Distribution of Gibberellins A7 and A4 in Tobacco Proembryos Using Immunoelectron Microscopy

The ovules of Nicotiana tabacum var. macrophylla 8 days after pollination were fixed successively with 2% EDC (1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide) and a mixture solution of paraformaldehyde and glutaraldehyde, then slightly postfixed in 0.5 % osmium solution. UI- trathin sections of 6-, 9- or 12- celled proembryos embedded in Epon 812 resin were stained in an anti-GA MAb and sheep anti-mouse IgG-colloidal gold (10 nm). This MAb specifically recognizes methyl esters of GA7 and GAn. Therefore, it could be used as a probe to localize GA7 and GAn in cells after EDC fixation. The 12-celled proembryo is composed of a 9-celled embryo and a 3-celled suspensor. Wide distribution of GA7 and GAn was observed in all proembryo cells and most organelles at subeellular level, including walls, plasmodesmata, plasma membrane, cytoplasmic matrix, mitochondria, plastids, vacuoles, endoplasmic reticulum and nuclei. Clusters of gold granules were found in nuclear envelope, nucleomatrix, nucleolus, chromosome, cytoplamic matrix. In a region composed of cytoplasmic matrix, a vacuole and a mitochondrium, such concentrated gold granules were particularly obviously observed. There appeared a gradient distribution of GA7 and Gan from embryo cells decreasingly to suspensor cells. GA7 and GAn could be translocated via intercellular walls, plasmodesmata and vesicles between embryo and suspensor. 1he authors suspect the direction of GA translocation in proembryo may be from suspensor to embryo. To author's knowledge, this is the first report to indicate subcellular and gradient distributions of bioactive gibberellins in plant proembryos.     

NanC Crystal Structure, a Model for Outer-Membrane Channels of the Acidic Sugar-Specific KdgM Porin Family

Sialic acids are acidic sugars present mostly on vertebrate cell surfaces, which can be metabolized by bacteria and act as an inflammation signal. N-Acetylneuraminic acid, the most abundant sialic acid, can enter into Escherichia coli K12 through NanC, an N-acetylneuraminic acid-inducible outer-membrane channel. With its 215 residues, NanC belongs to the family of small monomeric KdgM-related porins. KdgM homologues are found in gammaproteobacteria, including major plant and human pathogens, and together they define a large family of putative acidic sugar/oligosaccharide transporters, which are as yet poorly characterized. Here, we present the first high-resolution structure of a KdgM family member. NanC folds into a 28-Å-high, 12-stranded β-barrel, resembling the β-domain of autotransporter NalP and defining an open pore with an average radius of 3.3 Å. The channel is lined by two strings of basic residues facing each other across the pore, a feature that appears largely conserved within the KdgM family and is likely to facilitate the diffusion of acidic oligosaccharides.