Gibberella Fujikuroi Research Articles

PIRA-PCR for Detection of Fusarium fujikuroi Genotypes with Carbendazim-Resistance Alleles.

Carbendazim, a methyl benzimidazole carbamate (MBC)-group fungicide, has been used to control rice bakanae disease, caused by Fusarium fujikuroi (teleomorph: Gibberella fujikuroi), for decades in China. Previous research revealed that point mutations (E198V, GAG to GTG at codon 198, and F200Y, TTC to TAC at codon 200) of the β2-tubulin gene conferred resistance of F. fujikuroi to MBC. In this study, primer-introduced restriction analysis polymerase chain reaction (PIRA-PCR) was developed to determine genotypes with resistance of F. fujikuroi to MBC. A PCR template of each strain was created by an outer primer pair. Fragments with 177 bp (for mutation at codon 235) and 146 bp (for E198V) were amplified by nested PCR, with two inner primer pairs designed and synthesized according to the nucleotide sequence of β2-tubulin for further enzyme digestion validation, respectively. AccII and PmaCI restriction enzyme recognition sites were introduced artificially by inner primers to differentiate MBC-sensitive and -resistant strains, respectively. The sensitivity of each strain to MBC was indirectly determined by analyzing electrophoresis patterns of the resulting amplified fragments after simultaneous digestion by both AccII and PmaCI. PIRA-PCR produced the same result as conventional methods in 6% of the time. PIRA-PCR is a sensitive and effective method for genotyping resistance alleles of F. fujikuroi strains to MBC.

Characterization and effects of two algicidal isolates on antioxidase activities of Chlorella pyrenoidosa

With increasing water pollution, algal bloom becomes more and more frequent. To manage algal blooms, the exploitation and study of novel algae‐lysing strains are extremely urgent. In this study, two algicidal isolates (NP23 and AM11) against Chlorella pyrenoidosa were obtained from the Baiguishan reservoir in China. Phylogenetic analysis of the 16S rDNA sequences revealed that isolates NP23 and AM11 were closely related to Enterobacter cloacae and Gibberella moniliformis, respectively. Moreover, to determine the effects of these two algicidal isolates on antioxidase activities of C. pyrenoidosa, the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were investigated in cultures of their host algae. The results showed that NP23 and AM11 were able to promote the activity of these three antioxidases soon after inoculation (1–3 days) but significantly inhibited antioxidase activities after the third day. These results provide a strong foundation for further research of the algicidal mechanism. © 2015 American Institute of Chemical Engineers Environ Prog, 34: 1647–1651, 2015

Endophytic fungi from medicinal plant Bauhinia forficata: Diversity and biotechnological potential.

Bauhinia forficata is native to South America and used with relative success in the folk medicine in Brazil. The diversity, antibacterial activity, and extracellular hydrolytic enzymes of endophytic fungi associated with this plant were studied. Plant samples, which included leaves, sepals, stems, and seeds, were used. Ninety-five endophytic fungal were isolated (18 from leaves, 22 from sepals, 46 from stems, and nine from seeds), comprising 28 species. The most frequently isolated species were Acremonium curvulum (9.5%), Aspergillus ochraceus (7.37%), Gibberella fujikuroi (10.53%), Myrothecium verrucaria (10.53%) and Trichoderma piluliferum (7.37%). Diversity and species richness were higher in stem tissues, and Sorensen’s index of similarity between the tissues was low. Eleven fungi showed antibacterial activity. Aspergillus ochraceus , Gibberella baccata , Penicillium commune , and P. glabrum were those with the greatest antibacterial activity against Staphylococcus aureus and/or Streptococcus pyogenes . Thirteen species showed proteolytic activity, particularly Phoma putaminum . Fourteen species were cellulase positive, particularly the Penicillium species and Myrmecridium schulzeri . All isolates tested were xylanase positive and 10 showed lipolytic activity, especially Penicillium glabrum . It is clear that the endophytic fungi from B. forficata have potential for the production of bioactive compounds and may be a source of new therapeutic agents for the effective treatment of diseases in humans, other animals, and plants. To our knowledge, this is the first study of endophytic fungi from different tissues of B. forficata and their biotechnological potential.

Use of silver nanoparticles for managing Gibberella fujikuroi on rice seedlings

It is of great interest for rice producers to develop novel seed disinfection methods with effective, less toxic, and environment-friendly active ingredients. This study evaluated silver nanoparticles (AgNP) as an alternative agent for seed treatment since elemental silver possesses strong antimicrobial properties and a low toxicity to humans and animals. AgNP (∼7.5 nm in diameter) solution was chemically synthesized and evaluated for antifungal activity against Gibberella fujikuroi as representing a seedborne fungal pathogen in rice (Oryza sativa). AgNP reduced viability of G. fujikuroi conidia by 50% when directly exposed to concentrations ranging from 0.015 to 1.5 μg mL−1 for 1 to 20 min. Rice seeds infested with G. fujikuroi and treated with 150 μg mL−1 of AgNP significantly decreased colony-forming units (CFU) of G. fujikuroi on the seed surface when treated for 10 min and up to 24 h. Seed treatment with 150 μg mL−1 AgNP for 12 or 24 h significantly improved seedling emergence and height of G. fujikuroi-infested seeds. Adverse effects on germination rate and seedling growth were not observed with any of the AgNP treatments (≤150 μg mL−1 for up to 48-h exposure) tested in this study. The antifungal effect of AgNP against G. fujikuroi attested in this study suggests that AgNP is a new antifungal ingredient that can be used for managing important seedborne fungal pathogens of rice.

Inhibitory effects of antimicrobial agents against Fusarium species.

We investigated the inhibitory effects of antibacterial, biocidal, and antifungal agents against Fusarium spp. Seven Fusarium spp: four F. falciforme (Fusarium solani species complex), one Fusarium spp, one Fusarium spp. (Fusarium incarnatum-equiseti species complex), and one F. napiforme (Gibberella fujikuroi species complex), isolated from eyes with fungal keratitis were used in this study. Their susceptibility to antibacterial agents: flomoxef, imipenem, gatifloxacin, levofloxacin, moxifloxacin, gentamicin, tobramycin, and Tobracin® (contained 3,000 μg/ml of tobramycin and 25 μg/ml of benzalkonium chloride (BAK), a biocidal agent: BAK, and antifungal agents: amphotericin B, pimaricin (natamycin), fluconazole, itraconazole, miconazole, voriconazole, and micafungin, was determined by broth microdilution tests. The half-maximal inhibitory concentration (IC50), 100% inhibitory concentration (IC100), and minimum inhibitory concentration (MIC) against the Fusarium isolates were determined. BAK had the highest activity against the Fusarium spp. except for the antifungal agents. Three fluoroquinolones and two aminoglycosides had inhibitory effects against the Fusarium spp. at relatively high concentrations. Tobracin® had a higher inhibitory effect against Fusarium spp. than tobramycin alone. Amphotericin B had the highest inhibitory effect against the Fusarium spp, although it had different degrees of activity against each isolate. Our findings showed that fluoroquinolones, aminoglycosides, and BAK had some degree of inhibitory effect against the seven Fusarium isolates, although these agents had considerably lower effect than amphotericin B. However, the inhibitory effects of amphotericin B against the Fusarium spp. varied for the different isolates. Further studies for more effective medications against Fusarium, such as different combinations of antibacterial, biocidal, and antifungal agents are needed.

In-vitro antibacterial, antifungal, antioxidant and functional properties of Bacillus amyloliquefaciens.

BackgroundFood born pathogenic bacteria and filamentous fungi are able to grow on most foods, including natural foods, processed foods, and fermented foods and create considerable economic loss. The aim of this study was to determine the antibacterial, antifungal, antioxidant and functional properties of Bacillus amyloliquefaciens recovered from silage.MethodsMinimum Inhibitory Concentration (MIC) of the compounds was assessed by using broth micro dilution method. The 1,1-diphenyl–2-picrylhydrazyl (DPPH)-radical scavenging and hydroxyl radical-scavenging abilities were measured to evaluate antioxidant activity of the strain.ResultsPrimary antimicrobial compound production screening revealed that B. amyloliquefaciens exhibited significant activity against all the tested bacteria and fungi compared to other strains. The 16S rRNA and gyrase A gene sequence analysis determined using molecular biological tools confirmed that the strain was 99% similarity towards B. amyloliquefaciens. The Minimum Inhibitory Concentration (MIC) of ethyl acetate extract against Bacillus subtilis, Enterococcus cloacae and Staphylococcus aureus were 25.0 μg ml−1, and S, epidermidis were 12.5 μg ml−1, respectively. Filamentous fungi Aspergillus clavatus, A. fumigates, A. niger and Gibberella moniliformis showed 25 μg ml−1. VJ-1 was able to survive the gastrointestinal conditions simulating the stomach and duodenum passage with the highest percentage of hydrophobicity. In addition, its resistance to hydrogen peroxide and highest hydroxyl radical and 2, 2-diphenyl–1-picrylhydrazyl (DPPH) scavenging activities, with inhibition rates of 56.84% and 67.12% respectively, were its advantage. An antimicrobial susceptibility pattern was an intrinsic feature of this strain, and thus, consumption does not represent a health risk to humans.ConclusionBacillus amyloliquefaciens might be a promising candidate for new pharmaceutical agents and probiotics.

Novel taxa in the Fusarium fujikuroi species complex from Pinus spp.

The pitch canker pathogen Fusarium circinatum has caused devastation to Pinus spp. in natural forests and non-natives in commercially managed plantations. This has drawn attention to the potential importance of Fusarium species as pathogens of forest trees. In this study, we explored the diversity of Fusarium species associated with diseased Pinus patula, P. tecunumanii, P. kesiya and P. maximinoi in Colombian plantations and nurseries. Plants displaying symptoms associated with a F. circinatum-like infection (i.e., stem cankers and branch die-back on trees in plantations and root or collar rot of seedlings) were sampled. A total of 57 isolates were collected and characterised based on DNA sequence data for the translation elongation factor 1-α and β-tubulin gene regions. Phylogenetic analyses of these data allowed for the identification of more than 10 Fusarium species. These included F. circinatum, F. oxysporum, species within the Fusarium solani species complex and seven novel species in the Fusarium fujikuroi species complex (formerly the Gibberella fujikuroi species complex), five of which are described here as new. Selected isolates of the new species were tested for their pathogenicity on Pinus patula and compared with that of F. circinatum. Of these, F. marasasianum, F. parvisorum and F. sororula displayed levels of pathogenicity to P. patula that were comparable with that of F. circinatum. These apparently emerging pathogens thus pose a significant risk to forestry in Colombia and other parts of the world.

In silico Design and Analysis of G. Moniliformis Class II Hydrophobin Inhibitor as a Potential Antifungal Drug

Gibberella moniliformis (anamorph: Fusarium verticillioides) is a fungal pathogen destructive to over 11,000 plant species. It is known to cause severe grain loss in maize and rice, as well as carcinogenic diseases in both humans and animals upon ingestion of the infested grains. Such a significant economic impact of G. moniliformis can be credited to class II hydrophobins generally secreted and localized on the outer surface of conidia and of the hyphal wall. These small related surface-active proteins (+/- 100 amino acids long and 10 kDa weight) can self assemble at hydrophilic: hydrophobic interfaces into different forms of nanometric biofilms about 10nm wide. Based on this unique property, class II hydrophobins are involved in signalling and attachment of aerial structures (like mycelia, hyphae, and spores) to hydrophobic surfaces of host plant during fungal pathogenesis. This paper thus seeks to comparatively model the 3D structure of class II hydrophobins, to computationally determine their surfactant-protein interactions ability to a potential inhibitor using molecular docking techniques, and to statistically predict the ADME-T properties of the lead molecule.

イプコナゾール上市前後における<i>Fusarium fujikuroi</i>種複合体の菌種構成，ジベレリン生産性および同剤に対する感受性

Sensitivities to ipconazole of 50 isolates of the Fusarium (Gibberella) fujikuroi species complex (Ff complex) obtained from rice in Japan before 1994 (group A: before the launch of ipconazole) and 65 isolates after 1994 (group B) were investigated. Most isolates were identified as Fusarium fujikuroi, while 5 isolates in group A and 4 isolates in group B were identified as Fusarium proliferatum with species diagnostic PCR-RFLPs. The minimum inhibitory concentration (MIC) range, profiles in sensitivity to ipconazole and gibberellin production were similar in both groups. Gibberellin production was detected in the F. fujikuroi isolates with higher sensitivity to ipconazole but not in the isolates with lower sensitivity and F. proliferatum. High efficacy of ipconazole against rice “Bakanae” disease was confirmed by using the gibberellin-producing F. fujikuroi isolates in group B. No shift in sensitivity to ipconazole was verified and this may contribute to the stable efficacy against rice “Bakanae” disease.

벼 키다리병 방제를 위한 차아염소산나트륨 이용

For application of sodium hypochlorite as a seed disinfectant to the control of bakanae disease caused by Gibberella fujikuroi in rice, we investigated the effects of sodium hypochlorite for antifungal activity, eliminating fungus from seeds and reducing disease occurrence in vitro and greenhouse. The viability of the pathogen was significantly reduced at 80 ml/l concentration of sodium hypochlorite, and the pathogens did not grow at over 100 ml/l concentration of sodium hypochlorite. The effect of eliminating fungus was 90% at treatment of 0.3% sodium hypochlorite solution to infected rice seeds for eight hours. When the rice seeds were soaked into 0.5% and 0.3% sodium hypochlorite solutions for twelve hours, the disease incidences of rice seedling were remarkably reduced to 4.3% and 4.7%, respectively, compared to 97.3% of non-treatment control. The rates of seedling stand were 29.1% and 26.9% higher with the sodium hypochlorite treatment than that of non-treatment control. When prochloraz and sodium hypochlorite was treated to naturally severely infested rice seeds with bakanae disease, the disinfection effect was higher than that of prochloraz alone treatment. When the seeds were soaked in sodium hypochlorite before or after prochloraz, the rate of seed contamination was low as 4.0% or 6.3%, respectively, compared to prochloraz alone as 13.7%. The disease incidence was low as 3.7% or 8.3%, respectively, compared to prochloraz alone as 14.3%. The disinfection effect of treatment with prochloraz after sodium hypochlorite was higher than that of treatment with prochloraz before sodium hypochlorite.

Genome-wide macrosynteny among Fusarium species in the Gibberella fujikuroi complex revealed by amplified fragment length polymorphisms.

The Gibberella fujikuroi complex includes many Fusarium species that cause significant losses in yield and quality of agricultural and forestry crops. Due to their economic importance, whole-genome sequence information has rapidly become available for species including Fusarium circinatum, Fusarium fujikuroi and Fusarium verticillioides, each of which represent one of the three main clades known in this complex. However, no previous studies have explored the genomic commonalities and differences among these fungi. In this study, a previously completed genetic linkage map for an interspecific cross between Fusarium temperatum and F. circinatum, together with genomic sequence data, was utilized to consider the level of synteny between the three Fusarium genomes. Regions that are homologous amongst the Fusarium genomes examined were identified using in silico and pyrosequenced amplified fragment length polymorphism (AFLP) fragment analyses. Homology was determined using BLAST analysis of the sequences, with 777 homologous regions aligned to F. fujikuroi and F. verticillioides. This also made it possible to assign the linkage groups from the interspecific cross to their corresponding chromosomes in F. verticillioides and F. fujikuroi, as well as to assign two previously unmapped supercontigs of F. verticillioides to probable chromosomal locations. We further found evidence of a reciprocal translocation between the distal ends of chromosome 8 and 11, which apparently originated before the divergence of F. circinatum and F. temperatum. Overall, a remarkable level of macrosynteny was observed among the three Fusarium genomes, when comparing AFLP fragments. This study not only demonstrates how in silico AFLPs can aid in the integration of a genetic linkage map to the physical genome, but it also highlights the benefits of using this tool to study genomic synteny and architecture.

Seed treatment with an aqueous extract of Agave sisalana improves seed health and seedling growth of sorghum

An aqueous extract of the sisal agave, A. sisalana, was prepared from dried stem tissue by boiling in water. The sisal extract (SE) displayed a broad inhibition of the seed-borne mycoflora when applied to farmer-saved seeds of Sorghum bicolor. Species of Fusarium (belonging to the Gibberella fujikuroi complex) were inhibited >75 % on seeds. Accordingly, seed-borne crown rot, associated with Fusarium thapsinum was significantly (p < 10−7) and dose-dependently reduced in plants from SE treated seeds. The SE treatment furthermore stimulated seedling vigour and increased dry weight of 4-week-old plants (p < 10−4) with 35 %. The growth promoting activity of SE was found to be similar to what has been reported for the extract of another agave, Yucca schidigera. Both extracts were found to reduce seed-borne Fusarium and crown rot more efficiently than fungicides based on tebuconazole, enilconazole and fludioxonil (p < 0.0042). The stem of the sisal agave is a waste product in fiber production. From our findings A. sisalana stem appears to represent an abundant source of antifungal activity, simple to extract, easy to store and with a promising potential in biological seed treatment.

Identification and Characterization of Lactobacillus brevis P68 with Antifungal, Antioxidant and Probiotic Functional Properties

In the present study, Lactobacillus pentosus, L. plantarum, L. fermentum, L. brevis, L. paraplantarum, L. buchneri, and L. acidipiscis, which are strains capable of producing antifungal metabolites against food-spoilage fungi, were isolated and identified based on 16S rRNA gene sequencing from different traditional pickles. L. brevis P68 exhibited significant antifungal activity, and it’s in vitro antioxidant and probiotic properties were investigated. The antifungal compound was characterized based on 13C nuclear magnetic resonance (NMR), 1H NMR, infrared, and mass spectral data. The minimum inhibitory concentration (MIC) of the compounds was assessed using the broth micro-dilution technique. The MIC of the compounds against Penicillium chrysogenum and P. roqueforti was 2.5 mg mL−1 and that against Gibberella moniliformis and Aspergillus clavatus was 5.0 mg mL−1. In addition, the H2O2 (1.0 mM) hydroxyl radical, and DPPH scavenging activity inhibition rates were 32.76 and 48.63 %, respectively, and the activities toward the glutathione peroxidase and superoxide dismutase enzymes were high. This strain tolerated low pH and bile salt, exhibited bile salt hydrolase and extracellular enzyme activities, and was sensitive to common antibiotics with high hydrophobicity. This study revealed that the antifungal, antioxidant, and probiotic properties of L. brevis P68 confirmed its application to the food industry.

Identification and Characterization of Fusarium Species Associated with Wilt of Eleocharis dulcis (Chinese water chestnut) in China.

Fusarium wilt is one of the most important diseases of Eleocharis dulcis (Chinese water chestnut) in China. In order to characterize the pathogens responsible, 69 Fusarium isolates were collected from diseased plants in E. dulcis production areas of the Chinese provinces Anhui, Fujian, Hubei, Hunan, Jiangsu, and Zhejiang. These were then identified based on morphological and molecular characteristics. F. commune was the most common species (92.8%) and was widely distributed in the six provinces. A novel species within the Gibberella fujikuroi species complex (GFSC) was found in Hubei and Zhejiang provinces (5.8%), and an unidentified Fusarium sp. was found only in Hubei province (1.4%). Thirty F. commune isolates from different provinces and four GFSC isolates were selected for sequence analyses of the translation elongation factor 1-α (EF-1α), the mitochondrial small subunit (mtSSU) ribosomal DNA, and the nuclear ribosomal intergenic spacer region (IGS). Maximum parsimony and Bayesian analyses of the multilocus sequence data of these two species plus other taxa showed that the two species formed two distinct, well-supported clades among the three individual and combined gene genealogies. Isolates from different locations were scattered, with no evidence of geographic specialization. Pathogenicity assays showed that the two Fusarium spp., including the unidentified Fusarium sp., were pathogenic to E. dulcis 'Tuanfeng seven'. There was no relationship between the source of isolates and their pathogenicity. This is the first description of F. commune, a novel species within the GFSC, and an unidentified Fusarium sp. as causal agents of Fusarium wilt of E. dulcis in China.

Dysbiosis of Gut Fungal Microbiota is Associated With Mucosal Inflammation in Crohn’s Disease

We aim to characterize the fungal microbiota in the intestinal mucosa and feces in patients with Crohn's disease (CD). Fungi represent a diverse microbial community in the human intestine and might play a role in the pathogenesis of CD; however, little is known about the structure and composition of the fungal microbiota especially adhering to the intestinal mucosa in CD patient. Nineteen patients with active CD and 7 healthy individuals were recruited in this study. The mucosa-associated and fecal fungal microbiotas in CD patients were analyzed using culture-independent community fingerprint techniques. The fungal richness and diversity were significantly elevated in the inflamed mucosa compared with the noninflamed mucosa. The predominant fungal composition in the inflamed mucosa was strikingly altered, mainly characterized by expansion in the proportions of Candida spp., Gibberella moniliformis, Alternaria brassicicola, and Cryptococcus neoformans. The fecal fungal community was perturbed in CD patients as accompanied by increased fungal diversity and prevalence in Candida albicans, Aspergillus clavatus, and C. neoformans. The species richness and diversity of the mucosal fungal community were associated with the expression of TNF-α, IFN-γ, or IL-10 (P<0.05). The diversity of the fecal fungal microbiota positively correlated with serum C-reactive protein and CD activity index (P<0.05). This study first demonstrates that the fungal microbiota in the inflamed mucosa is distinguishable from that of the noninflamed area. Shifts of gut fungal microbiota composition may be associated with mucosal inflammation and disease activity of CD. Our data would provide novel insights into understanding the potential of gut fungal microbiota in the pathogenesis of CD.

Treatment of hydrophobic VOCs in trickling bed air biofilter: Emphasis on long-term effect of initial alternate use of hydrophilic VOCs and microbial species evolution

The main research objective of this study is to enhance the removal of recalcitrant compounds that are not readily bioavailable due to limiting mass transfer rate between the liquid and gas phases. Four trickle-bed air biofilters (TBABs), loaded with pelletized diatomaceous earth support media, were run at an empty bed residence time (EBRT) of 120 sec. After an acclimation period at constant loading rate (LR) of n-hexane (13.2 g m−3 hr−1) and intermittent feeding of methanol, n-hexane influent LR was then increased in step-wise fashion to 47.7 g m−3 hr−1 for biofilters receiving acidic nutrients (pH 4), and to 36.3 g m−3 hr−1 for biofilters receiving nutrient at pH 7. The results have shown that for TBABs receiving nutrient at pH 4, greater elimination capacities were obtained as compared to TBABs working at pH 7. n-Hexane removal efficiency of more than 84% at LR up to 47.7 g m−3 hr−1 was obtained for pH 4 nutrient-fed biofilters, while for biofilters with nutrients fed at pH 7, the removal efficiency did not exceed 64% for n-hexane LR of 36.3 g m−3 hr−1. The microbial analysis revealed that no fungal community was detected in TBABs run at neutral pH. The fungi communities that were initially acclimating TBABs run at pH 4, namely, Aspergillus niger and Fusarium solani, were not detected at the end of the experiment, while Gibberella moniliformis (Fusarium verticillioides) genus became the dominant species. Gibberella moniliformis (Fusarium verticillioides) was present along all the biofilter media and sustained very high n-hexane elimination at steady-state condition.Implications:With growing apprehension about sustainability and environmental protection, with limited resources available, and with the passage of the 1990 Amendments to the Clean Air Act, there is more need for using air pollution control techniques that are sound economically and proven environmentally friendly. Biofiltration systems, namely, trickle-bed air biofilters, were for decades recognized as efficient in treating air pollutants. Thus, the application of this technique over a wide industrial spectrum would certainly contribute to reduction of hazardous gas emissions.

First report and characterization of Fusarium circinatum, the causal agent of pitch canker in Brazil

Pitch canker is one of the most important diseases of pine trees worldwide, including South America. The causal agent of this disease is Fusarium circinatum, a member of the Gibberella fujikuroi complex (GFC). In South America, the species is reported from Colombia, Uruguay and Chile, but is considered a quarantine organism in Brazil. In this study we characterized isolates obtained from symptomatic pine seedlings intercepted in a nursery in Santa Catarina State using phylogenetic analyses, crossings and morphological markers. The Brazilian isolates grouped with reference material in an unique clade and formed fertile perithecia when crossed with reference tester strains. The detailed characterization given here and the availability of tester strains will allow for reliable identification and support monitoring of this important plant pathogen.

Gibberella moniliformis AH13 with antitumor activity, an endophytic fungus strain producing triolein isolated from Adlay (Coix lacryma-jobi: poaceae).

In this study, the isolation of an endophytic fungus from the leaves of the medicinal herb adlay (Coix lacryma-jobi L. var. ma-yuen Stapf) is reported for the first time. The fungus produced Triolein (trioleoylglycerol), a major constituent of triacylglycerols (TAGs) of adlay, in rice medium under shake-flask and bench-scale fermentation conditions. The fungus was identified as Gibberella moniliformis (Fusarium verticillioides) by its morphology and authenticated by ITS analysis (ITS1 and ITS2 regions and the intervening 5.8S rDNA region). Triolein was identified by HPLC-ELSD coupled with APCI-MS and confirmed through comparison with authentic standard. The concentration of triolein produced by G. moniliformis AH13 reached 2.536 ± 0.006 mg/g dry weight of mycelium. Moreover, the EtOAc extract of G. moniliformis AH13 showed strong antitumor activity against four types of tumor cells (A549, HCT116, MDA-MB-231, and SW1990). These results suggest that G. moniliformis AH13 in adlay has significant scientific and industrial potential to meet the pharmaceutical demands and sustainable energy requirements for TAGs in a cost-effective, easily accessible, and reproducible way and is also a potential novel source of natural antitumor bioactive agents.

European Confederation of Medical Mycology (ECMM) epidemiological survey on invasive infections due to Fusarium species in Europe.

In order to better understand the epidemiology of fusariosis in Europe, a survey collecting information on the clinical characteristics of the patients infected by Fusarium as well as on the infecting isolates was launched. A total of 76 cases of invasive fusariosis occurring from January 2007 to June 2012 were collected and Fusarium isolates were identified by sequencing the translation elongation factor 1α (TEF) gene. Also, antifungal susceptibility was tested by broth microdilution according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the Etest. Disseminated disease was considered proven in 46 cases and probable in 17 cases. Localised infection was seen in 13 cases. Gibberella fujikuroi species complex (SC), including Fusarium verticillioides and F. proliferatum, and F. solani SC were the most frequent aetiology of disseminated and localised infections, respectively. The crude mortality rate was 46 %, the highest associated with F. solani SC (67 %) and F. proliferatum (62.5 %). A wide range of antifungal susceptibilities was observed. Amphotericin B was the most potent antifungal in vitro, and itraconazole the least effective. The azoles exhibited lower minimum inhibitory concentrations (MICs) against F. verticillioides strains, with posaconazole having a slightly better performance, while F. solani SC isolates were resistant to all three azoles tested. The essential agreement between the Etest and the EUCAST method was 100 % for itraconazole and voriconazole, and 96 % for amphotericin B and posaconazole. In conclusion, we confirm that fusariosis is a rare but severe event in Europe, that G. fujikuroi SC is the predominant cause of deep infections and that different species have different antifungal in vitro susceptibility patterns.

Genome distribution and validation of novel microsatellite markers of Fusarium verticillioides and their transferability to other Fusarium species

Improved population studies in the fungus Fusarium verticillioides require the development of reliable microsatellite markers. Here we report a set of ten microsatellite loci that can be used for genetic diversity analyses in F. verticillioides, and are equally applicable to other fungi, especially those belonging to the Gibberella fujikuroi clade.