Suchergebnisse

The excited-state relaxation within bacteriochlorophyll (BChl) e and a in chlorosomes of Chlorobium phaeobacteroides has been studied by femtosecond transient absorption spectroscopy at room temperature. Singlet-singlet annihilation was observed to strongly influence both the isotropic and anisotropic decays. Pump intensities in the order of 10(11) photons x pulse(-1) x cm(-2) were required to obtain annihilation-free conditions. The most important consequence of applied very low excitation doses is an observation of a subpicosecond process within the BChl e manifold (similar to200-500 fs), manifesting itself as a rise in the red part of the Q(y) absorption band of the BChl e aggregates. The subsequent decay of the kinetics measured in the BChl e region and the corresponding rise in the baseplate BChl a is not single-exponential, and at least two components are necessary to fit the data, corresponding to several BChl e-->BChl a transfer steps. Under annihilation-free conditions, the anisotropic kinetics show a generally slow decay within the BChl e band (10-20 ps) whereas it decays more rapidly in the BChl a region (similar to1 ps). Analysis of the experimental data gives a detailed picture of the overall time evolution of the energy relaxation and energy transfer processes within the chlorosome. The results are interpreted within an exciton model based on the proposed structure. ; This research was supported by the Swedish Natural Science Research Council, the European Union Training and Mobility of Researchers project "Green Bacterial Photosynthesis" (grant FMRX-CT96-0081), and the Kempe Foundation. J. Pšenčik also thanks the Swedish Institute and Grant Agency of the Czech Republic (grant 206/02/0942) for financial support. ; Peer reviewed

Sensitization of singlet delta oxygen (O2(1Δg)) by bacteriochlorophyll e (BChle) has been investigated to gain a better understanding of the photoprotection mechanism(s) operating in chlorosomes of green photosynthetic bacteria. The sensitization process has been studied in media where BChle forms monomers (acetone and aqueous solutions containing 0.5% Triton X-100 [TX]) and in systems where BChle aggregates, namely, aqueous solutions containing 0.003% monogalactosyl diglyceride (MGDG) and chlorosomes (control as well as hexanol perturbed) from Chlorobium phaeobacteroides strain CL1401. In Ar-purged acetone, BChle triplets (BChle†) have a lifetime of a few tens of microseconds; however, in air-saturated acetone, quenching of BChle† by ground-state oxygen (O2(3Σ−g)) and formation of O2(1Δg) take place. The O2(1Δg) so formed is susceptible to quenching by BChle0, a ground-state BChle molecule. A Stern–Volmer analysis reveals a linear fit between the decay rate of O2(1Δg) and the BChle concentration. The rate constants for the quenching of O2(1Δg) by BChle0 and for the deactivation of O2(1Δg) by the solvent come out to be kq= (1.4 ± 0.1) × 10^9 M−1 s−1 and k0= (18.5 ± 0.7) × 10^3 s−1, respectively. The absolute quantum yield of O2(1Δg) sensitization by BChle monomers is 0.65 ± 0.15 in air-saturated acetone. In aqueous phase, the triplet lifetime of BChle aggregates in native or hexanol-perturbed chlorosomes shortens by more than two orders of magnitude when compared with the triplet lifetime of BChle monomers in 0.5% TX solution (a few hundreds of microseconds). Quenching by carotenoids (Car) makes only a minor contribution to the decay of BChle† in aggregates. Because O2(1Δg) sensitization by BChle† could be detected neither in MGDG aggregates nor in chlorosomes (control as well as hexanol perturbed), it is concluded that (1) this process is highly likely when BChle is present as a monomer but not when it is tightly packed in artificial aggregates or in chlorosomes; and (2) Car, though vital for the baseplate BChla, are dispensable for BChle. ; We acknowledge financial support from the Research Council of Norway and the European Union (contract FMRX-CT96-0081). ; Peer reviewed

15 páginas, 8 figuras, 1 tabla ; SpainFestuca rubra subsp. pruinosa is a perennial grass that inhabits sea cliffs, a habitat where salinity and low nutrient availability occur. These plants have a rich fungal microbiome, and particularly common are their associations with Epichloë festucae in aboveground tissues and with Fusarium oxysporum and Periconia macrospinosa in roots. In this study, we hypothesized that these fungi could affect the performance of F. rubra plants under salinity, being important complements for plant habitat adaptation. Two lines of F. rubra, each one consisting of Epichloë-infected and Epichloë-free clones, were inoculated with the root endophytes (F. oxysporum and P. macrospinosa) and subjected to a salinity treatment. Under salinity, plants symbiotic with Epichloë had lower Na+ content than non-symbiotic plants, but this effect was not translated into plant growth. P. macrospinosa promoted leaf and root growth in the presence and absence of salinity, and F. oxysporum promoted leaf and root growth in the presence and absence of salinity, plus a decrease in leaf Na+ content under salinity. The growth responses could be due to functions related to improved nutrient acquisition, while the reduction of Na+ content might be associated with salinity tolerance and plant survival in the long term. Each of these three components of the F. rubra core mycobiome contributed with different functions, which are beneficial and complementary for plant adaptation to its habitat in sea cliffs. Although our results do not support an obvious role of Epichloë itself in FRP salt tolerance, there is evidence that Epichloë can interact with root endophytes, affecting host plant performance ; This study has received funding from the European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement no. 676480 and from the project "CLU-2019-05 – IRNASA/CSIC Unit of Excellence," funded by the Junta de Castilla y León and co-financed by the European Union (ERDF "Europe drives our growth"). ; Peer reviewed

19 páginas, 4 figuras, 2 tablas ; Global warming will inevitably affect crop development and productivity, increasing uncertainty regarding food production. The exploitation of genotypic variability can be a promising approach for selecting improved crop varieties that can counteract the adverse effects of future climate change. We investigated the natural variation in yield performance under combined elevated CO2 and high-temperature conditions in a set of 60 bread wheat genotypes (59 of the 8TH HTWSN CIMMYT collection and Gazul). Plant height, biomass production, yield components and phenological traits were assessed. Large variations in the selected traits were observed across genotypes. The CIMMYT genotypes showed higher biomass and grain yield when compared to Gazul, indicating that the former performed better than the latter under the studied environmental conditions. Principal component and hierarchical clustering analyses revealed that the 60 wheat genotypes employed different strategies to achieve final grain yield, highlighting that the genotypes that can preferentially increase grain and ear numbers per plant will display better yield responses under combined elevated levels of CO2 and temperature. This study demonstrates the success of the breeding programs under warmer temperatures and the plants' capacity to respond to the concurrence of certain environmental factors, opening new opportunities for the selection of widely adapted climate-resilient wheat genotypes. ; This research was supported by the Spanish National R&D&I Plan of the Ministry of Economy and Competitiveness (Projects AGL2013-41363-R and AGL2016-79589-R) and Ministry of Science and Innovation (Project PID2019-107154RB-100), as well as the regional government, the Junta de Castilla y León (Projects CSI083U16 and CSI260P20), with European Regional Development Fund, ERDF. Project "CLU-2019-05—IRNASA/CSIC Unit of Excellence", funded by the Junta de Castilla y León and co-financed by the European Union (ERDF "Europe drives our growth"). E.L. Marcos-Barbero was the recipient of a pre-doctoral contract from the Junta de Castilla y León (E-37-2017-0066125) ; Peer reviewed

23 páginas, 6 tablas, 3 figuras ; The progressive rise in atmospheric CO2 concentrations and temperature associated with climate change is predicted to have a major impact on the productivity and quality of food crops. Therefore, food security is highly dependent on climate change. Following a survey with 60 bread wheat genotypes, here we investigated the genetic variation in grain yield and nutritional quality among 10 of these genotypes grown under elevated CO2 and temperature. With this purpose, the biomass production, grain yield-related traits, the grain concentration of starch, total protein, phenolic compounds, and mineral nutrients, together with the total antioxidant capacity, were determined. Variation among genotypes was found for almost all the studied traits. Higher grain and ear numbers were associated with increased grain yield but decreased grain total protein concentration and minerals such as Cu, Fe, Mg, Na, P, and Zn. Mineral nutrients were mainly associated with wheat biomass, whereas protein concentration was affected by plant biomass and yield-related traits. Associations among different nutrients and promising nutrient concentrations in some wheat genotypes were also found. This study demonstrates that the exploration of genetic diversity is a powerful approach, not only for selecting genotypes with improved quality, but also for dissecting the effect of the environment on grain yield and nutritional composition ; This research was supported by the Spanish National R&D&I Plan of the Ministry of Economy and Competitiveness (Projects AGL2013-41363-R and AGL2016-79589-R) and the regional government, the Junta de Castilla y León (Projects CSI083U16 and CSI260P20), with European Regional Development Fund, ERDF. E.L. Marcos-Barbero was the recipient of a pre-doctoral contract from the Junta de Castilla y León (E-37-2017-0066125). ; Peer reviewed

We examined the microenvironment of the single tryptophan and the tyrosine residues of PsbQ, one of the three main extrinsic proteins of green algal and higher plant photosystem II. On the basis of this information and the previous data on secondary structure [Balsera, M., Arellano, J.B., Gutiérrez, J.R., Heredia, P., Revuelta, J.L. & De Las Rivas, J. (2003) Biochemistry42, 1000–1007], we screened structural models derived by combining various threading approaches. Experimental results showed that the tryptophan residue is partially buried in the core of the protein but still in a polar environment, according to the intrinsic fluorescence emission of PsbQ and the fact that fluorescence quenching by iodide was weaker than that by acrylamide. Furthermore, quenching by cesium suggested that a positively charged barrier shields the tryptophan microenvironment. Comparison of the absorption spectra in native and denaturing conditions indicated that one or two out of six tyrosines of PsbQ are buried in the core of the structure. Using threading methods, a 3D structural model was built for the C-terminal domain of the PsbQ protein family (residues 46–149), while the N-terminal domain is predicted to have a flexible structure. The model for the C-terminal domain is based on the 3D structure of cytochrome b562, a mainly α-protein with a helical up/down bundle folding. Despite the large sequence differences between the template and PsbQ, the structural and energetic parameters for the explicit model are acceptable, as judged by the corresponding tools. This 3D model is compatible with the experimentally determined environment of the tryptophan residue and with published structural information. The future experimental determination of the 3D structure of the protein will offer a good validation point for our model and the technology used. Until then, the model can provide a starting point for further studies on the function of PsbQ. ; This study was supported by funds from the Spanish Ministry of Science and Technology (project PB1998-0480) and the V European Union Framework Programme. M. Balsera holds a fellowship from the Spanish Ministry of Science and Technology. ; Peer reviewed

The role of carotenoids in chlorosomes of the green sulfur bacterium Chlorobium phaeobacteroides, containing bacteriochlorophyll (BChl) e and the carotenoid (Car) isorenieratene as main pigments, was studied by steady-state fluorescence excitation, picosecond single-photon timing and femtosecond transient absorption (TA) spectroscopy. In order to obtain information about energy transfer from Cars in this photosynthetic light-harvesting antenna with high spectral overlap between Cars and BChls, Car-depleted chlorosomes, obtained by inhibition of Car biosynthesis by 2-hydroxybiphenyl, were employed in a comparative study with control chlorosomes. Excitation spectra measured at room temperature give an efficiency of 60-70% for the excitation energy transfer from Cars to BChls in control chlorosomes. Femtosecond TA measurements enabled an identification of the excited state absorption band of Cars and the lifetime of their S-1 state was determined to be similar to10 ps. Based on this lifetime, we concluded that the involvement of this state in energy transfer is unlikely. Furthermore, evidence was obtained for the presence of an ultrafast (<100 fs) energy transfer process from the S-2 state of Cars to BChls in control chlorosomes. Using two time-resolved techniques, we further found that the absence of Cars leads to overall slower decay kinetics probed within the Qy band of BChl e aggregates, and that two time constants are generally required to describe energy transfer from aggregated BChl e to baseplate BChl a. ; Swedish Natural Science Research Council. European Union (Contract FMRX-CT96-0081). Kempe Foundation ; Peer reviewed

8 pages, 2 figures, 2 tables. ; Isolated chlorosomes of several species of filamentous anoxygenic phototrophic bacteria (FAPB) and green sulfur bacteria (GSB) were examined by atomic force microscopy (AFM) to characterize their topography and biometry. Chlorosomes of Chloroflexus aurantiacus, Chloronema sp., and Chlorobium (Chl.) tepidum exhibited a smooth surface, whereas those of Chl. phaeobacteroides and Chl. vibrioforme showed a rough one. The potential artifactual nature of the two types of surfaces, which may have arisen because of sample manipulation or AFM processing, was ruled out when AFM images and transmission electron micrographs were compared. The difference in surface texture might be associated with the specific lipid and polypeptide composition of the chlorosomal envelope. The study of three-dimensional AFM images also provides information about the size and shape of individual chlorosomes. Chlorosomal volumes ranged from ca. 35000 nm3 to 247000 nm3 for Chl. vibrioforme and Chl. phaeobacteroides, respectively. The mean height was about 25 nm for all the species studied, except Chl. vibrioforme, which showed a height of only 14 nm, suggesting that GSB have 1–2 layers of bacteriochlorophyll (BChl) rods and GFB have ~4. Moreover, the average number of BChl molecules per chlorosome was estimated according to models of BChl rod organisation. These calculations yielded upper limits ranging from 34000 BChl molecules in Chl. vibrioforme to 240000 in Chl. phaeobacteroides, values that greatly surpass those conventionally accepted. ; This study was supported by The European Union through the TMR Program, Contract No. FMRXCT96‐ 0081. ; Peer reviewed

We have studied the effect of the absence of carotenoids on the organization of bacteriochlorophylls (BChls) in chlorosomes of Chlorobium (Chl.) phaeobacteroides strain CL1401. Carotenoid-depleted chlorosomes were obtained by means of 2-hydroxybiphenyl–supplemented cultures. In the presence of the inhibitor, isorenieratene (Isr) and β-Isr biosynthesis were inhibited to more than 95%, leading to an accumulation of the colorless precursor phytoene inside the chlorosomes. In addition, there was a 30–40% decrease in the baseplate BChl a content. The absorption spectrum of the carotenoid-depleted chlorosomes showed a 10 nm blue shift in the BChl e Qy absorption peak. Under reducing conditions, a decrease in the BChl a/BChl e fluorescence emission ratio was observed in carotenoid-depleted chlorosomes relative to that in control chlorosomes, caused mainly by the decrease in the BChl a content. The steady-state fluorescence emission anisotropy in the BChl e region dropped from ;0.24 for native chlorosomes to ;0.14 for carotenoid-depleted ones, indicating reorganization of BChl e. The circular dichroism (CD) signal of the carotenoid-depleted chlorosomes was increased two times in the BChl e Qy region. A simple model based on the structure proposed was used to explain the observed effects. Carotenoids might affect the angle between the direction of the BChl e Qy transition and the axis of the rod. The orientation of BChl a in the baseplate remains unchanged in carotenoid-depleted chlorosomes, although there is a partial loss of BChl a as a consequence of a decrease in the baseplate size. The carotenoids are most likely rather close to the BChls and appear to be important for the aggregate structure in Chl. phaeobacteroides. ; European Union (Contract FMRX-CT96-0081). Spanish Ministry of Education and Science (Ref. BIO96-1229-002-01). ; Peer reviewed

12 páginas, 3 figuras, 1 tabla ; A rapid and high throughput protocol to measure the catalase activity in vitro has been designed. Catalase is an enzyme with unusual kinetic properties because it does not follow the standard Michaelis–Menten model and is inactivated by H2O2. This makes the analysis of the two rate equations of the second‐ordered reactions of the kinetic model rather complex. A two‐degree polynomial fitting of the experimental data is proposed after transforming the exponential form of the integrated rate equation of the [H2O2] into a polynomial using the Taylor series. The fitting is validated by establishing an experimental linear relationship between the initial rate of the H2O2 decomposition and the protein concentration, regardless of the suicide inactivation that catalase might undergo beyond t > 0. In addition, experimental considerations are taken into account to avoid statistical bias in the analysis of the catalase activity. ANOVA analyses show that the proposed protocol can be utilized to measure the initial rate of the H2O2 decomposition by catalase in 32 samples in triplicates if kept below 8 mM min−1 in the microplate wells. These kinetic and statistical analyses can pave the way for other antioxidant enzyme activity assays in microplate readers at small scale and low cost. ; This research was funded by the Spanish national plan for Scientific and Technical Research and Innovation, grant number AGL2016-79589-R, and the regional government of Castilla y León, grant number CSI260P20. The Project "CLU-2019-05—IRNASA/CSIC Unit of Excellence" funded by the Junta de Castilla y León and co-financed by the European Union (ERDF "Europe drives our growth") is also acknowledged ; Peer reviewed

The steady state kinetic mechanism of the H2O2-supported oxidation of different organic substrates by peroxidase from leaves of Chamaerops excelsa palm trees (CEP) has been investigated. An analysis of the initial rates vs. H2O2 and reducing substrate concentrations is consistent with a substrate-inhibited Ping-Pong Bi Bi reaction mechanism. The phenomenological approach expresses the peroxidase Ping-Pong mechanism in the form of the Michaelis–Menten equation and leads to an interpretation of the effects in terms of the kinetic parameters K(H2O2, m), K(AH2, m), k(cat), K(H2O2, SI), K(AH2, SI) and of the microscopic rate constants k1 and k3 of the shared three-step catalytic cycle of peroxidases. ; Funding from Consejeria de Educación (projects SA129A07 and SA052A10-2) and Consejeria de Agricultura y Ganaderia (Project SA06000) of the Regional Government of Castilla and León (Junta de Castilla y León, Spain) is acknowledged. ; Peer reviewed