Subcellular analysis of starch metabolism in developing barley seeds using a non-aqueous fractionation method

Compartmentation of metabolism in developing seeds is poorly understood due to the lack of data on metabolite distributions at the subcellular level. In this report, a non-aqueous fractionation method is described that allows subcellular concentrations of metabolites in developing barley endosperm to be calculated. (i) Analysis of subcellular volumes in developing endosperm using micrographs shows that plastids and cytosol occupy 50.5% and 49.9% of the total cell volume, respectively, while vacuoles and mitochondria can be neglected. (ii) By using non-aqueous fractionation, subcellular distribution between the cytosol and plastid of the levels of metabolites involved in sucrose degradation, starch synthesis, and respiration were determined. With the exception of ADP and AMP which were mainly located in the plastid, most other metabolites of carbon and energy metabolism were mainly located outside the plastid in the cytosolic compartment. (iii) In developing barley endosperm, the ultimate precursor of starch, ADPglucose (ADPGlc), was mainly located in the cytosol (80–90%), which was opposite to the situation in growing potato tubers where ADPGlc was almost exclusively located in the plastid (98%). This reflects the different subcellular distribution of ADPGlc pyrophosphorylase (AGPase) in these tissues. (iv) Cytosolic concentrations of ADPGlc were found to be close to the published Km values of AGPase and the ADPGlc/ADP transporter at the plastid envelope. Also the concentrations of the reaction partners glucose-1-phosphate, ATP, and inorganic pyrophosphate were close to the respective Km values of AGPase. (v) Knock-out of cytosolic AGPase in Riso16 mutants led to a strong decrease in ADPGlc level, in both the cytosol and plastid, whereas knock-down of the ADPGlc/ADP transporter led to a large shift in the intracellular distribution of ADPGlc. (v) The thermodynamic structure of the pathway of sucrose to starch was determined by calculating the mass–action ratios of all the steps in the pathway. The data show that AGPase is close to equilibrium, in both the cytosol and plastid, whereas the ADPGlc/ADP transporter is strongly displaced from equilibrium in vivo. This is in contrast to most other tissues, including leaves and potato tubers. (vi) Results indicate transport rather than synthesis of ADPGlc to be the major regulatory site of starch synthesis in barley endosperm. The reversibility of AGPase in the plastid has important implications for the regulation of carbon partitioning between different biosynthetic pathways

A morphometric method for correcting phytoplankton cell volume estimates

Cell volume calculations are often used to estimate biomass of natural phytoplankton assemblages. Such estimates may be questioned due to morphological differences in the organisms present. Morphometric analysis of 8 species representative of phytoplankton types found in the Great Lakes shows significant differences in cell constituent volumes. Volume of physiologically inert wall material ranges from nil, in some flagellates, to over 20% of the total cell volume in certain diatoms. Likewise, “empty” vacuole may comprise more than 40% of the total cell volume of some diatoms, but less than 3% of the volume of some flagellates. In the organisms investigated, the total carbon containing cytoplasm ranged from 52% to 98% of the total cell volume and the metabolizing biovolume ranged from 30% to 82%. Although these differences complicate direct biomass estimation, morphometric analysis at the ultrastructural level may provide ecologically valuable insights.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41732/1/709_2005_Article_BF01275650.pd

An adaptative filtering method to evalute normal vectors and surface areas of 3d objects. Application to snow images from X-ray tomography

International audienceEstimating the normal vector field on the boundary of discrete 3D objects is essential for rendering and image measurement problems. Most of the existing algorithms do not provide an accurate determination of the normal vector field for shapes that present edges. We propose here a new and simple computational method in order to obtain accurate results on all types of shapes whatever their local convexity degree is. The presented method is based on the gradient vector field analysis of the object distance map. This vector field is adaptively filtered around each surface voxel using angle and symmetry criteria, so that as many relevant contributions as possible are accounted for. This optimizes the smoothing of digitization effects while preserving relevant details of the processed numerical object. Thanks to the precise normal field obtained, a projection method can be proposed to derive immediately the surface area from a raw discrete object. An empirical justification of the validity of such an algorithm in the continuous limit is also provided. Some results on simulated data and snow images from X-ray tomography are presented, compared to the Marching Cubes and Convex Hull results and discussed

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L’inscription de l’altérité dans un texte peut se faire de diverses manières, puisque, appartenant à des systèmes de référence « autres », qui construisent des représentations prises dans une histoire, un espace, une culture et un système linguistique propres qui mettent en place un monde auquel l’accès n’est pas totalement dégagé. Comment les obstacles à cette entrée libre dans les textes traduits peuvent-ils se manifester ? Grâce à divers procédés somme toute assez banals, mais pas toujours..