The effects of different potato dextrose agar media on secondary metabolite production in Fusarium.

Potatoes contain several nutrients essential for fungal growth, making them an excellent component of media such as the popular Potato Dextrose Agar (PDA) medium. Commercially, PDA is available from multiple retailers offering virtually the same product. These media, however, could contain small differences in composition of nutrients affecting the expression of secondary metabolites. This study aims to investigate the use of four PDA media from different manufacturers (Fluka, Oxoid, Sigma, and VWR) and their effect on the metabolite profile of four species of Fusarium (F. fujikuroi, F. graminearum, F. pseudograminearum and F. avenaceum). Secondary metabolites were analysed using HPLC-HRMS, from which statistically significant differences in intensities were observed for 9 out of 10 metabolites.

Pearl millet populations characterized by Fusarium prevalence, morphological traits, phenolic content, and antioxidant potential.

BACKGROUND: Pearl millet (Pennisetum glaucum L.) has become increasingly attractive due to its health benefits. It is grown as food for human consumption and fodder for livestock in Africa and Asia. This study focused on five pearl millet populations from different agro-ecological zones from Tunisia, and on characterization by morphological traits, total phenolic and flavonoid content, antioxidant activity, and occurrence of Fusarium. RESULTS: Analysis of variance revealed highly significant differences between populations for the quantitative traits. The highest grain weights occurred in the pearl millet cultivated in Zaafrana and Gergis of Tunisia. Early flowering and early maturing populations cultivated in the center (Zaafrana, Rejiche) and south (Gergis) of Tunisia tended to have a higher grain yield. The Zaafrana population showed the highest value of green fodder potentiel (number and weight of leaves/cultivar and the weight of tillers and total plant/cultivar) followed by Gergis and Rejiche. The Kelibia population showed the highest total phenolic and flavonoid content. Rejiche exhibited the greatest antioxidant activity. Trans-cinnamic, protocatechuic, and hydroxybenzoic acids were the major phenolic compounds in all the extracts. Three Fusarium species were identified in Tunisian pearl millet populations based on morphologic and molecular characterization. Fusarium graminearum and Fusarium culmorum occurred most frequently. The average incidence of the three Fusarium species was relatively low (<5%) in all populations. The lowest infection rate (0.1%) was recorded in the samples from Zaafrana. CONCLUSION: Chemometric analysis confirmed the usefulness of the above traits for discrimination of pearl millet populations, where a considerable variation according to geographical origin and bioclimatic conditions was observed. © 2020 Society of Chemical Industry.

[Identification of Coptis chinensis root rot disease pathogenic Fusarium spp. fungi].

Root rot disease is vital disease of Coptis chinensis, it has bursted in most producing area in recent years, and has caused severe damage. To identify the pathogenic fungi, Fusarium spp. fungi were isolated from rot root, of which the pathogenic fungi were screened with inoculation on C. chinensis root and plant, and identified with molecular and morphological method. The 20 Fusarium spp. fungi were obtained, of which 5 displayed high pathogenicity. It was deduced that F. oxysporum, F. solani and F. tricinctum were the pathogen, possibly pioneer pathogen of C. chinensis root rot disease. Among which F. oxysporum was dominant and deserved to pay more attention. High temperature and high humidity can increase pathogenicity of Fusarium spp. So the global climate warming may lead to temperature rising of C. chinensis producing area and favor the pathogen fungi, which may be one of the main factors leading to bursting of C. chinensis root rot disease. To control the root rot, beside developing and using pesticide, producing base should be moved to a high altitude area.

Identification of a Huperzine A-producing endophytic fungus from Phlegmariurus taxifolius.

Highly prized huperzine A (Hup A), a natural alkaloid formerly isolated from the Chinese medicinal plant Huperzia serrata, has been widely used for the treatment of Alzheimer disease, inspiring us to search for endophytic fungi that produce this compound. In this study, we obtained the C17 fungus isolate from the Mexican club moss Phlegmariurus taxifolius, which produced a yield of 3.2 µg/g Hup A in mycelial dry weight, when cultured in potato dextrose broth medium. The C17 isolate was identified as belonging to the genus Fusarium with reference to the colony´s morphological characteristics and the presence of macroconidia and microconidia structures; and this was confirmed by DNA-barcoding analysis, by amplifying and sequencing the ribosomal internal transcribed spacer (rITS).

Comparative analysis uncovers the limitations of current molecular detection methods for Fusarium oxysporum f. sp. cubense race 4 strains.

Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4) is threatening banana production worldwide. Despite quarantine efforts, the pathogen continues to spread; thus, early diagnosis plays an essential role for the proper execution of contingency plans. Here, we assess the accuracy of four PCR-based molecular methods described in the literature for the identification and detection of race 4 strains, including Subtropical (Foc STR4) and Tropical Race 4 causing Fusarium wilt of banana. We screened a total of 302 isolates using these four markers, and performed phylogenetic analyses, Vegetative Compatibility Group (VCG) testing, sequence comparison, and pathogenicity tests for selected isolates. Our results show that three out of the four markers tested are not reliable for identification of Foc STR4 and TR4, as DNA from isolates from Ecuador, pathogenic and nonpathogenic to banana, obtained from different banana cultivars, displayed cross-reaction with these methods; that is, false positives can occur during the diagnostic process for race 4. Phylogenetic analyses, VCG testing, sequence comparison, and pathogenicity tests suggest the presence of non-target F. oxysporum isolates that share genomic regions with pathogenic strains but lack true pathogenicity to banana. The findings of this work are of foremost importance for international regulatory agencies performing surveillance tests in pathogen-free areas using the current diagnostic methods. We suggest the use of a genetic locus possibly related to virulence, previously identified by T-DNA, and amplified with primers W2987F/ W2987R, for diagnosis of Foc TR4 as the most reliable alternative. We urge the adoption of a more holistic view in the study of F. oxysporum as a plant pathogen that considers the biology and diversity of the species for the development of better diagnostic tools.

Biofilm formation and regulation of salicylic acid-inducible genes expression in Arabidopsis by Algerian indigenous bacteria from wheat and potatoes rhizospheres in semi-arid Sétif region.

45 bacterial isolates from potatoes and wheat rhizospheres near Sétif (Algeria) pre-selected for their antagonistic activity against three fungal plant pathogens, two necrotrophic Fusarium solani var. coeruleum and Phytophtora infestans, and a systemic F. oxysporum f. sp. albedinis. Molecular typing of the isolates showed abundance of Bacillus compared to Pseudomonas. Some of the tested strains have shown very high biofilm formation. Among the 24 Gram-positive bacilli screened for four cyclic lipopeptides genes, some isolates harbor two or more genes, while others have a single gene or have none. Four selected isolates were able to regulate the expression of six defense-related genes in Arabidopsis and produce salicylic acid. Upon the features assessed in this study, strain B. amyloliquefaciens A16 was selected for a subsequent use as seed treatment and biocontrol agent in semi-arid region fields. This strain showed important biofilm formation, regulation of Arabidopsis defenses, and harbored three cLPs genes.

Identification and Characterization of Fusarium proliferatum, a New Species of Fungi that Cause Fungal Keratitis.

Fusarium proliferatum (F. proliferatum) is known as a pathogen of corn and other crops, but its role in fungal keratitis has not been well investigated. Among 877 Fusarium isolates, we identified 155 (17.7%) stains as F. proliferatum according to their morphological features and partial DNA sequencing of translation elongation factor-[Formula: see text] (EF-[Formula: see text]) in this study. In vitro antifungal susceptibility tests showed that the F. proliferatum strains were sensitive to natamycin and vorionazole but resistant to amphotericin B, fluconazol, ketoconazole and itaconazole. Most of the F. proliferatum-positive keratitis patients (44/155,28.4%) were aged 51-60 years old. The main cause of infection was injury by a plant (51/155, 32.9%). A combination of 1% amphotericin B and 3% ketoconazole cured 45.2% (14/31) and a combination of 0.5% natamycin and 0.5% voriconazole cured 59.1% (13/22) of F. proliferatum-positive patients. The date suggests that F. proliferatum identified through EF-1ɑ DNA sequencing is an important new species that causes fungal keratitis. Based on antifungal susceptibility, treatment with a combination of 0.5% natamycin and 0.5% voriconazole improves the therapeutic efficacy in F. prolifertum-positive patients.

Complete cure of Fusarium solani sp. complex onychomycosis with Qs NdYAG treatment.

The incidence of non dermatophytic mould (NDM) onychomycosis (OM) has been steadily increasing Fusarium spp is the most common cause of NDM OM in most geographical locations. Fusarium spp and other NDMs are largely resistant to commonly used anti-fungals. The successful use of laser and light based devices has been demonstrated in dermatophytic OM, but there is no previous report of their successful use in any NDM OM. We describe a patient with OM caused by Fusarium solani spp, who was clinically (with a normal appearing nail) and mycologically (with negative microscopy and culture on repeated samples) cured of her infection following treatment with 2 sessions of Qs NdYAG (532nm and 1064nm) given 1 month apart.

Variability of composition and effects of essential oils from Rhanterium adpressum Coss. & Durieu against mycotoxinogenic Fusarium strains.

The antifungal potency of the essential oils of Rhanterium adpressum was evaluated against four mycotoxigenic strains of the genus Fusarium. The essential oils were obtained, separately, by hydro-distillation of the aerial parts of R. adpressum (leaves and flowers). The parts were collected during the period of bloom (3 months) for 3 years. The GC-MS analysis revealed thirty-six compounds for the essential oils, divided into four classes of chemical compounds, with variable percentages according to the month of extraction. The monoterpene hydrocarbons form the main class in these oils. On the other hand, the highest percentages of the oxygenated compounds are observed in the samples collected during the month of May. The direct contact method was used to evaluate the antifungal activity of the essential oils. The activity can be attributed to their relatively high composition of oxygenated monoterpenes. Flowers extract showed strong inhibitory activity, with very interesting concentrations of IC50 and MIC for both tests on solid and liquid medium. The effect of these oils on the production of type B trichothecenes (TCTBs) was evaluated, showing a significant inhibitory effect on TCTBs production, for both extracts (leaves and flowers). The rates of inhibition were 66-97 and 76-100% of FX, 3-ADON and 15-ADON, respectively. The inhibition of fungal biomass and the production of TCTBs depended on the used concentration of the essential oils. These results suggest that the essential oils from R. adpressum are able to control the growth of the tested strains and their subsequent production of TCTB mycotoxins.

Identification of a New Fungal Pathogen Causing White Villous Disease on the Fruiting Body of the Culinary-Medicinal Mushroom Auricularia auricula-judae (Agaricomycetes) in China.

Auricularia auricula-judae is an edible and medicinal fungus ranking fourth in production among the edible fungi cultivated worldwide. White villous disease is rampant in Northeast China; it infects the fruiting bodies of A. auricula-judae by forming a white mycelial layer on its ventral side. The disease not only causes an unacceptable morphological appearance and a poor-quality product, but it also significantly reduces the yield. In this study, based on fungal morphology, ribosomal DNA internal transcribed spacer sequences, identification of species-specific primers, and the pathogenicity of the mycelia and spores, 2 fungal pathogens were isolated and identified as Fusarium equiseti and F. sporotrichioides.

Antifungal activity, yield, and composition of Ocimum gratissimum essential oil.

Ocimum gratissimum L. or clove basil, belongs to the Lamiaceae family, has various desirable uses and applications. Beyond its aromatic, seasoning, and medicinal applications, this plant also has antimicrobial activity. This study was aimed at assessing the antifungal activity, yield, and composition of the essential oil (EO) of O. gratissimum. The species was cultivated in garden beds with dystrophic red latosol soil type containing high organic-matter content. The EO was obtained by hydrodistillation of dried leaves in a modified Clevenger apparatus, followed by determination of its content. Chemical characterization was carried out by gas chromatography-mass spectrometry (GC-MS). Microbial activity was assessed using the broth microdilution method, by determining the minimum inhibitory concentration (MIC), in order to compare the antimicrobial effect of EO in 10 isolates-Fusarium oxysporum f. sp tracheiphilum (CMM-0033), F. oxysporum f. sp. cubense (CMM-0813 and CMM-2819), F. oxysporum f. sp lycopersici (CMM-1104), F. solani (CMM-3828), Rhizoctonia solani (CMM-3274), and Macrophomina phaseolina (CMM-2715, CMM-3875, CMM-3615, and CMM-3650). The EO was a highly effective inhibitor of the studied phytopathogenic fungi, with MICs varying from 31.25 to 125 µg/mL. F. oxysporum f. sp lycopersici and R. solani were the most sensitive; both were inhibited at an MIC of 31.25 µg/mL. The EO content in the plant extract was 0.18%. Thirty chemical compounds were detected via GC-MS, with linalool (32.9%) being the major compound followed by 1,8-cineole (21.9%), both oxygenated monoterpenes. It can be concluded that clove basil EO is a highly effective antifungal agent, and therefore, a potential alternative for the control of plant pathogenic diseases.

Discovery of anti-microbial and anti-tubercular molecules from Fusarium solani: an endophyte of Glycyrrhiza glabra.

AIMS: Glycyrrhiza glabra is a high-value medicinal plant thriving in biodiversity rich Kashmir Himalaya. The present study was designed to explore the fungal endophytes from G. glabra as a source of bioactive molecules. METHODS AND RESULTS: The extracts prepared from the isolated endophytes were evaluated for anti-microbial activities using broth micro-dilution assay. The endophytic strain coded as A2 exhibiting promising anti-bacterial as well as anti-tuberculosis activity was identified as Fusarium solani by ITS-5.8S ribosomal gene sequencing technique. This strain was subjected to large-scale fermentation followed by isolation of its bioactive compounds using column chromatography. From the results of spectral data analysis and comparison with literature, the molecules were identified as 3,6,9-trihydroxy-7-methoxy-4,4-dimethyl-3,4-dihydro-1H-benzo[g]isochromene-5,10-dione (1), fusarubin (2), 3-O-methylfusarubin (3) and javanicin (4). Compound 1 is reported for the first time from this strain. All the four compounds inhibited the growth of various tested bacterial strains with MIC values in the range of <1 to 256 µg ml-1 . Fusarubin showed good activity against Mycobacterium tuberculosis strain H37Rv with MIC value of 8 µg ml-1 , whereas compounds 1, 3 and 4 exhibited moderate activity with MIC values of 256, 64, 32 µg ml-1 , respectively. CONCLUSIONS: To the best of our knowledge, this is the first study that reports significant anti-tuberculosis potential of bioactive molecules from endophytic F. solani evaluated against the virulent strain of M. tuberculosis. This study sets background towards their synthetic intervention for activity enhancement experiments in anti-microbial drug discovery programme. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to the chemoprofile variation of same endophyte with respect to source plant and ecoregions, further studies are required to explore endophytes of medicinal plants of all unusual biodiversity rich ecoregions for important and or novel bioactive molecules.

[Fusarium species associated with basal rot of garlic in North Central Mexico and its pathogenicity]. / Patogenicidad de especies de Fusarium asociadas a la pudrición basal del ajo en el centro norte de México.

Garlic in Mexico is one of the most profitable vegetable crops, grown in almost 5,451ha; out of which more than 83% are located in Zacatecas, Guanajuato, Sonora, Puebla, Baja California and Aguascalientes. Blossom-end rot caused by Fusarium spp is widely distributed worldwide and has been a limiting factor in onion and garlic production regions, not only in Mexico but also in other countries. The presence of Fusarium oxysporum has been reported in Guanajuato and Aguascalientes. Fusarium culmorum has been reported in onion cultivars of Morelos; and Fusarium proliferatum, Fusarium verticillioides, Fusarium solani and Fusarium acuminatum have been previously reported in Aguascalientes. The goal of this work was identifying the Fusarium species found in Zacatecas, Guanajuato and Aguascalientes, to assess their pathogenicity. Plants with disease symptoms were collected from hereinabove mentioned States. The samples resulted in the identification of: F. oxysporum, F. proliferatum, F. verticillioides, F. solani and F. acuminatum species; out of which Aguascalientes AGS1A (F. oxysporum), AGS1B (F. oxysporum) and AGSY-10 (F. acuminatum) strains showed higher severity under greenhouse conditions.

Patogenicidad de especies de Fusarium asociadas a la pudrición basal del ajo en el centro norte de México / Fusarium species associated with basal rot of garlic in North Central Mexico and its pathogenicity

El ajo en México es uno de los cultivos de hortalizas más rentables, más del 83% de esta superficie es aportada por los estados de Zacatecas, Guanajuato, Sonora, Puebla, Baja California y Aguascalientes. La pudrición basal ocasionada por Fusarium spp. se encuentra ampliamente distribuida a nivel mundial; esta enfermedad se ha convertido en una limitante en zonas productoras de cebolla y ajo, no solo en México, sino también en otros países, En México, se ha informado la presencia de Fusarium oxysporum en plantas en Guanajuato y en semillas de ajo en Aguascalientes. En el estado de Morelos se ha reportado la presencia de Fusarium culmorum en cultivares de cebolla. Asimismo, en Aguascalientes se tienen antecedentes de otras especies como Fusarium proliferatum, Fusarium verticillioides, Fusarium solani y Fusarium acuminatum. Para este trabajo se planteó como objetivo identificar las especies de Fusarium encontradas en los estados de Zacatecas, Guanajuato y Aguascalientes, y evaluar su patogenicidad. Se realizaron recolectas de plantas con síntomas de la enfermedad en los estados antes mencionados. De los muestreos realizados se identificaron las especies F. oxysporum, F. proliferatum, F. verticillioides, F. solani y F. acuminatum; las cepas de Aguascalientes identificadas como AGS1A (F. oxysporum), AGS1B (F. oxysporum) y AGSY-10 (F. acuminatum) fueron las que presentaron bajo condiciones de invernadero un mayor índice de severidad.

Garlic in Mexico is one of the most profitable vegetable crops, grown in almost 5,451 ha; out of which more than 83% are located in Zacatecas, Guanajuato, Sonora, Puebla, Baja California and Aguascalientes. Blossom-end rot caused by Fusarium spp is widely distributed worldwide and has been a limiting factor in onion and garlic production regions, not only in Mexico but also in other countries. The presence of Fusarium oxysporum has been reported in Guanajuato and Aguascalientes. Fusarium culmorum has been reported in onion cultivars of Morelos; and Fusarium proliferatum, Fusarium verticillioides, Fusarium solani and Fusarium acuminatum have been previously reported in Aguascalientes. The goal of this work was identifying the Fusarium species found in Zacatecas, Guanajuato and Aguascalientes, to assess their pathogenicity. Plants with disease symptoms were collected from hereinabove mentioned States. The samples resulted in the identification of: F. oxysporum, F. proliferatum, F. verticillioides, F. solani and F. acuminatum species; out of which Aguascalientes AGS1A (F. oxysporum), AGS1B (F. oxysporum) and AGSY-10 (F. acuminatum) strains showed higher severity under greenhouse conditions.

Epitypification of Fusisporium (Fusarium) solani and its assignment to a common phylogenetic species in the Fusarium solani species complex.

Fusisporium solani was described as the causal agent of a dry rot of potato in Germany in the mid 19th century. As Fusarium solani, the species became known as a plurivorous plant pathogen, endophyte, decomposer, and opportunistic pathogen of humans and nutritional symbiont of insects. In parallel, it became evident that the morphologically defined species F. solani represents a phylogenetically and biologically complex group of often morphologically cryptic species that has come to be known in part as the F. solani species complex (FSSC), accommodating several formae speciales and mating populations/biological species. The FSSC currently includes more than 60 phylogenetic species. Several of these have been named, but the majority remains unnamed and the identity of F. solani sensu stricto is unclear. To promote further taxonomic developments in the FSSC, lectoand epitypification is proposed for Fusisporium solani Although no type material for F. solani is known to exist, the species was abundantly illustrated in the protologue. Thus, a relevant illustration provided by von Martius is selected as the lectotype. The epitype selected here originates from a rotting potato collected in a field in Slovenia. This strain causes a dry rot of artificially inoculated potatoes. It groups in the heretofore unnamed phylogenetic species 5, which is nested within clade 3 of the FSSC (FSSC 5). Members of this phylogenetic species have a wide geographic distribution and include soil saprotrophs and plant and opportunistic human pathogens. This typification is consistent with the original description of Fusisporium solani and the concept of F. solani as a widely distributed soil inhabitant and pathogen.

Taxonomic analysis of the microbial community in stored sugar beets using high-throughput sequencing of different marker genes.

Post-harvest colonization of sugar beets accompanied by rot development is a serious problem due to sugar losses and negative impact on processing quality. Studies on the microbial community associated with rot development and factors shaping their structure are missing. Therefore, high-throughput sequencing was applied to describe the influence of environment, plant genotype and storage temperature (8°C and 20°C) on three different communities in stored sugar beets, namely fungi (internal transcribed spacers 1 and 2), Fusarium spp. (elongation factor-1α gene fragment) and oomycetes (internal transcribed spacers 1). The composition of the fungal community changed during storage mostly influenced by the storage temperature followed by a weak environmental effect. Botrytis cinerea was the prevalent species at 8°C whereas members of the fungal genera Fusarium and Penicillium became dominant at 20°C. This shift was independent of the plant genotype. Species richness within the genus Fusarium also increased during storage at both temperatures whereas the oomycetes community did not change. Moreover, oomycetes species were absent after storage at 20°C. The results of the present study clearly show that rot development during sugar beet storage is associated with pathogens well known as causal agents of post-harvest diseases in many other crops.

Potencial do óleo essencial de erva-luísa (Aloysia citriodora Palau) no controle de Fusarium sp. in vitro / Essential oil potential herb-luisa (Aloysia citriodora Palau) in the control of Fusarium sp. in vitro

RESUMO O presente trabalho teve como objetivo principal avaliar o potencial do uso do óleo essencial de Aloysia citriodora no controle in vitro de Fusarium sp., isolado de plântulas de beterraba infectadas com o mesmo. O trabalho foi realizado por meio de dois experimentos: um sobre o efeito do óleo essencial no crescimento micelial, e outro sobre o efeito na germinação de conídios do fungo. No primeiro trabalho, avaliou-se em delineamento inteiramente ao acaso, o efeito das concentrações 0,0155%; 0,0315%; 0,0625%; 0,125%; 0,250% e 0,500% do óleo essencial de A. citriodora em placas de Petri® com meio de cultivo BDA, mais a testemunha, com meio BDA puro. Cada placa foi considerada uma repetição, as quais foram incubadas a 24ºC±1ºC e submetidas a fotoperíodo de doze horas. Avaliou-se o crescimento radial do patógeno em função do crescimento micelial do patógeno quando este atingia as bordas da primeira placa. No segundo experimento, as mesmas concentrações foram testadas, nas mesmas condições de incubação, no entanto, em lâminas de microscopia com meio BD. Utilizou-se delineamento inteiramente ao acaso, sendo considerada como unidade experimental cada lâmina utilizada. Avaliou-se 24 horas após a incubação, em microscópio óptico, a germinação dos primeiros vinte conídios visualizados a partir do canto esquerdo superior para o direito. Os resultados do segundo experimento foram expressos em porcentagem de germinação de conídios. Os resultados obtidos permitiram concluir que este óleo essencial possui efeito fungistático e fungicida sobre o crescimento micelial e na germinação de conídios de Fusarium sp.. Além disso este efeito é maior em função do aumento da concentração do óleo essencial.

ABSTRACT This study aimed to evaluate the potential use of Aloysia citriodora essential oilisolated from beet seedlings infected with it. The study was performed by conducting two experiments evaluating the effect of the essential oil on mycelial growth and fungus conidia germination. In the first study, in a completely randomized design, the effect concentrations (0.0155%; 0.0315%; 0.0625%; 0.125%; 0.250% and 0.500%) of essential oil of A. citriodorawas evaluated on Petri® dish with a PDA cultivation medium, plus the control, with half pure PDA. Each plate was taken as a repeat, and incubated at 24°C±1°C and a photoperiod of twelve hours. The radial growth of the pathogen, when the first plate was hit by the mycelial growth of the pathogen on its edges was evaluated. In the second experiment, the same concentrations were tested under the same incubationconditions, however, on microscope slides with half PD medium. The design was used completely randomized, each microscope slide used was considered as one experimental unit. Germination of the first 2 conidia strains, viewed from the upper left to the right was evaluated24 hours after incubation, using an optical microscope. The results of the second experiment were expressed as a percentage of conidia germination. The results obtained showed that this essential oil has fungistatic and fungicidal effect on the mycelial growth and at the conidia germination of Fusarium sp., which has a greater effect with increasing concentrations.

Fungal Endophyte Diversity and Bioactivity in the Indian Medicinal Plant Ocimum sanctum Linn.

Endophytic mycopopulation isolated from India's Queen of herbs Tulsi (Ocimum sanctum) were explored and investigated for their diversity and antiphytopathogenic activity against widespread plant pathogens Botrytis cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani and Fusarium oxysporum. 90 fungal isolates, representing 17 genera were recovered from 313 disease-free and surface sterilised plant segments (leaf and stem tissues) from three different geographic locations (Delhi, Hyderabad and Mukteshwar) during distinct sampling times in consequent years 2010 and 2011 in India. Fungal endophytes were subjected to molecular identification based on rDNA ITS sequence analysis. Plant pathogens such as F. verticillioides, B. maydis, C. coarctatum, R. bataticola, Hypoxylon sp., Diaporthe phaseolorum, Alternaria tenuissima and A. alternata have occurred as endophyte only during second sampling (second sampling in 2011) in the present study. Bi-plot generated by principal component analysis suggested tissue specificity of certain fungal endophytes. Dendrogram revealed species abundance as a function of mean temperature of the location at the time of sampling. Shannon diversity in the first collection is highest in Hyderabad leaf tissues (H' = 1.907) whereas in second collection it was highest from leaf tissues of Delhi (H' = 1.846). Mukteshwar (altitude: 7500 feet) reported least isolation rate in second collection. Nearly 23% of the total fungal isolates were considered as potent biocontrol agent. Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml. Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in antiphytopathogenic activity of hexane extract. The production of 2H-pyran-2-one, 5,6-dihydro-6-pentyl from M. phaseolina, an endophytic fungus is being reported for the first time.

Fusarium redolens 6WBY3, an endophytic fungus isolated from Fritillaria unibracteata var. wabuensis, produces peimisine and imperialine-3ß-D-glucoside.

The major biological active ingredients of Bulbus Fritillariae cirrhosae (BFC) are steroidal alkaloids, such as peimisine, imperialine-3ß-D-glucoside, and peimine. The bulbus of Fritillaria unibracteata var. wabuensis (FUW) was officially recorded in the National Pharmacopoeia of China (2010 edition) as one of the sources of BFC because of its positive therapeutic effects and few side effects. The endophytic fungus strain 6WBY3 was isolated from the fresh bulbus of FUW that had been cultivated for six years. Based on morphological methods and the phylogenetic analysis of internal transcribed spacer (ITS) sequences, this strain was identified as Fusarium redolens. Using color reaction analysis, high performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD), and mass spectrometry (MS), it was demonstrated that F. redolens 6WBY3 could produce peimisine and imperialine-3ß-D-glucoside, similar to its host plant. The yields of peimisine and imperialine-3ß-D-glucoside were 16.0 µg·l(-1) and 18.8 µg·l(-1), respectively, in one week of culture. These results indicate that F. redolens 6WBY3 is a promising candidate for the large scale production of peimisine and imperialine-3ß-D-glucoside. In addition, the results from the strain 6WBY3 lay the foundation for further study into the mechanism of Fritillaria alkaloids biosynthesis in fungi.

Fatal disseminated infection with Fusarium petroliphilum.

Members of the Fusarium solani species complex (FSSC) are causing the majority of the fusariosis in humans. Disseminated fusariosis has a high mortality and is predominantly observed in patients with leukemia. Here, we present the case of a fatal infection by a Fusarium strain with a degenerated phenotype, in a patient with acute lymphatic leukemia. Multiple nasal and skin biopsies as well as blood cultures yielded fungal growth, while in direct and histopathological examination of biopsy material septate hyphae were visible. Initial colonies were white with slimy masses with microconidia reminiscent of Fusarium/Acremonium, but with conidiospore production directly on the hyphae. Multi-locus sequence typing discerned a pionnotal-morphologically degenerated-colony of the recently recognized F. petroliphilum as etiological agent. The culture returned to a typical F. solani species complex morphology only after several weeks of growth in culture. Antifungal susceptibility tests indicate amphotericin B as best drug for this FSSC member rather than any of the azoles or echinocandins.