Increased urea availability promotes adjustments in C/N metabolism and lipid content without impacting growth in Chlamydomonas reinhardtii.

INTRODUCTION: The use of urea as a nitrogen (N) source by Chlorophytes usually enhances biomass and lipid production when compared to ammonium (NH4+). However, the metabolic shifts displayed by Chlamydomonas reinhardtii growing with this organic N source are not known. OBJECTIVES: This study aimed: (i) to characterize the metabolism of C. reinhardtii cultivated in media containing only urea as N source as well as combined with different NH4+ ratios; (ii) to understand how metabolism respond to urea availability. METHODS: Specific quantification of metabolites using 96-well microplates, and high-performance liquid chromatography combined with non-targeted metabolite profiling by gas chromatography (GC)-time-of-flight (TOF)-mass spectrometry (MS) were used in this study. In addition, GC analysis was used to determine fatty acid profiling. RESULTS: The use of urea did not alter the growth rate in comparison with NH4+. Interestingly, the cell number decreased and the cell size increased proportionally with urea availability. Furthermore, chlorophyll, protein and lipid contents increased with the amount of urea. Regarding the fatty acid profile, oleic acid (C18:1 w8) decreased with amount of urea, while linoleic acid (C18:2 w6) doubled in urea-containing medium. CONCLUSIONS: These results indicate that urea promotes remarkable adjustments in metabolism, without drastic changes in biomass, promoting changes in carbohydrate and amino acid metabolism, as well as in lipids production and fatty acid profile.

Abundance, distribution and potential impact of transposable elements in the genome of Mycosphaerella fijiensis.

BACKGROUND: Mycosphaerella fijiensis is a ascomycete that causes Black Sigatoka in bananas. Recently, the M. fijiensis genome was sequenced. Repetitive sequences are ubiquitous components of fungal genomes. In most genomic analyses, repetitive sequences are associated with transposable elements (TEs). TEs are dispersed repetitive DNA sequences found in a host genome. These elements have the ability to move from one location to another within the genome, and their insertion can cause a wide spectrum of mutations in their hosts. Some of the deleterious effects of TEs may be due to ectopic recombination among TEs of the same family. In addition, some transposons are physically linked to genes and can control their expression. To prevent possible damage caused by the presence of TEs in the genome, some fungi possess TE-silencing mechanisms, such as RIP (Repeat Induced Point mutation). In this study, the abundance, distribution and potential impact of TEs in the genome of M. fijiensis were investigated. RESULTS: A total of 613 LTR-Gypsy and 27 LTR-Copia complete elements of the class I were detected. Among the class II elements, a total of 28 Mariner, five Mutator and one Harbinger complete elements were identified. The results of this study indicate that transposons were and are important ectopic recombination sites. A distribution analysis of a transposable element from each class of the M. fijiensis isolates revealed variable hybridization profiles, indicating the activity of these elements. Several genes encoding proteins involved in important metabolic pathways and with potential correlation to pathogenicity systems were identified upstream and downstream of transposable elements. A comparison of the sequences from different transposon groups suggested the action of the RIP silencing mechanism in the genome of this microorganism. CONCLUSIONS: The analysis of TEs in M. fijiensis suggests that TEs play an important role in the evolution of this organism because the activity of these elements, as well as the rearrangements caused by ectopic recombination, can result in deletion, duplication, inversion and translocation. Some of these changes can potentially modify gene structure or expression and, thus, facilitate the emergence of new strains of this pathogen.