Metabolic profiling of photoautotrophic and photomixotrophic potato plantlets (<Emphasis Type="Italic">Solanum tuberosum</Emphasis>) provides new insights into acclimatization

Conventional photomixotrophic micropropagation systems are inefficient due to the high rates of mortality upon the transfer of plantlets from in vitro to ex vitro conditions. Exogenous medium sugar has been suggested to be the major cause of this problem. The aim of this study was to investigate the role of sucrose supply on the metabolic profile of in vitro-grown potato plantlets subjected to different tissue culture conditions consisting of Murashige and Skoog medium and without sucrose [photoautotrophic (PAT) condition] or with 3% sucrose [photomixotrophic (PMT) condition]. Using gas chromatography–mass spectrometry, we identified a set of 51 different metabolites in leaf tissues during the rooting phase. Most growth parameters, such as shoot length, leaf fresh weight, leaf number, and leaf area/plant, were significantly lower under PMT than under PAT conditions. Moreover, photosynthesis was inhibited due to partial stomatal closure under PMT conditions. The metabolomic profiles along with principal component analysis and hierarchical cluster analysis revealed that the two treatments were characterized by distinct metabolic signatures. PAT leaves were characterized by the accumulation of urea and erythritol. In comparison, PMT leaves were characterized by the accumulation of metabolites belonging to the primary metabolism and catecholamines as well as compounds related to abiotic stress conditions, such as proline, hydroxyproline, asparagine, γ-aminobutyric acid (GABA), soluble sugars, and myo-inositol.     

Trichothecene biosynthesis in Fusarium species: chemistry, genetics, and significance.

Several species of the genus Fusarium and related fungi produce trichothecenes which are sesquiterpenoid epoxides that act as potent inhibitors of eukaryotic protein synthesis. Interest in the trichothecenes is due primarily to their widespread contamination of agricultural commodities and their adverse effects on human and animal health. In this review, we describe the trichothecene biosynthetic pathway in Fusarium species and discuss genetic evidence that several trichothecene biosynthetic genes are organized in a gene cluster. Trichothecenes are highly toxic to a wide range of eukaryotes, but their specific function, if any, in the survival of the fungi that produce them is not obvious. Trichothecene gene disruption experiments indicate that production of trichothecenes can enhance the severity of disease caused by Fusarium species on some plant hosts. Understanding the regulation and function of trichothecene biosynthesis may aid in development of new strategies for controlling their production in food and feed products.     

Effects of sucrose on photosynthesis and phosphoenolpyruvate carboxylase activity of in vitro cultured strawberry plantlets

Photosynthesis and phosphoenolpyruvate carboxylase activity were investigated in 5, 10 and 28 day-old micropropagated strawberry plantlets (Fragaria x ananassa Duch. cv Kent) rooted in vitro with different levels of sucrose (0, 1, 3 and 5%) on cellulose plugs (Sorbarods). The photosynthetic capability was influenced by the level of sucrose in the culture medium with the largest rates of photosynthesis corresponding to the cultures with 0 and 1% sucrose. The apparent quantum yield and the ratio of variable fluorescence to maximum fluorescence were also reduced in plantlets cultured with 3 or 5% sucrose as compared to those with 0 or 1%. Phosphoenolpyruvate carboxylase activity was largest 5 and 10 days after the onset of culture and decreased in the absence of sucrose in the culture medium. At 28 days after the onset of culture, the activity of this carboxylating enzyme was lower than at the beginning of culture and independent of the concentration of sucrose in the culture medium. Phosphoenolpyruvate carboxylase appears to be an important carboxylating enzyme in micropropagated plantlets.     

Substrate cleavage analysis of furin and related proprotein convertases. A comparative study

We present the data and the technology, a combination of which allows us to determine the identity of proprotein convertases (PCs) related to the processing of specific protein targets including viral and bacterial pathogens. Our results, which support and extend the data of other laboratories, are required for the design of effective inhibitors of PCs because, in general, an inhibitor design starts with a specific substrate. Seven proteinases of the human PC family cleave the multibasic motifs R-X-(R/K/X)-R downward arrow and, as a result, transform proproteins, including those from pathogens, into biologically active proteins and peptides. The precise cleavage preferences of PCs have not been known in sufficient detail; hence we were unable to determine the relative importance of the individual PCs in infectious diseases, thus making the design of specific inhibitors exceedingly difficult. To determine the cleavage preferences of PCs in more detail, we evaluated the relative efficiency of furin, PC2, PC4, PC5/6, PC7, and PACE4 in cleaving over 100 decapeptide sequences representing the R-X-(R/K/X)-R downward arrow motifs of human, bacterial, and viral proteins. Our computer analysis of the data and the follow-on cleavage analysis of the selected full-length proteins corroborated our initial results thus allowing us to determine the cleavage preferences of the PCs and to suggest which PCs are promising drug targets in infectious diseases. Our results also suggest that pathogens, including anthrax PA83 and the avian influenza A H5N1 (bird flu) hemagglutinin precursor, evolved to be as sensitive to PC proteolysis as the most sensitive normal human proteins.     

ER-mediated phagocytosis: a new membrane for new functions

Genomics and other high-throughput approaches, such as proteomics, are changing the way we study complex biological systems. In the past few years, these approaches have contributed markedly to improving our understanding of phagocytosis. Indeed, the ability to study biological systems by monitoring hundreds of proteins provides a level of resolution that is not attainable by the usual 'one protein at a time' approach. In this article, I discuss how proteomic approaches have revealed surprising findings that enable us to revisit established concepts, such as the origin of the phagosome membrane, and to propose new models of cell organization and the link between innate and adaptive immunity.     

Endoplasmic reticulum-mediated phagocytosis is a mechanism of entry into macrophages

Phagocytosis is a key aspect of our innate ability to fight infectious diseases. In this study, we have found that fusion of the endoplasmic reticulum (ER) with the macrophage plasmalemma, underneath phagocytic cups, is a source of membrane for phagosome formation in macrophages. Successive waves of ER become associated with maturing phagosomes during phagolysosome biogenesis. Thus, the ER appears to possess unexpectedly pluripotent fusion properties. ER-mediated phagocytosis is regulated in part by phosphatidylinositol 3-kinase and used for the internalization of inert particles and intracellular pathogens, regardless of their final trafficking in the host. In neutrophils, where pathogens are rapidly killed, the ER is not used as a major source of membrane for phagocytosis. We propose that intracellular pathogens have evolved to adapt and exploit ER-mediated phagocytosis to avoid destruction in host cells.     

Altered coronary and cardiac adrenergic response in the failing hamster heart: role of cyclooxygenase derivatives

Although the influence of the adrenergic system has been studied in the presence of heart failure, controversies still exist. Since cyclooxygenase derivatives appear to modulate coronary and cardiac adaptation in the failing heart, we hypothesized that cyclooxygenase derivatives may participate in the altered adrenergic responses in this situation. Isolated hearts from cardiomyopathic (UM-X7.1 subline) and normal hamsters, aged > 240 days, were utilized. Coronary and cardiac response to alpha1-, beta1-, and beta2-adrenergic stimulations was observed before and after pretreatment with indomethacin, a cyclooxygenase inhibitor. Reduction of coronary flow elicited by alpha1-adrenergic stimulation was unchanged in the presence of heart failure, while beta1- and beta2-induced vasodilatations were reduced. Inotropic response to alpha1 and beta1 stimulations were also reduced in failing hearts, while beta2-adrenergic action was unchanged. Pretreatment with indomethacin exacerbated coronary flow reduction observed with alpha1 stimulation in failing hearts only. Beta2-induced coronary vasodilatation and inotropic response to alpha1 and beta2 stimulations were impaired similarly in the presence of indomethacin in normal and failing hearts. The results suggest a complex interaction between adrenergic and cyclooxygenase activation.     

Isometric virus-like particles from citrus red mites, Panonychus citri

Virus-like spherical particles found in laboratory-reared citrus red mites but not in field populations were independent of the pathogenic virus affecting this species. Three sizes of particles are present: 18-nm spheres occuring in crystalline array, and 30- and 37-nm spheres. The particles possess antigenic properties and contain RNA.