Long‐term Preservation of <i>Podosphaera fusca</i> Using Silica Gel

Pérez-Garcı́a, Alejandro; Mingorance, E.; Rivera, María Eugenia Sánchez Díaz de; Pino, Daniel Del; Romero, Diego; Torés, J. A.; Vicente, Antonio de

doi:10.1111/j.1439-0434.2006.01086.x

Cited by 35 publications

(24 citation statements)

References 5 publications

(7 reference statements)

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“…Hair samples were transported to the lab in sterile Falcon tubes which had been half filled with silica gel, a desiccant that is effective for storing fungal tissue [31] [32]. Every piece of sloth hair placed on agar yielded multiple fungal isolates representing a variety of morphotypes.…”

Section: Resultsmentioning

confidence: 99%

Sloth Hair as a Novel Source of Fungi with Potent Anti-Parasitic, Anti-Cancer and Anti-Bacterial Bioactivity

et al. 2014

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The extraordinary biological diversity of tropical forests harbors a rich chemical diversity with enormous potential as a source of novel bioactive compounds. Of particular interest are new environments for microbial discovery. Sloths – arboreal mammals commonly found in the lowland forests of Panama – carry a wide variety of micro- and macro-organisms on their coarse outer hair. Here we report for the first time the isolation of diverse and bioactive strains of fungi from sloth hair, and their taxonomic placement. Eighty-four isolates of fungi were obtained in culture from the surface of hair that was collected from living three-toed sloths (Bradypus variegatus, Bradypodidae) in Soberanía National Park, Republic of Panama. Phylogenetic analyses revealed a diverse group of Ascomycota belonging to 28 distinct operational taxonomic units (OTUs), several of which are divergent from previously known taxa. Seventy-four isolates were cultivated in liquid broth and crude extracts were tested for bioactivity in vitro. We found a broad range of activities against strains of the parasites that cause malaria (Plasmodium falciparum) and Chagas disease (Trypanosoma cruzi), and against the human breast cancer cell line MCF-7. Fifty fungal extracts were tested for antibacterial activity in a new antibiotic profile screen called BioMAP; of these, 20 were active against at least one bacterial strain, and one had an unusual pattern of bioactivity against Gram-negative bacteria that suggests a potentially new mode of action. Together our results reveal the importance of exploring novel environments for bioactive fungi, and demonstrate for the first time the taxonomic composition and bioactivity of fungi from sloth hair.

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Section: Resultsmentioning

confidence: 99%

Sloth Hair as a Novel Source of Fungi with Potent Anti-Parasitic, Anti-Cancer and Anti-Bacterial Bioactivity

et al. 2014

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“…2001). For long‐term storage, the isolate was preserved at −80°C using silica gel (Pérez‐García et al. 2006).…”

Section: Methodsmentioning

confidence: 99%

Effect of lipopeptides of antagonistic strains ofBacillus subtilison the morphology and ultrastructure of the cucurbit fungal pathogenPodosphaera fusca

Romero

Vicente

Olmos

et al. 2007

Journal of Applied Microbiology

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111

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Aims: To analyse the morphological and ultrastructural effects of lipopeptides of cell‐free liquid cultures from the antagonistic Bacillus subtilis strains, UMAF6614 and UMAF6639, on the cucurbit powdery mildew fungus, Podosphaera fusca, conidial germination. Methods and Results: Butanolic extracts from cell‐free supernatants of B. subtilis cultures were tested for their ability to arrest P. fusca conidial germination using the zucchini cotyledon disc method. Previously, the occurrence of lipopeptide antibiotics fengycin, iturin/bacillomycin and surfactin in the extracts was verified by diverse chromatographic approaches. Conidial germination was strongly reduced by antifungal extracts obtained from liquid cultures of both B. subtilis strains. Scanning electron microscopy analysis showed morphological damage in conidia characterized by the presence of large depressions and loss of turgidness. Transmission electron microscopy analysis revealed severe modifications in the plasma membrane and disorganization of the P. fusca cell cytoplasm. Conclusions: The lipopeptides produced by the two strains of B. subtilis are able to reduce cucurbit powdery mildew disease by arresting conidial germination, which seems to result from the induction of important cytological alterations. Significance and Impact of the Study: We elucidated the mechanisms employed by these antagonistic strains of B. subtilis to suppress cucurbit powdery mildew disease and delineate the ultrastructural damages responsible for their suppressive effect.

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“…Single-spore isolates were then identified based on characteristics of the conidial stage (Braun, 1987). After identification, isolates were kept at )80°C (Pe´rez-Garcı´a et al, 2006) until use in fungicide sensitivity tests. A total of 250 isolates of P. fusca from this survey were used.…”

Section: Pathogen Isolatesmentioning

confidence: 99%

Occurrence and distribution of resistance to QoI fungicides in populations of Podosphaera fusca in south central Spain

Fernández-Ortuño¹,

Pérez-Garcı́a

Lopez-Ruiz³

et al. 2006

Eur J Plant Pathol

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Cucurbit powdery mildew caused by Podosphaera fusca limits crop production in Spain. Since its management is strongly dependent on chemicals, the rational design of control programmes requires a good understanding of the fungicide resistance phenomenon in field populations. Fifty single-spore isolates of P. fusca were tested for sensitivity to three quinone-outside inhibiting (QoI) fungicides: azoxystrobin, kresoxim-methyl and trifloxystrobin. Minimum inhibitory concentration (MIC) values for QoI-sensitive isolates were found to range from 0.25 to 10 lg ml )1 for azoxystrobin to 5-25 lg ml )1 for kresoximmethyl, using a leaf disc-based bioassay. High levels of cross-resistance to QoI fungicides were found. Eleven isolates showed resistance to the three QoI fungicides tested with MIC and EC 50 values >500 lg ml )1 resulting in RF values as high as >715 and >1000 for trifloxystrobin and azoxystrobin, respectively. A survey of P. fusca QoI resistance was carried out in different provinces located in the south central area of Spain during the cucurbit growing seasons in 2002, 2003 and 2004. Examination of a collection of 250 isolates for QoI resistance revealed that 32% were resistant to the three fungicides tested; the provinces of Ciudad Real, Co´rdoba and Murcia being the locations with the highest frequencies of resistance (44-74%). By contrast, no resistance was found in Badajoz, and relatively low frequencies were observed in Almerı´a and Valencia (10-13%). Nearly 50% of resistant isolates were collected from melon plants. Based on these data, recommendations about the use of QoI fungicides for cucurbit powdery mildew management in the sampled areas are made.

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Long‐term Preservation of Podosphaera fusca Using Silica Gel

Cited by 35 publications

References 5 publications

Sloth Hair as a Novel Source of Fungi with Potent Anti-Parasitic, Anti-Cancer and Anti-Bacterial Bioactivity

Sloth Hair as a Novel Source of Fungi with Potent Anti-Parasitic, Anti-Cancer and Anti-Bacterial Bioactivity

Effect of lipopeptides of antagonistic strains ofBacillus subtilison the morphology and ultrastructure of the cucurbit fungal pathogenPodosphaera fusca

Occurrence and distribution of resistance to QoI fungicides in populations of Podosphaera fusca in south central Spain

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