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1.
Plant Cell Physiol ; 62(4): 678-692, 2021 Sep 24.
Article in English | MEDLINE | ID: mdl-33570567

ABSTRACT

The force of gravity is a constant environmental factor. Plant shoots respond to gravity through negative gravitropism and gravity resistance. These responses are essential for plants to direct the growth of aerial organs away from the soil surface after germination and to keep an upright posture above ground. We took advantage of the effect of brassinosteroids (BRs) on the two types of graviresponses in Arabidopsis thaliana hypocotyls to disentangle functions of cell wall polymers during etiolated shoot growth. The ability of etiolated Arabidopsis seedlings to grow upward was suppressed in the presence of 24-epibrassinolide (EBL) but enhanced in the presence of brassinazole (BRZ), an inhibitor of BR biosynthesis. These effects were accompanied by changes in cell wall mechanics and composition. Cell wall biochemical analyses, confocal microscopy of the cellulose-specific pontamine S4B dye and cellular growth analyses revealed that the EBL and BRZ treatments correlated with changes in cellulose fibre organization, cell expansion at the hypocotyl base and mannan content. Indeed, a longitudinal reorientation of cellulose fibres and growth inhibition at the base of hypocotyls supported their upright posture whereas the presence of mannans reduced gravitropic bending. The negative effect of mannans on gravitropism is a new function for this class of hemicelluloses. We also found that EBL interferes with upright growth of hypocotyls through their uneven thickening at the base.


Subject(s)
Arabidopsis/physiology , Brassinosteroids/metabolism , Cellulose/metabolism , Hypocotyl/physiology , Mannans/metabolism , Arabidopsis/cytology , Arabidopsis/drug effects , Brassinosteroids/pharmacology , Cell Wall/chemistry , Cell Wall/drug effects , Cellulose/chemistry , Gravitropism/physiology , Hypocotyl/chemistry , Mannans/chemistry , Plant Shoots/drug effects , Plant Shoots/physiology , Polysaccharides/chemistry , Steroids, Heterocyclic/metabolism , Steroids, Heterocyclic/pharmacology , Time-Lapse Imaging
2.
Cell ; 162(6): 1353-64, 2015 Sep 10.
Article in English | MEDLINE | ID: mdl-26343580

ABSTRACT

Abiotic stress, such as salinity, drought, and cold, causes detrimental yield losses for all major plant crop species. Understanding mechanisms that improve plants' ability to produce biomass, which largely is constituted by the plant cell wall, is therefore of upmost importance for agricultural activities. Cellulose is a principal component of the cell wall and is synthesized by microtubule-guided cellulose synthase enzymes at the plasma membrane. Here, we identified two components of the cellulose synthase complex, which we call companion of cellulose synthase (CC) proteins. The cytoplasmic tails of these membrane proteins bind to microtubules and promote microtubule dynamics. This activity supports microtubule organization, cellulose synthase localization at the plasma membrane, and renders seedlings less sensitive to stress. Our findings offer a mechanistic model for how two molecular components, the CC proteins, sustain microtubule organization and cellulose synthase localization and thus aid plant biomass production during salt stress. VIDEO ABSTRACT.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/cytology , Arabidopsis/physiology , Cellulose/biosynthesis , Glucosyltransferases/metabolism , Microtubule-Associated Proteins/metabolism , Microtubules/metabolism , Arabidopsis/enzymology , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/genetics , Biomass , Cell Wall/metabolism , Glucosyltransferases/genetics , Microtubule-Associated Proteins/chemistry , Microtubule-Associated Proteins/genetics , Salinity , Stress, Physiological
3.
J Vis Exp ; (87)2014 05 16.
Article in English | MEDLINE | ID: mdl-24894952

ABSTRACT

The global demand for food, feed, energy and water poses extraordinary challenges for future generations. It is evident that robust platforms for the exploration of renewable resources are necessary to overcome these challenges. Within the multinational framework MultiBioPro we are developing biorefinery pipelines to maximize the use of plant biomass. More specifically, we use poplar and tobacco tree (Nicotiana glauca) as target crop species for improving saccharification, isoprenoid, long chain hydrocarbon contents, fiber quality, and suberin and lignin contents. The methods used to obtain these outputs include GC-MS, LC-MS and RNA sequencing platforms. The metabolite pipelines are well established tools to generate these types of data, but also have the limitations in that only well characterized metabolites can be used. The deep sequencing will allow us to include all transcripts present during the developmental stages of the tobacco tree leaf, but has to be mapped back to the sequence of Nicotiana tabacum. With these set-ups, we aim at a basic understanding for underlying processes and at establishing an industrial framework to exploit the outcomes. In a more long term perspective, we believe that data generated here will provide means for a sustainable biorefinery process using poplar and tobacco tree as raw material. To date the basal level of metabolites in the samples have been analyzed and the protocols utilized are provided in this article.


Subject(s)
Biomass , Nicotiana/genetics , Nicotiana/metabolism , Populus/genetics , Populus/metabolism , Animal Feed , Biofuels , Chromatography, Liquid , Gas Chromatography-Mass Spectrometry , Metabolomics , Transcription, Genetic
4.
PLoS One ; 5(6): e11342, 2010 Jun 28.
Article in English | MEDLINE | ID: mdl-20596537

ABSTRACT

BACKGROUND: Calreticulin (CRT) is a ubiquitous ER protein involved in multiple cellular processes in animals, such as protein folding and calcium homeostasis. Like in animals, plants have evolved divergent CRTs, but their physiological functions are less understood. Arabidopsis contains three CRT proteins, where the two CRTs AtCRT1a and CRT1b represent one subgroup, and AtCRT3 a divergent member. METHODOLOGY/PRINCIPAL FINDINGS: Through expression of single Arabidopsis family members in CRT-deficient mouse fibroblasts we show that both subgroups have retained basic CRT functions, including ER Ca2+-holding potential and putative chaperone capabilities. However, other more general cellular defects due to the absence of CRT in the fibroblasts, such as cell adhesion deficiencies, were not fully restored. Furthermore, in planta expression, protein localization and mutant analyses revealed that the three Arabidopsis CRTs have acquired specialized functions. The AtCRT1a and CRT1b family members appear to be components of a general ER chaperone network. In contrast, and as recently shown, AtCRT3 is associated with immune responses, and is essential for responsiveness to the bacterial Pathogen-Associated Molecular Pattern (PAMP) elf18, derived from elongation factor (EF)-Tu. Whereas constitutively expressed AtCRT1a fully complemented Atcrt1b mutants, AtCRT3 did not. CONCLUSIONS/SIGNIFICANCE: We conclude that the physiological functions of the two CRT subgroups in Arabidopsis have diverged, resulting in a role for AtCRT3 in PAMP associated responses, and possibly more general chaperone functions for AtCRT1a and CRT1b.


Subject(s)
Arabidopsis/physiology , Calreticulin/physiology , Amino Acid Sequence , Animals , Calcium/metabolism , Calreticulin/chemistry , Calreticulin/genetics , Endoplasmic Reticulum/metabolism , Fluorescent Antibody Technique , Mice , Molecular Chaperones/metabolism , Molecular Sequence Data , Mutation , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Sequence Homology, Amino Acid , Subcellular Fractions/metabolism
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