Membranes Of Higher Plants Research Articles

Suberin- A potential source of renewable chemicals for industrial applications

Suberin is a lipid-based biopolymer present in the plasma membrane in higher plants and acts as a hydrophobic barrier. It is present in roots, leaves, outer bark and periderm of the tuber plants. This polymer protects plants from environmental stress including drought, salinity, pathogen attack and water loss. The present review has explored the current aspects of suberin structure, its biosynthesis, depolymerization methods and Suberin-derived monomers application. Different chemicals, as well as biological processes, are present to recover these monomers and these suberin based monomers have shown several industrial applications. De-polymerisation of suberin is an essential step for extracting these valuable added macromolecules including di-hydroxy acids, monohydroxy fatty acids, phenolics and glycerol. The importance of suberin monomers in polymer, pharmaceutical and food industries is also highlighted. Hence, waste from agro-food industries rich in suberin components could be a promising source for these renewable chemicals extraction.

Overexpression of Rice Monogalactosyldiacylglycerol Synthase OsMGD Leads to Enhanced Salt Tolerance in Rice

Monogalactosyldiacylglycerol (MGDG) is the most abundant lipid on earth and constitutes about 50% of the polar lipids in the thylakoid membrane of higher plants. MGDG synthase (MGD) is the key enzyme responsible for MGDG synthesis in plants. However, so far, the function of the MGD gene in plant stress responses is still unknown, especially in crop plants. In this study, the performances of wild-type rice (Oryza sativa L. Nipponbare), OsMGD overexpression, and RNA interference lines were analyzed under salt stress to clarify the role of OsMGD in rice under salt stress. The results showed that, after 6 days of NaCl treatment, OsMGD-overexpressing rice seedlings had higher biomass than the wild-type rice, with significantly lower Na+ content and Na+ to K+ ratio in the shoots. Meanwhile, overexpression lines showed a significant increase in the contents of chlorophyll a and carotenoid and photosynthetic capabilities, including the net photosynthetic rate, maximal quantum yield (Fv/Fm), effective PSII quantum yield (ΦPSII), electron transport rate (ETR), and coefficient of photochemical quenching (qP), compared with the wild-type line. Also, OsMGD overexpression lines contained higher MGDG and 18:3 unsaturated fatty acid levels under salt stress. These results indicate that OsMGD participates in the regulation of improving salt tolerance through protecting photosynthetic capabilities and ion balance in rice.

Genome-Wide Analysis of AAT Genes and Their Expression Profiling during Fiber Development in Cotton.

Amino acid transporters (AATs) are a kind of membrane proteins that mediate the transport of amino acids across cell membranes in higher plants. The AAT proteins are involved in regulating plant cell growth and various developmental processes. However, the biological function of this gene family in cotton fiber development is not clear. In this study, 190, 190, 101, and 94 full-length AAT genes were identified from Gossypium hirsutum, G. barbadense, G. arboreum, and G. raimondii. A total of 575 AAT genes from the four cotton species were divided into two subfamilies and 12 clades based on phylogenetic analysis. The AAT genes in the four cotton species were distributed on all the chromosomes. All GhAAT genes contain multiple exons, and each GhAAT protein has multiple conserved motifs. Transcriptional profiling and RT qPCR analysis showed that four GhATT genes tend to express specifically at the fiber initiation stage. Eight genes tend to express specifically at the fiber elongation and maturity stage, and four genes tend to express specifically at the fiber initiation and elongation stages. Our results provide a solid basis for further elucidating the biological function of AAT genes related to cotton fiber development and offer valuable genetic resources for crop improvement in the future.

Special issue in honour of Prof. Reto J. Strasser - Thylakoid membrane reorganization, induced by growth light intensity, affects the plants susceptibility to drought stress

One of the major questions in photosynthesis research is related to the role of the ultrastructure of thylakoid membranes in higher plants for their functional performance at optimal and stress environmental conditions. In this work, we compared the drought stress response of pea plants grown at moderate and low light conditions, based on our recent findings that they differ in the extent of grana stacking and in the thermal stability of the major light-harvesting complex of PSII that plays important regulatory role for plant survival. Our data demonstrate that low light-adapted plants are structurally and functionally less affected by the water deficit, strongly suggesting a major role of thylakoid structural organization for drought stress response.

Nanophotonics of higher-plant photosynthetic membranes

The thylakoid membrane inside chloroplasts hosts the light-dependent reactions of photosynthesis. Its embedded protein complexes are responsible for light harvesting, excitation energy transfer, charge separation, and transport. In higher plants, when the illumination conditions vary, the membrane adapts its composition and nanoscale morphology, which is characterized by appressed and non-appressed regions known as grana and stroma lamellae, respectively. Here we investigate the nanophotonic regime of light propagation in chloroplasts of higher plants and identify novel mechanisms in the optical response of the thylakoid membrane. Our results indicate that the relative contributions of light scattering and absorption to the overall optical response of grana strongly depend on the concentration of the light-harvesting complexes. For the pigment concentrations typically found in chloroplasts, the two mechanisms have comparable strengths, and their relative value can be tuned by variations in the protein composition or in the granal diameter. Furthermore, we find that collective modes in ensembles of grana significantly increase light absorption at selected wavelengths, even in the presence of moderate biological disorder. Small variations in the granal separation or a large disorder can dismantle this collective response. We propose that chloroplasts use this mechanism as a strategy against dangerously high illumination conditions, triggering a transition to low-absorbing states. We conclude that the morphological separation of the thylakoid membrane in higher plants supports strong nanophotonic effects, which may be used by chloroplasts to regulate light absorption. This adaptive self-organization capability is of interest as a model for novel bioinspired optical materials for artificial photosynthesis, imaging, and sensing.

The DnaJ-Like Zinc-Finger Protein HCF222 Is Required for Thylakoid Membrane Biogenesis in Plants.

To understand the biogenesis of the thylakoid membrane in higher plants and to identify auxiliary proteins required to build up this highly complex membrane system, we have characterized the allelic nuclear mutants high chlorophyll fluorescence222-1 (hcf222-1) and hcf222-2 and isolated the causal gene by map-based cloning. In the ethyl methanesulfonate-induced mutant hcf222-1, the accumulation of the cytochrome b6f (Cytb6f) complex was reduced to 30% compared with the wild type. Other thylakoid membrane complexes accumulated to normal levels. The T-DNA knockout mutant hcf222-2 showed a more severe defect with respect to thylakoid membrane proteins and accumulated only 10% of the Cytb6f complex, accompanied by a reduction in photosystem II, the photosystem II light-harvesting complex, and photosystem I. HCF222 encodes a protein of 99 amino acids in Arabidopsis (Arabidopsis thaliana) that has similarities to the cysteine-rich zinc-binding domain of DnaJ chaperones. The insulin precipitation assay demonstrated that HCF222 has disulfide reductase activity in vitro. The protein is conserved in higher plants and bryophytes but absent in algae and cyanobacteria. Confocal fluorescence microscopy showed that a fraction of HCF222-green fluorescent protein was detectable in the endoplasmic reticulum but that it also could be recognized in chloroplasts. A fusion construct of HCF222 containing a plastid transit peptide targets the protein into chloroplasts and was able to complement the mutational defect. These findings indicate that the chloroplast-targeted HCF222 is indispensable for the maturation and/or assembly of the Cytb6f complex and is very likely involved in thiol-disulfide biochemistry at the thylakoid membrane.

The Interactions of Aquaporins and Mineral Nutrients in Higher Plants.

Aquaporins, major intrinsic proteins (MIPs) present in the plasma and intracellular membranes, facilitate the transport of small neutral molecules across cell membranes in higher plants. Recently, progress has been made in understanding the mechanisms of aquaporin subcellular localization, transport selectivity, and gating properties. Although the role of aquaporins in maintaining the plant water status has been addressed, the interactions between plant aquaporins and mineral nutrients remain largely unknown. This review highlights the roles of various aquaporin orthologues in mineral nutrient uptake and transport, as well as the regulatory effects of mineral nutrients on aquaporin expression and activity, and an integrated link between aquaporins and mineral nutrient metabolism was identified.

Plastoglobules in algae: A comprehensive comparative study of the presence of major structural and functional components in complex plastids

Plastoglobules (PG) are lipophilic droplets attached to thylakoid membranes in higher plants and green algae and are implicated in prenyl lipid biosynthesis. They might also represent a central hub for integration of plastid signals under stress and therefore the adaptation of the thylakoid membrane under such conditions. In Arabidopsis thaliana, PG contain around 30 specific proteins of which Fibrillins (FBN) and Activity of bc1 complex kinases (ABC1K) represent the majority with respect to both number and protein mass. However, nothing is known about the presence of PG in most algal species, which are responsible for about 50% of global primary production. Therefore, we searched the genomes of publicly available algal genomes for components of PG and the associated functional network in order to predict their presence and potential evolutionary conservation of physiological functions. We could identify homologous sequences for core components of PG, like FBN and ABC1K, in most investigated algal species. Furthermore, proteins at central and interesting positions within the PG functional coexpression network were identified. Phylogenetic sequence analysis revealed diversity within FBN and ABC1K sequences among algal species with complex plastids of the red lineage and large differences compared with green lineage species. Two types of FBN were detected that differ in their isoelectric point which seems to correlate with subcellular localization. Subgroups of FBN were shared between many investigated species and modeling of their 3D-structure implied a conserved structure. FBN and ABC1K are essential structural and functional components of PG. Their occurrence in investigated algal species suggests presence of PG therein and functions in prenyl lipid metabolism and adaptation of the thylakoid membrane that are conserved during evolution.

A bestrophin-like protein modulates the proton motive force across the thylakoid membrane in Arabidopsis.

During photosynthesis, photosynthetic electron transport generates a proton motive force (pmf) across the thylakoid membrane, which is used for ATP biosynthesis via ATP synthase in the chloroplast. The pmf is composed of an electric potential (ΔΨ) and an osmotic component (ΔpH). Partitioning between these components in chloroplasts is strictly regulated in response to fluctuating environments. However, our knowledge of the molecular mechanisms that regulate pmf partitioning is limited. Here, we report a bestrophin‐like protein (AtBest), which is critical for pmf partitioning. While the ΔpH component was slightly reduced in atbest, the ΔΨ component was much greater in this mutant than in the wild type, resulting in less efficient activation of nonphotochemical quenching (NPQ) upon both illumination and a shift from low light to high light. Although no visible phenotype was observed in the atbest mutant in the greenhouse, this mutant exhibited stronger photoinhibition than the wild type when grown in the field. AtBest belongs to the bestrophin family proteins, which are believed to function as chloride (Cl−) channels. Thus, our findings reveal an important Cl− channel required for ion transport and homeostasis across the thylakoid membrane in higher plants. These processes are essential for fine‐tuning photosynthesis under fluctuating environmental conditions.

Les onychodystrophies en consultation externe de dermatologie au CHU/YO

The structure of the major protein constituent of photosynthetic membranes in higher plants, the chlorophyll a/b-light harvesting complex (LHC), was studied by x-ray diffraction and electron microscopy. The LHC was purified from Triton X-100 solubilized thylakoid membranes of the pea, and contained 6 mol of chlorophylls a and b per mole of a polypeptide of 27,000 molecular weight. X-ray diffraction showed that in the presence of 10 mM MgCl2, purified LHC forms planar aggregates that stack with a period of 51 A. Within each layer, LHC molecules pack with a center-to-center distance of 85 A but without long-range order. However, when LHC is incorporated into single-walled vesicles of plant lecithin, the addition of NaCl above 10 mM, or MgCl2 above 2 mM, led to the formation of plaques of hexagonal lattices, with a lattice constant of 125 A. The large domain size and high degree of order in the plane of the membrane are evident from the sharp lattice lines observed to 7 A resolution on the equator of the x-ray pattern. Freeze-fracture electron micrographs demonstrated an aligned stacking of the lattices in adjacent membranes, resulting in crystallinity in the third dimension over short distances. Micrographs of negatively stained membranes revealed a hexagonal lattice of the same lattice constant, formed by surface-exposed parts of the LHC molecules which are probably responsible for the ordered stacking of lattices. In both the LHC aggregates and in the reconstituted membrane lattices the diffracted x-ray intensities at 10-A spacing on the equator indicate that the LHC molecule contains paralled alpha-helices or beta-sheets that are oriented perpendicular to the planar arrays.

Hamartome folliculo-sébacé kystique : à propos de quatre cas

The structure of the major protein constituent of photosynthetic membranes in higher plants, the chlorophyll a/b-light harvesting complex (LHC), was studied by x-ray diffraction and electron microscopy. The LHC was purified from Triton X-100 solubilized thylakoid membranes of the pea, and contained 6 mol of chlorophylls a and b per mole of a polypeptide of 27,000 molecular weight. X-ray diffraction showed that in the presence of 10 mM MgCl2, purified LHC forms planar aggregates that stack with a period of 51 A. Within each layer, LHC molecules pack with a center-to-center distance of 85 A but without long-range order. However, when LHC is incorporated into single-walled vesicles of plant lecithin, the addition of NaCl above 10 mM, or MgCl2 above 2 mM, led to the formation of plaques of hexagonal lattices, with a lattice constant of 125 A. The large domain size and high degree of order in the plane of the membrane are evident from the sharp lattice lines observed to 7 A resolution on the equator of the x-ray pattern. Freeze-fracture electron micrographs demonstrated an aligned stacking of the lattices in adjacent membranes, resulting in crystallinity in the third dimension over short distances. Micrographs of negatively stained membranes revealed a hexagonal lattice of the same lattice constant, formed by surface-exposed parts of the LHC molecules which are probably responsible for the ordered stacking of lattices. In both the LHC aggregates and in the reconstituted membrane lattices the diffracted x-ray intensities at 10-A spacing on the equator indicate that the LHC molecule contains paralled alpha-helices or beta-sheets that are oriented perpendicular to the planar arrays.

The outer mitochondrial membrane in higher plants

The acquisition and integration of intracellular organelles, such as mitochondria and plastids, were important steps in the emergence of complex multicellular life. Although the outer membranes of these organelles have lost many of the functions of their free-living bacterial ancestor, others were acquired during organellogenesis. To date, the biological roles of these proteins have not been systematically characterized. In this review, we discuss the evolutionary origins and functions of outer membrane mitochondrial (OMM) proteins in Arabidopsis thaliana. Our analysis, using phylogenetic inference, indicates that several OMM proteins either acquired novel functional roles or were recruited from other subcellular localizations during evolution in Arabidopsis. These observations suggest the existence of novel communication routes and functions between organelles within plant cells.

The Arabidopsis pex12 and pex13 mutants are defective in both PTS1‐ and PTS2‐dependent protein transport to peroxisomes

Peroxisome biogenesis requires various complex processes including organelle division, enlargement and protein transport. We have been studying a number of Arabidopsis apm mutants that display aberrant peroxisome morphology. Two of these mutants, apm2 and apm4, showed green fluorescent protein fluorescence in the cytosol as well as in peroxisomes, indicating a decrease of efficiency of peroxisome targeting signal 1 (PTS1)-dependent protein transport to peroxisomes. Interestingly, both mutants were defective in PTS2-dependent protein transport. Plant growth was more inhibited in apm4 than apm2 mutants, apparently because protein transport was more severely decreased in apm4 than in apm2 mutants. APM2 and APM4 were found to encode proteins homologous to the peroxins PEX13 and PEX12, respectively, which are thought to be involved in transporting matrix proteins into peroxisomes in yeasts and mammals. We show that APM2/PEX13 and APM4/PEX12 are localized on peroxisomal membranes, and that APM2/PEX13 interacts with PEX7, a cytosolic PTS2 receptor. Additionally, a PTS1 receptor, PEX5, was found to stall on peroxisomal membranes in both mutants, suggesting that PEX12 and PEX13 are components that are involved in protein transport on peroxisomal membranes in higher plants. Proteins homologous to PEX12 and PEX13 have previously been found in Arabidopsis but it is not known whether they are involved in protein transport to peroxisomes. Our findings reveal that APM2/PEX13 and APM4/PEX12 are responsible for matrix protein import to peroxisomes in planta.

Non-Radiative Dissipation of Absorbed Excitation Energy Within Photosynthetic Apparatus of Higher Plants

The review deals with thermal dissipation of absorbed excitation energy within pigment-protein complexes of thylakoid membranes in higher plants. We focus on the de-excitation regulatory processes within photosystem 2 (PS2) that can be monitored as non-photochemical quenching of chlorophyll (Chl) a fluorescence consisting of three components known as energy-dependent quenching (qE), state-transition quenching (qT), and photoinhibitory quenching (qI). We summarize the role of thylakoid lumen pH, xanthophylls, and PS2 proteins in qE mechanism. Further, both the similarity between qE and qI and specific features of qI are described. The other routes of thermal energy dissipation are also mentioned, that is dissipation within photosystem 1 and dissipation through the triplet Chl pathway. The significance of the individual de-excitation processes in protection against photo-oxidative damage to the photosynthetic apparatus under excess photon supply is stretched.

Electron microscopy in structural studies of Photosystem II.

Various techniques of electron microscopy (EM) such as ultrathin sectioning, freeze-fracturing, freeze-etching, negative staining and (cryo-)electron crystallography of two-dimensional crystals have been employed, since now, to obtain much of the structural information of the Photosystem II (PS II) pigment-protein complex at both low and high resolution. This review summarizes information about the structure of this membrane complex as well as its arrangement and interactions with the antenna proteins in thylakoid membranes of higher plants and cyanobacteria obtained by means of EM. Results on subunit organization, with the emphasis on the proteins of the oxygen-evolving complex (OEC), are compared with the data obtained by X-ray crystallography of cyanobacterial PS II.

A remarkable constancy in the relative amount of stroma lamellae of chloroplasts from plants, and its functional significance

A model is presented which gives a quantitative picture of the distribution of the photosynthetic components in the thylakoid membrane of higher plants. The model is based on fragmentation and separation analysis together with microscopy. A salient feature of the model is that over 80 per cent of the pigments are located in the grana where the two photosystems 1 and 2 produce oxygen, NADPH and ATP by the linear electron transport pathway while the stroma lamellae, which harbour less than 20 per cent of the pigments, carry out Photosystem 1 mediated cyclic electron transport coupled to ATP production. This arrangement derives from the observation that more pigments are associated with Photosystem 1 which therefore captures more quanta than Photosystem 2. The excess pigments associated with Photosystem 1 are suggested to be located in the stroma lamellae. Tentative numbers of the different photosystem and cytochrome b.f. complexes located in the different domains of the thylakoid membrane will be presented.

Organization of the pigment molecules in the chlorophyll a/b/c containing alga Mantoniella squamata (Prasinophyceae) studied by means of absorption, circular and linear dichroism spectroscopy

In order to obtain information on the organization of the pigment molecules in chlorophyll (Chl) a/b/c-containing organisms, we have carried out circular dichroism (CD), linear dichroism (LD) and absorption spectroscopic measurements on intact cells, isolated thylakoids and purified light-harvesting complexes (LHCs) of the prasinophycean alga Mantoniella squamata. The CD spectra of the intact cells and isolated thylakoids were predominated by the excitonic bands of the Chl a/b/c LHC. However, some anomalous bands indicated the existence of chiral macrodomains, which could be correlated with the multilayered membrane system in the intact cells. In the red, the thylakoid membranes and the LHC exhibited a well-discernible CD band originating from Chl c, but otherwise the CD spectra were similar to that of non-aggregated LHC II, the main Chl a/b LHC in higher plants. In the Soret region, however, an unusually intense (+) 441 nm band was observed, which was accompanied by negative bands between 465 and 510 nm. It is proposed that these bands originate from intense excitonic interactions between Chl a and carotenoid molecules. LD measurements revealed that the Q Y dipoles of Chl a in Mantoniella thylakoids are preferentially oriented in the plane of the membrane, with orientation angles tilting out more at shorter than at longer wavelengths (9° at 677 nm, 20° at 670 nm and 26° at 662 nm); the Q Y dipole of Chl c was found to be oriented at 29° with respect to the membrane plane. These data and the LD spectrum of the LHC, apart from the presence of Chl c, suggest an orientation pattern of dipoles similar to those of higher plant thylakoids and LHC II. However, the tendency of the Q Y dipoles of Chl b to lie preferentially in the plane of the membrane (23° at 653 nm and 30° at 646 nm) is markedly different from the orientation pattern in higher plant membranes and LHC II. Hence, our CD and LD data show that the molecular organization of the Chl a/b/c LHC, despite evident similarities, differs significantly from that of LHC II.

Molecular and functional characterization of a novel low-affinity cation transporter (LCT1) in higher plants.

The transport of cations across membranes in higher plants plays an essential role in many physiological processes including mineral nutrition, cell expansion, and the transduction of environmental signals. In higher plants the coordinated expression of transport mechanisms is essential for specialized cellular processes and for adaptation to variable environmental conditions. To understand the molecular basis of cation transport in plant roots, a Triticum aestivum cDNA library was used to complement a yeast mutant deficient in potassium (K+) uptake. Two genes were cloned that complemented the mutant: HKT1 and a novel cDNA described in this report encoding a cation transporter, LCT1 (low-affinity cation transporter). Analysis of the secondary structure of LCT1 suggests that the protein contains 8-10 transmembrane helices and a hydrophilic amino terminus containing sequences enriched in Pro, Ser, Thr, and Glu (PEST). The transporter activity was assayed using radioactive isotopes in yeast cells expressing the cDNA. LCT1 mediated low-affinity uptake of the cations Rb+ and Na+, and possibly allowed Ca2+ but not Zn2+ uptake. LCT1 is expressed in low abundance in wheat roots and leaves. The precise functional role of this cation transporter is not known, although the competitive inhibition of cation uptake by Ca2+ has parallels to whole plant and molecular studies that have shown the important role of Ca2+ in reducing Na+ uptake and ameliorating Na+ toxicity. The structure of this higher plant ion transport protein is unique and contains PEST sequences.

Isolation of Thylakoid Membrane Vesicles ofChlamydomonas reinhardiiChloroplasts That Are Able to Integrate and Importin VitroSynthesized Precursor Proteins

Once imported into the stroma, nuclear encoded proteins of the chloroplast have to be routed to their final compartment, e.g. the thylakoid membranes. Four different pathways have been reported for the translocation of precursor proteins across and for the integration of mature proteins into the thylakoid membranes in higher plants. To study the sorting of precursor proteins in chloroplasts of higher plants the generation of anin vitrosystem using isolated intact thylakoid membrane vesicles was of major importance. Here we report the isolation of intact thylakoid membrane vesicles of the green algaeChlamydomonas reinhardiifor the generation of a similar algal system. Further we show successful transport of severalChlamydomonasprecursor proteins into isolated thylakoids: Lumenal precursors were translocated into the vesicles resulting in the accumulation of their mature, thermolysin-insensitive forms and thylakoid membrane proteins were specifically integrated into isolatedChlamydomonasthylakoid membranes.

Molecular approaches towards an understanding of loading and unloading of assimilates in higher plants

The combination of two sets of molecular tools, namely yeast expression cloning and the possibility of constructing transgenic plants, has allowed analysis of the transport processes occurring at the plasma membrane in higher plants. To date, more than 30 different plant genes for plasma membrane transporters of sugars and amino acids have been identified, mainly by expression cloning. Furthermore, the functional expression of genes in Schizosaccharomyces pombe, Saccharomyces cerevisiae and Xenopus oocytes has been applied to obtain detailed information on the biochemical properties of the transporters. The expression systems have also allowed the purification of the proteins for structural analysis and to study structure-function relationship using mutagenesis approaches. A number of mutants and transgenic plants defective in certain transport properties are available and these will help in understanding the physiology of the long-distance transport of assimilates.