scholarly journals A partial C4 photosynthetic biochemical pathway in rice

2020 ◽  
Author(s):  
Hsiang-Chun Lin ◽  
Stéphanie Arrivault ◽  
Robert A Coe ◽  
Shanta Karki ◽  
Sarah Covshoff ◽  
...  
Keyword(s):  
Carbon Flux ◽  
Malic Enzyme ◽  
Pep Carboxylase ◽  
Pyruvate Phosphate Dikinase ◽  
Kranz Anatomy ◽  
Bundle Sheath Cells ◽  
Phenotype Characteristic ◽  
A Cell ◽  
First Time ◽  
H Protein

AbstractIntroduction of a C4 photosynthetic pathway into C3 rice (Oryza sativa) requires installation of a biochemical pump that concentrates CO2 at the site of carboxylation in modified bundle sheath cells. To investigate the feasibility of this, we generated a quadruple line that simultaneously expresses four of the core C4 photosynthetic enzymes from the NADP-malic enzyme subtype, phosphoenolpyruvate carboxylase (ZmPEPC), NADP-malate dehydrogenase (ZmNADP-MDH), NADP-malic enzyme (ZmNADP-ME) and pyruvate phosphate dikinase (ZmPPDK), in a cell-specific manner. This led to enhanced enzyme activity but was largely neutral in its effects on photosynthetic rate and growth. Measurements of the flux of 13CO2 through photosynthetic metabolism revealed a significant increase in the incorporation of 13C into malate, consistent with increased fixation of 13CO2 via PEP carboxylase in lines expressing the maize PEPC enzyme. We also showed 13C labelling of aspartate indicating additional 13CO2 fixation into oxaloacetate by PEPC and conversion to aspartate by the endogenous aspartate aminotransferase activity. However, there were no significant differences in labelling of 3-phosphoglycerate (3PGA) or phosphoenolpyruvate (PEP) indicating limited carbon flux through C4 enzymes into the Calvin-Benson cycle. Crossing the quadruple line with a line with reduced glycine decarboxylase H-protein (OsGDCH) abundance led to a photosynthetic phenotype characteristic of the reduced OsGDCH line and higher labelling of malate, aspartate and citrate. While Kranz anatomy or other anatomical modifications have not yet been installed in these plants to enable a fully functional C4 cycle, these results demonstrate for the first-time flux through the carboxylation phase of C4 metabolism in transgenic rice containing the key metabolic steps in the C4 pathway.

scholarly journals Characterization of a Novel Fructosyltransferase from Lactobacillus reuteri That Synthesizes High-Molecular-Weight Inulin and Inulin Oligosaccharides

2002 ◽  
Vol 68 (9) ◽  
pp. 4390-4398 ◽  
Author(s):  
S. A. F. T. van Hijum ◽  
G. H. van Geel-Schutten ◽  
H. Rahaoui ◽  
M. J. E. C. van der Maarel ◽  
L. Dijkhuizen
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Lactobacillus Reuteri ◽  
Bacterial Strains ◽  
Potential Health ◽  
Isolation And Characterization ◽  
A Cell ◽  
Encoding Gene ◽  
First Time

ABSTRACT Fructosyltransferase (FTF) enzymes produce fructose polymers (fructans) from sucrose. Here, we report the isolation and characterization of an FTF-encoding gene from Lactobacillus reuteri strain 121. A C-terminally truncated version of the ftf gene was successfully expressed in Escherichia coli. When incubated with sucrose, the purified recombinant FTF enzyme produced large amounts of fructo-oligosaccharides (FOS) with β-(2→1)-linked fructosyl units, plus a high-molecular-weight fructan polymer (>107) with β-(2→1) linkages (an inulin). FOS, but not inulin, was found in supernatants of L. reuteri strain 121 cultures grown on medium containing sucrose. Bacterial inulin production has been reported for only Streptococcus mutans strains. FOS production has been reported for a few bacterial strains. This paper reports the first-time isolation and molecular characterization of (i) a Lactobacillus ftf gene, (ii) an inulosucrase associated with a generally regarded as safe bacterium, (iii) an FTF enzyme synthesizing both a high molecular weight inulin and FOS, and (iv) an FTF protein containing a cell wall-anchoring LPXTG motif. The biological relevance and potential health benefits of an inulosucrase associated with an L. reuteri strain remain to be established.

scholarly journals Macrophage Autophagy and Oxidative Stress: An Ultrastructural and Immunoelectron Microscopical Study

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Ida Perrotta ◽  
Valentina Carito ◽  
Emilio Russo ◽  
Sandro Tripepi ◽  
Saveria Aquila ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Death ◽  
Programmed Cell Death ◽  
Defense Mechanisms ◽  
Ultrastructural Localization ◽  
Microscopical Study ◽  
Beclin 1 ◽  
Cell Organelles ◽  
A Cell ◽  
First Time

The word autophagy broadly refers to the cellular catabolic processes that lead to the removal of damaged cytosolic proteins or cell organelles through lysosomes. Although autophagy is often observed during programmed cell death, it may also serve as a cell survival mechanism. Accumulation of reactive oxygen species within tissues and cells induces various defense mechanisms or programmed cell death. It has been shown that, besides inducing apoptosis, oxidative stress can also induce autophagy. To date, however, the regulation of autophagy in response to oxidative stress remains largely elusive and poorly understood. Therefore, the present study was designed to examine the ratio between oxidative stress and autophagy in macrophages after oxidant exposure (AAPH) and to investigate the ultrastructural localization of beclin-1, a protein essential for autophagy, under basal and stressful conditions. Our data provide evidence that oxidative stress induces autophagy in macrophages. We demonstrate, for the first time by immunoelectron microscopy, the subcellular localization of beclin-1 in autophagic cells.

scholarly journals Selection and Characterization of Rupintrivir-Resistant Norwalk Virus Replicon Cells In Vitro

2018 ◽  
Vol 62 (5) ◽  
Author(s):  
Mitsutaka Kitano ◽  
Myra Hosmillo ◽  
Edward Emmott ◽  
Jia Lu ◽  
Ian Goodfellow
Keyword(s):  
Reverse Genetics ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Murine Norovirus ◽  
Norwalk Virus ◽  
Irreversible Inhibitor ◽  
Antiviral Compounds ◽  
A Cell ◽  
First Time

ABSTRACT Human norovirus (HuNoV) is a major cause of nonbacterial gastroenteritis worldwide, yet despite its impact on society, vaccines and antivirals are currently lacking. A HuNoV replicon system has been widely applied to the evaluation of antiviral compounds and has thus accelerated the process of drug discovery against HuNoV infection. Rupintrivir, an irreversible inhibitor of the human rhinovirus 3C protease, has been reported to inhibit the replication of the Norwalk virus replicon via the inhibition of the norovirus protease. Here we report, for the first time, the generation of rupintrivir-resistant human Norwalk virus replicon cells in vitro . Sequence analysis revealed that these replicon cells contained amino acid substitutions of alanine 105 to valine (A105V) and isoleucine 109 to valine (I109V) in the viral protease NS6. The application of a cell-based fluorescence resonance energy transfer (FRET) assay for protease activity demonstrated that these substitutions were involved in the enhanced resistance to rupintrivir. Furthermore, we validated the effect of these mutations using reverse genetics in murine norovirus (MNV), demonstrating that a recombinant MNV strain with a single I109V substitution in the protease also showed reduced susceptibility to rupintrivir. In summary, using a combination of different approaches, we have demonstrated that, under the correct conditions, mutations in the norovirus protease that lead to the generation of resistant mutants can rapidly occur.

Light as a Life Tool

2020 ◽  
pp. 58-81
Author(s):  
John Parrington
Keyword(s):  
Fluorescent Protein ◽  
Green It ◽  
Live Animal ◽  
New Science ◽  
Gene Coding ◽  
A Cell ◽  
Electron Microscopes ◽  
First Time ◽  
Protein Channels ◽  
The Brain

Visual light, and radiation of other frequencies, are highly important for scientific research. The first light microscopes made it possible for the first time to see that organisms from plants to humans are composed of cells. Electron microscopes have allowed scientists to study the structural components of cells in great detail, and even determine the shapes of individual proteins. Many lifeforms also use light to attract a mate or prey, or deter an attacker. Following the identification of the gene coding for the fluorescent protein that makes certain jellyfish glow green it has become possible to use this to genetically label proteins in a living cell, or even a live animal. This means that now the location of proteins in a cell can be determined exactly. A major recent step forward in neuroscience came with the discovery of protein channels in algae that conduct ions in response to light. By creating transgenic mice that have these proteins in their brain neurons, it is now possible to modulate the activity of these neurons by shining light into the brain though microscopic fibre optic cables. This new science of optogenetics allows neurons to be switched on or off experimentally. The optogenetic approach has been used to uncover the neural circuits involved in memory, pain and pleasure. In the future this technique might be used to treat physical pain or depression in people. Controversially, it might be also be misused, to supress memories, or even create completely false ones in people’s heads.

Electrophysiological and Anatomical Characterization of PDF-Positive Clock Neurons in the Intact Adult Drosophila Brain

2006 ◽  
Vol 95 (6) ◽  
pp. 3955-3960 ◽  
Author(s):  
Demian Park ◽  
Leslie C. Griffith
Keyword(s):  
Biological Rhythms ◽  
Cell Group ◽  
Resting Membrane Potential ◽  
Electrophysiological Properties ◽  
Drosophila Brain ◽  
A Cell ◽  
Locomotor Rhythms ◽  
Firing Properties ◽  
First Time

Daily biological rhythms in both prokaryotes and eukaryotes are controlled by circadian clocks. In Drosophila, there is a good basic understanding of both the molecular and anatomical components of the clock. In this study we directly measure, for the first time, electrophysiological properties and anatomy of individual filled large lateral PDF-positive clock neurons, a cell group believed to be involved in synchronization of the clock in constant conditions. We find that the large PDF-positive neurons are morphologically homogeneous and that their resting membrane potential is modulated both by the clock and by light inputs. Expression of a leak channel, dORK-ΔC, which has been shown to disrupt circadian locomotor rhythms, hyperpolarizes these neurons, and blocks firing. These data imply that the firing properties of large PDF neurons are both regulated by and critical for clock function.

Phenolic Deposits and Kranz Syndrome in Leaf Tissues of Spotted (Euphorbia maculata) and Prostrate (Euphorbia supina) Spurge

Weed Science ◽  
1983 ◽  
Vol 31 (1) ◽  
pp. 131-136 ◽  
Author(s):  
C. Dennis Elmore ◽  
Rex N. Paul
Keyword(s):  
Electron Microscopy ◽  
Phenolic Compounds ◽  
Light And Electron Microscopy ◽  
Bundle Sheath ◽  
Fixation Technique ◽  
Mesophyll Cells ◽  
Kranz Anatomy ◽  
Bundle Sheath Cells ◽  
Leaf Tissues ◽  
Sheath Cells

Spotted spurge (Euphorbia maculataL.) and prostrate spurge (E. supinaRaf.), both in subgenusChamesyce,were examined by light and electron microscopy using a caffeine - fixation technique to sequester the phenolic pools intercellularly. Both species have typical dicotyledon-type Kranz anatomy. Sequestered phenolic pools were located in vacuoles in epidermal and mesophyll cells. Only in spotted spurge, however, were additional phenolic pools formed in bundle - sheath cells. This study was undertaken because allelopathy has been demonstrated in prostrate spurge and because phenolic compounds have been implicated in allelopathy. These results would indicate that spotted spurge should also be allelopathic.

scholarly journals Fluvial organic carbon flux from an eroding peatland catchment, southern Pennines, UK

2008 ◽  
Vol 12 (2) ◽  
pp. 625-634 ◽  
Author(s):  
R. R. Pawson ◽  
D. R. Lord ◽  
M. G. Evans ◽  
T. E. H. Allott
Keyword(s):  
Organic Carbon ◽  
Carbon Flux ◽  
Relative Importance ◽  
Carbon Export ◽  
Event Scale ◽  
Organic Carbon Flux ◽  
Poc Export ◽  
Event Based ◽  
First Time ◽  
Annual Flux

Abstract. This study investigates for the first time the relative importance of dissolved organic carbon (DOC) and particulate organic carbon (POC) in the fluvial carbon flux from an actively eroding peatland catchment in the southern Pennines, UK. Event scale variability in DOC and POC was examined and the annual flux of fluvial organic carbon was estimated for the catchment. At the event scale, both DOC and POC were found to increase with discharge, with event based POC export accounting for 95% of flux in only 8% of the time. On an annual cycle, exports of 35.14 t organic carbon (OC) are estimated from the catchment, which represents an areal value of 92.47 g C m−2 a−1. POC was the most significant form of organic carbon export, accounting for 80% of the estimated flux. This suggests that more research is required on both the fate of POC and the rates of POC export in eroding peatland catchments.

scholarly journals Functional Role of the PE Domain and Immunogenicity of the Mycobacterium tuberculosis Triacylglycerol Hydrolase LipY

2007 ◽  
Vol 76 (1) ◽  
pp. 127-140 ◽  
Author(s):  
Kanhu C. Mishra ◽  
Chantal de Chastellier ◽  
Yeddula Narayana ◽  
Pablo Bifani ◽  
Alistair K. Brown ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Humoral Response ◽  
Host Immune System ◽  
Functional Roles ◽  
Triacylglycerol Hydrolase ◽  
Immunodominant Antigen ◽  
Surface Localization ◽  
A Cell ◽  
First Time

ABSTRACT PE and PPE proteins appear to be important for virulence and immunopathogenicity in mycobacteria, yet the functions of the PE/PPE domains remain an enigma. To decipher the role of these domains, we have characterized the triacylglycerol (TAG) hydrolase LipY from Mycobacterium tuberculosis, which is the only known PE protein expressing an enzymatic activity. The overproduction of LipY in mycobacteria resulted in a significant reduction in the pool of TAGs, consistent with the lipase activity of this enzyme. Unexpectedly, this reduction was more pronounced in mycobacteria overexpressing LipY lacking the PE domain [LipY(ΔPE)], suggesting that the PE domain participates in the modulation of LipY activity. Interestingly, Mycobacterium marinum contains a protein homologous to LipY, termed LipYmar, in which the PE domain is substituted by a PPE domain. As for LipY, overexpression of LipYmar in Mycobacterium smegmatis significantly reduced the TAG pool, and this was further pronounced when the PPE domain of LipYmar was removed. Fractionation studies and Western blot analysis demonstrated that both LipY and LipY(ΔPE) were mainly present in the cell wall, indicating that the PE domain was not required for translocation to this site. Furthermore, electron microscopy immunolabeling of LipY(ΔPE) clearly showed a cell surface localization, thereby suggesting that the lipase may interact with the host immune system. Accordingly, a strong humoral response against LipY and LipY(ΔPE) was observed in tuberculosis patients. Together, our results suggest for the first time that both PE and PPE domains can share similar functional roles and that LipY represents a novel immunodominant antigen.

Photosynthetic Enzyme Activities in the C3-C4 Intermediate Neurachne minor S. T . Blake (Poaceae)

1986 ◽  
Vol 13 (3) ◽  
pp. 399 ◽  
Author(s):  
PW Hattersley ◽  
NE Stone
Keyword(s):  
Malic Enzyme ◽  
Acid Decarboxylase ◽  
Decarboxylase Activity ◽  
Carboxylase Activity ◽  
Pep Carboxylase ◽  
C3 Plant ◽  
Bisphosphate Carboxylase ◽  
C3 Species ◽  
Species Activity ◽  
Pep Carboxykinase

The activities of eight key photosynthetic enzymes were measured in leaf blade extracts of the C3-C4 intermediate Neurachne minor S. T. Blake, its C3 and C4 relatives, C3-C4 Panicum milioides Nees ex Trin., and controls (all Poaceae). Phosphoenolpyruvate (PEP) carboxylase (PEPC) activity in N. minor (5.46 �mol mg Chl-1 min-1) is higher than previously reported for any other C3-C3 plant, and the ratio of ribulose-1,5-bisphosphate carboxylase activity to PEPC activity is lower than for P. milioides or C3 species. Activity of pyruvate,PI dikinase (up to 0.88 �mol mg Chl-1 min-1) is 3-5 times higher than in P. milioides. Assays of NADP-malic enzyme (NADP-ME), NAD-malic enzyme (NAD-ME) and PEP carboxykinase (PCK) show Paraneurachne muelleri (Hack.) S. T. Blake and Neurachne munroi (F. Muell.) F. Muell., N. minor's two close C4 relatives, to be NADP-ME type, as predicted from leaf anatomy. Aspartate and alanine aminotransferase activities in these species are higher than expected, however. N. minor (C3-C4) exhibits higher C4 acid decarboxylase activity than C3 species or P. milioides, for NADP-ME only (up to 1.07 �mol mg Chl-1 min-1). Our results suggest that N. minor possesses a limited C4 acid cycle, and that it is the most C4-like C3-C4 intermediate grass currently identified, comparable with some of the known C3-C4 Flaveria (Asteraceae) species.

Photosynthesis in Gomphrena celosioides and Its Classification Amongst C4-pathway Plants

1976 ◽  
Vol 3 (6) ◽  
pp. 863 ◽  
Author(s):  
E Repo ◽  
MD Hatch
Keyword(s):  
Malate Dehydrogenase ◽  
Malic Enzyme ◽  
Bundle Sheath ◽  
Mesophyll Cells ◽  
C4 Species ◽  
Bundle Sheath Cells ◽  
Major Route ◽  
Gomphrena Celosioides ◽  
A Minor ◽  
Sheath Cells

Monocotyledonous C4 species classified as NADP-ME-type transfer malate from mesophyll to bundle sheath cells where this acid is decarboxylated via NADP malic enzyme (EC 1.1.1.40) to yield pyruvate and CO2. The dicotyledon G. celosioides is most appropriately classified in thls group on the basis of high leaf activities of NADP malic enzyme and NADP malate dehydrogenase (EC 1.1.1.82). However, this species contains high aspartate aminotransferase (EC 2.6.1.1) and alanine aminotransferase (EC 2.6.1.2) activities and centripetally located bundle sheath chloroplasts, features more typical of other groups of C4 species that cycle aspartate and alanine between mesophyll and bundle sheath cells. During the present study, we found that these aminotransferases and NADP malate dehydrogenase were predominantly located in mesophyll cells, that malate was the major C4 acid labelled when leaves were exposed to 14CO2, and that label was initially lost most rapidly from the C-4 of malate during a chase in 12CO2. These results are consistent with the major route of photosynthetic metabolism being the same as that operative in other NADP-ME-type species, although this may be supplemented by a minor route utilizing aspartate. In contrast to monocotyledonous NADP-ME-type C4 species, isolated bundle sheath cells from G. celosioides were capable of rapid photoreduction of NADP as judged by products formed during assimilation of 14CO2 and their capacity for light-dependent oxygen evolution. This was related to a relatively high frequency of single unstacked granum in the chloroplasts of these cells.

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