BackgroundThe industrially important yeast Blastobotrys (Arxula) adeninivorans is an asexual hemiascomycete phylogenetically very distant from Saccharomyces cerevisiae. Its unusual metabolic flexibility allows it to use a wide range of carbon and nitrogen sources, while being thermotolerant, xerotolerant and osmotolerant.ResultsThe sequencing of strain LS3 revealed that the nuclear genome of A. adeninivorans is 11.8 Mb long and consists of four chromosomes with regional centromeres. Its closest sequenced relative is Yarrowia lipolytica, although mean conservation of orthologs is low. With 914 introns within 6116 genes, A. adeninivorans is one of the most intron-rich hemiascomycetes sequenced to date. Several large species-specific families appear to result from multiple rounds of segmental duplications of tandem gene arrays, a novel mechanism not yet described in yeasts. An analysis of the genome and its transcriptome revealed enzymes with biotechnological potential, such as two extracellular tannases (Atan1p and Atan2p) of the tannic-acid catabolic route, and a new pathway for the assimilation of n-butanol via butyric aldehyde and butyric acid.ConclusionsThe high-quality genome of this species that diverged early in Saccharomycotina will allow further fundamental studies on comparative genomics, evolution and phylogenetics. Protein components of different pathways for carbon and nitrogen source utilization were identified, which so far has remained unexplored in yeast, offering clues for further biotechnological developments. In the course of identifying alternative microorganisms for biotechnological interest, A. adeninivorans has already proved its strengthened competitiveness as a promising cell factory for many more applications.

Eimeria tenella is a coccidian parasite of great economical importance for poultry industry. The surface of Eimeria invasive agents, sporozoites and merozoites, is coated with a family of developmentally regulated glycosylphosphatidylinositol (GPI)-linked surface antigens (SAGs), some of them involved in the initiation of the infection process. Using 2D gel electrophoresis followed by mass spectrometry, an antigenic surface protein EtSAG1 (TA4) of E. tenella sporozoites has been identified as a target of neutralizing monoclonal antibody 2H10E3. To clarify the mechanism of invasion inhibition caused by the EtSAG1-specific antibodies, a structural model of EtSAG1 was generated. It appears that "EtSAG fold" does not bear an evolutionary relationship to any known protein structure. The intra- and interchain disulfide bonds could be assigned to certain pairs of six conserved cysteines found in members of the EtSAG protein family. The outward-facing surface of the antigen was found to comprise an expanded positively charged patch, thus suggesting that the parasite invasion process may be initiated by sporozoite attachment to negatively charged sulfated proteoglycans on the surface of the host cell.

Background: Coccidiosis caused by protozoans of genus Eimeria is a chicken parasitic disease of great economical importance. Conventional disease control strategies depend on vaccination and prophylactic use of anticoccidial drugs. Alternative solution to prevent and treat coccidiosis could be provided by passive immunization using orally delivered neutralizing antibodies. We investigated the possibility to mitigate the parasitic infection by feeding poultry with antibody expressing transgenic crop seeds.

The organ specificity of four promoters that are known to direct seed-specific gene expression was tested. Whereas the phaseolin (phas)- and legumin B4 (leB4)-promoters were from genes encoding 7S and 11S globulins from Phaseolus vulgaris and Vicia faba, respectively, the usp- and the sbp-promoters were from non-storage protein genes of V. faba. The expression of different promoter-reporter gene fusions was followed either by RT-PCR or by registering the reporter enzyme activity in organs of transgenic tobacco, pea, narbon bean, or linseed. In addition to seeds, the promoters directed reporter gene expression in pollen and in seed coats. USP-, vicilin- and legumin-mRNA were detected by RT-PCR in pollen of Pisum sativum and V. faba. Expression during microsporogenesis and embryogenesis seems to be a general character of various seed protein genes.

SUMMARYThe transition of pea embryos from pre-storage to maturation is partially controlled by abscisic acid (ABA). Immunomodulation in pea embryos specifically reduces free ABA levels during transition stages. Such seeds are, therefore, suitable models for studying ABA deficiency by global transcript and metabolite analysis. Compared with the wild type, anti-ABA seeds are smaller, contain fewer globulins and show lower dry matter accumulation and delayed differentiation. Free sugars are decreased, indicating lower uptake and/or elevated mobilisation. Lower levels of trans-zeatins suggest that ABA reduction influences rates of cytokinin synthesis and/or its level of accumulation. Abscisic acid deficiency leads to a general downregulation of gene expression related to transcription and translation. At the transcriptional level, anti-ABA embryos reveal a wide-range repression of carbohydrate oxidation, downregulated sucrose mobilisation, glycolysis and the tricarboxylic acid cycle/Krebs cycle (TCA cycle). Genes related to starch, amino acid and storage protein biosynthesis are downregulated, indicating a general decrease in metabolic fluxes. We conclude that during embryo differentiation ABA triggers broad upregulation of gene activity and genetic reprogramming, involving regulated protein degradation via the ubiquitin/proteasome system. Abscisic acid deficiency affects gene expression associated with transport processes and stimulation of membrane energisation. Our study identified mediators and downstream signalling elements of ABA during embryo differentiation, such as the transcription factor FUSCA3, SnRK1 kinase and Ca 2+ signalling processes. This suggests that ABA interacts with SnRK1 complexes, thus connecting SnRK1, sugar and stress signalling with ABA. Certain protein kinases/ phosphatases known to negatively respond to ABA are upregulated in the modulated line, whilst those which respond positively are downregulated, pointing to a highly coordinated response of the gene network to ABA levels.

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