Slavica Janevska

Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute | HKI · (Epi-)Genetic Regulation of Fungal Virulence

The high therapeutic potential of psilocybin, a prodrug of the psychotropic psilocin, holds great promise for the treatment of mental disorders such as therapy‐refractory depression, alcohol use disorder and anorexia nervosa. Psilocybin has been designated a ‘Breakthrough Therapy’ by the US Food and Drug Administration, and therefore a sustainable...

Background Fusarium fujikuroi is a pathogen of rice causing diverse disease symptoms such as ‘bakanae’ or stunting, most likely due to the production of various natural products (NPs) during infection. Fusaria have the genetic potential to synthesize a plethora of these compounds with often diverse bioactivity. The capability to synthesize NPs exce...

Sphingofungins are sphinganine analog mycotoxins acting as inhibitors of serine palmitoyl transferases, enzymes responsible for the first step in the sphingolipid biosynthesis. Eukaryotic cells are highly organized with various structures and organelles to facilitate cellular processes and chemical reactions, including the ones occurring as part of...

It has been discovered that plant pathogens produce effectors that spread via plasmodesmata (PD) to allow modulation of host processes in distal uninfected cells. Fusarium oxysporum f. sp. lycopersici ( Fol) facilitates effector translocation by expansion of the size-exclusion limit of PD using the Six5/Avr2 effector pair. How other fungal pathogen...

Sphingofungins belong to a group of structurally related sphingolipid inhibitors produced by fungi, which specifically inhibit serine palmitoyl transferases, enzymes catalyzing the initial step during sphingolipid biosynthesis. Sphingolipids are integral parts of the eukaryotic cell membrane, and disturbances in their homeostasis have been linked t...

The biosynthesis of fungal natural products is highly regulated not only in terms of transcription and translation but also regarding the cellular localization of the biosynthetic pathway. In all eukaryotes, the endoplasmic reticulum (ER) is involved in the production of organelles, which are subject to cellular traffic or secretion. Here, we show...

Dimethylallyltryptophan synthases catalyze the regiospecific transfer of (oligo)-prenylpyrophosphates to aromatic sub-strates like tryptophan derivatives. These reactions are key steps in many biosynthetic pathways of fungal and bacterial secondary metabolites. In vitro investigations on recombinant DMATS1Ff from Fusarium fujikuroi identified the e...

Aspergillus fumigatus is the most important airborne human pathogenic fungus, causing thousands of deaths per year. Its lethality is due to late and often inaccurate diagnosis and the lack of efficient therapeutics. The failure of efficient prophylaxis and therapy is based on the ability of this pathogen to activate numerous salvage pathways that a...

Fig. S1. Western blot analysis of the Δkdm5 mutant. The western blot was done using the H3K4me3 and H3K4me2 antibodies. Indicated strains were grown in liquid culture (ICI + 60 mM Gln) for 3 days prior to protein extraction. 15 μg of the protein extract was loaded on to the gel, and an unspecific band served as loading control. Fig. S2. Microarray...

Here we present the identification and characterisation of the H3K4‐specific histone methyltransferase Set1 and its counterpart, the Jumonji C demethylase Kdm5, in the rice pathogen Fusarium fujikuroi. While Set1 is responsible for all detectable H3K4me2/me3 in this fungus, Kdm5 antagonises the H3K4me3 mark. Notably, deletion of both SET1 and KDM5...

The fungus Fusarium fujikuroi causes “bakanae” disease of rice due to its ability to produce gibberellins (GAs), a family of plant hormones. Recent genome sequencing revealed the genetic capacity for the biosynthesis of 46 additional secondary metabolites besides the industrially produced GAs. Among them are the pigments bikaverin and fusarubins, a...

The fungus Fusarium fujikuroi causes bakanae disease of rice due to its ability to produce the plant hormones, the gibberellins. The fungus is also known for producing harmful mycotoxins (e.g., fusaric acid and fusarins) and pigments (e.g., bikaverin and fusarubins). However, for a long time, most of these well-known products could not be linked to...

In this work, we present a comprehensive analysis of the H3K36 histone methyltransferases Set2 and Ash1 in the filamentous ascomycete Fusarium fujikuroi In Saccharomyces cerevisiae, one single methyltransferase, Set2, confers all H3K36 methylation, while there are two members of the Set2 family in filamentous fungi, and even more H3K36 methyltransf...

Fusarium fujikuroi causes bakanae (“foolish seedling”) disease of rice which is characterized by hyper-elongation of seedlings resulting from production of gibberellic acids (GAs) by the fungus. This plant pathogen is also known for production of harmful mycotoxins, such as fusarins, fusaric acid, apicidin F and beauvericin. Recently, we generated...

PCR validation of telomere-proximal genes in all strains according to reference sequence of strain IMI 58289 (Primer list see S6 Table). (TIF)

Shoot and root growth of rice seedlings after inoculation with B20 mutants. Rice seedlings (A-C) and roots (D) inoculated with the gene deletion strains derived from the F. fujikuroi B20 strain. Mock: no fungal inoculation; Δfum1, Δfub1, and Δcps/ks–deletion strains for the fumonisin, fusaric acid and giberellic acid key genes, respectively. (TIF)

Biosynthesis of gibberellic acids (GA3, GA4, GA7) and fumonisins (FB1, FB2) of stunting-type F. fujikuroi isolates compared to bakanae strains under in vitro (A, B) and in planta (C, D) conditions. GA (A) and fumonisin (B) production levels after 7 days of growth in synthetic medium with 6 mM glutamine. The strains were grown in triplicates. GA (C)...

MRM transitions for liquid culture analysis in negative ionization mode, depicting also transition specific variables. (DOCX)

MRM transitions for in planta analysis in positive ionization mode, depicting also transition specific variables. (DOCX)

Shoot (A) and root (B) growth of rice seedlings 7 days after inoculation of high fumonisin-producing F. verticillioides strains. Mock: no fungal inoculation; B14: the F. fujikuroi B14 strain; FvOS35: the F. verticilliodies OS35 strain; FvOS40: the F. verticilliodies OS40 strain. (TIF)

BUSCO single-copy analysis, performed in gene set (protein) assessment mode on the library Sordariomyceta_odb9. (DOCX)

Presence of secondary metabolite gene clusters in the analyzed Fusarium strains. (XLSX)

Rice seed germination assay. (DOCX)

(A) Expression data and list of all genes with a transcription factor specific InterPro ID in the corresponding protein sequence. (B) Expression data and list of 37 genes which are present in most of the 9 genomes and which are specifically up-regulated during infection of rice. (C) Expression data and list of 28 B14 strain-specific TFs which are n...

Northern blot analysis for comparison of bikaverin (BIK2) and fusarubins (FSR2) gene expression. The strains were grown for 3 days in synthetic medium with either 6 mM glutamine (bikaverin) or 6 mM NaNO3 (fusarubins) as nitrogen source. (TIF)

Content and arrangement of genes in gene clusters with variations in the single isolates. (A) The apicidin F (NRPS31) gene cluster is present in most isolates but missing in strain B14. (B) The fumonisin (PKS11) gene cluster is present in most of the strains, but several genes of the cluster are missing in strain B20. Arrows in blue represent genes...

Rice seedlings inoculated with F. fujikuroi wild-type and mutant strains with or without addition of culture fluids to the pathosystem. (A) Shoot growth of rice seedlings inoculated with B14 (wild type) and the B14 deletion and complemented strains 9 days after inoculation. (B, C, D) Shoot growth of rice seedlings inoculated with strain B14 (B), B2...

Differentiation of field isolates for their B14-like or B20-like pathotype by PCR and phylogenetic analysis. (A) Diagnostic PCR amplification using the primer pairs derived from PKS51 (unknown product, 382 bp) and NRPS31 (apicidin F, 434 bp), respectively. (B) Phylogenetic tree constructed by the NJ method using the nucleotide sequences of combined...

Distribution of the mating type loci among the F. fujikuroi isolates. (XLSX)

Relative transcript levels for the key genes of GA (CPS/KS) and fumonisin (FUM1) biosynthesis. The transcript levels of CPS/KS (A) and FUM1 (B) were determined by qPCR using total RNA from several F. fujikuroi field isolates grown in ICI liquid medium containing 6 mM glutamine for 7 d. Amplification levels of CPS/KS and FUM1 in the B14 strain were...

Confirmation of the gene deletions (A-F) or complementations (G and H) by PCR. Left panel in each figure: gene deletion or complementation schemes, right panel: PCR gel picture. The genomic positions of the primer pairs (S6 Table) used in the PCR amplification and expected size of PCR products (designated a or b) are indicated in the deletion schem...

PCR primers used in this work. (DOCX)

MRM transitions for liquid culture analysis in positive ionization mode, depicting also transition specific variables. (DOCX)

MRM transitions for in planta analysis in negative ionization mode, depicting also transition specific variables. (DOCX)

The PKS-NRPS-derived tetramic acid equisetin and its N-desmethyl derivative trichosetin exhibit remarkable biological activities against a variety of organisms, including plants and bacteria, e.g., Staphylococcus aureus. The equisetin biosynthetic gene cluster was first described in Fusarium heterosporum, a species distantly related to the notoriou...

The range of secondary metabolites (SMs) produced by the rice pathogen Fusarium fujikuroi is quite broad. Several polyketides, nonribosomal peptides and terpenes have been identified. However, no products of dimethylallyltryptophan synthases (DMATSs) have been elucidated so far, though two putative DMATS genes are present in the F. fujikuroi genome...

In the two fungal pathogens Fusarium fujikuroi and Fusarium graminearum, secondary metabolites (SMs) are fitness and virulence factors and there is compelling evidence that the coordination of SM gene expression is under epigenetic control. Here, we characterized Ccl1, a subunit of the COMPASS complex responsible for methylating lysine 4 of histone...

The 2H-pyran-2-one gibepyrone A and its oxidized derivatives gibepyrones B-F have been isolated from the rice pathogenic fungus Fusarium fujikuroi already more than 20 years ago. However, these products have not been linked to the respective biosynthetic genes and therefore, their biosynthesis has not been analyzed on a molecular level so far. Feed...

Fusaric acid (FSA) is a mycotoxin produced by several fusaria, including the rice pathogen Fusarium fujikuroi. Genes involved in FSA biosynthesis were previously identified as a cluster containing a polyketide synthase (PKS)-encoding (FUB1) and four additional genes (FUB2-FUB5). However, the biosynthetic steps leading to FSA as well as the origin o...

The plant pathogenic fungus Fusarium fujikuroi is the causal agent of bakanae disease on rice due to its ability to produce gibberellins. Besides these phytohormones, F. fujikuroi is able to produce several other secondary metabolites (SMs). Although much progress has been made in the field of secondary metabolism, the transcriptional regulation of...

The fungus F. fujikuroi is well known for its production of gibberellins causing the 'bakanae' disease of rice. Besides these plant hormones, it is able to produce other secondary metabolites (SMs), such as pigments and mycotoxins. Genome sequencing revealed altogether 45 potential SM gene clusters, most of which are cryptic and silent. In this stu...