La marchitez tardía del maíz (Zea mays L.) causada por Cephalosporium maydis en la Península Ibérica, y otros hongos asociados

Las especies de hongos de suelo asociadas a Cephalosporium maydis como agente causal de la marchitez tardía del maíz en la Península Ibérica se identificaron muestreando 19 campos con síntomas de marchitez en las principales zonas de cultivo entre 2011 y 2012. En el 47% de los campos no se identificó C. maydis, pero sí Fusarium graminearum, F. verticillioides, F. equiseti, F. proliferatum, Macrophomina phaseolina, Rhizoctonia solani y Trichoderma harzianum infectando las plantas de maíz. En los campos restantes, junto a C. maydis se identificaron otros hongos de suelo en porcentajes apreciables: F. verticillioides (19%), F. proliferatum (19%), F.equiseti (9%), F. oxysporum (9%) y Pythium oligandrum (9%). El crecimiento vascular de C. maydis y de otras especies fúngicas en plantas de maíz se confirmó analizando plantas con marchitez procedentes de tres campos diferentes. Tanto C. maydis como F. graminearum, F. equiseti, F. proliferatum y T. harzianum se aislaron de la inserción entre la raíz y tallo y a 10 cm de altura en el tallo de las plantas. El efecto de la infección por C. maydis sobre la producción de las plantas de maíz se cuantificó en macetas y condiciones seminaturales en el 2011. En plantas inoculadas se obtuvo una reducción del peso de las mazorcas del 54%, además de pesos de raíz y de parte aérea (tallo y hojas) significativamente menores en comparación con el control no inoculado, lo que sugiere el gran impacto económico que puede tener la marchitez tardía en condiciones naturales. Asimismo este trabajo pone de manifiesto el grado de complejidad de la etiología de la marchitez tardía, que debería ser estudiado mediante la confirmación de la patogenicidad de los hongos de suelo identificados en maíz, con el fin de determinar el papel que puede jugar cada una de estas especies en el desarrollo de la enfermedad y/o severidad de los síntomas.

Atividade antif��ngica de extratos de alho e capim-santo sobre o desenvolvimento de Fusarium proliferatum isolado de gr��os de milho

Dentre os pat��genos veiculados pelas sementes e gr��os do milho (Zea mays L.), Fusarium proliferatum causa tombamento e morte de pl��ntulas, podrid��o de ra��zes, de colmo, de espiga e gr��os. Assim, o tratamento de sementes mostra-se como importante medida para preservar a qualidade. O objetivo do presente trabalho foi avaliar a atividade antif��ngica de diferentes concentra����es dos extratos de alho (Allium sativum L.) e capim-santo (Cymbopogon citratus Stapf.) visando o controle de F. proliferatum em sementes de milho e observar os efeitos dos tratamentos sobre a germina����o de sementes, sanidade e desenvolvimento das pl��ntulas. A partir de gr��os de milho, foi feito o isolamento de F. proliferatum, que foi cultivado em meio BDA. Foram avaliados os efeitos dos extratos de alho e capim santo sobre crescimento micelial, nas concentra����es 0,5%, 1,0%, 2,5%, 5,0% e 10,0%, medindo-se os di��metros das col��nias do fungo durante oito dias. Esporos de F. proliferatum foram imersos em solu����es dos extratos, nas concentra����es mencionadas, e avaliados, quanto �� germina����o de con��dios, ��s 6, 12, 18 e 24 horas de imers��o. Sementes de milho foram tratadas em solu����es dos extratos e inoculadas com F. proliferatum, sendo avaliada quanto �� incid��ncia do fungo, percentagem de germina����o e incid��ncia de tombamento e podrid��o do colmo das pl��ntulas. Os extratos empregados reduziram a taxa de crescimento micelial e a germina����o dos esporos, como tamb��m a incid��ncia de F. proliferatum em gr��os de milho. O extrato de alho, a partir da concentra����o 2,5%, mostrou maior efici��ncia em rela����o aos demais tratamentos. Os produtos vegetais aumentaram a germina����o das sementes e tamb��m controlaram o tombamento e a podrid��o do colmo das pl��ntulas de milho.

La podredumbre del ajo causada por Fusarium proliferatum

La producci��n anual de ajo en Espa��a es de 154.587 t y su cultivo ocupa unas 15.900 ha distribuidas principalmente en Castilla La Mancha (53,1%), Andaluc��a (27,4%) y Castilla y Le��n (9,1%). En Castilla La Mancha, la principal Comunidad productora, se ha cultivado en la presente campa��a un 7 % m��s que en la campa��a 2009/2010, siendo Albacete la provincia con mayor superficie cultivada de ajos con 4.750 ha, seguida de Cuenca con 2.200 ha. Las plagas y enfermedades en el cultivo del ajo suponen elevadas p��rdidas econ��micas cada a��o en nuestro pa��s. En 2008, distintos agricultores de varios municipios de Castilla y Le��n detectaron bulbos de ajo del cultivar ���Blancomor de Vallelado��� que presentaban s��ntomas de podredumbre h��meda durante el almacenamiento. Posteriormente, en el a��o 2009, esta misma podredumbre se observ�� tambi��n en las provincias de Albacete y Cuenca en el cultivar ���Morado de Pedro��eras���. Estudios sobre esta nueva enfermedad se est��n llevando a cabo en colaboraci��n con productores castellanomanchegos pertenecientes a Coopaman SCL

Divergence of the IGS rDNA in Fusarium proliferatum and Fusarium globosum reveals two strain specific non-orthologous types

A phylogenic analysis of Fusarium proliferatum and closely related species was performed using the most variable part within the intergenic spacer of the nuclear ribosomal DNA (IGS) and compared with a previously reported phylogeny performed in the same group of samples with a partial region of the nuclear single copy gene encoding the elongation factor 1�� (EF-1��). The phylogenies from both genomic sequences were not concordant and revealed the presence of two nonorthologous IGS types, named types I and II, in F. proliferatum and Fusarium globosum. Two specific PCR assays designed to amplify either IGS type I or type II revealed that only one IGS type was present in each individual in these two species. The presence of both IGS types at the species level indicates that homogenization has not been achieved yet. This might be retarded if panmictic sexual reproduction was affected by certain levels of clonal reproduction and/or by the diverse hosts that these species are able to colonize. This study indicates that taxonomic studies carried out with the IGS rDNA, which has been widely used in Fusarium, should be undertaken with caution.

Fusarium proliferatum isolated from garlic in Spain: Identification, toxigenic potential and pathogenicity on related Allium species

Fusarium proliferatum has been reported on garlic in the Northwest USA, Spain and Serbia, causing water-soaked tan-colored lesions on cloves. In this work, Fusarium proliferatum was isolated from 300 symptomatic garlic bulbs. Morphological identification of Fusarium was confirmed using species-specific PCR assays and EF-1�� sequencing. Confirmation of pathogenicity was conducted with eighteen isolates. Six randomly selected F. proliferatum isolates from garlic were tested for specific pathogenicity and screened for fusaric acid production. Additionally, pathogenicity of each F. proliferatum isolate was tested on healthy seedlings of onion (Allium cepa), leek (A. porrum), scallions (A. fistulosum), chives (A. schoenoprasum) and garlic (A. sativum). A disease severity index (DSI) was calculated as the mean severity on three plants of each species with four test replicates. Symptoms on onion and garlic plants were observed three weeks after inoculation. All isolates tested produced symptoms on all varieties inoculated. Inoculation of F. proliferatum isolates from diseased garlic onto other Allium species provided new information on host range and pathogenicity. The results demonstrated differences in susceptibility with respect to host species and cultivar. The F. proliferatum isolates tested all produced fusaric acid (FA); correlations between FA production and isolate pathogenicity are discussed. Additionally, all isolates showed the presence of the FUM1 gene suggesting the ability of Spanish isolates to produce fumonisins.

The effects of storage duration, temperature and cultivar on the severity of garlic clove rot caused by Fusarium proliferatum.

Diseases that affect garlic during storage can lead to severe economic losses for farmers worldwide. One causal agent of clove rot is Fusarium proliferatum. Here, the progress of clove rot caused by F. proliferatum and its dependence on different storage conditions and cultivar type were studied. The effect of temperature on mycelial growth, conidial viability, and fungal survival during garlic commercial storage was documented. Samples of 50 bulbs from a randomized field trial with three different clonal generations for purple garlic (F3, F4 and F5) and the F4 clonal generation for white garlic were labeled and stored for two months (short-term storage). In addition, another sample of the F5 clonal generation of purple garlic was stored for 6 months after harvest (long-term storage). The presence of the pathogen and the percentage of symptomatic cloves were evaluated. A notable difference in the rot severity index (RSI) of different garlic varieties was observed. In all studied cases, clove rot increased with storage time at 20 ��� C, and the white garlic variety had a higher index of rot severity after two months of storage. Additionally, there were clear differences between the growth rates of F. proliferatum isolates. Studies conducted on the temperature responses of the pathogen propagules showed that expo- sure for at least 20 min at 50 ��� C was highly effective in significantly reducing the viability of fungal conidia. Pathogenicity studies showed that the fungus is pathogenic in all commercial varieties. However, there were significant differences in varietal susceptibility between Chinese and white garlic type cultivars (81.84 �� 16.44% and 87.5 �� 23.19% symptomatic cloves, respectively) and purple cultivars (49.06 �� 13.42% symptomatic cloves)

Infec����o por Fusarium graminearum e Fusarium verticillioides em sementes de milho

The previous knowledge of the infection process and pathogens behavior, for evaluating the physiological potential of maize seeds, is essential for decision making on the final destination of lots that can endanger sowing. This research was carried out in order to study the minimum period required for maize seeds contamination by Fusarium graminearum Schwabe and Fusarium verticillioides (Sacc.) Nirenberg, as well as these pathogens influence on seed germination and vigor, by using the cold test. Three maize seeds hybrids, kept in contact with the pathogens for different periods, were evaluated with and without surface disinfection. After determining the most suitable period, new samples were contaminated by F. graminearum and F. verticillioides, under different infection levels, and subjected to germination tests in sand. The cold test was conducted with healthy and contaminated seeds, at different periods, in a cold chamber. The contact of maize seeds with F. graminearum and F. verticillioides for 16 hours was enough to cause infection. F. graminearum and F. verticillioides did not affect the maize seeds germination, however, F. graminearum reduced the vigor of seeds lots.

Identifica����o, caracteriza����o e avalia����o da patogenicidade de diferentes isolados de fusarium spp. para o controle de thaumastocoris peregrinus (hemiptera: thaumastocoridae)

P��s-gradua����o em Agronomia (Prote����o de Plantas) - FCA

The Virome of two Fusarium spp. Collections and its Potential for Controlling Fusarium Disease

This research aims to discover the virome diversity and composition in Fusarium poae and Fusarium proliferatum collections, characterize the mycovirus that may have an effect on host pathogenicity to provide potential materials for the biological control of Fusarium spp. pathogens. Next-Generation Sequencing (NGS) analysis of 30 F. poae isolates revealed an extreme diversity of mycoviruses. Bioinformatic analysis shows that contigs associated with viral genome belong to the families: Hypoviridae, Mitoviridae, Partitiviridae, Polymycoviridae, proposed Alternaviridae, proposed Fusagraviridae, proposed Fusariviridae, proposed Yadokariviridae, and Totiviridae. The complete genomes of 12 viruses were obtained by assembling contigs and overlapping cloning sequences. Moreover, all the F. poae isolates analyzed are multi-infected. Fusarium poae partitivirus 1 appears in all the 30 strains, followed by Fusarium poae fusagravirus 1 (22), Fusarium poae mitovirus 2 (18), Fusarium poae partitivirus 3 (16), and Fusarium poae mitovirus 2 and 3 (11). Using the same approach, the virome of F. proliferatum collections resulted in lower diversity and abundance. The identified mycoviruses belong to the family Mitoviridae and Mymonaviridae. Interestingly, most F. proliferatum isolates are not multi-infected. The complete genomes of four viruses were obtained by assembling contigs and overlapping cloning sequences. By multiple liner regression of the virome composition and growth rate of 30 F. poae, Fusarium poae mitovirus 3 is significantly correlated with the growth rate among F. poae collection. Furthermore, the principal component analysis of the virome composition from 30 F. poae showed that the presence of Fusarium poae mitovirus 3 and other two viruses could increase the F. poae growth rate. The curing experiment and pathogenicity test in Petri indicated that Fusarium poae hypovirus 1 might be associated with the host hypovirulence phenotype, while Fusarium poae fusagravirus 1 and Fusarium poae partitivirus 3 may have some beneficial effect on host pathogenicity.

Controle biol��gico de thaumastocoris peregrinus (Hemiptera: thaumastocoridae) com fungos entomopatog��nicos

P��s-gradua����o em Agronomia (Prote����o de Plantas) - FCA

Biological control of the bronze bug, Thaumastocoris peregrinus, in eucalyptus plantations in Brazil

The bronze bug Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae) was detected in Brazil in 2008 and infested >180 000 ha of eucalyptus plantations in 2011. The bronze bug can cause a reduction of 10���15% in wood productivity after 2 years of heavy infestation. Although there is not an effective control method known, biological control is the main control strategy studied. An exotic egg parasitoid, Cleruchoides noackae (Hymenoptera: Mymaridae), was imported from Australia in 2012, reared in a laboratory and released in three Brazilian regions. Parasitoids were recovered at release points after 20���30 d. In 2013, preliminary evaluations demonstrated parasitoid establishment in these areas, and the parasitoid was recovered in adjacent areas after 1 year of release. Bioassays confi rmed egg parasitism of 15���20% by C. noackae. Other native natural enemies were studied. We found green lacewing Chrysoperla externa and predatory bugs Supputius cincticeps and Atopozelus opsimus preying on nymphs and/or adults of T. peregrinus. Another promising possibility is entomopathogenic fungi. Commercial formulations of Beauveria bassiana were tested with success in lab and fi eld conditions. Fusarium proliferatum and Paecilomyces cateniannulatus caused mortality of T. peregrinus in natural epizooties. After 5 years of research, it is possible to develop an integrated pest management system (IPM) for eucalyptus plantations based on biocontrol strategy for bronze bug.

Microbiota asociada a la enfermedad de la punta negra del trigo duro. Efectos del riego, el abonado nitrogenado y la variedad cultivada en la incidencia de la enfermedad

La enfermedad f��ngica de la punta negra del trigo se caracteriza por provocar en las semillas afectadas un oscurecimiento de la zona del embri��n que en ocasiones puede extenderse hacia la hendidura central dejando el grano completamente ennegrecido. Su incidencia en el cultivo de trigo es extremadamente variable y depende en gran medida de las condiciones ambientales, as�� pues, condiciones de alta humedad pueden incrementar a punta negra. Su presencia en el trigo duro repercute en el rendimiento semolero. Adem��s la pasta elaborada a partir de semilla enferma presenta manchas negras y adquiere color y olor desagradable. Esta enfermedad es poco conocida a nivel europeo, sin embargo existen numerosos estudios para conocer su etiolog��a y los factores que afectan a su aparici��n en pa��ses como Nueva Zelanda, Australia, Estados Unidos o Canad��. El presente trabajo pretende dar a conocer esta patolog��a en Espa��a, determinando la influencia del riego, el abonado nitrogenado y la variedad cultivada en la incidencia de la enfermedad. Para ello se ha contado con un dise��o experimental basado en 10 cultivares sembrados en parcelas con dos tratamientos de riego y dos de abonado nitrogenado. El an��lisis de las semillas infectadas en c��mara h��meda y medios de cultivo PDA y K revel�� 12 g��neros f��ngicos diferentes, de los cuales Alternaria alternata y Fusarium proliferatum estaban presentes en todas las muestras. El estudio del riego y abonado nitrogenado mostr�� diferencias significativas en la incidencia de punta negra pero fueron los 10 cultivares incluidos en el ensayo los que mayor importancia cobraron desde el punto de vista de la aparici��n de la enfermedad. El genotipo result�� determinante a la hora de establecer los niveles de afectaci��n ya que las muestras encuadradas bot��nicamente como Triticum turgidum subsp. Turgidum convar. Turgidum presentaron una mayor susceptibilidad. las prueas de patogenicidd con los tres principales hongos asociados a la punta negra dieron resultados negativos para la germinaci��n-nascencia de las pl��ntulas de trigo duro inoculadas.

Enfermedades y fisiopat��as que afectan a la calidad del ajo en postcosecha

Las enfermedades y fisiopat��as que afectan al ajo pueden llegar a ocasionar importantes p��rdidas en postcosecha, de vital importancia cuando el producto se destina a la exportaci��n a mercados que cada d��a son m��s exigentes. En 2008, distintos agricultores de varios municipios de Castilla y Le��n detectaron bulbos de ajo que presentaban s��ntomas de podredumbre h��meda durante el almacenamiento. Posteriormente, en el a��o 2009, esta misma podredumbre se observ�� tambi��n en las provincias de Albacete y Cuenca y en muestras procedentes de C��rdoba. Hasta la fecha se ha identificado a Fusarium proliferatum como el agente causal de la podredumbre del diente.

Structure Variation And Dynamics of the Nuclear Ribosomal Intergenic Spacer Region in Key Gibberella Fujikuroi Complex Species

The intergenic spacer (IGS) region of the ribosomal DNA was cloned and sequenced in eight species within the Gibberella fujikuroi species complex with anamorphs in the genus Fusarium , a group that includes the most relevant toxigenic species. DNA sequence analyses revealed two categories of repeated elements: long repeats and short repeats of 125 and 8 bp, respectively. Long repeats were present in two copies and were conserved in all the species analyzed, whereas different numbers of short repeat elements were observed, leading to species-specific IGS sequences with different length. In Fusarium subglutinans and Fusarium nygamai , these differences seemed to be the result of duplication and deletion events. Here, we propose a model based on unequal crossing over that can explain these processes. The partial IGS sequence of 22 Fusarium proliferatum isolates was also obtained to study variation at the intraspecific level. The results revealed no differences in terms of number or pattern of repeated elements and detected frequent gene conversion events. These results suggest that the homogenization observed at the intraspecific level might not be achieved primarily by unequal crossing-over events but rather by processes associated with recombination such as gene conversion events.

Carbon and nitrogen limitation increase chitosan antifungal activity in Neurospora crassa and fungal human pathogens

Chitosan permeabilizes plasma membrane and kills sensitive filamentous fungi and yeast. Membrane fluidity and cell energy determine chitosan sensitivity in fungi. A five-fold reduction of both glucose (main carbon (C) source) and nitrogen (N) increased 2-fold Neurospora crassa sensitivity to chitosan. We linked this increase with production of intracellular reactive oxygen species (ROS) and plasma membrane permeabilization. Releasing N. crassa from nutrient limitation reduced chitosan antifungal activity in spite of high ROS intracellular levels. With lactate instead of glucose, C and N limitation increased N. crassa sensitivity to chitosan further (4-fold) than what glucose did. Nutrient limitation also increased sensitivity of filamentous fungi and yeast human pathogens to chitosan. For Fusarium proliferatum, lowering 100-fold C and N content in the growth medium, increased 16-fold chitosan sensitivity. Similar results were found for Candida spp. (including fluconazole resistant strains) and Cryptococcus spp. Severe C and N limitation increased chitosan antifungal activity for all pathogens tested. Chitosan at 100 ��g ml-1 was lethal for most fungal human pathogens tested but non-toxic to HEK293 and COS7 mammalian cell lines. Besides, chitosan increased 90% survival of Galleria mellonella larvae infected with C. albicans. These results are of paramount for developing chitosan as antifungal.