Transcriptional response of Fusarium oxysporum and Neocosmospora solani challenged with amphotericin B or posaconazole.

Some species of fusaria are well-known pathogens of humans, animals and plants. Fusarium oxysporum and Neocosmospora solani (formerly Fusarium solani) cause human infections that range from onychomycosis or keratitis to severe disseminated infections. In general, these infections are difficult to treat due to poor therapeutic responses in immunocompromised patients. Despite that, little is known about the molecular mechanisms and transcriptional changes responsible for the antifungal resistance in fusaria. To shed light on the transcriptional response to antifungals, we carried out the first reported high-throughput RNA-seq analysis for F. oxysporum and N. solani that had been exposed to amphotericin B (AMB) and posaconazole (PSC). We detected significant differences between the transcriptional profiles of the two species and we found that some oxidation-reduction, metabolic, cellular and transport processes were regulated differentially by both fungi. The same was found with several genes from the ergosterol synthesis, efflux pumps, oxidative stress response and membrane biosynthesis pathways. A significant up-regulation of the C-22 sterol desaturase (ERG5), the sterol 24-C-methyltransferase (ERG6) gene, the glutathione S-transferase (GST) gene and of several members of the major facilitator superfamily (MSF) was demonstrated in this study after treating F. oxysporum with AMB. These results offer a good overview of transcriptional changes after exposure to commonly used antifungals, highlights the genes that are related to resistance mechanisms of these fungi, which will be a valuable tool for identifying causes of failure of treatments.

Effect of bioactive compounds extracted from Cordyceps nidus ANDES-F1080 on laccase activity of Pleurotus ostreatus ANDES-F515.

Laccases are ligninolytic enzymes produced by different microorganisms, especially by fungi such as the white-rot fungus Pleurotus ostreatus. Chemical inductors have been used to promote laccase secretion due to the application of these enzymes in lignocellulosic biomass pretreatment. Cordyceps nidus ANDES-F1080 was previously described as a source of bioactive compounds that could influence the enzymatic production system of other fungi. For that reason, this study evaluates the effect of C. nidus' ANDES-F1080 extracts on the laccase activity of P. ostreatus ANDES-F515. To achieve this objective, C. nidus ANDES-F1080 was grown in four different substrates: two artificial-based and two natural-based culture media. Metabolites were extracted from C. nidus ANDES-F1080 using water and methanol as solvents. Biochemical characterization of these extracts was performed to complement the analysis of their effect on laccase activity. Our results revealed an enhancement on the laccase activity of P. ostreatus ANDES-F515 grown in natural-based cultures when C. nidus' ANDES-F1080 extracts were supplemented. The best laccase activities registered values around 10,575 ±â€¯813 U·L-1.

Genome-Wide Association Study Identifies Single Nucleotide Polymorphism Markers Associated with Mycelial Growth (at 15, 20, and 25°C), Mefenoxam Resistance, and Mating Type in Phytophthora infestans.

Phenotypic diversity among individuals defines the potential for evolutionary selection in a species. Phytophthora infestans epidemics are generally thought to be favored by moderate to low temperatures, but temperatures in many locations worldwide are expected to rise as a result of global climate change. Thus, we investigated variation among individuals of P. infestans for relative growth at different temperatures. Isolates of P. infestans came from three collections: (i) individual genotypes recently dominant in the United States, (ii) recently collected individuals from Central Mexico, and (iii) progeny of a recent sexual recombination event in the northeastern United States. In general, these isolates had optimal mycelial growth rates at 15 or 20°C. However, two individuals from Central Mexico grew better at higher temperatures than did most others and two individuals grew relatively less at higher temperatures than did most others. The isolates were also assessed for mefenoxam sensitivity and mating type. Each collection contained individuals of diverse sensitivities to mefenoxam and individuals of the A1 and A2 mating type. We then searched for genomic regions associated with phenotypic diversity using genotyping-by-sequencing. We found one single nucleotide polymorphism (SNP) associated with variability in mycelial growth at 20°C, two associated with variability in mycelial growth at 25°C, two associated with sensitivity to mefenoxam, and one associated with mating type. Interestingly, the SNPs associated with mefenoxam sensitivity were found in a gene-sparse region, whereas the SNPs associated with growth at the two temperatures and mating type were found both at more gene-dense regions.

Multiscale design of a dairy beverage model composed of Candida utilis single cell protein supplemented with oleic acid.

One of the main challenges in the food industry is to design strategies for the successful incorporation of natural sources of bioactive compounds. Recently, yogurts and other fermented dairy beverages have been proposed as ideal carriers of such bioactive compounds such as fatty acids and antioxidants that could improve consumers' health. However, the incorporation of new ingredients causes functional and structural modifications that may affect the consumers' preferences. In this work, a dairy beverage model supplemented with oleic acid has been designed by partial substitution of milk by Candida utilis single-cell protein extract. The changes in the structural properties of this new beverage were evaluated by following the fermentation process, pH, aggregate size, microstructure, and changes in rheological properties. Furthermore, molecular dynamics simulations were carried out to analyze the interaction between its main components. Our data revealed that samples with a percentage of milk substitution of 30% showed a higher viscosity as compared with the other percentages and less viscosity than the control (no substitution). These samples were then selected for fortification by incorporating oleic acid microcapsules. A concentration of 1.5 g/100 g was shown to be the optimal quantity of microcapsules for oleic acid supplementation. Molecular dynamic simulations revealed glutathione as an important component of the micro-gel structure. The present study forms the basis for novel studies where Candida utilis single-cell protein and microencapsulated essential oils could be used to design innovative bioproducts.

The effect of cyclic heat treatment on the physicochemical properties of bio hydroxyapatite from bovine bone.

This paper focus on physicochemical changes in bio-hydroxyapatite (BIO-HAp) from bovine femur obtained by calcination at high temperatures: 520-620 (each 20 °C) at 7.4 °C/min and from 700 to 1100 °C (each 100 °C) at three heating rates: 7.4, 9.9, and 11.1 °C/min. BIO-HAp samples were obtained using a multi-step process: cleaning, milling, hydrothermal process, calcination in an air atmosphere, and cooling in furnace air. Inductively Couple Plasma (ICP) showed that the presence of Mg, K, S, Ba, Zn, and Na, is not affected by the annealing temperature and heating rate. While Scanning Electron Microscopy (SEM) images showed the continuous growth of the HAp crystals during the calcination process due to the coalescence phenomenon, and the Full Width at the Half Maximum for the X-ray patterns for temperatures up to 700 is affected by the annealing temperature and the heating rate. Through X-ray diffraction, thermal, and calorimetric analysis (TGA-DSC), a partial dehydroxylation of hydroxyapatite was found in samples calcined up to 900 °C for the three heating rates. Also, Ca/P molar ratio decreased for samples calcined up to 900 °C as a result of the dehydroxylation process. NaCaPO4, CaCO3, Ca3(PO4)2, MgO, and Ca(H2PO4)2 are some phases identified by X-ray diffraction; some of them are part of the bone and others were formed during the calcination process as a function of annealing temperature and heating rate, as it is the case for MgO.

Description of Alicyclobacillus montanus sp. nov., a mixotrophic bacterium isolated from acidic hot springs.

Three morphologically similar thermo-acidophilic strains, USBA-GBX-501, USBA-GBX-502 and USBA-GBX-503T, were isolated from acidic thermal springs at the National Natural Park Los Nevados (Colombia). All isolates were spore-forming, Gram-stain-positive and motile, growing aerobically at 25-55 °C (optimum ~45 °C) and at pH 1.5-4.5 (optimum pH ~3.0). Phylogenetic analysis of the 16S rRNA gene sequences of these isolates showed an almost identical sequence (99.0 % similarity) and they formed a robust cluster with the closest relative Alicyclobacillus tolerans DSM 16297T with a sequence similarity of 99.0 %. Average similarity to other species of the genus Alicyclobacillus was 93.0 % and average similarity to species of the genus Effusibacillus was 90 %. In addition, the level of DNA-DNA hybridization between strain USBA-GBX-503T and Alicyclobacillus tolerans DSM 16297T was 31.7 %. The genomic DNA G+C content of strain USBA-GBX-503T was 44.6 mol%. The only menaquinone was MK-7 (100.0 %). No ω-alicyclic fatty acids were detected in strain USBA-GBX-503T, and the major cellular fatty acids were C18 : 1ω7c, anteiso-C17 : 0 and iso-C17 : 0. Based on phenotypic and chemotaxonomic characteristics, phylogenetic analysis and DNA-DNA relatedness values, along with low levels of identity at the whole genome level (ANIb and ANIm values of <67.0 and <91.0 %, respectively), it can be concluded that strain USBA-GBX-503T represents a novel species of the genus Alicyclobacillus, for which the name Alicyclobacillus montanus sp. nov. is proposed. The type strain is USBA-GBX-503T (=CMPUJ UGB U503T=CBMAI1927T).

Phytophthora betacei, a new species within Phytophthora clade 1c causing late blight on Solanum betaceum in Colombia.

Over the past few years, symptoms akin to late blight disease have been reported on a variety of crop plants in South America. Despite the economic importance of these crops, the causal agents of the diseases belonging to the genus Phytophthora have not been completely characterized. In this study, a new Phytophthora species was described in Colombia from tree tomato (Solanum betaceum), a semi-domesticated fruit grown in northern South America. Comprehensive phylogenetic, morphological, population genetic analyses, and infection assays to characterize this new species, were conducted. All data support the description of the new species, Phytophthora betacei sp. nov. Phylogenetic analyses suggest that this new species belongs to clade 1c of the genus Phytophthora and is a close relative of the potato late blight pathogen, P. infestans. Furthermore, it appeared as the sister group of the P. andina strains collected from wild Solanaceae (clonal lineage EC-2). Analyses of morphological and physiological characters as well as host specificity showed high support for the differentiation of these species. Based on these results, a complete description of the new species is provided and the species boundaries within Phytophthora clade 1c in northern South America are discussed.

Publisher's Note: Driven Open Quantum Systems and Floquet Stroboscopic Dynamics [Phys. Rev. Lett. 117, 250401 (2016)].

This corrects the article DOI: 10.1103/PhysRevLett.117.250401.

Description of a new anaerobic thermophilic bacterium, Thermoanaerobacterium butyriciformans sp. nov.

Strain USBA-019T, an anaerobic and thermophilic strain, was identified as a new member of the genus Thermoanaerobacterium. USBA-019T cells are gram-positive, strictly anaerobic, thermophilic, chemoorganotrophic, moderately acidophilic, non-motile, endospore-forming, slightly curved, and rod-shaped. Cells measure 0.4×3.0-7.0µm. Optimal growth occurs at 50-55°C (35-65°C). Optimum pH is 5.0-5.5 (4.0-8.5). Thiosulfate, elemental sulfur and nitrate were utilized as electron acceptors. Fermentation of glucose, lactose, cellobiose, galactose, arabinose, xylose, starch and xylan primarily produced acetate and butyrate. Xylan, starch and cellobiose produced ethanol and starch, cellobiose, galactose, arabinose and mannose produced lactic acid. Phylogenetic analyses based on 16S rRNA gene sequence comparison and genomic relatedness indices show the close relation of USBA-019T to Thermoanaerobacterium thermostercoris and Thermoanaerobacterium aotearoense (similarity value: 99%). Hybridization of USBA-019T, Th. thermostercoris DSM22141T and Th. aotearoense DMS10170T found DNA-DNA relatedness of 33.2% and 18.2%, respectively. Based on phenotypic, chemotaxonomic and phylogenetic evidence, along with low identity at whole genome level, USBA-019T is a novel species of the genus Thermoanaerobacterium which we propose to name Thermoanaerobacterium butyriciformans sp. nov. The type strain is USBA-019T (=CMPUJ U-019T=DSM 101588T).

Highly efficient transformation system for Malassezia furfur and Malassezia pachydermatis using Agrobacterium tumefaciens-mediated transformation.

Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M. furfur and M. pachydermatis by combining the transformation protocols of Agaricus bisporus and Cryptococcus neoformans. Optimal temperature and co-cultivation time for transformation were 5 and 7days at 19°C and 24°C, respectively. Transformation efficiency was 0.75-1.5% for M. furfur and 0.6-7.5% for M. pachydermatis. Integration of the hpt resistance cassette and gfp was verified using PCR and fluorescence microscopy, respectively. The T-DNA was mitotically stable in approximately 80% of the transformants after 10 times sub-culturing in the absence of hygromycin. Improving transformation protocols contribute to study the biology and pathophysiology of Malassezia.

Driven Open Quantum Systems and Floquet Stroboscopic Dynamics.

We provide an analytic solution to the problem of system-bath dynamics under the effect of high-frequency driving that has applications in a large class of settings, such as driven-dissipative many-body systems. Our method relies on discrete symmetries of the system-bath Hamiltonian and provides the time evolution operator of the full system, including bath degrees of freedom, without weak-coupling or Markovian assumptions. An interpretation of the solution in terms of the stroboscopic evolution of a family of observables under the influence of an effective static Hamiltonian is proposed, which constitutes a flexible simulation procedure of nontrivial Hamiltonians. We instantiate the result with the study of the spin-boson model with time-dependent tunneling amplitude. We analyze the class of Hamiltonians that may be stroboscopically accessed for this example and illustrate the dynamics of system and bath degrees of freedom.

Caracterización de una población con discapacidad visual (baja visión y ceguera) atendida en dos instituciones prestadoras de salud de Medellín / Characterization of a population with visual disabilities (low vision and blindness) attended in two health institutions from Medellín / Caracterização de uma população com incapacidade visual (baixa visão e cegueira) atendida em duas instituições prestadoras de saúde de Medellín

Objetivo: caracterizar, en los ámbitos clínico y sociodemográfico, una población de pacientes con discapacidad visual atendidos en dos instituciones de salud de la ciudad de Medellín (departamento de Antioquia/Colombia), con énfasis en la etiología del déficit visual irreversible. Metodología: estudio observacional descriptivo. Estudio macro sobre deficiencias visuales unilaterales y bilaterales en 1 742 registros de historias clínicas para identificar pacientes con baja visión o ceguera. Aplicación de un formato de investigación orientado a validar los pacientes con discapacidad visual y se seleccionaron 107 historias clínicas. Resultados: el 56.6% presenta discapacidad visual tipo baja visión y el 43.4% discapacidad visual tipo ceguera. El déficit visual responsable de la discapacidad visual fue del 39% por causas oftalmológicas, 20% por alteraciones neuro-oftalmológicas y 17% por trastornos neurológicos de cortezas visuales. Además de la agudeza visual, se encontraron otras deficiencias de la función visual: atrofia óptica, alteración electrofisiológica de la conducción visual y encefalomalacia en cortezas visuales. El 82% de los pacientes tiene al menos una comorbilidad no oftalmológica. Conclusiones: es fundamental un adecuado registro de las características biológicas, sociales, psicológicas y de las actividades de rehabilitación de los pacientes con baja visión y ceguera, para entender en forma integral no sólo la discapacidad sino el impacto que produce.

Objective: to characterize the clinical and socio-demographical characteristics of a population of patients with visual impairment attended at two health institutions in Medellín (Antioquia, Colombia), with an emphasis on the etiology of irreversible vision loss. Methodology: Observational, descriptive study of unilateral and bilateral visual impairment in 1 742 medical records to identify patients with low vision or blindness. A research form was used to validate patients with visual impairment, and 107 medical records were selected. Results: 56.6% presented low vision and 43.4% presented blindness. Vision loss was due to ophthalmic causes in 39% of cases, 20% were caused by neuro-ophthalmic alterations and 17% by neurological disorders of the visual cortex. In addition to visual acuity, other visual impairments were found, such as optic atrophy, electrophysiological alteration of the visual pathway, and encephalomalacia in visual cortices. 82% of patients had at least one nonophthalmic comorbidity. Conclusions: Adequate registration of rehabilitation activity, biological, social, and psychological characteristics of patients with low vision and blindness is essential in order to fully understand both the impairment and its impact.

Objetivo: caracterizar, nos âmbitos clínico e sócio-demográfico, uma população de pacientes com incapacidade visual atendidos em duas instituições de saúde da cidade de Medellín (departamento de Antioquia/Colômbia), com énfase na etiologia do déficit visual irreversível. Metodologia: estudo observacional descritivo. Estudo macro sobre deficiências visuais unilaterais e bilaterais em 1 742 registros de histórias clínicas para identificar pacientes com baixa visão ou cegueira. Aplicação de um formato de investigação orientado a validar os pacientes com incapacidade visual e se selecionaram 107 histórias clínicas. Resultados: 56.6% apresenta incapacidade visual tipo baixa visão e 43.4% incapacidade visual tipo cegueira. O déficit visual responsável da incapacidade visual foi de 39% por causas oftalmológicas, 20% por alterações neuro-oftalmológicas e 17% por transtornos neurológicos de córtex visual. Ademais da agudeza visual, se encontraram outras deficiências da função visual: atrofia óptica, alteração eletrofisiológica da condução visual e encefalomalácia em córtex visual. 82% dos pacientes têm pelo menos uma comorbilidade não oftalmológica. Conclusões: é fundamental um adequado registro das características biológicas, sociais, psicológicas e das atividades de reabilitação dos pacientes com baixa visão e cegueira, para entender em forma integral não só a incapacidade senão o impacto que produz.

Characterization of the first Batrachochytrium dendrobatidis isolate from the Colombian Andes, an amphibian biodiversity hotspot.

The pathogenic chytrid fungus, Batrachochytrium dendrobatidis (Bd), constitutes a significant threat to more than 790 amphibian species occurring in Colombia. To date there is no molecular or morphological description of strains infecting Colombian populations. Here we report the genetic and morphological characterization of the first Colombian isolate of Bd (strain EV001). Our goals were threefold: (1) to characterize the morphology of EV001 using light and scanning electron microscopy, (2) to genotype this strain by direct sequencing of 17 polymorphic nuclear markers developed previously, and (3) to compare our findings with published reports on strains from other areas of the globe. We found that EV001 is morphologically consistent with previously described strains. Multi-locus genotyping suggested that EV001 is grouped genetically with Panamanian strains and is most similar to strain JEL203 isolated from a captive individual. This finding fills an important gap in our knowledge of Neotropical strains of Bd and provides a baseline for further evolutionary and functional analyses.

Estudio de la mdduración de carne de cordero empleando electroforesis sds-page / Study of lamb meat ageing using electrophoresis sds-pgee / Estudo domaturaçção cordeiros usando eletroforese sds-page

La maduración de la carne es causada por reacciones metabólicas que producen cambios en su calidad, mejorando las características organolépticas como la terneza, la jugosidad y el color. El presente estudio tuvo como objetivo encontrar los cambios producidos en las proteínas sarcoplasmáticas y miofibrilares durante la instauración del rigor mortis y la maduración en carne de cordero semiestabulado (AS) y cordero alimentado por pastoreo (AP). Para tal fin se tomaron muestras del músculo Longissimus dorsi de corderos de raza Dorper x criollo ¾ de diez semanas de edad. Las proteínas sarcoplasmáticas se extrajeron con agua y solución salina diluida, y las proteínas miofibrilares con solución salina concentrada. La cuantificación se hizo por el método de Bradford y la separación por electroforesis SDS-PAGE. En las proteínas sarcoplasmáticas se observó la aparición de una banda de 146 kDa a los cinco y trece días para los corderos AS y AP, respectivamente. En las proteínas miofibrilares se encontró evidencia del rigor mortis a las 16 h y una intensificación drástica de una banda de 28 kDa a las 16 h en el cordero AS. Esta banda también se observó en el cordero AP a las 14 h. En todo el proceso se pudo ver la degradación gradual de la DESMINA, ACTINA Y TROPONINA T.

The ageing of meat is caused by metabolic reactions than result in changes in its quality, improving its organoleptics characteristics such as the tenderness, the juiciness and the color. This study had the aim to find changes in sarcoplasmic and myofibrillar proteins during the establishment of rigor mortis and the ageing in lamb meat: semi feedloot (SF) and lamb feeding for pasture (FP). For this purpose, were sampled Longissimus dorsi muscle of lambs bred Dorper x Criollo ¾ and 10 weeks old. Sarcoplasmic proteins were extracted with water or saline solution diluted and the myofibrillar protein with saline solution concentrate. The Quantification was done by the Bradford method and the separation by SDS-PAGE electrophoresis. In the sarcoplasmic proteins observed the appearance of a band of 146 kDa in the samples of five and thirteen days for lamb SB and AP, respectively. In the myofibrillar proteins found evidence of the establishment of rigor mortis at 16 hours and a drastic intensification of a band of 27 kDa to 16 hours in the lamb SB. This band was also observed in the lamb FP at 14 hours. In the whole process could see the gradual degradation of Desmin, Actin and Troponin T.

A maturação da carne é causada por reações metabólicas que resultam em alterações na qualidade, melhorando as características organolépticas, tais como a suculência, ternura e cor. Este estudo teve como objetivo encontrar alterações nas proteínas miofibrilares sarcoplasmático e durante o início do rigor mortis e maturação na carne: cordeiro Semiestabulado (AS) e pastagem-fed cordeiro (AP). Para o efeito, amostras do músculo Longissimus dorsi de cordeiros Dorper x criollo ¾ de 10 semanas de idade. As proteínas sarcoplasmático foram extraídas com água ou soro fisiológico e diluído com concentrado de proteína miofibrilar salina. Quantificação foi feita pelo método de Bradford e separação por electroforese SDS-PAGE. Nas proteínas sarcoplasmático observado o aparecimento de uma banda de 146 kDa cinco e 13 dias de cordeiro AS e AP, respectivamente. As proteínas miofibrilares encontraram evidências de rigor mortis em 16 horas e uma intensificação drástica de uma banda de 28 kDa a 16 horas no cordeiro AS. Esta banda também foi observada na AP cordeiro em 14 horas. Durante todo o processo podia ver a degradação gradual da Desmina, Actina e Troponina T.

Amatoxin and phallotoxin composition in species of the genus Amanita in Colombia: a taxonomic perspective.

Some species in the genus Amanita have a great variety of toxic secondary metabolites. They are characterized macroscopically by having a white spore print and free gills, and microscopically by the presence of a divergent hymenophoral trama. Some species of Amanita present in Colombia were chemically characterized by analyzing their toxin composition using HPLC. Samples were collected in oak (Quercus humboldtii) and pine (Pinus radiata) forests. Twelve species were recovered, Amanita fuligineodisca, Amanita xylinivolva, Amanita flavoconia, Amanita rubescens, Amanita bisporigera, Amanita muscaria, Amanita humboldtii, Amanita sororcula, Amanita brunneolocularis, Amanita colombiana, Amanita citrina, Amanita porphyria as well as two unreported species. Results showed that most of the analyzed species have α -amanitin in concentrations ranging from 50 ppm to 6000 ppm. Concentrations of α-amanitin in the pileus were significantly greater than in the stipe. Phalloidin and phallacidin were only present in A. bisporigera. Chromatographic profiles are proposed as an additional taxonomic tool since specific peaks with similar retention times were conserved at the species level.

Differential PbP27 expression in the yeast and mycelial forms of the Paracoccidioides brasiliensis species complex.

p27 is an antigenic protein produced by Paracoccidioides brasiliensis, the etiologic agent of paracoccidioidomycosis (PCM). Despite its unknown function, it has been suggested as a putative virulence factor, proposed as a suitable target for the design of diagnostic tools and vaccines, and considered as an enhancer in antifungal treatment of PCM. We evaluated sequence polymorphisms of PbP27 gene sequence among isolates, finding some polymorphisms associated with the isolates' phylogenetic origin. In order to determine if there was a differential expression pattern between morphological states and among isolates, we also evaluated PbP27 expression, at transcriptional and translational levels, in mycelia and yeast cultures in 14 isolates belonging to the P. brasiliensis species complex (S1, PS2, PS3, and "Pb01-like", proposed to be named Paracoccidioides lutzii) by two techniques, real time RT-PCR (RT-qPCR) and protein dot blot. For the latter, four protein extracts from different cell localizations (SDS or ß-mercaptoethanol, cytoplasmic and extracellular proteins) were analyzed for each isolate. p27 was present in the four extracts evaluated, mainly in the SDS extract, corresponding to an extract containing proteins loosely attached to the cell wall. This information correlates with immunohistochemical analysis, where positive staining of the yeasts' cell wall was observed. We found that p27 was present in all isolates, mainly in the yeast form. This pattern was corroborated by RT-qPCR results, with higher expression levels found in the yeast form for most of the isolates. The results provide new insights into the expression patterns of this protein, and further characterize it in view of potential uses as a diagnostic and/or therapeutic tool.

Selection of antagonistic bacteria isolated from the Physalis peruviana rhizosphere against Fusarium oxysporum.

AIMS: Cape gooseberries (Physalis peruviana) have become increasingly important in Colombia for both domestic consumption and the international export market. Vascular wilting caused by Fusarium oxysporum is the most damaging disease to P. peruviana crops in Colombia. The control of this pathogen is mainly carried out by chemical and cultural practices, increasing production costs and generating resistance. Therefore, the objectives of this study were to test rhizobacteria isolates from P. peruviana rhizosphere against F. oxysporum under in vitro and in vivo conditions. METHODS AND RESULTS: Over 120 strains were isolated, and five were selected for their high inhibition of F. oxysporum growth and conidia production under in vitro conditions. These strains inhibited growth by 41-58% and reduced three- to fivefold conidia production. In the in vivo assays, all the tested isolates significantly reduced fungal pathogenicity in terms of virulence. Isolate B-3.4 was the most efficient in delaying the onset of the first symptoms. All isolates were identified as belonging to the genus Pseudomonas except for A-19 (Bacillus sp.). CONCLUSIONS: Our results confirmed that there are prospective rhizobacteria strains that can be used as biological control agents; some of them being able to inhibit in vitro F. oxysporum growth and sporulation. SIGNIFICANCE AND IMPACT OF THE STUDY: Incorporating these bacteria into biological control strategies for the disease that causes high economical losses in the second most exported fruit from Colombia would result in a reduced impact on environment and economy.

Phylogeography and molecular epidemiology of Papaya ringspot virus.

Papaya ringspot virus (PRSV) is the most important virus affecting papaya and cucurbit plants in tropical and subtropical areas. PRSV isolates are divided into biotypes P and W: both the P and W types naturally infect plants in the family Cucurbitaceae, whereas the P type naturally infects papaya (Carica papaya). Understanding the origin and nature of the PRSV genetic diversity and evolution is critical for the implementation of control strategies based on cross-protection and the deployment of transgenic plants that show resistance to virus isolates highly similar to the transgene. The molecular epidemiology of PRSV was evaluated by analyzing the nucleotide sequence of the capsid protein (CP) and helper component-proteinase (HC-Pro) genes of isolates from around the world, including newly characterized ones from Colombia and Venezuela, using a relaxed molecular clock-based approach and a phylogeographic study. Our results confirm previous estimates on the origin of PRSV around 400 years ago and suggest distinct dispersion events from the Indian Peninsula to the rest of Asia, via Thailand, and subsequently to the Americas. A historical reconstruction of the P- and W-type characters in the phylogenetic study supports the need to revise the hypothesis that PRSV-P derives from PRSV-W since our results suggest that the ancestral state could be either of the two biotypes. Moreover, estimates of epidemic growth predict an increasing genetic diversity of the virus over time that has direct implications for control strategies of PRSV based on cross-protection and the use of transgenic plants.

First Report of Phytophthora infestans Causing Late Blight on Solanum viarum in Colombia.

Solanum viarum Dunal (tropical soda apple) belongs to the section Acanthophora in the genus Solanum, which includes nearly 20 neotropical species of herbs and small shrubs (2). The species in this section are sometimes called the 'spiny Solanums' (2) and are adapted mainly to highly disturbed habitats and open secondary forests. The center of diversity is eastern Brazil (3). Since the early 1990s, S. viarum has been a problematic weed in Florida where it was listed as a noxious weed in 1993, followed in 1994 by its addition to the Federal Noxious Weed List of the USDA. On 17 April 2010, 12 plants of S. viarum located close to a S. betaceum crop (tree tomato) in the province of Caldas (Department of Antioquia, central northwestern Colombia) were found with symptoms similar to late blight caused by Phytophthora infestans on S. tuberosum (potato). Fifteen leaves from 12 plants with blackish, water-soaked lesions showing a white sporulation on the abaxial side were collected and processed within 3 days. The leaves were placed in a humid chamber and incubated in darkness at room temperature (18°C mean temperature) until sporulation was observed. Microscopic characteristics were consistent with Phytophthora spp. Only one axenic culture was obtained by successive subcultures in rye B agar plates. After an incubation period of 8 days, plates were washed with distilled water and ovoid, semipapillate caduceus sporangia ranging from 38 to 41 µm long (average 39; N = 86) and 23 to 29 µm wide (average 26; N = 86) were observed. To fulfill Koch's postulates and test the isolate for the ability to infect potato as well as Solanum spp. associated with potato crops in Colombia, triplicate pathogenicity tests were carried out on three detached leaves of S. viarum, S. tuberosum, and S. americanum (American nightshade). A 1 × 104 sporangia/ml suspension of the Phytophthora isolate, estimated using a haemocytometer, was obtained from 8-day-old cultures grown on rye B agar. The suspension was incubated at 4°C for 2 h to induce zoospore release. The leaves were then inoculated by spraying them until runoff. After an incubation period of 5 days at 18°C in a humidity chamber, mycelia, sporangia, and brownish lesions, similar to those described above, were observed in the leaves of all three hosts, indicating pathogenicity. DNA extraction was performed from the P. infestans isolate (4). Four nuclear loci, ITS, ß-tubulin, Ras, and Avr3a, as well as one mitochondrial gene, cytochrome c oxidase 1 (Cox1), were amplified and sequenced. Sequences were compared with GenBank databases using Blastn. In all cases, the best hits corresponded to P. infestans (GenBank Accession No. HQ639930 for Avr3A, HQ639931 for ß-tubulin, HQ639932 for Cox1, HQ639933 for iRas, HQ639934 for Ras, and JF419363 for ITS). Reports of P. infestans causing typical late blight symptoms on wild solanaceous plants are becoming more frequent and have been made from other countries such as Peru (1). To our knowledge, this is the first time that P. infestans has been observed and isolated from S. viarum in Colombia, introducing the possibility of this wild solanaceous weed as another late blight host. References: (1) G. Garry et al. Eur. J. Plant Pathol. 113:71, 2005. (2) R. Levin et al. Am. J. Bot. 92:603, 2005. (3) M. Nee. A Revision of Solanum Section Acanthophora. Ph.D. diss. University of Wisconsin, Madison, 1979. (4) A. M. Vargas et al. Phytopathology 99:82, 2009.

Physiological and molecular characterization of atypical isolates of Malassezia furfur.

The species constituting the genus Malassezia are considered to be emergent opportunistic yeasts of great importance. Characterized as lipophilic yeasts, they are found in normal human skin flora and sometimes are associated with different dermatological pathologies. We have isolated seven Malassezia species strains that have a different Tween assimilation pattern from the one typically used to differentiate M. furfur, M. sympodialis, and M. slooffiae from other Malassezia species. In order to characterize these isolates of Malassezia spp., we studied their physiological features and conducted morphological and molecular characterization by PCR-restriction fragment length polymorphism and sequencing of the 26S and 5.8S ribosomal DNA-internal transcribed spacer 2 regions in three strains from healthy individuals, four clinical strains, and eight reference strains. The sequence analysis of the ribosomal region was based on the Blastn algorithm and revealed that the sequences of our isolates were homologous to M. furfur sequences. To support these findings, we carried out phylogenetic analyses to establish the relationship of the isolates to M. furfur and other reported species. All of our results confirm that all seven strains are M. furfur; the atypical assimilation of Tween 80 was found to be a new physiological pattern characteristic of some strains isolated in Colombia.