NADPH c.f. NADH to prevent cross- contaminating catabolic & anabolic pathways

PSI and PSII work together in the “Z-scheme” - a.k.a. “non-cyclic photophosphorylation” General idea: ∆ redox potential from H2O to NADP+ is so great that must boost energy of H2O e- in 2 steps

PSI and PSII work together in the “Z-scheme” General idea: ∆ redox potential from H2O to NADP+ is so great that must boost energy of H2O e- in 2 steps each step uses a photon

PSI and PSII work together in the “Z-scheme” General idea: ∆ redox potential from H2O to NADP+ is so great that must boost energy of H2O e- in 2 steps each step uses a photon 2 steps = 2 photosystems

PSI and PSII work together in the “Z-scheme” 1) PSI reduces NADP+

PSI and PSII work together in the “Z-scheme” 1) PSI reduces NADP+ e- are replaced by PSII

PSI and PSII work together in the “Z-scheme” 2) PSII gives excited e- to ETS ending at PSI

PSI and PSII work together in the “Z-scheme” 2) PSII gives excited e- to ETS ending at PSI Each e- drives cyt b6/f

PSI and PSII work together in the “Z-scheme” 2) PSII gives excited e- to ETS ending at PSI Each e- drives cyt b6/f Use PMF to make ATP

PSI and PSII work together in the “Z-scheme” 2) PSII gives excited e- to ETS ending at PSI Each e- drives cyt b6/f Use PMF to make ATP PSII replaces e- from H2O forming O2

PSI and PSII work together in the “Z-scheme” Light absorbed by PS II makes ATP Light absorbed by PS I makes reducing power

Ultimate e- source None water O2 released? No yes cyclic non-cyclic Ultimate e- source None water O2 released? No yes Terminal e- acceptor None NADP+ Form in which energy is ATP ATP & temporarily captured NADPH Photosystems required PSI PSI & PSII

Z-scheme energetics

Physical organization of Z-scheme PS II consists of: P680 (a dimer of chl a) ~ 30 other chl a & a few carotenoids > 20 proteins D1 & D2 bind P680 & all e- carriers

Physical organization of Z-scheme PSII has 2 groups of closely associated proteins 1) OEC (oxygen evolving complex) on lumen side, near rxn center Ca2+, Cl- & 4 Mn2+

Physical organization of Z-scheme PSII also has two groups of closely associated proteins 1) OEC (oxygen evolving complex) on lumen side, near rxn center Ca2+, Cl- & 4 Mn2+ 2) variable numbers of LHCII complexes

Physical organization of Z-scheme D1 & D2 bind P680 & all e- carriers Synechoccous elongatus associates phycobilisomes cf LHCII with PSII

Physical organization of Z-scheme D1 & D2 bind P680 & all e- carriers Synechoccous elongatus associates phycobilisomes cf LHCII with PSII

Physical organization of Z-scheme 2 mobile carriers plastoquinone : lipid similar to ubiquinone

Physical organization of Z-scheme 2 mobile carriers 1) plastoquinone : lipid similar to ubiquinone “headgroup” alternates between quinone & quinol

Physical organization of Z-scheme 2 mobile carriers 1) plastoquinone : lipid similar to ubiquinone “headgroup” alternates between quinone & quinol Carries 2 e- & 2 H+

Physical organization of Z-scheme 2 mobile carriers 1) plastoquinone : hydrophobic molecule like ubiquinone “headgroup” alternates between quinone and quinol Carries 2 e- & 2 H+ diffuses within bilayer

Physical organization of Z-scheme 2 mobile carriers 1) plastoquinone 2) plastocyanin (PC) : peripheral membrane protein of thylakoid lumen

Physical organization of Z-scheme 2) plastocyanin (PC) : peripheral membrane protein of thylakoid lumen Cu is alternately oxidized & reduced carries 1 e- & 1 H+

Physical organization of Z-scheme 3 protein complexes (visible in EM of thylakoid) 1) PSI 2) PSII 3) cytochrome b6/f 2 cytochromes & an Fe/S protein

Physical organization of Z-scheme 2 mobile carriers 1) plastoquinone 2) plastocyanin (PC) 3 protein complexes 1) PSI 2) PSII 3) cytochrome b6/f ATP synthase (CF0-CF1 ATPase) is also visible in E/M

Physical organization of Z-scheme 3 protein complexes (visible in EM of thylakoid) 1) PSI 2) PSII 3) cytochrome b6/f 2 cytochromes & an Fe/S protein

Physical organization of Z-scheme 2 mobile carriers 1) plastoquinone 2) plastocyanin (PC) 3 protein complexes 1) PSI 2) PSII 3) cytochrome b6/f ATP synthase (CF0-CF1 ATPase) is also visible in E/M

Physical organization of Z-scheme Complexes are arranged asymmetrically! PSII is in appressed regions of grana

Physical organization of Z-scheme Complexes are arranged asymmetrically! PSII is in appressed regions of grana PSI and ATP synthase are found in exposed regions (ends & margins of grana, and stromal lamellae)

Physical organization of Z-scheme Complexes are arranged asymmetrically! PSII is in appressed regions of grana PSI and ATP synthase are in exposed regions cytochrome b6/f, PC and PQ are evenly dispersed

Physical organization of Z-scheme Complexes are arranged asymmetrically! PSII is in appressed regions of grana PSI and ATP synthase in exposed regions cytochrome b6/f, PC and PQ are evenly dispersed why PC and PQ must be mobile

Physical organization of Z-scheme Complexes are arranged asymmetrically! PSII is in appressed regions of grana PSI and ATP synthase in exposed regions cytochrome b6/f, PC and PQ are evenly dispersed why PC and PQ must be mobile why membrane must be very fluid

PSII Photochemistry 1) LHCII absorbs a photon 2) energy is transferred to P680

PSII Photochemistry 3) P680* reduces pheophytin ( chl a with 2 H+ instead of Mg2+) = primary electron acceptor

PSII Photochemistry 3) P680* reduces pheophytin ( chl a with 2 H+ instead of Mg2+) = primary electron acceptor charge separation traps the electron

PSII Photochemistry 4) pheophytin reduces PQA (plastoquinone bound to D2) moves electron away from P680+ & closer to stroma

PSII Photochemistry 5) PQA reduces PQB (forms PQB- )

PSII Photochemistry 6) P680+ acquires another electron , and steps 1-4 are repeated

PSII Photochemistry 7) PQA reduces PQB - -> forms PQB2-

PSII Photochemistry 8) PQB2- acquires 2 H+ from stroma forms PQH2 (and adds to ∆pH)

PSII Photochemistry 9) PQH2 diffuses within bilayer to cyt b6/f - is replaced within D1 by an oxidized PQ

Photolysis: Making Oxygen 1) P680+ oxidizes tyrZ ( an amino acid of protein D1)

Photolysis: Making Oxygen 2) tyrZ + oxidizes one of the Mn atoms in the OEC Mn cluster is an e- reservoir

Photolysis: Making Oxygen 2) tyrZ + oxidizes one of the Mn atoms in the OEC Mn cluster is an e- reservoir Once 4 Mn are oxidized replace e- by stealing them from 2 H2O

Shown experimentally that need 4 flashes/O2

Shown experimentally that need 4 flashes/O2 Mn cluster cycles S0 -> S4 Reset to S0 by taking 4 e- from 2 H2O

Electron transport from PSII to PSI 1) PQH2 diffuses to cyt b6/f 2) PQH2 reduces cyt b6 and Fe/S, releases H+ in lumen since H+ came from stroma, transports 2 H+ across membrane (Q cycle)

Electron transport from PSII to PSI 3) Fe/S reduces plastocyanin via cyt f cyt b6 reduces PQ to form PQ-

Electron transport from PSII to PSI 4) repeat process, Fe/S reduces plastocyanin via cyt f cyt b6 reduces PQ- to form PQH2

Electron transport from PSII to PSI 4) PC (Cu+) diffuses to PSI, where it reduces an oxidized P700

Electron transport from PSI to Ferredoxin 1) LHCI absorbs a photon 2) P700* reduces A0 3) e- transport to ferredoxin via A1 & 3 Fe/S proteins

Electron transport from Ferredoxin to NADP+ 2 Ferredoxin reduce NADP reductase

Electron transport from Ferredoxin to NADP+ 2 Ferredoxin reduce NADP reductase reduces NADP+

Electron transport from Ferredoxin to NADP+ 2 Ferredoxin reduce NADP reductase reduces NADP+ this also contributes to ∆pH

Overall reaction for the Z-scheme 8 photons + 2 H2O + 10 H+stroma + 2 NADP+ = 12 H+lumen + 2 NADPH + O2