Phosphatidic acid phosphohydrolase modulates glycerolipid synthesis in Marchantia polymorpha and is crucial for growth under both nutrient-replete and -deficient conditions.

The phosphatidic acid phosphohydrolase of Marchantia polymorpha modulates plastid glycolipid synthesis through the ER pathway and is essential for normal plant development regardless of nutrient availability. Membrane lipid remodeling is one of the strategies plant cells use to secure inorganic phosphate (Pi) for plant growth, but many aspects of the molecular mechanism and its regulation remain unclear. Here we analyzed membrane lipid remodeling using a non-vascular plant, Marchantia polymorpha.

Crystal structures of photosystem II from a cyanobacterium expressing psbA in comparison to psbA reveal differences in the D1 subunit.

Three psbA genes (psbA, psbA, and psbA) encoding the D1 subunit of photosystem II (PSII) are present in the thermophilic cyanobacterium Thermosynechococcus elongatus and are expressed differently in response to changes in the growth environment. To clarify the functional differences of the D1 protein expressed from these psbA genes, PSII dimers from two strains, each expressing only one psbA gene (psbA or psbA), were crystallized, and we analyzed their structures at resolutions comparable to previously studied PsbA1-PSII. Our results showed that the hydrogen bond between pheophytin/D1 (Pheo) and D1-130 became stronger in PsbA2- and PsbA3-PSII due to change of Gln to Glu, which partially explains the increase in the redox potential of Pheo observed in PsbA3.

Genome editing with removable TALEN vectors harboring a yeast centromere and autonomous replication sequence in oleaginous microalga.

Algal lipids are expected to become a basis for sustainable fuels because of the highly efficient lipid production by photosynthesis accompanied by carbon dioxide assimilation. Molecular breeding of microalgae has been studied to improve algal lipid production, but the resultant gene-modified algae containing transgenes are rarely used for outdoor culture because the use of genetically modified organisms (GMOs) is strictly restricted under biocontainment regulations. Recently, it was reported that plasmids containing yeast centromere and autonomous replication sequence (CEN/ARS) behaved as episomes in Nannochloropsis species.

Recycling of the major thylakoid lipid MGDG and its role in lipid homeostasis in Chlamydomonas reinhardtii.

Monogalactosyldiacylglycerol (MGDG), the most abundant lipid in thylakoid membranes, is involved in photosynthesis and chloroplast development. MGDG lipase has an important role in lipid remodeling in Chlamydomonas reinhardtii. However, the process related to turnover of the lysogalactolipid that results from MGDG degradation, monogalactosylmonoacylglycerol (MGMG), remains to be clarified.

Methods of Lipid Analyses for Microalgae: Charophytes, Eustigmatophytes, and Euglenophytes.

Algae are ecologically important organisms and are widely used for basic research, with a focus on for example photosynthesis, evolution, and lipid metabolism. Many biosynthetic pathways of algal lipids have been deciphered using available genomic information. Here we describe methods for lipid analyses from three representative algae, including Archaeplastida, the SAR lineage (Stramenopiles, Alveolata, Rhizaria), and Excavata.

Efficient and multiplexable genome editing using Platinum TALENs in oleaginous microalga, Nannochloropsis oceanica NIES-2145.

Algae accumulate large amounts of lipids produced by photosynthesis, and these lipids are expected to be utilized as feedstocks for sustainable new energies, known as biodiesels. Nannochloropsis species are eukaryotic microalgae that produce high levels of lipids. However, since the production costs of algal biodiesels are higher than those of fossil fuels, the improved productivity of algal lipids by molecular breeding of algae is required for practical use.

MYB-like transcription factor NoPSR1 is crucial for membrane lipid remodeling under phosphate starvation in the oleaginous microalga Nannochloropsis oceanica.

Membrane lipid remodeling under phosphate (Pi) limitation, a process that replaces structural membrane phospholipids with nonphosphorus lipids, is a widely observed adaptive response in plants and algae. Here, we identified the transcription factor phosphorus starvation response 1 (NoPSR1) as an indispensable player for regulating membrane lipid conversion during Pi starvation in the microalga Nannochloropsis oceanica. Knocking out NoPSR1 scarcely perturbed membrane lipid composition under Pi-sufficient conditions but significantly impaired dynamic alteration in membrane lipids during Pi starvation.

Lipid remodeling regulator 1 (LRL1) is differently involved in the phosphorus-depletion response from PSR1 in Chlamydomonas reinhardtii.

The elucidation of lipid metabolism in microalgae has attracted broad interest, as their storage lipid, triacylglycerol (TAG), can be readily converted into biofuel via transesterification. TAG accumulates in the form of oil droplets, especially when cells undergo nutrient deprivation, such as for nitrogen (N), phosphorus (P), or sulfur (S). TAG biosynthesis under N-deprivation has been comprehensively studied in the model microalga Chlamydomonas reinhardtii, during which TAG accumulates dramatically.

Lysyl oxidase-like protein secreted from an acidophilic red alga, .

is a primitive acidophilic red alga which grown optimally at pH 1-3. When the alga was cultured at pH 6, which is the upper limit of acidity for its survival, most of the algal cells became large cells with four endospores which did not split into daughter cells. This suggests that the alga survives in the endospore state at pH 6 to protect against nutrient uptake deficiency due to low pH gradient across the cell membranes.

Photocurrent Generation of Reconstituted Photosystem II on a Self-Assembled Gold Film.

Photosystem II (PSII)-modified gold electrodes were prepared by the deposition of PSII reconstituted with platinum nanoparticles (PtNPs) on Au electrodes. PtNPs modified with 1-[15-(3,5,6-trimethyl-1,4-benzoquinone-2-yl)]pentadecyl disulfide ((TMQ(CH)S)) were incorporated into the Q site of PSII isolated from thermophilic cyanobacterium Thermosynechococcus elongatus. The reconstitution was confirmed by Q-reoxidation measurements.

Manipulation of oil synthesis in Nannochloropsis strain NIES-2145 with a phosphorus starvation-inducible promoter from Chlamydomonas reinhardtii.

Microalgae accumulate triacylglycerols (TAGs) under conditions of nutrient stress. Phosphorus (P) starvation induces the accumulation of TAGs, and the cells under P starvation maintain growth through photosynthesis. We recently reported that P starvation-dependent overexpression of type-2 diacylglycerol acyl-CoA acyltransferase (CrDGTT4) from Chlamydomonas reinhardtii using a sulfoquinovosyldiacylglycerol synthase 2 (SQD2) promoter, which has increased activity during P starvation, enhances TAG accumulation in C.

Usefulness of Adalimumab for Treating a Case of Intestinal Behçet's Disease With Trisomy 8 Myelodysplastic Syndrome.

Behçet's disease (BD) is a systemic vasculitis, while myelodysplastic syndrome (MDS) is a heterogeneous group of clonal hematologic disorders characterized by ineffective hematopoiesis. Some studies suggest a relationship between MDS and BD, especially intestinal BD, and trisomy 8 seems to play an important role in both diseases. There are several reports on patients with BD comorbid with MDS involving trisomy 8 that frequently have intestinal lesions refractory to conventional medical therapies.

Enhancement of extraplastidic oil synthesis in Chlamydomonas reinhardtii using a type-2 diacylglycerol acyltransferase with a phosphorus starvation-inducible promoter.

When cultivated under stress conditions, many plants and algae accumulate oil. The unicellular green microalga Chlamydomonas reinhardtii accumulates neutral lipids (triacylglycerols; TAGs) during nutrient stress conditions. Temporal changes in TAG levels in nitrogen (N)- and phosphorus (P)-starved cells were examined to compare the effects of nutrient depletion on TAG accumulation in C.

Klebsormidium flaccidum genome reveals primary factors for plant terrestrial adaptation.

The colonization of land by plants was a key event in the evolution of life. Here we report the draft genome sequence of the filamentous terrestrial alga Klebsormidium flaccidum (Division Charophyta, Order Klebsormidiales) to elucidate the early transition step from aquatic algae to land plants. Comparison of the genome sequence with that of other algae and land plants demonstrate that K.

Roles of PsbI and PsbM in photosystem II dimer formation and stability studied by deletion mutagenesis and X-ray crystallography.

PsbM and PsbI are two low molecular weight subunits of photosystem II (PSII), with PsbM being located in the center, and PsbI in the periphery, of the PSII dimer. In order to study the functions of these two subunits from a structural point of view, we crystallized and analyzed the crystal structure of PSII dimers from two mutants lacking either PsbM or PsbI. Our results confirmed the location of these two subunits in the current crystal structure, as well as their absence in the respective mutants.

Functional characterization and quantification of the alternative PsbA copies in Thermosynechococcus elongatus and their role in photoprotection.

The D1 protein (PsbA) of photosystem II (PSII) from Thermosynechococcus elongatus is encoded by a psbA gene family that is typical of cyanobacteria. Although the transcription of these three genes has been studied previously (Kós, P. B.

Binding and functional properties of five extrinsic proteins in oxygen-evolving photosystem II from a marine centric diatom, Chaetoceros gracilis.

Oxygen-evolving photosystem II (PSII) isolated from a marine centric diatom, Chaetoceros gracilis, contains a novel extrinsic protein (Psb31) in addition to four red algal type extrinsic proteins of PsbO, PsbQ', PsbV, and PsbU. In this study, the five extrinsic proteins were purified from alkaline Tris extracts of the diatom PSII by anion and cation exchange chromatographic columns at different pH values. Reconstitution experiments in various combinations with the purified extrinsic proteins showed that PsbO, PsbQ', and Psb31 rebound directly to PSII in the absence of other extrinsic proteins, indicating that these extrinsic proteins have their own binding sites in PSII intrinsic proteins.

Topological analysis of the extrinsic PsbO, PsbP and PsbQ proteins in a green algal PSII complex by cross-linking with a water-soluble carbodiimide.

The close association of the extrinsic PsbO, PsbP and PsbQ proteins with PSII core subunits in oxygen-evolving PSII complexes from a green alga, Chlamydomonas reinhardtii, was examined by cross-linking experiments with a water-soluble carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). The green algal PSII complexes treated with EDC were washed with alkaline Tris to remove the non-cross-linked extrinsic proteins, and then applied to Blue-Native-PAGE to prepare PSII core complexes. The extrinsic proteins cross-linked with PSII core complexes were detected by immunoblotting analysis using antibodies against extrinsic proteins and PSII core subunits.

The PsbK subunit is required for the stable assembly and stability of other small subunits in the PSII complex in the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1.

PsbK is a small membrane protein of the PSII core complex and is highly conserved from cyanobacteria to plants. Here, we studied its role in the thermophilic cyanobacterium, Thermosynechococcus elongatus BP-1, by focusing on a psbK disruptant with hexahistidine-tagged CP47. The psbK disruptant showed photoautotrophic growth comparable with that of the wild type under a wide range of light conditions.

Structural and functional studies on Ycf12 (Psb30) and PsbZ-deletion mutants from a thermophilic cyanobacterium.

Ycf12 (Psb30) and PsbZ are two low molecular weight subunits of photosystem II (PSII), with one and two trans-membrane helices, respectively. In order to study the functions of these two subunits from a structural point of view, we constructed deletion mutants lacking either Ycf12 or PsbZ from Thermosynechococcus elongatus, and purified, crystallized and analyzed the structure of PSII dimer from the two mutants. Our results showed that Ycf12 is located in the periphery of PSII, close to PsbK, PsbZ and PsbJ, and corresponded to the unassigned helix X1 reported previously, in agreement with the recent structure at 2.

Photosensor based on an FET utilizing a biocomponent of photosystem I for use in imaging devices.

We have investigated a photosensor that consists of a field emission transistor (FET) utilizing the biocomponent of the photosystem I (PSI) protein complex for use in an imaging device. The PSI was immobilized on a gold electrode via the self-assembling monolayer (SAM) of 3-mercapto-1-propanesulfonic acid sodium salt to obtain a PSI-modified gold electrode. As for the PSI-modified gold electrode, the basic photoresponses originating from the excitation of PSI, including the photocurrent (106 nA) and the photoresponse of the open-circuit voltage (photo-Voc: 28.

Is the photosystem II complex a monomer or a dimer?

It is widely believed that the photosystem II (PSII) complex may function as a dimer in the thylakoid membrane. Here, we report experimental conversion from the monomeric PSII to the dimeric form by treatment with high concentrations of n-dodecyl-beta-D-maltopyranoside (DM). The content of the PSII monomer in a PsbTc deletion mutant was much higher than in the wild type when solubilized with low concentrations of DM.

Surface immobilization of PSI using vitamin K1-like molecular wires for fabrication of a bio-photoelectrode.

We present a bio-photoelectrode made of photosystem I complex (PSI) of cyanobacteria, Thermosynechococcus elongatus, and molecular wires. The newly designed and synthesized molecular wire similar to vitamin K1 (VK1) has a naphthoquinone moiety that can connect to PSI, and a terpyridine moiety for connection to a Co(II) ion. Surface immobilization of PSI and molecular wires was performed by a self-assembled monolayer (SAM) formation process at indium tin oxide (ITO) followed by stepwise coordination reactions.

Chlorogenic acid facilitates root hair formation in lettuce seedlings.

Root hairs, which arise from root epidermal cells, are tubular structures that increase the efficiency of water absorption and nutrient uptake. A low pH (pH 4) medium induced root hair formation in lettuce (Lactuca sativa L.) seedlings, and the decapitation of shoots inhibited root hair formation.