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The oomycete pathogen Phytophthora cactorum causes crown rot, a major disease of cultivated strawberry. We report the draft genome of P. cactorum isolate 10300, isolated from symptomatic Fragaria x ananassa tissue. Our analysis revealed that there are a large number of genes encoding putative secreted effectors in the genome, including nearly 200 RxLR domain containing effectors, 77 Crinklers (CRN) grouped into 38 families, and numerous apoplastic effectors, such as phytotoxins (PcF proteins) and necrosis inducing proteins. As in other Phytophthora species, the genomic environment of many RxLR and CRN genes differed from core eukaryotic genes, a hallmark of the two-speed genome. We found genes homologous to known Phytophthora infestans avirulence genes including Avr1, Avr3b, Avr4, Avrblb1 and AvrSmira2 indicating effector sequence conservation between Phytophthora species of clade 1a and clade 1c. The reported P. cactorum genome sequence and associated annotations represent a comprehensive resource for avirulence gene discovery in other Phytophthora species from clade 1 and, will facilitate effector informed breeding strategies in other crops.

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The proportion of Norwegian wheat used for food has varied significantly during the recent decade, mainly because of the instability of factors that are essential to baking quality (i.e. protein content and gluten functionality). During the same period, serious contamination of Fusarium spp. and mycotoxins was observed in some grain lots [1, 2]. A project was established to generate greater knowledge of the interface between gluten functionality and effects of Fusarium species and other microorganisms on Norwegian wheat quality. Instances of severe degradation of gluten proteins that resulted in an almost complete loss of gluten functionality were observed in some lots of Norwegian wheat. The degradation of the gluten appeared to be caused by exogenous proteases. Metabarcoding of fungi and bacteria in these grain lots identified fungi within the Fusarium Head Blight complex, as well as one bacterial species, as candidate species for influencing gluten functionality. Some of these candidates were inoculated on wheat during flowering [3]. Analysis of baking quality of the flour from this experiment revealed a reduced proportion of un-extractable polymeric proteins (%UPP) and severe reductions in the gluten’s resistance to stretching (RMAX) in wheat flour from plants inoculated with Fusarium graminearum. Flour from wheat inoculated with Fusarium avenaceum was generally less infested, and showed minimal or no reduction in gluten functionality and %UPP compared to flour from the F. graminearum infested samples. Flour from wheat inoculated with Michrodochium majus is yet to be analysed. References 1. Koga, S., et al., Investigating environmental factors that cause extreme gluten quality deficiency in winter wheat (Triticum aestivum L.). Acta Agriculturae Scandinavica, Section B—Soil & Plant Science, 2016. 66(3): p. 237-246. 2. Hofgaard, I., et al., Associations between Fusarium species and mycotoxins in oats and spring wheat from farmers’ fields in Norway over a six-year period. World Mycotoxin Journal, 2016. 9(3): p. 365-378. 3. Nielsen, K.A.G., Effect of microorganisms on gluten quality in wheat., in Faculty of Biosciences. 2017, Norwegian University of Life Sciences: Ås.

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Helminthosporium solani causes silver scurf, which affects the quality of potato. The biocontrol agent Clonostachys rosea greatly limited the severity of silver scurf symptoms and amount of H. solani genomic DNA in laboratory experiments. Transcriptomic analysis during interaction showed that H. solani gene expression was highly reduced when coinoculated with the biocontrol agent C. rosea, whereas gene expression of C. rosea was clearly boosted as a response to the pathogen. The most notable upregulated C. rosea genes were those encoding proteins involved in cellular response to oxidative stress, proteases, G-protein signaling, and the methyltransferase LaeA. The most notable potato response to both fungi was downregulation of defense-related genes and mitogen-activated protein kinase kinase kinases. At a later stage, this shifted, and most potato defense genes were turned on, especially those involved in terpenoid biosynthesis when H. solani was present. Some biocontrol-activated defense-related genes in potato were upregulated during early interaction with C. rosea alone that were not triggered by H. solani alone. Our results indicate that the reductions of silver scurf using C. rosea are probably due to a combination of mechanisms, including mycoparasitism, biocontrol-activated stimulation of plant defense mechanisms, microbial competition for nutrients, space, and antibiosis.

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Production of chrysogine has been reported from several fungal genera including Penicillium, Aspergillus, and Fusarium. Anthranilic acid and pyruvic acid, which are expected precursors of chrysogine, enhance production of this compound. A possible route for the biosynthesis using these substrates is via a nonribosomal peptide synthetase (NRPS). Through comparative analysis of the NRPSs from genome-sequenced producers of chrysogine we identified a candidate NRPS cluster comprising five additional genes named chry2–6. Deletion of the two-module NRPS (NRPS14 = chry1) abolished chrysogine production in Fusarium graminearum, indicating that the gene cluster is responsible for chrysogine biosynthesis. Overexpression of NRPS14 enhanced chrysogine production, suggesting that the NRPS is the bottleneck in the biosynthetic pathway.

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The genus Microbacterium contains bacteria that are ubiquitously distributed in various environments and includes plant-associated bacteria that are able to colonize tissue of agricultural crop plants. Here, we report the 3,508,491 bp complete genome sequence of Microbacterium sp. strain BH-3-3-3, isolated from conventionally grown lettuce (Lactuca sativa) from a field in Vestfold, Norway. The nucleotide sequence of this genome was deposited into NCBI GenBank under the accession CP017674.

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In fungi, distribution of secondary metabolite (SM) gene clusters is often associated with host- or environment-specific benefits provided by SMs. In the plant pathogen Alternaria brassicicola (Dothideomycetes), the DEP cluster confers an ability to synthesize the SM depudecin, a histone deacetylase inhibitor that contributes weakly to virulence. The DEP cluster includes genes encoding enzymes, a transporter, and a transcription regulator. We investigated the distribution and evolution of the DEP cluster in 585 fungal genomes and found a wide but sporadic distribution among Dothideomycetes, Sordariomycetes, and Eurotiomycetes. We confirmed DEP gene expression and depudecin production in one fungus, Fusarium langsethiae. Phylogenetic analyses suggested 6–10 horizontal gene transfers (HGTs) of the cluster, including a transfer that led to the presence of closely related cluster homologs in Alternaria and Fusarium. The analyses also indicated that HGTs were frequently followed by loss/pseudogenization of one or more DEP genes. Independent cluster inactivation was inferred in at least four fungal classes. Analyses of transitions among functional, pseudogenized, and absent states of DEP genes among Fusarium species suggest enzyme-encoding genes are lost at higher rates than the transporter (DEP3) and regulatory (DEP6) genes. The phenotype of an experimentally-induced DEP3 mutant of Fusarium did not support the hypothesis that selective retention of DEP3 and DEP6 protects fungi from exogenous depudecin. Together, the results suggest that HGT and gene loss have contributed significantly to DEP cluster distribution, and that some DEP genes provide a greater fitness benefit possibly due to a differential tendency to form network connections.

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The aim of this study was to evaluate the natural occurrence of Beauveria spp. in soil, from infections in the stink bug Piezodorus guildinii, an important pest of common bean (Phaseolus vulgaris) and as endophytes in bean plant tissue. Twelve conventional and 12 organic common bean fields in the Villa Clara province, Cuba were sampled from September 2014 to April 2015. One hundred and fifty Beauveria isolates were obtained from soil samples, bean plant parts and stink bugs. The overall frequency of occurrence of Beauveria isolates in conventional fields (8.4%) was significantly lower than that in organic fields (23.6%). Beauveria were also obtained significantly more frequently from bean roots in organic fields (15.0%) compared to bean roots in conventional fields (3.3%). DNA sequencing of the intergenic Bloc region was performed for Beauveria species identification. All isolates where characterized as Beauveria bassiana (Balsamo-Crivelli) Vuillemin, and clustered with isolates of neotropical origin previously described as AFNEO_1. The Cuban B. bassiana isolates formed five clusters in the phylogeny. Isolates of two clusters originated from all four locations, organic and conventional fields, as well as soil, plants and stink bugs. Organic fields contained isolates of all five clusters while conventional fields only harbored isolates of the two most frequent ones. Mating type PCR assays revealed that mating type distribution was skewed, with MAT1/MAT2 proportion of 146/4, indicating limited potential for recombination. The present study is the first to report of B. bassiana as a naturally occurring endophyte in common bean. Further, it shows that B. bassiana occurs naturally in diverse environments of common bean fields, and constitutes a potential reservoir of natural enemies against pest insects particularly in organic fields.

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The genus Pectobacterium, which belongs to the bacterial family Enterobacteriaceae, contains numerous species that cause soft rot diseases in a wide range of plants. The species Pectobacterium carotovorum is highly heterogeneous, indicating a need for re-evaluation and a better classification of the species. PacBio was used for sequencing of two soft-rot-causing bacterial strains (NIBIO1006T and NIBIO1392), initially identified as P. carotovorum strains by fatty acid analysis and sequencing of three housekeeping genes (dnaX, icdA and mdh). Their taxonomic relationship to other Pectobacterium species was determined and the distance from any described species within the genus Pectobacterium was less than 94% average nucleotide identity (ANI). Based on ANI, phylogenetic data and genome-to-genome distance, strains NIBIO1006T, NIBIO1392 and NCPPB3395 are suggested to represent a novel species of the genus Pectobacterium, for which the name Pectobacterium polaris sp. nov. is proposed. The type strain is NIBIO1006T (=DSM 105255T=NCPPB 4611T).

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Putative proton coupled di-peptide transporters, PTR2s, are found in filamentous fungi in different numbers and their function during fungal development and plant infection is unresolved. In Fusarium graminearum, the cause of head blight in cereals, we identified four putative PTR2 transporters (FgPTR2A-D). The genes did not cluster together in phylogenetic analyses and only FgPTR2A and FgPTR2C were able to complement a PTR2 deficient yeast mutant in uptake of di-peptides. All FgPTR2s are continuously expressed throughout the fungal lifecycle, although at different levels. In silico analyses of existing expression-data show that FgPTR2B is found at higher levels than the others in planta and during sexual development. Deletion mutants of FgPTR2A, FgPTR2C, and FgPTR2D had a higher production of deoxynivalenol (DON) and zearalenone and lower production of fusarielin H than the wild type. Perithecium development was reduced in these mutants but unaffected by deletion of FgPTR2B. Conidia production was reduced in the FgPTR2B mutant and unaffected by deletion of the other PTR2 transporters. Sexual development and secondary metabolite production are known to be linked at the regulatory level and the results suggest that PTR2s are active in nitrogen turnover and thereby influence signal processes.

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Biosynthesis of the black perithecial pigment in the filamentous fungus Fusarium graminearum is dependent on the polyketide synthase PGL1 (oPKS3). A seven-membered PGL1 gene cluster was identified by over-expression of the cluster specific transcription factor pglR. Targeted gene replacement showed that PGL1, pglJ, pglM and pglV were essential for the production of the perithecial pigment. Over-expression of PGL1 resulted in the production of 6-O-demethyl-5-deoxybostrycoidin (1), 5-deoxybostrycoidin (2), and three novel compounds 5-deoxybostrycoidin anthrone (3), 6-O-demethyl-5-deoxybostrycoidin anthrone (4) and purpurfusarin (5). The novel dimeric bostrycoidin purpurfusarin (5) was found to inhibit the growth of Candida albicans with an IC50 of 8.0 +/− 1.9 μM. The results show that Fusarium species with black perithecia have a previously undescribed form of 5-deoxybostrycoidin based melanin in their fruiting bodies.

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Fusarium langsethiae is a widespread pathogen of small grain cereals, causing problems with T-2 and HT-2 toxin contamination in grains every year. In an effort to better understand the biology of this fungus, we present a draft genome sequence of F. langsethiae Fl201059 isolated from oats in Norway. The assembly was fragmented, but reveals a genome of approximately 37.5 Mb, with a GC content around 48%, and 12,232 predicted protein-coding genes. Focusing on secondary metabolism we identified candidate genes for 12 polyketide synthases, 13 non-ribosomal peptide synthetases, and 22 genes for terpene/isoprenoid biosynthesis. Some of these were found to be unique compared to sequence databases. The identified putative Tri5 cluster was highly syntenic to the cluster reported in F. sporotrichioides. Fusarium langsethiae Fl201059 produces a high number of secondary metabolites on Yeast Extract Sucrose (YES) agar medium, dominated by type A trichothecenes. Interestingly we found production of glucosylated HT-2 toxin (Glu-HT-2), previously suggested to be formed by the host plant and not by the fungus itself. In greenhouse inoculations of F. langsethiae Fl201059 on barley and oats, we detected the type A trichothecenes: neosolaniol, HT-2 toxin, T-2 toxin, Glu-HT-2 and numerous derivatives of these.

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The mycotoxin enniatin B, a cyclic hexadepsipeptide produced by the plant pathogen Fusarium, is prevalent in grains and grain-based products in different geographical areas. Although enniatins have not been associated with toxic outbreaks, they have caused toxicity in vitro in several cell lines. In this study, the cytotoxic effects of enniatin B were assessed in relation to cellular energy metabolism, cell proliferation, and the induction of apoptosis in Balb 3T3 and HepG2 cells. The mechanism of toxicity was examined by means of whole genome expression profiling of exposed rat primary hepatocytes. Enniatin B altered cellular energy metabolism and reduced cell proliferation in Balb 3T3 and HepG2 cell lines. Furthermore, the proportion of apoptotic cell populations of Balb 3T3 cells slightly increased. On the other hand, enniatin B caused necrotic cell death in primary hepatocytes. Gene expression studies revealed the alteration of energy metabolism due to effects on mitochondrial organization and function and the assembly of complex I of the electron transport chain.

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The proportion of Norwegian wheat used for food has recently been dramatically lower due to both reduced production and poor quality. Furthermore, the Norwegian milling and baking industries have experienced major challenges in utilizing Norwegian wheat due to the instability of factors, such as protein content and gluten functionality, that are of major importance for baking quality. The variation in the wheat quality can itself cause economic losses for the milling and baking industry due to uncertainty in the marketplace. In the same period as a large variation in baking quality was reported in Norwegian wheat, serious contamination of Fusarium spp. and mycotoxins were observed in some grain lots. We have revealed the severe degradation of gluten proteins in some Norwegian wheat samples leading to an almost complete loss in the gluten functionality. The degradation of the gluten appears to be caused by exogenous proteases, and was associated with the presence of Fusarium spp., and their metabolites, and other microorganisms in the wheat. Increased knowledge is needed to establish the cause of the poor gluten functionality and to develop control measures to reduce the amount of poor quality wheat entering the food value chain. In 2014, a new project was established to generate deeper knowledge in the interface between gluten functionality and effects of Fusarium spp. and other microorganism on wheat quality, and to better utilize Norwegian wheat grown in this challenging environment. A metagenomic analysis, designed to identify microorganisms associated with reduced baking quality, has been undertaken. To study the influence of the identified microorganisms and their metabolites on gluten functionality, wheat plants were inoculated with microorganisms, selected based upon the results of the metagenomics study. Fusarium species are among those microorganisms being tested.

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The aim of this study was to evaluate the effect of conventional versus organic common bean (Phaseolus vulgaris) production on natural occurrence of Beauveria spp. as entophytes in bean plant tissue, from soil and as infections in stink bugs (Hemiptera: Pentatomidae), an important pest of bean in Cuba. Twenty-four organic and conventionally managed bean fields were sampled from September 2014 to April 2015 and Beauveria spp. were isolated and DNA extracted. PCR amplification of the intergenic Bloc region was performed for the identification of Beauveria species. Eighty-seven isolates were obtained from the soil samples by using the Galleria mellonella baiting technique. Further, 45 isolates were obtained from endophytic tissues of bean plant parts and 18 isolates were acquired from stink bugs. Only Beauveria bassiana was identified by DNA sequencing in this material. B. bassiana was more prevalent in soil, plant and stink bugs sampled from organic fields (41% soil, 22% plant, 9% bugs) compared to conventional fields (17% soil, 8% plant, 2% bugs). All plant parts were colonized by B. bassiana, but a significantly higher occurrence of this fungus was found in roots (9%) compared to stems (6%), leaves (4%) and pods (2%) in organic fields. In conventional fields there was a significantly higher occurrence of B. bassiana acquired from root (4%) and stem (3%) compared to leaves (1%) and pods (1%). Mating type PCR assays revealed that each of the isolates carried single mating types, with frequencies of 146/150 (MAT1) and 4/150 (MAT2), indicating limited potential for recombination. Our findings show that B. bassiana occur naturally as endophytes in bean fields in Cuba and contribute to a better ecological understanding of B. bassiana in agriculture.

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In this letter, we advocate recognizing the genus Fusarium as the sole name for a group that includes virtually all Fusarium species of importance in plant pathology, mycotoxicology, medicine, and basic research. This phylogenetically guided circumscription will free scientists from any obligation to use other genus names, including teleomorphs, for species nested within this clade, and preserve the application of the name Fusarium in the way it has been used for almost a century. Due to recent changes in the International Code of Nomenclature for algae, fungi, and plants, this is an urgent matter that requires community attention. The alternative is to break the longstanding concept of Fusarium into nine or more genera, and remove important taxa such as those in the F. solani species complex from the genus, a move we believe is unnecessary. Here we present taxonomic and nomenclatural proposals that will preserve established research connections and facilitate communication within and between research communities, and at the same time support strong scientific principles and good taxonomic practice.

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Fusarium langsethiae er et stort problem i nordisk kornproduksjon. Den forekommer i hovedsak på havre, men finnes også på bygg og i svært liten grad i hvete. Når kornet blir angrepet av Fusarium- sopper kan man vanligvis se skadene fra soppen og dermed vurdere hvor stort angrepet er. Dette er ikke tilfelle for F. langsethiae, som opptrer uten symptomer på havre. Dette kan gi en falsk trygghet, fordi akkurat denne arten produserer de mest giftige av Fusarium trichothecenene, T-2 og HT-2 toksin. For å forstå mer om F. langsethiae har vi studert genuttrykket av soppen under tre forskjellige betingelser, 1) på havrekorn, 2) på blomstrende havreaks og 3) på et næringsrikt vekstmedium. Den tredje prøven ble brukt som sammenligningsgrunnlag. Fra disse tre eksperimentene har vi sekvensert de aktive soppgenene. Vi har benyttet en ny metode (454 pyrosekvensering) som gir ufattelige mengder data, i dette tilfellet 1,3 millioner sekvenser. Man bruker deretter bioinformatiske verktøy for å finne ut hvilke biologiske mekanismer som fungerer under de forskjellige betingelsene. Vi vil presentere noen resultater fra dette studiet. F. langsethiae er foreslått å være i slektskap med F. sporotrichioides, som produserer de samme toksinene, HT-2 og T-2. Denne vurderingen er gjort på bakgrunn av sekvensering av enkelte gener, sammenligning av metabolske profiler, morfologisk karakter mm. Resultater fra vår studie, som er basert på store menger sekvensdata, viser at F. langsethiae har mer likhet med en annen viktig sopp, F. graminearum. Denne Fusarium-arten produserer toksinet deoxynivalenol (DON), og er vanlig å finne i norsk korn. Dette viser at selv om to arter produserer samme toksiner, trenger de ikke være i nært slektskap. En annen egenskap som er funnet hos F. langsethiae, er sannsynlig produksjon av en ny forbindelse som til nå ikke er kjent i norsk korn. Ingen andre Fusarium arter er kjent for å produsere denne metabolitten. På korn-mediet er alle genene som behøves for syntesen av dette stoffet, høyt uttrykt. Neste skritt er å se om vi finner stoffet i kornprøver. Giftigheten til dette stoffet er fremdeles ukjent, men det kan vise seg å utgjøre et nytt problem hvis det er vanlig forekommende i havre. Dette viser at med bioinformatiske metoder kan man avdekke nye egenskaper som i utgangspunktet er skjulte hos soppene.

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Fusarium langsethiae er et stort problem i nordisk kornproduksjon. Den forekommer i hovedsak på havre, men finnes også på bygg og i svært liten grad i hvete. Når kornet blir angrepet av Fusarium- sopper kan man vanligvis se skadene fra soppen og dermed vurdere hvor stort angrepet er. Dette er ikke tilfelle for F. langsethiae, som opptrer uten symptomer på havre. Dette kan gi en falsk trygghet, fordi akkurat denne arten produserer de mest giftige av Fusarium trichothecenene, T-2 og HT-2 toksin. For å forstå mer om F. langsethiae har vi studert genuttrykket av soppen under tre forskjellige betingelser, 1) på havrekorn, 2) på blomstrende havreaks og 3) på et næringsrikt vekstmedium. Den tredje prøven ble brukt som sammenligningsgrunnlag. Fra disse tre eksperimentene har vi sekvensert de aktive soppgenene. Vi har benyttet en ny metode (454 pyrosekvensering) som gir ufattelige mengder data, i dette tilfellet 1,3 millioner sekvenser. Man bruker deretter bioinformatiske verktøy for å finne ut hvilke biologiske mekanismer som fungerer under de forskjellige betingelsene. Vi vil presentere noen resultater fra dette studiet. F. langsethiae er foreslått å være i slektskap med F. sporotrichioides, som produserer de samme toksinene, HT-2 og T-2. Denne vurderingen er gjort på bakgrunn av sekvensering av enkelte gener, sammenligning av metabolske profiler, morfologisk karakter mm. Resultater fra vår studie, som er basert på store menger sekvensdata, viser at F. langsethiae har mer likhet med en annen viktig sopp, F. graminearum. Denne Fusarium-arten produserer toksinet deoxynivalenol (DON), og er vanlig å finne i norsk korn. Dette viser at selv om to arter produserer samme toksiner, trenger de ikke være i nært slektskap. En annen egenskap som er funnet hos F. langsethiae, er sannsynlig produksjon av en ny forbindelse som til nå ikke er kjent i norsk korn. Ingen andre Fusarium arter er kjent for å produsere denne metabolitten. På korn-mediet er alle genene som behøves for syntesen av dette stoffet, høyt uttrykt. Neste skritt er å se om vi finner stoffet i kornprøver. Giftigheten til dette stoffet er fremdeles ukjent, men det kan vise seg å utgjøre et nytt problem hvis det er vanlig forekommende i havre. Dette viser at med bioinformatiske metoder kan man avdekke nye egenskaper som i utgangspunktet er skjulte hos soppene.

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Fusarium graminearum causes head blight disease in wheat, and to help understand the infection process we studied global gene expression of F. graminearum in a time series of 24 to 196 hours after inoculation, compared to a water control. The infection is rapid and already after 48h over 4000 genes is expressed. The number of genes expressed increased over time up to 96h (>8000 genes), and went down again 144h and 192h post inoculation. After subtraction of genes found expressed on complete media, during C and N starvation, and on barley, only 355 were found exclusively expressed in wheat, mostly ones with unknown function (72,6%). These were mainly found in SNP enriched islands on the chromosomes, suggesting a higher evolutionary selection pressure of wheat specific genes. The annotated genes found were enriched in functional groups involved in allantoin and allantoate transport, detoxification, nitrogen, sulfur and selenium metabolism, secondary metabolism, carbohydrate metabolism and degradation of polysaccharides and ester compounds. Several putative secreted virulence factors were also found expressed in wheat.

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Fusarium graminearum causes head blight disease in wheat and barley. To help understand the infection process on wheat we studied global gene expression of F. graminearum in a time series from 24 to 196 hours after inoculation, compared to a non-inoculated control. The infection was rapid and after 48h over 4000 fungal genes were expressed. The number of genes expressed increased over time up to 96h (>8000 genes), and then declined at the 144h and 192h post inoculation time points. After subtraction of genes found expressed on complete medium, during carbon or nitrogen starvation, and on barley, only 355 were found exclusively expressed in wheat, mostly ones with unknown function (72.6%). These genes were mainly found in single-nucleotide polymorphism enriched islands on the chromosomes, suggesting a higher evolutionary selection pressure. The annotated genes were enriched in functional groups predicted to be involved in allantoin and allantoate transport, detoxification, nitrogen, sulfur and selenium metabolism, secondary metabolism, carbohydrate metabolism and degradation of polysaccharides and ester compounds. Several putative secreted virulence factors were also found expressed in wheat.

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Fusarium graminearum causes head blight disease in wheat and barley. To help understand the infection process on wheat we studied global gene expression of F. graminearum in a time series from 24 to 196 hours after inoculation, compared to a non-inoculated control. The infection was rapid and after 48h over 4000 fungal genes were expressed. The number of genes expressed increased over time up to 96h (>8000 genes), and then declined at the 144h and 192h post inoculation time points. After subtraction of genes found expressed on complete medium, during carbon or nitrogen starvation, and on barley, only 355 were found exclusively expressed in wheat, mostly ones with unknown function (72.6%). These genes were mainly found in single-nucleotide polymorphism enriched islands on the chromosomes, suggesting a higher evolutionary selection pressure. The annotated genes were enriched in functional groups predicted to be involved in allantoin and allantoate transport, detoxification, nitrogen, sulfur and selenium metabolism, secondary metabolism, carbohydrate metabolism and degradation of polysaccharides and ester compounds. Several putative secreted virulence factors were also found expressed in wheat.

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The estrogenic mycotoxin zearalenone (ZON) produced by some Fusarium spp. causes reproductive problems and hyperestrogenic syndromes in mammals. In an effort to elucidate the molecular pathways of ZON production, we present a comparative real-time quantitative polymerase chain reaction expression study of seven contiguous genes in the ZON biosynthetic cluster on sterile rice and during wheat and oat infection. Under ZON production on rice, the polyketide synthase (PKS) genes PKS4 and PKS13, alcohol oxidase FG12056 gene, and transcriptional regulator FG02398 gene showed similarly upregulated patterns, whereas the nonribosomal peptide synthetase (NPS) FG02394, the K+ channel beta subunit FG12015, and the protein kinase FG02399 displayed a variant pattern. During the same time period under wheat infection when no ZON was produced, the PKS genes and the NPS were downregulated relative to rice whereas the K+ channel beta subunit gene FG12015 was markedly upregulated, suggesting that it may play a role in the infection process. This is the first expression study of ZON biosynthetic genes in planta. The results give insight into the regulation and activities of the ZON gene cluster under different experimental systems and suggest a connection between ZON and a K+ channel that could reveal a novel function for ZON in Fusarium spp.

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Members of the APSES family of fungal proteins have been identified as key regulators of fungal development, controlling processes such as mating, sporulation and dimorphic growth. We deleted the FgStuA gene in Fusarium graminearum and show that the mutant is greatly impaired in spore development, pathogenicity and secondary metabolism. FgStuA is closely related to FoStuA in F. oxysporum, but unlike FoStuA mutants the FgStuA mutants were greatly reduced in pathogenicity both on wheat and apple slices. The lack of ability to cause disease on wheat heads may be due to lack of trichothecene accumulation in planta. The FgStuA mutant also had a white/yellow mycelial phenotype compared to the red pigmented (aurofusarin) wild-type, had reduced aerial mycelium, susceptibility to oxidative stress, and had a less hydrophobic surface. Microarray analysis showed that most phenotypes could be inferred from gene expression data, such as down-regulation of the trichothecene gene cluster in the mutant. In an attempt to separate primary and secondary effects of FgStuA deletion, we carefully examined gene expression data together with promoter analysis and comparative genomics. The genes flanking FgStuA are conserved and syntenous in other fungal genomes and contain a gene encoding a putative clock controlled protein. FgStuAp and other APSES proteins share significant homology with DNA-binding domains of transcription factors controlling the critical G1/S phase cell cycle transition in both S. cerevisiae and S. pombe. Genes within MIPS Functional Category (FunCat) 10 "Cell cycle and DNA processing" are enriched among those more highly expressed in the FgStuA mutant than wild-type. Aspergillus StuAp response elements (A/TCGCGT/ANA/C) also were found highly enriched in promoter sequences for FunCat 10 genes, compared to the genome as a whole. Our results suggests that FgStuAp may act primarily as a repressor involved in cell cycle regulation, and may act only secondarily on sporulation, pathogenicity, and secondary metabolism. 

Sammendrag

Members of the APSES family of fungal proteins have been identified as key regulators of fungal development, controlling processes such as mating, sporulation and dimorphic growth. We deleted the FgStuA gene in Fusarium graminearum and show that the mutant is greatly impaired in spore development, pathogenicity and secondary metabolism. FgStuA is closely related to FoStuA in F. oxysporum, but unlike FoStuA mutants the FgStuA mutants were greatly reduced in pathogenicity both on wheat and apple slices. The lack of ability to cause disease on wheat heads may be due to lack of trichothecene accumulation in planta. The FgStuA mutant also had a white/yellow mycelial phenotype compared to the red pigmented (aurofusarin) wild-type, had reduced aerial mycelium, susceptibility to oxidative stress, and had a less hydrophobic surface. Microarray analysis showed that most phenotypes could be inferred from gene expression data, such as down-regulation of the trichothecene gene cluster in the mutant. In an attempt to separate primary and secondary effects of FgStuA deletion, we carefully examined gene expression data together with promoter analysis and comparative genomics. The genes flanking FgStuA are conserved and syntenous in other fungal genomes and contain a gene encoding a putative clock controlled protein. FgStuAp and other APSES proteins share significant homology with DNA-binding domains of transcription factors controlling the critical G1/S phase cell cycle transition in both S. cerevisiae and S. pombe. Genes within MIPS Functional Category (FunCat) 10 "Cell cycle and DNA processing" are enriched among those more highly expressed in the FgStuA mutant than wild-type. Aspergillus StuAp response elements (A/TCGCGT/ANA/C) also were found highly enriched in promoter sequences for FunCat 10 genes, compared to the genome as a whole. Our results suggests that FgStuAp may act primarily as a repressor involved in cell cycle regulation, and may act only secondarily on sporulation, pathogenicity, and secondary metabolism. 

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I to veksthusforsøk ble hvete og havre sprayinokulert under blomstring med en enkelt eller en blanding av flere Fusarium arter. De artene som ble studert var F. graminearum, F. culmorum, F. avenaceum, F. poae og F. langsethiae. Vi studerte i hvilken grad samspillet mellom artene påvirket etableringen og veksten av Fusarium, og også hvilken effekt dette hadde på mykotoksinproduksjonen.

Sammendrag

I to veksthusforsøk ble hvete og havre sprayinokulert under blomstring med en enkelt eller en blanding av flere Fusarium arter. De artene som ble studert var F. graminearum, F. culmorum, F. avenaceum, F. poae og F. langsethiae. Vi studerte i hvilken grad samspillet mellom artene påvirket etableringen og veksten av Fusarium, og også hvilken effekt dette hadde på mykotoksinproduksjonen.

Sammendrag

Fusarium head blight (FHB) is a widespread and destructive disease of cereals caused by a number of Fusarium species. Under field conditions a mixture of Fusarium species exists. While FHB in wheat has been well studied, Fusarium infection of oats has not yet been characterized. Little is known about how the presence of a mixture of different Fusarium species in the same sample affects the mycotoxin production. During flowering plants of wheat and oats grown under greenhouse conditions were spray inoculated with single and multiple Fusarium species (F. graminearum, F. culmorum, F. avenaceum, F. poae and F. langsethiae). Chemical toxin analysis of harvested grain showed that the content of mycotoxins in oat were generally lower than in corresponding wheat samples. Neither T-2 nor HT-2 was detected in wheat or oat. Neither was it possible to detect F. langsethiae in the kernels when analysed by real-time TaqMan PCR. All wheat samples inoculated with F. graminearum contained relative high levels of deoxynivalenol. Samples infected with F. culmorum contained nivalenol in addition to deoxynivalenol. Moniliformin was detected at levels below the quantification limit in one third of the samples. The inoculation experiment was repeated with an adjusted inoculation procedure for F. langsethiae and F. poae, resulting in good establishment of all Fusarium species. The amounts of the different Fusarium species and the level of the corresponding mycotoxins were determined. The interactions between the Fusarium species regarding establishment on the fungus on the developing kernels and the production of the mycotoxins, will be discussed.

Sammendrag

Fusarium head blight (FHB) is a widespread and destructive disease of cereals caused by a number of Fusarium species. Under field conditions a mixture of Fusarium species exists. While FHB in wheat has been well studied, Fusarium infection of oats has not yet been characterized. Little is known about how the presence of a mixture of different Fusarium species in the same sample affects the mycotoxin production. During flowering plants of wheat and oats grown under greenhouse conditions were spray inoculated with single and multiple Fusarium species (F. graminearum, F. culmorum, F. avenaceum, F. poae and F. langsethiae). Chemical toxin analysis of harvested grain showed that the content of mycotoxins in oat were generally lower than in corresponding wheat samples. Neither T-2 nor HT-2 was detected in wheat or oat. Neither was it possible to detect F. langsethiae in the kernels when analysed by real-time TaqMan PCR. All wheat samples inoculated with F. graminearum contained relative high levels of deoxynivalenol. Samples infected with F. culmorum contained nivalenol in addition to deoxynivalenol. Moniliformin was detected at levels below the quantification limit in one third of the samples. The inoculation experiment was repeated with an adjusted inoculation procedure for F. langsethiae and F. poae, resulting in good establishment of all Fusarium species. The amounts of the different Fusarium species and the level of the corresponding mycotoxins were determined. The interactions between the Fusarium species regarding establishment on the fungus on the developing kernels and the production of the mycotoxins, was discussed.

Sammendrag

To understand trichothecene accumulation and the infection cycle of the head light pathogen Fusarium gramineaum sesu stricto, fungal gene expression profiles were monitored during plant infection using the F. graminearum Affymetrix GeneChip. Strains containing mutations in genes for three transcription factors were found to control trichothecene accumulations in planta and pathogenicity. Expression profiles were compared between wildtype and these mutants during infection of wheat. Mutants deleted for the StuA gene were greatly decreased in sporulation and produced no perithecia in culture. Unlike stuA mutants in F. oxysporum, F. graminearum stuA mutants were greatly reduced in pathogenicity. Reducd pathogenicity may be due to decreased trichothecene levels in planta, which in the mutant were two-fold in either tri6 or tri10 deleted strains. A model for global regulation and cross pathway control of sporulation, mycotoxin biosynthesis and pathogenicity will be presented.

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Sammendrag

The Fusarium genus includes devastating plant pathogenic fungi that cause diseases in cereals around the world. They produce several mycotoxins, including the estrogenic compound zearalenone. To better understand the molecular mechanisms determining zearalenone production, we performed differential display RT-PCR under conditions where Fusarium graminearum and F. culmorum produced high amounts of zearalenone. We found 133 expressed sequence tags (ESTs) and 54 of these were considered to be up-regulated during high zearalenone production. Several of the ESTs were confirmed to be up-regulated by real-time qPCR, but none showed any significant down-regulation in the zearalenone negative mutant Delta PKS4-T9, or were similar to typical gene expression patterns of previously described zearalenone-related genes. Some of the up-regulated ESTs were similar to genes involved in secondary metabolite production, lipid metabolism, transcriptional activation, provision of precursors, signal transduction, transport or detoxification. Several of the ESTs were also located adjacent to one another in the genome and therefore might represent genes involved in the same biosynthetic pathway. Members of six such putative pathways could be found. All sequences were compared to the MIPS F. graminearum Genome Database to verify autocalled gene predictions experimentally and to introduce new exons and gene structures.

Sammendrag

Increasing levels of Fusarium toxins, particularly deoxynivalenol (DON), T-2 /HT-2 and moniliformin, have been recorded in Norwegian cereals during the last few years. Previously F. avenaceum, F. culmorum, F. poae and F. tricinctum were the most common Fusarium species found on cereals in Norway. However, more recently F. graminearum has occurred more frequently, and the T-2/HT-2 toxin producing species F. langsethiae has also been detected, especially in oats. Investigations were made to clarify if there has been a change in the composition of Fusarium species. We are aiming to establish a three-step screening system in order to identify grain lots with high levels of Fusarium toxins: 1- Identify `high-risk" fields/lots, based on information on cultivation practice and climatic conditions, through the use of a FHB-prediction model. 2- Analyze the `high-risk" lots using a rapid test selected for its capacity to detect Fusarium toxins in a large number of grain samples at low cost. 3- Forward selected samples (based on analyses in step 2) for chemical mycotoxin analyses. Since 2004, we have run chemical mycotoxin analyses on grain samples from oat and spring wheat grown in Norway under different climatic conditions. Prediction models will be developed to estimate the risk of Fusarium infection and mycotoxin development in cereal fields, based on data for weather and cultivation practices. Preliminary results from using the prediction models will be presented.

Sammendrag

Increasing levels of Fusarium toxins, particularly deoxynivalenol (DON), T-2 /HT-2 and moniliformin, have been recorded in Norwegian cereals during the last few years. Previously F. avenaceum, F. culmorum, F. poae and F. tricinctum were the most common Fusarium species found on cereals in Norway. However, more recently F. graminearum has occurred more frequently, and the T-2/HT-2 toxin producing species F. langsethiae has also been detected, especially in oats. Investigations were made to clarify if there has been a change in the composition of Fusarium species. We are aiming to establish a three-step screening system in order to identify grain lots with high levels of Fusarium toxins: 1- Identify `high-risk" fields/lots, based on information on cultivation practice and climatic conditions, through the use of a FHB-prediction model. 2- Analyze the `high-risk" lots using a rapid test selected for its capacity to detect Fusarium toxins in a large number of grain samples at low cost. 3- Forward selected samples (based on analyses in step 2) for chemical mycotoxin analyses. Since 2004, we have run chemical mycotoxin analyses on grain samples from oat and spring wheat grown in Norway under different climatic conditions. Prediction models will be developed to estimate the risk of Fusarium infection and mycotoxin development in cereal fields, based on data for weather and cultivation practices. Preliminary results from using the prediction models will be presented.

Sammendrag

We deleted the FgStuA gene in Fusarium graminearum and demonstrate its involvement in spore development, pathogenicity and secondary metabolism. The FgStuA protein is a member of the APSES family which regulate morphogenesis and virulence in ascomycetes. FgStuA is closely related to FoStuA in F. oxysporum and stuA in Aspergillus, but unlike FoStuA mutants, the FgStuA mutants were greatly reduced in pathogenicity on wheat and colonization of apple slices. The mutant produces no detectable trichothecenes on the wheat cultivar Bobwhite, and

Sammendrag

We deleted the FgStuA gene in Fusarium graminearum and demonstrate its involvement in spore development, pathogenicity and secondary metabolism. The FgStuA protein is a member of the APSES family which regulate morphogenesis and virulence in ascomycetes. FgStuA is closely related to FoStuA in F. oxysporum and stuA in Aspergillus, but unlike FoStuA mutants, the FgStuA mutants were greatly reduced in pathogenicity on wheat and colonization of apple slices. The mutant produces no detectable trichothecenes on the wheat cultivar Bobwhite, and

Sammendrag

The filamentous fungus Fusarium graminearum causes extensive losses on cereals world-wide and contaminates harvested grain with mycotoxins, whose levels in the food supply are strictly regulated. We deleted the FgStuA gene in Fusarium graminearum and demonstrate its involvement in several different processes, such as spore development, pathogenicity and secondary metabolism. The FgStuA protein is a members of the APSES family which regulate morphogenesis and virulence in ascomycetes. FgStuA is closely related to FoStuA in F. oxysporum and StuA in Aspergillus, but unlike FoStuA mutants, the FgStuA mutants were greatly reduced in pathogenicity both on wheat and apple slices. Reduced pathogenicity may be due to decreased levels of trichothecene mycotoxins (

Sammendrag

To understand mycotoxin accumulation and the infection cycle of the wheat head blight pathogen Fusarium graminearum sensu stricto,fungal gene expression profiles were monitored during plant infection. Strains containing mutations in genes for three transcription factors were found to control deoxynivalenol (DON) accumulation in planta and pathogenicity. Expression profiles were compared between wildtype and these mutants during infection of wheat. Mutants deleted for the StuA gene were greatly decreased in sporulation and produced no perithecia in culture. Unlike "stuA mutants in F. oxysporum, F. graminearum "stuA mutants were greatly reduced in pathogenicity. Reduced pathogenicity may be due to decreased DON levels in planta, which in the mutant were

Sammendrag

To understand mycotoxin accumulation and the infection cycle of the wheat head blight pathogen Fusarium graminearum sensu stricto,fungal gene expression profiles were monitored during plant infection. Strains containing mutations in genes for three transcription factors were found to control deoxynivalenol (DON) accumulation in planta and pathogenicity. Expression profiles were compared between wildtype and these mutants during infection of wheat. Mutants deleted for the StuA gene were greatly decreased in sporulation and produced no perithecia in culture. Unlike "stuA mutants in F. oxysporum, F. graminearum "stuA mutants were greatly reduced in pathogenicity. Reduced pathogenicity may be due to decreased DON levels in planta, which in the mutant were

Sammendrag

Fusarium-toksiner i korn kan utgjøre en helserisiko for mennesker og dyr. For å kunne redusere dette problemet er det viktig å forstå hvordan soppinfeksjonen skjer og når, hvordan og hvor mye soppene produserer av de forskjellige toksinene. Det er nødvendig å utvikle gode metoder for å kunne detektere og kvantifisere toksinene, samt modeller for å kunne forutsi sannsynligheten for at kornet er kontaminert med mykotoksiner.

Sammendrag

Fusarium-toksiner i korn kan utgjøre en helserisiko for mennesker og dyr. For å kunne redusere dette problemet er det viktig å forstå hvordan soppinfeksjonen skjer og når, hvordan og hvor mye soppene produserer av de forskjellige toksinene. Det er nødvendig å utvikle gode metoder for å kunne detektere og kvantifisere toksinene, samt modeller for å kunne forutsi sannsynligheten for at kornet er kontaminert med mykotoksiner.

Sammendrag

Fusarium-toksiner i korn kan utgjøre en helserisiko for mennesker og dyr. For å kunne redusere dette problemet er det viktig å forstå hvordan soppinfeksjonen skjer og når, hvordan og hvor mye soppen produserer av de forskjellige toksinene. Det er nødvendig å utvikle gode metoder for å kunne detektere og kvantifisere toksinene, samt modeller for å kunne forutsi sannsynligheten for at kornet er kontaminert med mykotoksiner.

Sammendrag

To understand trichothecene accumulation and the infection cycle of the head blight pathogen F. graminearum sensu stricto, fungal gene expression profiles were monitored during germination of ascospores and during plant infection. A total of 328 genes were determined to be specifically expressed in ascospores. Among genes highly up-regulated in ascospores was one most closely related to FoStuA of F. oxysporum and StuA in Aspergillus. Mutants deleted for this gene in F. graminearum (FgStuA) are greatly decreased in sporulation and produce no perithecia. Unlike FoStuA mutants in F. oxysporum, FgStuA mutants are greatly reduced in pathogenicity. Reduced pathogenicity may be due to decreased le vels of trichothecene toxins, which in the mutant are

Sammendrag

Flere av de soppartene som naturlig finnes i korn, kan produsere giftstoffer. Det er et mål å redusere mengden av slike sopptoksiner til et nivå som gjør at vi trygt kan benytte kornet som råstoff for produksjon av mat eller dyrefòr. Den viktigste kilden til mykotoksiner ute på kornåkeren er Fusarium. Ulike Fusarium-arter kan produsere flere forskjellige mykotoksiner. Kunnskap om hvordan denne produksjonen skjer kan bidra til å redusere dette problemet.Ved å studere prosesser i Fusarium-sopper som produserer store mengder av soppgiften zearalenon, har de klart å frambringe viktig kunnskap om hvordan denne soppgiften dannes i soppen. Zearalanon er en østrogenlignende forbindelse som virker som en hormonhermer i mennesker og dyr, og er et av de vanligste mykotoksinene man finner i korn. Flere soppgener som var mye mer aktive når zearalenon ble produsert enn når denne forbindelsen ikke ble laget, ble funnet og karakterisert. Det ble laget en Fusarium-mutant der ett av soppgenene, PKS4, var inaktivert. Denne mutanten hadde mistet sin evne til å produsere zearalenon. På denne måten kunne man sikkert identifisere ett av flere gener som var absolutt nødvendig for at Fusarium skulle kunne lage dette toksinet. Genet PKS4 kodet for en polyketidsyntase, og hvordan aktiviteten av dette og andre gener ble regulert når zearalenon ble laget, ble studert i mer detalj. Aktiviteten av de samme genene ble også studert i unge hvete- og havrekorn som var smittet med Fusarium. Zearalenon sin rolle i soppen var tidligere ukjent, men basert på resultater framkommet i dette arbeidet er man ett skritt nærmere en forståelse av denne prosessen.