Basic phenotypic analysis of six novel yeast genes reveals two essential genes and one which affects the growth rate

Phenotypic analysis was performed on six mutants of Saccharomyces cerevisiae deleted in one of the following open reading frames (ORFs), located on chromosome II: YBR254c, YBR255w, YBR257w, YBR258c, YBR259w and YBR266c. Disruption of the ORFs was carried out in the diploid strain FY1679 using the kanMX4 marker flanked by short sequences homologous to the target locus. Tetrad analysis following sporulation of the heterozygous disruptants showed that YBR254c and YBR257w are essential genes. YBR257w was later characterized and renamed POP4, its gene product being involved in 5.8S rRNA and tRNA processing (Chu et al., 1997). The tetrad analysis performed for the heterozygous disruptant for YBR2…

Characterisation of Bobal and Crujidera grape cultivars, in comparison with Tempranillo and Cabernet Sauvignon: Evolution of leaf macronutrients and berry composition during grape ripening

Abstract We present characterisation data for two Spanish autochthonous grapevines, Bobal and Crujidera, in comparison with the well-known cultivars Tempranillo and Cabernet Sauvignon. Microsatellite markers were used for the molecular characterisation of Crujidera grapevines. Leaf macronutrient contents of the four cultivars were evaluated, as well as their changes at different vine developmental stages, and veraison was seen as the most suitable time to evaluate the nutritional status. Quantitative changes in some physiological parameters and the phenolic composition of the four grape varieties were measured during the last month of ripening. Polyphenols, tannins and anthocyanins increase…

A singleFKShomologue inYarrowia lipolyticais essential for viability

The synthesis of β-1,3-glucan, the structural component of the yeast cell wall which gives shape to the cell, occurs at the plasma membrane and is the result of the activity of at least a two-component complex. Fks1p is the catalytic subunit directly responsible for the synthesis of β-1,3-glucan, whilst the second subunit, Rho1p, has a GTP-dependent regulatory role. FKS1 has been characterized in Saccharomyces cerevisiae, where its function is at least partially redundant with that of FKS2/GSC2. FKS homologues have also been identified in several other fungal species, including Candida albicans, Schizosaccharomyces pombe, Aspergillus nidulans, Cryptococcus neoformans and Paracoccidiodes bra…

First report of Diplodia fraxini causing dieback of Fraxinus angustifolia in Spain

RHO1(YlRHO1) is a non-essential gene inYarrowia lipolyticaand complementsrho1Δlethality inSaccharomyces cerevisiae

The synthesis of beta-1,3-glucan, the structural component of the yeast cell wall that gives shape to the cell, occurs at the plasma membrane and is the result of the activity of at least a two-component complex. Fks1p is the catalytic subunit directly responsible for the synthesis of beta-1,3-glucan, whilst the second subunit, Rho1p, has a GTP-dependent regulatory role (Yamochi et al., 1994). RHO1 has been characterized in Saccharomyces cerevisiae (Yamochi et al., 1994), and in several other fungal species. In this work, we have used degenerate oligonucleotides derived from the conserved regions of Rho1ps to isolate the RHO1 gene of Yarrowia lipolytica. The gene isolated in this way, which…

Isolation of theMNN9gene ofYarrowia lipolytica(YlMNN9) and phenotype analysis of a mutantylmnn9Δ strain

In this work we describe the isolation of the Yarrowia lipolytica homologue of Saccharomyces cerevisiae MNN9 gene, which we have named YlMNN9, and the phenotype analysis of a Y. lipolytica strain containing the disrupted YlMNN9 allele. YlMNN9 was cloned using degenerate consensus oligonucleotides to generate specific probes that were in turn used to screen mini-gene libraries. The gene is defined by a 1014 bp ORF predicted to encode a protein 337 amino acids long that shares significant homology with the Mnn9ps of S. cerevisiae, Candida albicans and Hansenula polymorpha, including a putative N-terminal transmembrane domain. Disruption of YlMNN9 leads to phenotypes such as resistance to sodi…