We recently reported that DAG (diacylglycerol) generated during sphingomyelin synthesis plays an important role in protein kinase C activation and cell proliferation in Madin-Darby canine kidney cells [Cerbon and Lopez-Sanchez (2003) Biochem. J. 373, 917-924]. In yeast cells, IPC (inositol phosphoceramide) synthase catalyses the transfer of phosphoinositol from phosphatidylinositol to ceramide to form IPC and generates DAG. In the present study, we found that, during the G1 to S transition after N2-starvation, there was a significant increase in the synthesis of IPC accompanied by a progressive increase (up to 6-fold) in the level of DAG. The increased DAG levels coincided with decrements in ceramide and sphingoid base levels, conditions that are adequate for the activation of putative protein kinase C required for the G1 to S transition and proliferation of yeast cells. To separate the role of DAG generated during IPC synthesis from that originating from other sources, we utilized beta-chloroalanine and myriocin, inhibitors of serine:palmitoyl-CoA transferase, the first committed step in sphingolipid synthesis, to avoid accumulation of sphingolipid intermediates. When the synthesis of sphingolipids was inhibited, DAG accumulation was significantly decreased and the G1 to S transition was blocked; such blockage was avoided by metabolic complementation with phytosphingosine. The DAG/ceramide ratio was 0.27 and it changed to 2.0 during growth re-initiation, suggesting that the synthesis of phosphosphingolipids could act to switch growth arrest (increased ceramide) to a mitogenic signal (increased DAG), and that this signalling process is preserved in yeast and mammalian cells.
The influence of chitosaccharides on the symbiotic interaction between Bradyrhizobium and soybean was examined. The results show that chitosaccharides either positively or negatively affect soybean nodulation or plant growth depending on their molecular weight, concentration and the application methods. When directly added to the in vitro culture media, chitosan of high molecular weight inhibit Bradyrhizobium viability in a dose dependent manner while chitooligosaccharides reduce slightly the bacteria viability only at concentration equal or higher than 50 mg•L −1. Chitooligosaccharides significantly enhance nodule formation and dry mass in soybean roots at doses between 10 and 100 mg•L −1. Both types of chitosaccharides, at the highest doses (>500 mg•L −1), negatively affect plant height and root size, whereas medium doses (50 to 100 mg•L −1) increase slightly leave number. Under field conditions, foliar application of both chitosaccharides enhances growth and nodulation of soybean plants. Nevertheless, using this application method, chitosan remains more effective than chitooligosaccharides.
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