Photosynthesis Research最新文献

Red algae are photosynthetic eukaryotes whose light-harvesting complexes (LHCs) associate with photosystem I (PSI). In this study, we examined characteristics of PSI-LHCI, PSI, and LHCI isolated from the red alga Galdieria sulphuraria NIES-3638. The PSI-LHCI supercomplexes were purified using anion-exchange chromatography followed by hydrophobic-interaction chromatography, and finally by trehalose density gradient centrifugation. PSI and LHCI were similarly prepared following the dissociation of PSI-LHCI with Anzergent 3-16. Polypeptide analysis of PSI-LHCI revealed the presence of PSI and LHC proteins, along with red-lineage chlorophyll a/b-binding-like protein (RedCAP), which is distinct from LHC proteins within the LHC protein superfamily. RedCAP, rather than LHC proteins, exhibited tight binding to PSI. Carotenoid analysis of LHCI identified zeaxanthin, β-cryptoxanthin, and β-carotene, with zeaxanthin particularly enriched, which is consistent with other red algal LHCIs. A Qy peak of chlorophyll a in the LHCI absorption spectrum was blue-shifted compared with those of PSI-LHCI and PSI, and a fluorescence emission peak was similarly shifted to shorter wavelengths. Based on these results, we discuss the diversity of LHC proteins and RedCAP in red algal PSI-LHCI supercomplexes.

Maize (Zea mays L.) performs highly efficient C₄ photosynthesis by dividing photosynthetic metabolism between mesophyll and bundle sheath cells. In vivo physiological measurements are indispensable for C₄ photosynthesis research as photosynthetic activities are easily interrupted by leaf section or cell isolation. Yet, direct in vivo observation regarding bundle sheath cells in the delicate anatomy of the C₄ leaf is still challenging. In the current work, we used two-photon fluorescence-lifetime imaging microscopy (two-photon-FLIM) to access the photosynthetic properties of bundle sheath cells on intact maize leaves. The results provide spectroscopic evidence for the diminished total PSII activity in bundle sheath cells at its physiological level and show that the single PSIIs could undergo charge separation as usual. We also report an acetic acid-induced chlorophyll fluorescence quenching on intact maize leaves, which might be a physiological state related to the nonphotochemical quenching mechanism.

The Orange Carotenoid Protein (OCP) is a unique water-soluble photoactive protein that plays a critical role in regulating the balance between light harvesting and photoprotective responses in cyanobacteria. The challenge in understanding OCP´s photoactivation mechanism stems from the heterogeneity of the initial configurations of its embedded ketocarotenoid, which in the dark-adapted state can form up to two hydrogen bonds to critical amino acids in the protein's C-terminal domain, and the extremely low quantum yield of primary photoproduct formation. While a series of experiments involving point mutations within these contacts helped us to identify these challenges, they did not resolve them. To overcome this, we shifted from classical mutagenesis to the translational introduction of non-canonical amino acid residues into the OCP structure. In this work, we demonstrate that replacing a single meta-hydrogen in tyrosine-201 with a halogen atom (chlorine, bromine, or iodine) leads to targeted modifications in the keto-carotenoid-protein matrix interaction network, both in the dark-adapted state and upon photoactivation. We found that such atomic substitutions allow us to effectively weaken key hydrogen bonds without disrupting protein folding, thereby increasing the yield of OCP photoactivation products. Such genetically encoded chemical modification of individual atoms and their systematic in situ variation in complex protein structures establishes a foundation for transforming OCP into a practical tool for optogenetics and other applications.

The perennially ice-covered Lake Bonney in Antarctica has been deemed a natural laboratory for studying life at the extreme. Photosynthetic algae dominate the lake food webs and are adapted to a multitude of extreme conditions including perpetual shading even at the height of the austral summer. Here we examine how the unique light environment in Lake Bonney influences the physiology of two Chlamydomonas species. Chlamydomonas priscui is found exclusively in the deep photic zone where it receives very low light levels biased in the blue part of the spectrum (400-500 nm). In contrast, Chlamydomonas sp. ICE-MDV is represented at various depths within the water column (including the bright surface waters), and it receives a broad range of light levels and spectral wavelengths. The psychrophilic character of both species makes them an ideal system to study the effects of light quality and quantity on chlorophyll biosynthesis and photosynthetic performance in extreme conditions. We show that the shade-adapted C. priscui exhibits a decreased ability to accumulate chlorophyll and severe photoinhibition when grown under red light compared to blue light. These effects are particularly pronounced under red light of higher intensity, suggesting a loss of capability to acclimate to varied light conditions. In contrast, ICE-MDV has retained the ability to synthesize chlorophyll and maintain photosynthetic efficiency under a broader range of light conditions. Our findings provide insights into the mechanisms of photosynthesis under extreme conditions and have implications on algal survival in changing conditions of Antarctic ice-covered lakes.

Pheophytin-a derivatives possessing plastoquinone and phylloquinone analogs in the peripheral 3-substituent were prepared by Friedel-Crafts reactions of a 3-hydroxymethyl-chlorin as one of the chlorophyll-a derivatives with benzo- and naphthohydroquinones, respectively, and successive oxidation of the 1,4-dihydroxy-aryl groups in the resulting dehydration products. The 3-quinonylmethyl-chlorins exhibited ultraviolet-visible absorption and circular dichroism spectra in acetonitrile, which were composed of those of the starting 3-hydroxymethyl-chlorin and the corresponding methylated benzo- and naphthoquinones. No intramolecular interaction between the chlorin and quinone π-systems was observed in the solution owing to the methylene spacer. The first reduction potentials of the quinone moieties in the synthetic conjugates were determined by cyclic voltammetry and shifted positively from those of the reference quinones. The former quinonyl groups were reduced more readily by approximately 0.1 V than the latter quinones, which was ascribable to the stabilization of the quinonyl anion radical by the nearby macrocyclic chlorin π-chromophore. This observation implied that the reduction potentials of quinones were regulated by the close pheophytin-a derivative by through-space interaction. Considering the charge shift from pheophytin-a anion radical to plastoquinone and phylloquinone in reaction centers of photosystems II and I, respectively, the reduction potentials of these quinones as a determinant factor of the rapid electron transfer process would be dependent on the pheophytin-a in the photosynthetic reaction centers of oxygenic phototrophs as well as on the neighboring peptides.

Seed priming and plant growth-promoting bacteria (PGPB) may alleviate salt stress effects. We exposed a salt-sensitive variety of melon to salinity following seed priming with NaCl and inoculation with Bacillus. Given the sensitivity of photosystem II (PSII) to salt stress, we utilized dark- and light-adapted chlorophyll fluorescence alongside analysis of leaf stomatal conductance of water vapour (G_sw). Priming increased total seed germination by 15.5% under salt-stress. NaCl priming with Bacillus inoculation (PB) increased total leaf area (LA) by 45% under control and 15% under stress. Under the control condition, priming (P) reduced membrane permeability (RMP) by 36% and PB by 55%, while under stress Bacillus (BS) reduced RMP by 10%. Although Bacillus inoculation (B) and priming (P) treatments did not show significant effects on some PSII efficiency parameters (F_V/F_M, ABS/RC, PI_ABS, F_M), the BS treatment induced a significantly higher quantum efficiency of PSII (ΦPSII) and increased G_sw by 159% in the final week of the experiment. The BS treatment reduced electron transport rate per reaction center (ET_O/RC) by 10% in comparison to the salt treatment, which showed less reaction centre damage. Bacillus inoculation and seed priming treatment under the stressed condition (PBS) induced an increase in electron transport rate of 40%. Salt stress started to show significant effects on PSII after 12 days, and adversely impacted all morphological and photosynthetic parameters after 22 days. Salt priming and PGPB mitigated the negative impacts of salt stress and may serve as effective tools in future-proofing saline agriculture.

The active site for water oxidation in photosystem II (PSII) comprises a Mn₄CaO₅ cluster adjacent to a redox-active tyrosine residue (Tyr_Z). During the water-splitting process, the enzyme transitions through five sequential oxidation states (S₀ to S₄), with O₂ evolution occurring during the S₃Tyr_Z· to S₀Tyr_Z transition. Chloride also plays a role in this mechanism. Using PSII from Thermosynechococcus vestitus, where Ca and Cl were replaced with Sr and Br to slow the S₃Tyr_Z· to S₀Tyr_Z + O₂ transition (t_1/2 ~ 5 ms at room temperature), it was observed that the recovery of a S₀ state, defined as the state able to progress to S₁, exhibits similar kinetics (t_1/2 ~ 5 ms). This suggests that in CaCl-PSII, the reformation of the functional S₀ state directly follows the S₃Tyr_Z· to S₀Tyr_Z + O₂ transition, with no additional delay required for the insertion of a new substrate water molecule (O5) and associated protons.

Phosphoenolpyruvate (PEP) carboxylase (PEPC) has an anaplerotic role in central plant metabolism but also initiates the carbon concentrating mechanism during C₄ photosynthesis. The C₄ PEPC has different binding affinities (K_m) for PEP (K_0.5PEP) and HCO₃^- (K_0.5HCO3), and allosteric regulation by glucose-6-phosphate (G6-P) compared to non-photosynthetic isoforms. These differences are linked to specific changes in amino acids within PEPC. For example, region II (residues 302-433, Zea mays numbering) has been identified as important for G6-P regulation and within this region residue 353 may be conserved in C₄ PEPC enzymes. Additionally, residue 780 influences the C₄ PEPC kinetic properties and may interact with region II as well as residue 353 to influence G6-P regulation. We test the hypothesis that variation within region II, including residue 353, and their interactions with residue 780 influence the kinetic and allosteric regulation by G6-P of two C₄ PEPC isozymes from two C₄ grasses. The data does not support a kinetic tradeoff between K_0.5HCO3 and K_0.5PEP in these PEPC isozymes. Additionally, these enzymes had different response to G6-P that was only partially attributed to region II, residue 353 and residue 780. This data provides new insights into factors influencing the kinetic variation of C₄ PEPC isozymes.

This study examined the impacts of different LED spectra on the growth of in vitro cultures of Musa acuminata cv. red banana and their biochemical profile, including the antioxidant enzymes catalase and ascorbate peroxidase, photosynthetic pigment and accumulation of total carbohydrate content. The far-red LEDs significantly increase shoot elongation (10.04 cm). The greatest number of shoots (2.97) and the greatest multiplication rate (80%) were obtained under the treatment with blue + red LEDs. The formation of microshoots were also enhanced by blue and white LED exposure in a range of 2-2.57 shoots per explant. Root formation was also stimulated by dichromatic blue + red (6.00) LED using MS medium with 2 µM indole-3-butyric acid (IBA). The enzymes catalase and ascorbate peroxidase were significantly up-regulated by irradiation with far-red (0.11 ± 0.02 CAT, 0.18 ± 0.04 APX U/mg) and blue (0.08 ± 0.01CAT, 0.10 ± 0.01APX U/mg) LED light. Total chlorophyll (0.45 to 0.80 mg/g) was elevated significantly by blue, blue + red and mint-white LED. On the other hand, carotenoids (12.08-14.61 mg/g) were significantly boosted by blue + red, red and mint-white LED light. Meanwhile, porphyrin (294.10-350.57 mg/g) was highly synthesised after irradiation with mint-white light. Irradiation with LED light significantly increased the accumulation of carbohydrates with the highest carbohydrate content under blue + red LED light (102.22 ± 2.46 mg/g) and blue light (91.69 ± 2.10 mg/g). In conclusion, these results confirm that the vegetative properties and biochemical profile of red banana in vitro are eustress response to LED spectra.