European larch (Larix decidua Mill.) and Norway spruce [Picea abies (L.) Karst.] synthesize chlorophyll (Chl) in darkness. This paper compares Chl accumulation in 14-d-old dark-grown seedlings of L. decidua and P. abies after shortterm (24 h) feeding with 5-aminolevulinic acid (ALA). We used two ALA concentrations (1 and 10 mM) fed to cotyledons of both species in darkness and in continuous light. The dark-grown seedlings of L. decidua accumulated Chl only in trace amounts and the seedlings remained etiolated. In contrast, P. abies seedlings grown in darkness were green and had significantly higher Chl content. After ALA feeding, higher protochlorophyllide (Pchlide) content was observed in L. decidua than in P. abies cotyledons incubated in darkness. Although short-term ALA feeding stimulated the synthesis of Pchlide, Chl content did not change significantly in cotyledons incubated in darkness. The Chl accumulation in cotyledons fed with ALA was similar to the rate of Chl accumulation in the controls. Higher Chl accumulation was reported in control samples after illumination: 86.9% in L. decidua cotyledons and 46.4% in P. abies cotyledons. The Chl content decreased and bleaching occurred in cotyledons incubated with ALA in light due to photooxidation. Analyses of Chlbinding proteins (D1 and LHCIIb) by Western blotting proved differences between Chl biosynthesis in L. decidua and P. abies seedlings in the dark and in the light. No remarkable increase was found in protein accumulation (D1 and LHCIIb) after ALA application. Our results showed interspecific difference in Chl synthesis between two gymnosperms. Shortterm ALA feeding did not stimulate Chl synthesis, thus ALA synthesis was not the rate-limiting step in Chl synthesis in the dark., N. Maximová, Ľ. Slováková., and Obsahuje bibliografii
After saturating irradiation for 3 h (SI), the original fluorescence F0 increased while the photosystem 2 (PS2) photochemical efficiency (Fv/Fm) declined significantly. These parameters could largely recover to the levels of dark-adapted leaves after 3 h of subsequent dark recovery. No net loss of the D1 proteins occurred after SI. Soybean and pumpkin leaves had different responses to SI. Low temperature fluorescence parameters, F685 and F685/F735, decreased significantly in soybean leaves but not in pumpkin leaves. Part of the light-harvesting complex LHC2 dissociated from PS2 complexes in soybean leaves but not in pumpkin leaves, as shown by sucrose density gradient centrifugation and SDS-PAGE. The photon-saturated PS2 electron transport activity declined significantly in pumpkin thylakoids but not in soybean thylakoids. In addition, a large amount of phosphorylated D1 proteins was found in dark-adapted soybean leaves but not in dark-adapted pumpkin leaves. Hence at excessive irradiance soybean and pumpkin have the same protective strategy against photo-damage, reversible down-regulation of PS2, but two different mechanisms, namely the reversible down-regulation is related to the dissociation of LHC2 in soybean leaves but not in pumpkin leaves. and Hai-Bo Zhang, Da-Quan Xu.
We investigated the effect of chromium (20-40 g m-3, 8-72 h) on the photosystem 2 (PS2) activities of Chlorella pyrenoidosa cells. By using chlorophyll fluorescence transients, thermoluminescence, oxygen polarography, and Western blot analysis for D1 protein we found that inhibition of PS2 can be accounted for by the enhanced photodestruction of the reaction centres in the cells cultivated in the presence of Cr(VI) at 25 °C in "white light" (18 W m-2). Hence photodestruction of D1 is caused by an enhanced oxidative stress and lipid peroxidation, as indicated by the appearance of a high-temperature thermoluminescence band. and Z. T. Hörcsik ... [et al.].
Thermal stability of thylakoid membranes isolated from acclimated and non-acclimated wheat (Triticum aestivum L. cv. HD 2329) leaves under irradiation was studied. Damage to the photosynthetic electron transport activity was more pronounced in thylakoid membranes isolated from non-acclimated leaves as compared to thylakoid membrane isolated from acclimated wheat leaves at 35 °C. The loss of D1 protein was faster in non-acclimated thylakoid membrane as compared to acclimated thylakoid membranes at 35 °C. However, the effect of elevated temperature on the 33 kDa protein associated with oxygen evolving complex in these two types of thylakoid membranes was minimal. Trypsin digestion of the 33 kDa protein in the thylakoid membranes isolated from control and acclimated seedlings suggested that re-organisation of 33 kDa protein occurs before its release during high temperature treatment. and A. K. Singh, G. S. Singhal.
Irradiation of thylakoid membranes at 40 °C resulted in complete inhibition of photosystem (PS) 2 activity measured as 2,6-dichlorophenol indophenol (DCIP) photoreduction either in the absence or presence of 1,5-diphenylcarbazide (DPC). Concomitant with the inactivation of PS2 activity, several thylakoid proteins were lost and high molecular mass cross-linking products appeared that cross-reacted with antibodies against proteins of PS2 but not with antibodies against proteins of other three complexes PS1, ATP synthase, and cytochrome b6f. Irradiation of thylakoid membranes suspended in buffer of basic pH or high concentration of Tris at 25 °C resulted in the formation of cross-linking products similar to those in thylakoid membranes irradiated at 40 °C. Presence of radical scavengers and DPC during the high temperature treatment prevented the formation of cross-linking products. These results suggest the involvement of oxygen evolving co mplex (OEC) in the formation of cross-linking between PS2 proteins in thylakoid membrane irradiated at high temperature. and Abhay K. Singh, G. S. Singhal.
Photoinhibition of photosynthesis was studied in young and mature detached sun needles of cypress under high irradiance (HI) of about 1 900 μmol m-2 s-1. The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll (Chl) fluorescence (Fv/Fm) and electron transport measurements. Compared with the mature needles, the young needles, containing about half the amount of Chl a+b per unit area, exhibited a higher proportion of total carotenoids (Car) as xanthophyll cycle pigments and had an increased ratio of Car/Chl a+b. The potential efficiency of photosystem (PS) 2, Fv/Fm, markedly declined in HI-treated young needles without significant increase of F0 level. In contrast, the Fv/Fm ratio declined with significant increase of F0 level in mature needles. In isolated thylakoids, the rate of whole chain and PS2 activity markedly decreased in young HI-needles in comparison with mature needles. A smaller inhibition of PS1 activity was observed in both needles. In the subsequent dark incubation, fast recovery was found in both needle Types that reached maximum PS2 efficiencies similar to those observed in non-photoinhibited needles. The artificial exogenous electron donors DPC, NH2OH, and Mn2+ failed to restore the HI-induced loss of PS2 activity in mature needles, while DPC and NH2OH significantly restored it in young needles. Hence, HI-inactivation was on the donor side of PS2 in young needles and on the acceptor side of PS2 in mature needles. Quantification of the PS2 reaction centre proteins D1 and 33 kDa protein of water splitting complex following HI-exposure of needles showed pronounced differences between young and mature needles. The large loss of PS2 activity in HI-needles was due to the marked loss of D1 protein of the PS2 reaction centre in mature needles and of the 33 kDa protein in young needles. and N. La Porta ... [et al.]
Seeds of Suaeda salsa were cultured in dark for 3 d and betacyanin accumulation in seedlings was promoted significantly. Then the seedlings with accumulated betacyanin (C+B) were transferred to 14/10 h light/dark and used for chilling treatment 15 d later. Photosystem 2 (PS2) photochemistry, D1 protein content, and xanthophyll cycle during the chilling-induced photoinhibition (exposed to 5 °C at a moderate photon flux density of 500 µmol m-2 s-1 for 3 h) and the subsequent restoration were compared between the C+B seedlings and the control (C) ones. The maximal efficiency of PS2 photochemistry (Fv/Fm), the efficiency of excitation energy capture by open PS2 centres (Fv'/Fm'), and the yield of PS2 electron transport (ΦPS2) of the C+B and C leaves both decreased during photoinhibition. However, smaller decreases in Fv/Fm, Fv'/Fm', and ΦPS2 were observed in the C+B leaves than in C ones. At the same time, the deepoxidation state of xanthophyll cycle, indicated by (A+Z)/(V+A+Z) ratio, increased rapidly but the D1 protein content decreased considerably during the photoinhibition. The increase in rate of (A+Z)/(V+A+Z) was higher but the D1 protein turnover was slower in C+B than C leaves. After photoinhibition treatment, the plants were transferred to a dim irradiation (10 µmol m-2 s-1) at 25 °C for restoration. During restoration, the chlorophyll (Chl) fluorescence parameters, D1 protein content, and xanthophyll cycle components relaxed gradually, but the rate and level of restoration in the C+B leaves was greater than those in the C leaves. The addition of betacyanins to the thylakoid solution in vitro resulted in similar changes of Fv/Fm, D1 protein content, and (A+Z)/(V+A+Z) ratio during the chilling process. Therefore, betacyanin accumulation in S. salsa seedlings may result in higher resistance to photoinhibition, larger slowing down of D1 protein turnover, and enhancement of non-radiative energy dissipation associated with xanthophyll cycle, as well as in greater restoration after photoinhibition than in the control when subjected to chilling at moderate irradiance. and C.-Q. Wang, T. Liu
Photoinhibition of photosynthesis was investigated in control (C) and chilling night (CN) leaves of grapevine under natural photoperiod at different sampling time in a day. The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll fluorescence (Fv/Fm) and photosynthetic electron transport measurements. When the potential efficiency of photosystem (PS) 2, Fv/Fm was measured at midday, it markedly declined with significant increase of F0 in CN leaves. In isolated thylakoids, the rate of whole chain and PS2 activity were markedly decreased in CN leaves than control leaves at midday. A smaller inhibition of PS1 activity was also observed in both leaf types. Later, the leaves reached maximum PS2 efficiencies similar to those observed in the morning during sampling at evening. The artificial exogenous electron donors diphenyl carbazide, NH2OH, and Mn2+ failed to restore the PS2 activity in both leaf types at midday. Thus CN enhanced inactivation on the acceptor side of PS2 in grapevine leaves. Quantification of the PS2 reaction centre protein D1 following midday exposure of leaves showed pronounced differences between C and CN leaves. The marked loss of PS2 activity in CN leaves noticed in midday samples was mainly due to the marked loss of D1 protein of the PS2 reaction centre. and M. Bertamini ... [et al.].
The degree of photoinhibition of sun and shade grown leaves of grapevine was determined by means of the ratio of variable to maximum chlorophyll (Chl) fluorescence (Fv/Fm) and electron transport measurements. The potential efficiency of photosystem 2 (PS2), Fv/Fm, markedly declined under high irradiance (HI) in shade leaves with less than 10 % of F0 level. In contrast, Fv/Fm ratio declined with about 20 % increase of F0 level in sun leaves. In isolated thylakoids, the rate of whole chain and PS2 activity in HI shade and sun leaves was decreased by about 60 and 40 %, respectively. A smaller inhibition of photosystem 1 (PS1) activity was also observed in both leaf types. In the subsequent dark incubation, fast recovery was observed in both leaf types that reached maximum PS2 efficiencies similar to non-photoinhibited control leaves. The artificial exogenous electron donors DPC, NH2OH, and Mn2+ failed to restore the HI-induced loss of PS2 activity in sun leaves, while DPC and NH2OH were significantly restored in shade leaves. Hence HI in shade leaves inactivates on the donor side of PS2 whereas it does at the acceptor side in sun leaves, respectively. Quantification of the PS2 reaction centre protein D1 and the 33 kDa protein of water splitting complex following HI-treatment of leaves showed pronounced differences between shade and sun leaves. The marked loss of PS2 activity in HI leaves was due to the marked loss of D1 protein of the PS2 reaction centre protein and the 33 kDa protein of the water splitting complex in sun and shade leaves, respectively. and M. Bertamini, K. Muthuchelian, N. Nedunchezhian.
The protective role of light-harvesting complex 2 (LHC2) dissociation from photosystem 2 (PS2) complex was explored by the 5'-p-fluorosulfonylbenzoyl adenosine (FSBA, an inhibitor of protein kinase) treatment at saturating irradiance (SI) in soybean leaves and thylakoids. The dissociation of some LHC2s from PS2 complex occurred after SI treatment, but FSBA treatment inhibited the dissociation as demonstrated by analysis of sucrose density gradient centrifugation of thylakoid preparation and low-temperature (77 K) chlorophyll (Chl) fluorescence. A significant increase in F0 and decrease in Fv/Fm occurred after SI, and the two parameters could largely recover to the levels of dark-adapted leaves after subsequent 3 h in the dark, but they could not recover in the FSBA-treated leaves at SI. Neither the electron transport activity of PS2 nor the D1 protein amount in vivo had significant change after SI without FSBA, whereas FSBA treatment at SI could result in significant decreases in both the PS2 electron transport activity and the D1 protein amount. When thylakoids instead of leaves were used, the PS2 electron transport activity and the D1 protein amount declined more after SI with FSBA than without FSBA. The phosphorylation level of PS2 core proteins increased, while the phosphorylation level of LHC2 proteins was reduced after SI. Also, the phosphorylation of PS2 core proteins could be greatly inhibited by the FSBA treatment at SI. Hence in soybean leaf the LHC2 dissociation is an effective strategy protecting PS2 reaction centres against over-excitation and photodamage by reducing the amount of photons transferred to the centres under SI, and the phosphorylation of PS2 core proteins plays an important role in the dissociation. and Hai-Bo Zhang, Da-Quan Xu.