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Summary: Woody plants display some photosynthetic activity in stems, but the biological role of stem photosynthesis and the specific contributions of bark and wood to carbon uptake and oxygen evolution remain poorly understood.We aimed to elucidate the functional characteristics of chloroplasts in stems of different ages in Fraxinus ornus. Our investigation employed diverse experimental approaches, including microsensor technology to assess oxygen production rates in whole stem, bark, and wood separately. Additionally, we utilized fluorescence lifetime imaging microscopy (FLIM) to characterize the relative abundance of photosystems I and II (PSI : PSII chlorophyll ratio) in bark and wood.Our findings revealed light‐induced increases in O2 production in whole stem, bark, and wood. We present the radial profile of O2 production in F. ornus stems, demonstrating the capability of stem chloroplasts to perform light‐dependent electron transport. Younger stems exhibited higher light‐induced O2 production and dark respiration rates than older ones.While bark emerged as the primary contributor to net O2 production under light conditions, our data underscored that wood chloroplasts are also photosynthetically active. The FLIM analysis unveiled a lower PSI abundance in wood than in bark, suggesting stem chloroplasts are not only active but also acclimate to the spectral composition of light reaching inner compartments. [ABSTRACT FROM AUTHOR]

Life on earth depends on photosynthetic primary producers that exploit sunlight to fix CO2 into biomass. Approximately half of global primary production is associated with microalgae living in aquatic environments. Microalgae also represent a promising source of biomass to complement crop cultivation, and they could contribute to the development of a more sustainable bioeconomy. Photosynthetic organisms evolved multiple mechanisms involved in the regulation of photosynthesis to respond to highly variable environmental conditions. While essential to avoid photodamage, regulation of photosynthesis results in dissipation of absorbed light energy, generating a complex trade-off between protection from stress and light-use efficiency. This work investigates the impact of the xanthophyll cycle, the light-induced reversible conversion of violaxanthin into zeaxanthin, on the protection from excess light and on biomass productivity in the marine microalgae of the genus Nannochloropsis. Zeaxanthin is shown to have an essential role in protection from excess light, contributing to the induction of non-photochemical quenching and scavenging of reactive oxygen species. On the contrary, the overexpression of zeaxanthin epoxidase enables a faster reconversion of zeaxanthin to violaxanthin that is shown to be advantageous for biomass productivity in dense cultures in photobioreactors. These results demonstrate that zeaxanthin accumulation is critical to respond to strong illumination, but it maylead to unnecessary energy losses in light-limiting conditions and accelerating its reconversion to violaxanthin provides an advantage for biomass productivity in microalgae. [ABSTRACT FROM AUTHOR]

Leaves are the most important photosynthetic organs in most woody plants, but chloroplasts are also found in organs optimized for other functions. However, the actual photosynthetic efficiency of these chloroplasts is still unclear. We analyzed bark and wood chloroplasts of Fraxinus ornus L. saplings. Optical and spectroscopic methods were applied to stem samples and compared with leaves. A sharp light gradient was detected along the stem radial direction, with blue light mainly absorbed by the outer bark, and far-red-enriched light reaching the underlying xylem and pith. Chlorophylls were evident in the xylem rays and the pith and showed an increasing concentration gradient toward the bark. The stem photosynthetic apparatus showed features typical of acclimation to a low-light environment, such as larger grana stacks, lower chlorophyll a/b and photosystem I/II ratios compared with leaves. Despite likely receiving very few photons, wood chloroplasts were photosynthetically active and fully capable of generating a light-dependent electron transport. Our data provide a comprehensive scenario of the functional features of bark and wood chloroplasts in a woody species and suggest that stem photosynthesis is coherently optimized to the prevailing micro-environmental conditions at the bark and wood level. [ABSTRACT FROM AUTHOR]

SUMMARY: Plants exposed to light fluctuations are protected from photodamage by non‐photochemical quenching (NPQ), a reversible mechanism that enables dissipation of excess absorbed energy as heat, which is essential for plant fitness and crop productivity. In plants NPQ requires the presence of the membrane protein PsbS, which upon activation interacts with antenna proteins, inducing their dissipative conformation. Here, we exploited base editing (BE) in the moss Physcomitrium patens to introduce specific amino acid changes in vivo and assess their impact on PsbS activity, targeting transmembrane regions to investigate their role in essential protein–protein interactions. This approach enabled the recognition of residues essential for protein stability and the identification of a hydrophobic cluster of amino acids impacting PsbS activity. This work provides new information on the molecular mechanism of PsbS while also demonstrating the potential of BE approaches for in planta gene function analysis. [ABSTRACT FROM AUTHOR]

Microalgae represent a sustainable source of biomass that can be exploited for pharmaceutical, nutraceutical, cosmetic applications, as well as for food, feed, chemicals, and energy. To make microalgae applications economically competitive and maximize their positive environmental impact, it is however necessary to optimize productivity when cultivated at a large scale. Independently from the final product, this objective requires the optimization of biomass productivity and thus of microalgae ability to exploit light for CO2 fixation. Light is a highly variable environmental parameter, continuously changing depending on seasons, time of the day, and weather conditions. In microalgae large scale cultures, cell self-shading causes inhomogeneity in light distribution and, because of mixing, cells move between different parts of the culture, experiencing abrupt changes in light exposure. Microalgae evolved multiple regulatory mechanisms to deal with dynamic light conditions that, however, are not adapted to respond to the complex mixture of natural and artificial fluctuations found in large-scale cultures, which can thus drive to oversaturation of the photosynthetic machinery, leading to consequent oxidative stress. In this work, the present knowledge on the regulation of photosynthesis and its implications for the maximization of microalgae biomass productivity are discussed. Fast mechanisms of regulations, such as Non-Photochemical-Quenching and cyclic electron flow, are seminal to respond to sudden fluctuations of light intensity. However, they are less effective especially in the 1–100 s time range, where light fluctuations were shown to have the strongest negative impact on biomass productivity. On the longer term, microalgae modulate the composition and activity of the photosynthetic apparatus to environmental conditions, an acclimation response activated also in cultures outdoors. While regulation of photosynthesis has been investigated mainly in controlled lab-scale conditions so far, these mechanisms are highly impactful also in cultures outdoors, suggesting that the integration of detailed knowledge from microalgae large-scale cultivation is essential to drive more effective efforts to optimize biomass productivity. How does microalgae photosynthesis respond to outdoor cultivation at industrial-scale? [ABSTRACT FROM AUTHOR]

Eukaryotic algae are photosynthetic organisms capable of exploiting sunlight to fix carbon dioxide into biomass with highly variable genetic and metabolic features. Information on algae metabolism from different species is inhomogeneous and, while green algae are, in general, more characterized, information on red algae is relatively scarce despite their relevant position in eukaryotic algae diversity. Within red algae, the best‐known species are extremophiles or multicellular, while information on mesophilic unicellular organisms is still lacunose. Here, we investigate the photosynthetic properties of a recently isolated seawater unicellular mesophilic red alga, Dixoniella giordanoi. Upon exposure to different illuminations, D. giordanoi shows the ability to acclimate, modulate chlorophyll content, and re‐organize thylakoid membranes. Phycobilisome content is also largely regulated, leading to almost complete disassembly of this antenna system in cells grown under intense illumination. Despite the absence of a light‐induced xanthophyll cycle, cells accumulate zeaxanthin upon prolonged exposure to strong light, likely contributing to photoprotection. D. giordanoi cells show the ability to perform cyclic electron transport that is enhanced under strong illumination, likely contributing to the protection of Photosystem I from over‐reduction and enabling cells to survive PSII photoinhibition without negative impact on growth. [ABSTRACT FROM AUTHOR]

During samplings aimed at isolating microalgal strains, a coccoid greyish-green alga was collected along the North Adriatic coasts (Mediterranean Sea, Italy) and grown in culture. The microalgal strain (named strain B1A) was then subjected to an integrative taxonomy approach in order to correctly identify it. Morphological and ultrastructural observations and phycobiliprotein content analysis were carried out, as well as molecular analyses based on the 18S rRNA, rbcL, psbA and plastid-encoded 23S rRNA genes. Phylogenetic placement and ultrastructural observations clearly indicated that strain B1A is a member of the red microalgal genus Dixoniella (Rhodellophyceae, Proteorhodophytina) and is distinct from the only species so far described for this taxon. Therefore, a new species was described to encompass the isolate from the North Adriatic Sea and another isolate from Japan (MF-G2), which was phylogenetically related to strain B1A: Dixoniella giordanoi sp. nov. [ABSTRACT FROM AUTHOR]

Summary: CRISPR‐Cas9 has proven to be highly valuable for genome editing in plants, including the model plant Physcomitrium patens. However, the fact that most of the editing events produced using the native Cas9 nuclease correspond to small insertions and deletions is a limitation.CRISPR‐Cas9 base editors enable targeted mutation of single nucleotides in eukaryotic genomes and therefore overcome this limitation. Here, we report two programmable base‐editing systems to induce precise cytosine or adenine conversions in P. patens.Using cytosine or adenine base editors, site‐specific single‐base mutations can be achieved with an efficiency up to 55%, without off‐target mutations. Using the APT gene as a reporter of editing, we could show that both base editors can be used in simplex or multiplex, allowing for the production of protein variants with multiple amino‐acid changes. Finally, we set up a co‐editing selection system, named selecting modification of APRT to report gene targeting (SMART), allowing up to 90% efficiency site‐specific base editing in P. patens.These two base editors will facilitate gene functional analysis in P. patens, allowing for site‐specific editing of a given base through single sgRNA base editing or for in planta evolution of a given gene through the production of randomly mutagenised variants using multiple sgRNA base editing. [ABSTRACT FROM AUTHOR]

Alternative electron pathways contribute to regulation of photosynthetic light reactions to adjust to metabolic demands in dynamic environments. The chloroplast NADH dehydrogenase-like (NDH) complex mediates the cyclic electron transport pathway around PSI in different cyanobacteria, algae, and plant species, but it is not fully conserved in all photosynthetic organisms. In order to assess how the physiological role of this complex changed during plant evolution, we isolated Physcomitrella patens lines knocked out for the NDHM gene that encodes a subunit fundamental for the activity of the complex. ndhm knockout mosses indicated high PSI acceptor side limitation upon abrupt changes in illumination. In P. patens , pseudo-cyclic electron transport mediated by flavodiiron proteins (FLVs) was also shown to prevent PSI over-reduction in plants exposed to light fluctuations. flva ndhm double knockout mosses had altered photosynthetic performance and growth defects under fluctuating light compared with the wild type and single knockout mutants. The results showed that while the contribution of NDH to electron transport is minor compared with FLV, NDH still participates in modulating photosynthetic activity, and it is critical to avoid PSI photoinhibition, especially when FLVs are inactive. The functional overlap between NDH- and FLV-dependent electron transport supports PSI activity and prevents its photoinhibition under light variations. [ABSTRACT FROM AUTHOR]

Oxygenic photosynthetic microorganisms are a focal point of research in the context of human space exploration. As part of the bioregenerative life-support systems, they could have a key role in the production of breathable O2, edible biomasses and in the regeneration of CO2 rich-atmospheres and wastewaters produced by astronauts. The test of the organism's response to simulated physico-chemical parameters of planetary bodies could also provide important information about their habitability potential. It is believed that the success of future planetary and space missions will require innovative technologies, developed on the base of preliminary experiments in custom-made laboratory facilities. In this context, simulation chambers will play a pivotal role by allowing the growth of the microorganisms under controlled conditions and the evaluation in real-time of their biomass productivity and impact on atmosphere composition. We here present a system capable of addressing these requirements with high replicability and low costs. The setup is composed by three main parts: 1) a Star Light Simulator, able to generate different light intensities and spectra, including those of non-solar stars; 2) an Atmosphere Simulator Chamber where cultures of photosynthetic microorganisms can be exposed to different gas compositions; 3) a reflectivity detection system to measure from remote the Normalized Difference Vegetation Indexes (NDVI). Such a setup allows us to monitor photosynthetic microorganism's growth and gas exchange performances under selected conditions of light quality and intensity, temperature, pressure, and atmospheres simulating non-terrestrial environments. All parameters are detected by remote sensing techniques, thus without interfering with the experiments and altering the environmental conditions set. We validated the setup by growing cyanobacteria liquid cultures under different light intensities of solar illumination, collecting data on their growth rate, photosynthetic activity, and gas exchange capacity. We utilized the reflectivity detection system to measure the reflection spectra of the growing cultures, obtaining their relative NDVI that was shown to correlate with optical density, chlorophyll content, and dry weight, demonstrating the potential application of this index as a proxy of growth. [ABSTRACT FROM AUTHOR]

In nature, photosynthetic organisms are exposed to highly dynamic environmental conditions where the excitation energy and electron flow in the photosynthetic apparatus need to be continuously modulated. Fluctuations in incident light are particularly challenging because they drive oversaturation of photosynthesis with consequent oxidative stress and photoinhibition. Plants and algae have evolved several mechanisms to modulate their photosynthetic machinery to cope with light dynamics, such as thermal dissipation of excited chlorophyll states (non-photochemical quenching, NPQ) and regulation of electron transport. The regulatory mechanisms involved in the response to light dynamics have adapted during evolution, and exploring biodiversity is a valuable strategy for expanding our understanding of their biological roles. In this work, we investigated the response to fluctuating light in Nannochloropsis gaditana , a eukaryotic microalga of the phylum Heterokonta originating from a secondary endosymbiotic event. Nannochloropsis gaditana is negatively affected by light fluctuations, leading to large reductions in growth and photosynthetic electron transport. Exposure to light fluctuations specifically damages photosystem I, likely because of the ineffective regulation of electron transport in this species. The role of NPQ, also assessed using a mutant strain specifically depleted of this response, was instead found to be minor, especially in responding to the fastest light fluctuations. [ABSTRACT FROM AUTHOR]

The regulation of photosynthesis is crucial to efficiently support the assimilation of carbon dioxide and to prevent photodamage. One key regulatory mechanism is the pseudo-cyclic electron flow (PCEF) mediated by class-C flavodiiron proteins (FLVs). These enzymes use electrons coming from Photosystem I (PSI) to reduce oxygen to water, preventing over-reduction in the acceptor side of PSI. FLVs are widely distributed among organisms performing oxygenic photosynthesis and they have been shown to be fundamental in many different conditions such as fluctuating light, sulfur deprivation and plant submersion. Moreover, since FLVs reduce oxygen they can help controlling the redox status of the cell and maintaining the microoxic environment essential for processes such as nitrogen fixation in cyanobacteria. Despite these important roles identified in various species, the genes encoding for FLV proteins have been lost in angiosperms where their activity could have been at least partially compensated by a more efficient cyclic electron flow (CEF). The present work reviews the information emerged on FLV function, analyzing recent structural data that suggest FLV could be regulated through a conformational change. [ABSTRACT FROM AUTHOR]