Los científicos del futuro están en la escuela

El proyecto de extensión universitaria “Los Científicos del Futuro están en la Escuela” se encuentra acreditado desde el año 2004 por la Facultad de Ciencias Exactas de la UNLP. El mismo propone la divulgación, la promoción del interés por la ciencia y la tecnología, y la complementación de la enseñanza de ciencias naturales a nivel primario.Centro de Investigación y Desarrollo en Ciencias Aplicada

Los científicos del futuro están en la escuela

El proyecto de extensión universitaria “Los Científicos del Futuro están en la Escuela” se encuentra acreditado desde el año 2004 por la Facultad de Ciencias Exactas de la UNLP. El mismo propone la divulgación, la promoción del interés por la ciencia y la tecnología, y la complementación de la enseñanza de ciencias naturales a nivel primario.Centro de Investigación y Desarrollo en Ciencias Aplicada

Los científicos del futuro están en la escuela

El proyecto de extensión universitaria “Los Científicos del Futuro están en la Escuela” se encuentra acreditado desde el año 2004 por la Facultad de Ciencias Exactas de la UNLP. El mismo propone la divulgación, la promoción del interés por la ciencia y la tecnología, y la complementación de la enseñanza de ciencias naturales a nivel primario.Centro de Investigación y Desarrollo en Ciencias Aplicada

Alkane-grown Beauveria bassiana produce mycelial pellets displaying peroxisome proliferation, oxidative stress, and cell surface alterations

The entomopathogenic fungus Beauveria bassiana is able to grow on insect cuticle18 hydrocarbons as the sole carbon source, inducing several enzymes involved in alkane19 assimilation and concomitantly increasing virulence against insect hosts. In this study, we20 describe some physiological and molecular processes implicated in growth, nutritional21 stress response, and cellular alterations found in alkane-grown fungi. The fungal cytology22 was investigated using light and transmission electron microscopy (TEM) while the surface23 topography was examined using atomic force microscopy (AFM). Fungal hydrophobicity24 was also measured on the cell surface. Additionally, the expression pattern of several genes25 associated with oxidative stress, peroxisome biogenesis, and hydrophobicity were analysed26 by qPCR. We found a novel type of growth in alkane-cultured B. bassiana similar to27 mycelial pellets described in other alkane-free fungi, which were able to germinate and28 produce viable conidia in media without a carbon source and to be pathogenic against29 larvae of the beetles Tenebrio molitor and Tribolium castaneum. Optical microscopy and30 TEM showed that pellets were formed by hyphae cumulates with high peroxidase activity,31 exhibiting peroxisome proliferation and an apparent surface thickening. Alkane-grown32 conidia appeared to be more hydrophobic and cell surfaces displayed different topography33 than glucose-grown cells, as it was observed by AFM. We also found a significant34 induction in several genes encoding for peroxins, catalases, superoxide dismutases, and35 hydrophobins. These results show that both morphological and metabolic changes are36 triggered in mycelial pellets derived from alkane-grown B. bassiana.Fil: Huarte Bonnet, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Santos Da Paixao, Flavia Regina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Ponce, Juan C. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Santana, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Prieto, Eduardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas; ArgentinaFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentin

Tenebrionid secretions and a fungal benzoquinone oxidoreductase form competing components of an arms race between a host and pathogen

Entomopathogenic fungi and their insect hosts represent a model system for examining invertebrate-pathogen coevolutionary selection processes. Here we report the characterization of competing components of an arms race consisting of insect protective antimicrobial compounds and evolving fungal mechanisms of detoxification. The insect pathogenic fungus Beauveria bassiana has a remarkably wide host range; however, some insects are resistant to fungal infection. Among resistant insects is the tenebrionid beetle Tribolium castaneum that produces benzoquinone-containing defensive secretions. Reduced fungal germination and growth was seen in media containing T. castaneum dichloromethane extracts or synthetic benzoquinone. In response to benzoquinone exposure, the fungus expresses a 1,4-benzoquinone oxidoreductase, BbbqrA, induced >40-fold. Gene knockout mutants (ΔBbbqrA) showed increased growth inhibition, whereas B. bassiana overexpressing BbbqrA (Bb::BbbqrAO) displayed increased resistance to benzoquinone compared with wild type. Increased benzoquinone reductase activity was detected in wild-type cells exposed to benzoquinone and in the overexpression strain. Heterologous expression and purification of BbBqrA in Escherichia coli confirmed NAD(P)H-dependent benzoquinone reductase activity. The ΔBbbqrA strain showed decreased virulence toward T. castaneum, whereas overexpression of BbbqrA increased mortality versus T. castaneum. No change in virulence was seen for the ΔBbbqrA or Bb::BbbqrAO strains when tested against the greater wax moth Galleria mellonella or the beetle Sitophilus oryzae, neither of which produce significant amounts of cuticular quinones. The observation that artificial overexpression of BbbqrA results in increased virulence only toward quinone-secreting insects implies the lack of strong selection or current failure of B. bassiana to counteradapt to this particular host defense throughout evolution.Instituto de Investigaciones Bioquímicas de La Plat

Riboflavin induces Metarhizium spp. to produce conidia with elevated tolerance to UV-B, and upregulates photolyases, laccases and polyketide synthases genes

Aims: The effect of nutritional supplementation of two Metarhizium species with riboflavin (Rb) during production of conidia was evaluated on (i) conidial tolerance (based on germination) to UV-B radiation and on (ii) conidial expression following UV-B irradiation, of enzymes known to be active in photoreactivation, viz., photolyase (Phr), laccase (Lac) and polyketide synthase (Pks). Methods and Results: Metarhizium acridum (ARSEF 324) and Metarhizium robertsii (ARSEF 2575) were grown either on (i) potato dextrose agar medium (PDA), (ii) PDA supplemented with 1% yeast extract (PDAY), (iii) PDA supplemented with Rb (PDA+Rb), or (iv) PDAY supplemented with Rb (PDAY+Rb). Resulting conidia were exposed to 866·7 mW m−2 of UV-B Quaite-weighted irradiance to total doses of 3·9 or 6·24 kJ m−2. Some conidia also were exposed to 16 klux of white light (WL) after being irradiated, or not, with UV-B to investigate the role of possible photoreactivation. Relative germination of conidia produced on PDA+Rb (regardless Rb concentration) or on PDAY and exposed to UV-B was higher compared to conidia cultivated on PDA without Rb supplement, or to conidia suspended in Rb solution immediately prior to UV-B exposure. The expression of MaLac3 and MaPks2 for M. acridum, as well as MrPhr2, MrLac1, MrLac2 and MrLac3 for M. robertsii was higher when the isolates were cultivated on PDA+Rb and exposed to UV-B followed by exposure to WL, or exposed to WL only. Conclusions: Rb in culture medium increases the UV-B tolerance of M. robertsii and M. acridum conidia, and which may be related to increased expression of Phr, Lac and Pks genes in these conidia. Significance and Impact of the Study: The enhanced UV-B tolerance of Metarhizium spp. conidia produced on Rb-enriched media may improve the effectiveness of these fungi in biological control programs.Fil: Pereira-Junior, R.A.. Universidade Federal de Goiás; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Huarte Bonnet, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina. Universidade Federal de Goiás; BrasilFil: Paixão, F.R.S.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina. Universidade Federal de Goiás; BrasilFil: Roberts, D.W.. State University of Utah; Estados UnidosFil: Luz, C.. Universidade Federal de Goiás; BrasilFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Fernandes, É.K.K.. Universidade Federal de Goiás; Brasi

Alkane-grown <i>Beauveria bassiana</i> produce mycelial pellets displaying peroxisome proliferation, oxidative stress, and cell surface alterations

The entomopathogenic fungus Beauveria bassiana is able to grow on insect cuticle hydrocarbons as the sole carbon source, inducing several enzymes involved in alkane assimilation and concomitantly increasing virulence against insect hosts. In this study, we describe some physiological and molecular processes implicated in growth, nutritional stress response, and cellular alterations found in alkane-grown fungi. The fungal cytology was investigated using light and transmission electron microscopy (TEM) while the surface topography was examined using atomic force microscopy (AFM). Fungal hydrophobicity was also measured on the cell surface. Additionally, the expression pattern of several genes associated with oxidative stress, peroxisome biogenesis, and hydrophobicity were analysed by qPCR. We found a novel type of growth in alkane-cultured B. bassiana similar to mycelial pellets described in other alkane-free fungi, which were able to germinate and produce viable conidia in media without a carbon source and to be pathogenic against larvae of the beetles Tenebrio molitor and Tribolium castaneum. Optical microscopy and TEM showed that pellets were formed by hyphae cumulates with high peroxidase activity, exhibiting peroxisome proliferation and an apparent surface thickening. Alkane-grown conidia appeared to be more hydrophobic and cell surfaces displayed different topography than glucose-grown cells, as it was observed by AFM. We also found a significant induction in several genes encoding for peroxins, catalases, superoxide dismutases, and hydrophobins. These results show that both morphological and metabolic changes are triggered in mycelial pellets derived from alkane-grown B. bassiana.Instituto de Investigaciones Bioquímicas de La PlataInstituto de Investigaciones Fisicoquímicas Teóricas y Aplicada

Puesta en valor de la extensión universitaria: curricularización para una integración real con investigación y docencia

This work is presented as the result of a complex, continuum and long reflexive process about our teaching practices along different workshops, meetings and collaborative exchanges in our teaching formation journey. The text includes our revisions about educational fragmentation and the importance of an integral, social and collectively constructed professional formation. We consider the importance of the articulation between teaching, research and extension practices for the social, political and historically situated teaching and learning processes, which promote a professional profile more committed to political-social problems, and we place the University as a fundamental institutional actor in the permanent exchange with the community. We also include concrete proposals for curricularisation of extension as an innovative pedagogical strategy to implement in university teaching practices in response to the problems addressed.Este trabajo se origina en respuesta a un proceso reflexivo complejo y continuo de nuestras prácticas docentes a lo largo de distintos encuentros, talleres e intercambios de la especialización en docencia universitaria de la Universidad Nacional de La Plata. A lo largo del texto plasmamos nuestras revisiones sobre la fragmentación en la educación y la necesidad de una formación integral, social y colectivamente construida. Consideramos la importancia de la articulación entre docencia, investigación y extensión para los procesos de enseñanza y aprendizaje social, política e históricamente situados, que promuevan un perfil profesional y ciudadano más comprometido con las problemáticas político-sociales, y situamos a la Universidad como actor institucional fundamental en el intercambio permanente con la comunidad. Incluimos también propuestas concretas de curricularización de la extensión como estrategia pedagógica innovadora para implementar en las prácticas docentes universitarias en respuesta a las problematizaciones abordadas

Tolerance to Abiotic Factors of Microsclerotia and Mycelial Pellets From Metarhizium robertsii, and Molecular and Ultrastructural Changes During Microsclerotial Differentiation

Metarhizium spp. fungi are able to produce resistant structures termed microsclerotia, formed by compacted and melanized threads of hyphae. These propagules are tolerant to desiccation and produce infective conidia, thus they are promising candidates to use in biological control programs. In this study, we investigated the tolerance to both UV-B radiation and heat of microsclerotia of M. robertsii strain ARSEF 2575. We also adapted the liquid medium and culture conditions to obtain mycelial pellets from the same isolate in order to compare these characteristics between both types of propagules. We followed the peroxisome biogenesis and studied the oxidative stress during differentiation from conidia to microsclerotia by transmission electron microscopy after staining with a peroxidase activity marker, and by the expression pattern of genes potentially involved in these processes. We found that despite their twice smaller size, microsclerotia exhibited higher dry biomass, yield, and conidial productivity than mycelial pellets, both with and without UV-B and heat stresses.. From the sixteen genes measured, we found an induction after 96-h differentiation in the oxidative stress marker genes MrcatA, MrcatP, and Mrgpx, the peroxisome biogenesis factors Mrpex5 and Mrpex14/17, and the photoprotection genes Mrlac1, Mrlac2, and Mrlac3. We concluded that an oxidative stress scenario is induced during microsclerotia differentiation in M. robertsii, and confirmed that due to its tolerance to desiccation, heat, and UV-B, this fungal structure could be an excellent candidate for use in biological control of pests under tropical and subtropical climates where heat and UV radiation are detrimental to entomopathogenic fungi survival and persistence.Fil: Paixão, Flávia R. S.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Huarte Bonnet, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Ribeiro-Silva, Cárita de S.. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Mascarin, Gabriel M.. Empresa Brasilera de Pesquisa Agropecuaria.; BrasilFil: Fernandes, Éverton. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

Targeting of insect epicuticular lipids by the entomopathogenic fungus Beauveria bassiana: Hydrocarbon oxidation within the context of a host-pathogen interaction

Broad host range entomopathogenic fungi such as Beauveria bassiana attack insect hosts via attachment to cuticular substrata and the production of enzymes for the degradation and penetration of insect cuticle. The outermost epicuticular layer consists of a complex mixture of non-polar lipids including hydrocarbons, fatty acids, and wax esters. Long chain hydrocarbons are major components of the outer waxy layer of diverse insect species, where they serve to protect against desiccation and microbial parasites, and as recognition molecules or as a platform for semiochemicals. Insect pathogenic fungi have evolved mechanisms for overcoming this barrier, likely with sets of lipid degrading enzymes with overlapping substrate specificities. Alkanes and fatty acids are substrates for a specific subset of fungal cytochrome P450 monooxygenases involved in insect hydrocarbon degradation. These enzymes activate alkanes by terminal oxidation to alcohols, which are further oxidized by alcohol and aldehyde dehydrogenases, whose products can enter β-oxidation pathways. B. bassiana contains at least 83 genes coding for cytochrome P450s (CYP), a subset of which are involved in hydrocarbon oxidation, and several of which represent new CYP subfamilies/families. Expression data indicated differential induction by alkanes and insect lipids and four CYP proteins have been partially characterized after heterologous expression in yeast. Gene knockouts revealed a phenotype for only one (cyp52X1) out of six genes examined to date. CYP52X1 oxidizes long chain fatty acids and participates in the degradation of specific epicuticular lipid components needed for breaching the insect waxy layer. Examining the hydrocarbon oxidizing CYP repertoire of pathogens involved in insect epicuticle degradation can lead to the characterization of enzymes with novel substrate specificities. Pathogen targeting may also represent an important co-evolutionary process regarding insect cuticular hydrocarbon synthesis.Instituto de Investigaciones Bioquímicas de La Plat