1. Target photosynthetic products and chlorophyll fluorescence Biophysics Department Biology faculty Moscow State University Andrey Rubin

2. h  h  h  CO 2 N 2 , P  pH ATP e - Chl  Fl

3. PS II e - e - General scheme of photosynthetic processes h ν H + H + Primary photosynthetic processes Nitrogen and sulfur deficiency e - e - Normal h ν PQ Protein synthesis Carbohydrate pool Carbohydrate synthesis ( Calvin cycle ) Nitrogen and sulfur deficiency Nitrogen deficiency e - PS I Nitrogen assimilation NADP 2 H 2 O 4 H + + O 2( газ ) Pool ATP Fd Hydrogen evolution Lipid synthesis biofuel CO 2 ATPase

4. Growth rate dependence on light intensity

5. General view of the annular reactors used for mass cultivation of Nannochloropsis sp. In the foreground, 50-cm annular reactors; in the background, 91-cm annular reactors ( Zittelli et al., 2002).

6. Light intensity dependence on the distance from the light source in a biophotoreacter

7. Scheme of primary photosynthetic processes 10 -2 с 10 -11 s 10 -6 s 10 -4 с

8. Test biophotoreactor. Early (left) and late (right) stages of cell growth

10. PQ PQH 2 PQ PQ H 2 O P 680 Q A 2H + 1/2O 2 Chl Chl 2H + 2H + 2H + Fd Pc b h b l FeS R P 700 FeS I Chl 3H + K + Cl - H + NADPH NADP + ADP + Pi ATP + + _ _ lumen stroma Thylakoid membrane h  h  fluorescence PS II PS I ATP- synthase bf Q-cycle R-COO - - OOC - OOC R-COO - f F 0 F F m 0 1 10 t, c Calvin cycle Photosynthetic pathways in chloroplasts  pH

11. P S D O F F o 0 0.05 0.7 Time (s) 5 Fluorescence induction curve Photosynthetic efficiency F m F v F o

12. H 2 O PSII Q a Q b PSI .. CO 2 … * * O 2 h v h v Chloro Respir. - ! F (ns) ! (-N 2 , -P) starvation O 2 * excess membrane destruction (DEATH) signal ATP Δ pH q N Car. cycle P quenching * photoprotection ? 8 20 day hours 3 2 1 0 F v /F m phytoplankton Photooxidation and defense mechanisms O 2 -

13. 10 -1 10 0 10 1 10 2 Production, mg · C · m -3 · day -1 Calculated production (from F 0 · F V · I) Correlation between phytoplankton productivities measured with fluorometer and radiocarbon method corr = 0.84717

14. 0 2 4 0 2 4 6 0 2 4 6 0 2 4 F m F 0 F m F m F 0 F 0 Dark-adapted fluorescence parameters F 0 and F m as a function of time after adding 0 (a), 0.39 (b), or 0.78 (c) μM Cu 2+ to Chlorella Time, h a b c Early diagnostics of heavy metal ions effects

15. Description of the states of complex C 1 C 2 Probabilities of the electron carriers С i states The initial probabilities p i (0)=b i , i=1,…, l . 

16. Scheme of the states of Photosystem 2 Cl-chlorophyll Phe-pheophytitn Q A ,Q b – quinone acceptors 7 z 1 z 2 z 3 z 4 z 5 z 6 z 7 g 1 g 2 g 3 g 4 g 5 g 6 g 7 2H s + PQH 2 2H s + PQH 2 2H s + PQH 2 2H s + PQH 2 2H s + PQH 2 2H s + PQH 2 2H s + PQH 2 Chl Phe Q A Q B - y 1 H l + x 1 x 2 x 3 x 4 x 5 x 6 x 7 5 6 33 32 31 30 29 28 15 16 17 18 19 20 21 22 23 24 25 26 27 34 35 36 37 38 39 40 H l + 1 8 11 H l + 1 4 4 1 Chl + Phe - Q A Q B - y 3 Chl + Phe Q A - Q B - y 4 Chl* Phe Q A - Q B - y 6 Chl Phe Q A Q B 2 - Chl* Phe Q A Q B 2 - Chl + Phe - Q A Q B 2 - Chl + Phe Q A - Q B 2 - Chl Phe Q A - Q B 2 - Chl* Phe Q A - Q B 2 - Chl + Phe - Q A - Q B 2 - Chl Phe Q A Chl* Phe Q A Chl + Phe - Q A Chl + Phe Q A - Chl Phe Q A - Chl* Phe Q A - Chl + Phe - Q A - PQ PQ PQ PQ PQ PQ PQ Chl + Phe - Q A - Q B - y 7 Chl Phe Q A - Q B - y 5 Chl Phe Q A Q B Chl* Phe Q A Q B Chl + Phe - Q A Q B Chl + Phe Q A - Q B Chl Phe Q A - Q B Chl* Phe Q A - Q B Chl + Phe - Q A - Q B Chl* Phe Q A Q B - y 2 2 3 4 H l + 1 2 9 10 13

17. PGA BPGA GAP Ru5P RuBP ATP CO 2 ATP P 8 P 6 P 0 X P 9 β NADPH Q P 5 PS1 P 4 ADP ATP PS2 P 3 P 2 H 2 O P 1 P 7 F α (z) (s) (u) (y) (x) days of growth α β γ γ β α 0 40 80 N 1

18. PGA BPGA GAP Ru5P RuBP ATP CO 2 ATP P 8 P 6 P 0 X P 9 β NADPH Q P 5 PS1 P 4 ADP ATP PS2 P 3 P 2 H 2 O P 1 P 7 F α (z) (s) (u) (y) (x) Δ pH [ATP] Δ pH = s 2 - electron flow rate dependence on Δ pH - fl uorescence

19. Interaction of the complex with the mobile electron carrier D - concentrations of the mobile carrier in the oxidized and reduced forms; - concentrations of the components of the complex; k i - bimolecular rate constants.

20. Transmembrane electrochemical potential To describe the ATP synthesis we used the expression based on the minimal kinetic scheme of ATP synthesis-hydrolysis reaction: Where  =F  /RT . The dependence of the proton leakage on the potential was considered according to the mechanism of ion transfer trough the three barrier channel : The similar expression was used to describe K + transfer : :

21. Kinetics of the variables of the model

22. Flow fluorimeter device

23. Phytoplankton concentration ( Fo ) (A) and photosynthetical activity ( Fv/Fm )(B), as well as water temperature (C) in the cross section of Issik-Kul Lake (Tamga-Grigor’evka). Data were obtained with using submersible fluorometer in July 1999. Investigation of the vertical distribution of phytoplankton in oligotrophic Issyk-Kul Lake showed a complex structure of phytoplankton, which is due to pronounced water stratification. The lowest values of the abundance and photosynthetic activity were found in the upper layer under conditions of a high solar irradiation and low content of mineral nutrients. High abundance and high activity of algal cells were found in the deep layers of the photic zone indicating the presence of active algae, adapted to low light condition.

24. Light intensity dependence on the distance from the light source in a biophotoreacter

25. Cyt С 2 M Photosynthetic reaction center Rb. sphaeroides L H

26. Electron transfer in reaction centers

28. Space distribution of protein electron carriers in a membrane

29. Scene of the direct model

30. Equipotential surfaces calculated according to Poisson-Boltzmann equations model Oxidesed Рс Reduced cyt f Ion strength - 1 0 0 mM , pH=7, ε р-ра = 80; ε белка =2 ; red -6.5 мВ, blue + 6.5 мВ ; green – atoms of molecules. Dotted lines connect residueson Pc and Cytf that were used by simulation for calculation the distance between proteins r1 r2 r4 r3

31. Rate constant dependence on the membrane thickness