Comparative chloroplast proteome analysis of exogenously supplied trehalose to wheat seedlings under heat stress

The aim of our study was to investigate the underlying molecular mechanisms of exogenously supplied trehalose affecting wheat photosynthesis under heat stress. The amount of ATP synthase (ATPase), oxygen-evolving enhancer protein (OEE), PsbP, Rubisco, chloroplast fructose-bisphosphate aldolase (FBPA), and ferredoxin-NADP(H) oxidoreductase (FNR) were downregulated, while PSI reaction center subunits were upregulated under heat stress. However, in the trehalose-pretreated groups, the amount of FNR, cytochrome b₆f complex, PSI reaction center subunits, ATPase, FBPA, and Rubisco were upregulated under normal growth conditions and heat stress. Besides, during the recovery period, the upregulation in CAB, PsbP, OEE2, and ATPase suggested that trehalose pretreatment might help to the recovery of PSII and PSI. These results indicate that trehalose pretreatment effectively regulates the levels of the photosynthesis-related proteins and relieves the damage of heat stress to wheat chloroplast.     

Membrane Polypeptides associated with Photochemical Systems

WE wish to report a specific relationship between certain chloroplast membrane polypeptides and functional properties of the chloroplast usually associated with one or the other of the two photochemical systems (designated PSI and PSII). We have known this by the analysis of the chloroplast membrane polypeptides of the wild type strain of the unicellular green alga Chlamydomonas reinhardi and of mutant strains derived from it which have lost the capacity to carry out normal photosynthesis. These mutants have been characterized by the loss of particular membrane-bound components of the photosynthetic electron transport chain^1,2. This relationship is supported by analysis of the membrane polypeptides obtained from chloroplast fractions of wild type C. reinhardi and spinach enriched for reactions characteristic of either PSI or PSII by fractionation of the membranes either with digitonin^3,4 or ‘Triton X-100’⁵.     

Effects of heat stress on photosynthetic electron transport in a marine cyanobacterium <Emphasis Type="Italic">Arthrospira</Emphasis> sp.

Arthrospira (Spirulina) is widely used as human health food and animal feed. In cultures grown outdoors in open ponds, Arthrospira cells are subjected to various environmental stresses, such as high temperature. A better understanding of the effects of high temperature on photosynthesis may help optimize the productivity of Arthrospira cultures. In this study, the effects of heat stress on photosynthetic rate, chlorophyll a fluorescence transients, and photosystem (PS) II, PSI activities in a marine cyanobacterium Arthrospira sp. were examined. Arthrospira cells grown at 25 °C were treated for 30 min at 25 (control), 30, 34, 37, or 40 °C in the dark. Heat stress (30–37 °C) enhanced net photosynthetic O₂ evolution rate. Heat stress caused over-reduction PSII acceptor side, damage of donor side of PSII, decrease in the energetic connectivity of PSII units, and decrease in the performance of PSII. When the temperature changed from 25 to 37 °C, PSII activity decreased, while PSI activity increased, the enhancement of photosynthetic O₂ evolution was synchronized with the increase in PSI activity. When temperature was further increased to 40 °C, it induced a decrease in photosynthetic O₂ evolution rate and a more severe decrease in PSII activity, but an increase in PSI activity. These results suggest that PSI activity was the decisive factor determining the change of photosynthetic O₂ evolution when Arthrospira was exposed to a temperature from 25 to 37 °C, but then, PSII activity became the decisive factor adjusting the change of photosynthetic O₂ evolution when the temperature was increased to 40 °C.     

Overexpression of <Emphasis Type="Italic">CCCH</Emphasis> zinc finger protein gene delays flowering time and enhances salt tolerance in Arabidopsis by increasing fatty acid unsaturation

CCCH-type zinc-finger proteins constitute a large family playing key roles during plant development and growth. In the present study, we investigated the involvement of the CCCH-type zinc finger protein of AtZFP1 (At2g25900) in flowering and salt stress response in Arabidopsis. Compared with the wild type (WT), bolting and flowering were delayed in AtZFP1-overexpressing plants. Real-time PCR analysis of floral regulating genes in overexpressing Arabidopsis revealed that enhanced expression of FLC decreased the expressions of FT and SOC1. The Fv/Fm of overexpressing Arabidopsis lines was unchanged under salt stress. In contrast, ΦPSII activity and PSI oxidoreduction decreased in WT, overexpressing and mutant strains under salt stress conditions, with the smallest reduction in these parameters observed in the overexpressing strains. These results suggest that the CCCH zinc-finger protein AtZFP1 primarily controls flowering time by changing the expression of flowering genes under long-day conditions. The overexpression of this protein delayed flowering and increased the content and double-bond index of unsaturated fatty acids. Elevation of unsaturated fatty acid content might play important role in protecting the photosynthetic apparatus and maintaining the membrane function at salt stress by alleviating PSII and PSI photoinhibition.     

Photosynthetic responses of the low intertidal macroalga Sargassum fusiforme (Sargassaceae) to saline stress

Sargassum fusiforme, a species of brown seaweed with economic importance, inhabits lower intertidal zones where algae are often exposed to various stresses. In this study, changes in the photosynthetic performance of S. fusiforme under saline stress were investigated. The PSII performance in S. fusiforme significantly improved, when the thalli were exposed to 0% salinity, and remained high with prolonging treatment time. In contrast, the PSII activity declined considerably under salinities of 4.5 and 6%. The PSI activity did not change remarkably under saline stress, thus demonstrating higher tolerance to saline stress than PSII. In addition, the PSI activity could be also restored after saline treatments, when PSII was inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea. It might be as a result of changes in the NAD(P)H content in the thalli under saline stress. Our results suggested that PSI was much more tolerant to different saline stress than PSII in S. fusiforme. We demonstrated that S. fusiforme was much more tolerant to hyposaline than to hypersaline stress.     

Photoinhibition of photosystem I in <Emphasis Type="Italic">Nephrolepis falciformis</Emphasis> depends on reactive oxygen species generated in the chloroplast stroma

We studied how high light causes photoinhibition of photosystem I (PSI) in the shade-demanding fern Nephrolepis falciformis, in an attempt to understand the mechanism of PSI photoinhibition under natural field conditions. Intact leaves were treated with constant high light and fluctuating light. Detached leaves were treated with constant high light in the presence and absence of methyl viologen (MV). Chlorophyll fluorescence and P700 signal were determined to estimate photoinhibition. PSI was highly oxidized under high light before treatments. N. falciformis showed significantly stronger photoinhibition of PSI and PSII under constant high light than fluctuating light. These results suggest that high levels of P700 oxidation ratio cannot prevent PSI photoinhibition under high light in N. falciformis. Furthermore, photoinhibition of PSI in N. falciformis was largely accelerated in the presence of MV that promotes the production of superoxide anion radicals in the chloroplast stroma by accepting electrons from PSI. From these results, we propose that photoinhibition of PSI in N. falciformis is mainly caused by superoxide radicals generated in the chloroplast stroma, which is different from the mechanism of PSI photoinhibition in Arabidopsis thaliana and spinach. Here, we provide some new insights into the PSI photoinhibition under natural field conditions.     

Disturbed structure and function of chloroplasts during blocked biosynthesis of 5-aminolevulinic acid in the light

Xantha-702 mutant of cotton (Gossypium hirsutum L.) proved to have blocked synthesis of 5-aminolevulinic acid in the light. Accordingly, mutant leaves accumulated 2–5% chlorophyll of baseline. Mutant plants demonstrated disturbed production of pigment-protein complexes of photosystems I (PSI) and II (PSII) and generation of the chloroplast membrane system blocked at the early stages, largely, at the stages of vesicles and single short thylakoid. The functional activity of the PSI and PSII reaction centers was close to zero. Only the chlorophyll a/b light-harvesting complexes of PSI and PSII with the chlorophyll fluorescence peaks at 728 and 681 nm, respectively, were produced in the xantha-702 mutant. We propose that the genetic block of 5-aminolevunilic acid biosynthesis in the light in the xantha-702 mutant disturbs the formation and activity of the complexes of the reaction centers of PS-I and PS-II and inhibits the development of the whole membrane system of chloroplasts.     

Photosynthetic responses of a wheat (Asakaze)–barley (Manas) 7H addition line to salt stress

The photosynthetic responses to salt stress were examined in a wheat (Triticum aestivum L. cv. Asakaze)–barley (Hordeum vulgare L. cv. Manas) 7H addition line having elevated salt tolerance and compared to the parental wheat genotype. For this purpose, increasing NaCl concentrations up to 300 mM were applied and followed by a 7-day recovery period. Up to moderate salt stress (200 mM NaCl), forcible stomatal closure, parallel with a reduction in the net assimilation rate (P _N), was only observed in wheat, but not in the 7H addition line or barley. Since the photosynthetic electron transport processes of wheat were not affected by NaCl, the impairment in P _N could largely be accounted for the salt-induced decline in stomatal conductance (g _s), accompanied by depressed intercellular CO₂ concentration and carboxylation efficiency. Both, P _N and nonstomatal limitation factors (L_ns) were practically unaffected by moderate salt stress in barley and in the 7H addition line due to the sustained g _s, which might be an efficient strategy to maintain the efficient photosynthetic activity and biomass production. At 300 mM NaCl, both P _N and g _s decreased significantly in all the genotypes, but the changes in P _N and L_ns in the 7H addition line were more favourable similar to those in wheat. The downregulation of photosynthetic electron transport processes around PSII, accompanied by increases in the quantum yield of regulated energy dissipation and of the donor side limitation of PSI without damage to PSII, was observed in the addition line and barley during severe stress. Incomplete recovery of P _N was observed in the 7H addition line as a result of declined PSII activity probably caused by enhanced cyclic electron flow around PSI. These results suggest that the better photosynthetic tolerance to moderate salt stress of barley can be manifested in the 7H addition line which may be a suitable candidate for improving salt tolerance of wheat.     

Assessment of the preventive effect of vermicompost on salinity resistance in tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis> cv. Ailsa Craig)

To determine the effects of vermicompost leachate (VCL) on resistance to salt stress in plants, young tomato seedlings (Solanum lycopersicum, cv. Ailsa Craig) were exposed to salinity (150 mM NaCl addition to nutrient solution) for 7 days after or during 6 mL L^??1 VCL application. Salt stress significantly decreased leaf fresh and dry weights, reduced leaf water content, significantly increased root and leaf Na⁺ concentrations, and decreased K⁺ concentrations. Salt stress decreased stomatal conductance (g_s), net photosynthesis (A), instantaneous transpiration (E), maximal efficiency of PSII photochemistry in the dark-adapted state (F_v/F_m), photochemical quenching (qP), and actual PSII photochemical efficiency (ΦPSII). VCL applied during salt stress increased leaf fresh weight and g_s, but did not reduce leaf osmotic potential, despite increased proline content in salt-treated plants. VCL reduced Na⁺ concentrations in leaves (by 21.4%), but increased them in roots (by 16.9%). VCL pre-treatment followed by salt stress was more efficient than VCL concomitant to salt stress, since VCL pre-treatment provided the greatest osmotic adjustment recorded, with maintenance of net photosynthesis and K⁺/Na⁺ ratios following salt stress. VCL pre-treatment also led to the highest proline content in leaves (50 µmol g^??1 FW) and the highest sugar content in roots (9.2 µmol g^??1 FW). Fluorescence-related parameters confirmed that VCL pre-treatment of salt-stressed plants showed higher PSII stability and efficiency compared to plants under concomitant VCL and salt stress. Therefore, VCL represents an efficient protective agent for improvement of salt-stress resistance in tomato.     

Increase in the rate of photosynthetic linear electron transport in cyanobacteruim <Emphasis Type="Italic">Synechocystis</Emphasis> sp. PCC 6803 lacking phycobilisomes

Compensating changes in the pigment apparatus of photosynthesis that resulted from a complete loss of phycobilisomes (PBS) were investigated in the cells of a PAL mutant of cyanobacterium Synechocystis sp. PCC 6803. The ratio PBS/chlorophyll calculated on the basis of the intensity of bands in the action spectra of photosynthetic activity of two photosystems in the wild strain was 1: 70 for PSII and 1: 300 for PSI. Taking into consideration the number of chlorophyll molecules per reaction center in each photosystem, these ratios could be interpreted as association of PBS with dimers of PSII and trimers of PSI as well as greater dependence of PSII as compared with PSI on light absorption by PBS. The ratio PSI/PSII determined by photochemical cross-section of the reactions of two photosystems was 3.5: 1.0 for wild strain of Synechocystis sp. PCC 6803 and 0.7: 1.0 for the PAL mutant. A fivefold increase in the relative content of PSII in pigment apparatus corresponds to a 5-fold increase in the intensity of bands at 685 and 695 nm as related to the band of PSI at 726 nm recorded in low-temperature fluorescence spectrum of the PAL mutant. Inhibition of PSII with diuron resulted in a pronounced stimulation of chlorophyll fluorescence in the PAL mutant as compared to the wild strain of Synechocystis sp. PCC 6803; these data suggested an activation of electron transfer between PSII and PSI in the mutant cells. Thus, the lack of PBS in the mutant strain of Synechocystis sp. PCC 6803 was compensated for by the higher relative content of PSII in the pigment apparatus of photosynthesis and by a rise in the rate of linear electron transport.     

The Involvement of the Complexes of Photosystems in the Organization of Chloroplast Ultrastructure in Pea Mutants

We studied the involvement of pigment-protein complexes of photosystems (PS) in the development and spatial arrangement of thylakoids in chloroplasts of pea (Pisum sativum L.) leaves. The initial line (cv. Torsdag) and its mutants, chlorotica 2004 displaying primary disturbances in the PSI reaction centers and chlorotica 2014 containing only 50% of chlorophyll and, as a sequence, the reduced amount of all pigment-protein complexes. A proportional decrease in the content of PSI and PSII complexes in the chlorotica 2014 mutant resulted in a partial reduction of the whole chloroplast membrane system, whereas grana and stroma thylakoid regions were well developed. In contrast, a loss of only 20% of chlorophyll and destruction of PSI complexes in the chlorotica 2004 mutant by 50% resulted in the destruction of stroma thylakoid regions and disturbed longitudinal thylakoid and grana orientation. It was concluded that protein-protein interactions in pigment-protein complexes played a key role in the structure of thylakoid membranes and their longitudinal orientation.     

Effects of arbuscular mycorrhizal fungi on photosystem II activity of three pistachio rootstocks under salt stress as probed by the OJIP-test

We applied chlorophyll a fluorescence as a biomarker to assess the growth response and PSII behavior and performance of three pistachio (Pistacia vera) rootstocks to different salt levels after inoculation with arbuscular mycorrhizal fungi Glomus mosseae and compared it with non-mycorrhizal plants (control). Our results confirmed the depressing effect of salt stress on mycorrhization extent and showed that the effect of salinity on colonization rate is completely under the influence of host plant. In this experiment, mycorrhizal symbiosis could enhance plants total dry mass (TDM), electron transfer on the donor and the acceptor side of PSII, decrease the energy dissipation and increase the comprehensive photosynthesis performance under salt stress as well as under normal conditions. We found that both donor and acceptor sides of PSII are the target sides under high salinity in pistachio rootstocks. We also found that performance index is the parameter that better reflects the responses of the studied rootstocks to progressive salt stress. Bane-baqi was less affected by salinity in terms of TDM followed by Sarakhs and Abareqi.     

The response of transgenic <Emphasis Type="Italic">Brassica</Emphasis> species to salt stress: a review

Salt stress is considered one of the main abiotic factors to limit crop growth and productivity by affecting morpho-physiological and biochemical processes. Genetically, a number of salt tolerant Brassica varieties have been developed and introduced, but breeding of such varieties is time consuming. Therefore, current focus is on transgenic technology, which plays an important role in the development of salt tolerant varieties. Various salt tolerant genes have been characterized and incorporated into Brassica. Therefore, such genetic transformation of Brassica species is a significant step for improvement of crops, as well as conferring salt stress resistance qualities to Brassica species. Complete genome sequencing has made the task of genetically transforming Brassica species easier, by identifying desired candidate genes. The present review discusses relevant information about the principles which should be employed to develop transgenic Brassica species, and also will recommend tools for improved tolerance to salinity.     

Connectivity between electron transport complexes and modulation of photosystem II activity in chloroplasts

In chloroplasts, photosynthetic electron transport complexes interact with each other via the mobile electron carriers (plastoquinone and plastocyanin) which are in surplus amounts with respect to photosystem I and photosystem II (PSI and PSII), and the cytochrome b ₆ f complex. In this work, we analyze experimental data on the light-induced redox transients of photoreaction center P₇₀₀ in chloroplasts within the framework of our mathematical model. This analysis suggests that during the action of a strong actinic light, even significant attenuation of PSII [for instance, in the result of inhibition of a part of PSII complexes by DCMU or due to non-photochemical quenching (NPQ)] will not cause drastic shortage of electron flow through PSI. This can be explained by “electronic” and/or “excitonic” connectivity between different PSII units. At strong AL, the overall flux of electrons between PSII and PSI will maintain at a high level even with the attenuation of PSII activity, provided the rate-limiting step of electron transfer is beyond the stage of PQH₂ formation. Results of our study are briefly discussed in the context of NPQ-dependent mechanism of chloroplast protection against light stress.     

Structural and functional organization of the photosynthetic apparatus in halophytes with different strategies of salt tolerance

The specific features of the structural and functional organisation of the photosynthetic apparatus (PSA) were studied in wild halophytes representing three strategies of salt tolerance: euhalophyte Salicornia perennans, crynohalophyte Limonium gmelinii, and glycohalophyte Artemisia santonica. The sodium content in aboveground parts of the plants corresponded to the strategy of salt tolerance. The photosynthetic cells of the euhalophyte were large and contained a higher number of chloroplasts than those in other species. In contrast, the number of cells per a leaf area unit was lower in S. perennans as compared to cryno- and glycohalophytes. Thereupon, the cell and chloroplast surface area per leaf area unit declined in the following sequence: A. santonica > L. gmelinii > S. perennans. However, the large cells of euhalophyte contained chloroplasts of larger sizes with 4- to 5-fold higher chlorophyll (Chl) content per chloroplast and Chl concentration in chloroplast volume unit. Also, chloroplasts of S. perennans were characterised by the higher content of glyco- and phospholipids. Qualitative composition of fatty acids (FA) in lipids isolated from the chloroplast-enriched fraction was similar in all three species; however, the index of unsaturation of FA was higher in glycohalophyte A. santonica than those in two other species. Under natural condition, PSA of all three halophytes showed high resistance to soil salinity. The results indicated tolerance of PSII to the photodamage in halophytes. The high rate of electron transport through PSII can be important to prevent oxidative damage of PSA in halophytes under strong light and hight temperature in vivo. Thus, the strategy of salt tolerance is provided by both the leaf anatomical structure and the ultrastructure of photosynthetic membranes, which is determined in particular by the specific composition of lipids.