Colonization of Onions by Endophytic Fungi and Their Impacts on the Biology of Thrips tabaci

Endophytic fungi, which live within host plant tissues without causing any visible symptom of infection, are important mutualists that mediate plant-herbivore interactions. Thrips tabaci (Lindeman) is one of the key pests of onion, Allium cepa L., an economically important agricultural crop cultivated worldwide. However, information on endophyte colonization of onions, and their impacts on the biology of thrips feeding on them, is lacking. We tested the colonization of onion plants by selected fungal endophyte isolates using two inoculation methods. The effects of inoculated endophytes on T. tabaci infesting onion were also examined. Seven fungal endophytes used in our study were able to colonize onion plants either by the seed or seedling inoculation methods. Seed inoculation resulted in 1.47 times higher mean percentage post-inoculation recovery of all the endophytes tested as compared to seedling inoculation. Fewer thrips were observed on plants inoculated with Clonostachys rosea ICIPE 707, Trichoderma asperellum M2RT4, Trichoderma atroviride ICIPE 710, Trichoderma harzianum 709, Hypocrea lixii F3ST1 and Fusarium sp. ICIPE 712 isolates as compared to those inoculated with Fusarium sp. ICIPE 717 and the control treatments. Onion plants colonized by C. rosea ICIPE 707, T. asperellum M2RT4, T. atroviride ICIPE 710 and H. lixii F3ST1 had significantly lower feeding punctures as compared to the other treatments. Among the isolates tested, the lowest numbers of eggs were laid by T. tabaci on H. lixii F3ST1 and C. rosea ICIPE 707 inoculated plants. These results extend the knowledge on colonization of onions by fungal endophytes and their effects on Thrips tabaci.

Alternativas botánica, biológica y química para el manejo de piojo del tomate (Halticus sp) (Hemíptera: Miridae) y mosca blanca (Bemisia tabaci, Gen) (Hemiptera: Aleyroridae) en el cultivo de tomate (Solanum lycopersicum L.), bajo condiciones de casa malla.

El piojo del tomate (Halticus sp.) y la mosca blanca (Bemisia tabaci) son los principales agentes de daño, causantes de problemas fitosanitarios severos para los productores de tomate (Solanum lycopersicum, L), bajo condiciones de casa malla. Esta plaga provoca importantes pérdidas económicas, disminuyendo así los rendimientos al afectar la calidad de los frutos lo cual incide en mayores costos de producción. En base a esta problemática en el CEVAT (Centro de Experimentación y Validación de Tecnologías), UNA, se realizó un estudio en el período comprendido entre los meses de Septiembre 2015 a Enero 2016, con el objetivo de evaluar insecticidas biológico, botánico y químico contra el piojo del tomate y mosca blanca. Los productos evaluados fueron: Engeo, Madero Negro y Metarhizium anisopliae. Las variables evaluadas fueron: número de Halticus sp por planta, incidencia del daño por Halticus sp por planta, numero de mosca blanca por planta, incidencia del daño de virosis por planta, severidad del daño de virosis por planta. De los tratamientos evaluados, el menor promedio de piojo del tomate por planta lo presentó el tratamiento Engeo y de igual manera en mosca blanca, seguido de madero negro. El menor porcentaje de incidencia y severidad lo presentó el tratamiento Engeo en comparación con los demás tratamientos evaluados. El análisis económico realizado determinó que los tratamientos que presentaron los mejores rendimientos fueron Engeo con 18,156.25 kg/ha, seguido por Madero Negro con 16,891 kg/ha y Metarhizium anisopliae que obtuvo 14,797 kg/ha. Engeo presentó el mayor beneficio neto con 16,642.92 US$/ha, seguido por Madero Negro con 15,240.86 US$/ha. En el análisis de la tasa de retorno marginal resultó que el tratamiento Madero Negro es el que obtuvo la mejor tasa de retorno marginal con 6,307.36 % es decir 63.07 US$ por cada dólar invertido.

Optimizing Western Flower Thrips Management on French Beans by Combined Use of Beneficials and Imidacloprid

Western flower thrips (WFT), Frankliniella occidentalis (Pergande), is an important pest of vegetable crops worldwide and has developed resistance to many insecticides. The predatory mites Neoseiulus (=Amblyseius) cucumeris (Oudemans), the entomopathogenic fungus Metarhizium anisopliae (Metsch.), and an insecticide (imidacloprid) were tested for their efficacy to reduce WFT population density and damage to French bean (Phaseolus vulgaris L.) pods under field conditions in two planting periods. Metarhizium anisopliae was applied as a foliar spray weekly at a rate of one litre spray volume per plot while imidacloprid was applied as a soil drench every two weeks at a rate of two litres of a mixture of water and imidacloprid per m(2). Neoseiulus cucumeris was released every two weeks on plant foliage at a rate of three mites per plant. Single and combined treatment applications reduced WFT population density by at least three times and WFT damage to French bean pods by at least 1.7 times compared with untreated plots. The benefit-cost ratios in management of WFT were profitable with highest returns realized on imidacloprid treated plots. The results indicate that M. anisopliae, N. cucumeris, and imidacloprid have the potential for use in developing an integrated pest management program against WFT on French beans.

Evaluación y producción masiva de hongos entomopatógenos para el manejo del psilido de la papa (Bactericera cockerelli Sulc) en cultivos de tomate (Solanum lycopersicum L.)y chile (Capsicum annuum L.) en invernadero

En la búsqueda de nuevas estrategias para el manejo de la Paratrioza, Bactericera cockerelli, se ha incluido recientemente el uso de hongos entomopatógenos, principalmente debido a su modo de acción. En este trabajo se evaluaron cuatro cepas de hongos entomopatógenos, dos pertencientes a Beauveria bassiana (BB09 y BB42) y dos a Metarhizium anisopliae (MA25 y MA28). Para las cuatro cepas se valoraron cuatro medios de cultivo (Agar Agua, Papa Dextrosa Agar, Papa Dextrosa Agar + 5% Levadura y Papa Dextrosa Agar + 5% Sacarosa) y tres temperaturas diferentes (20, 25 y 30°C); de igual manera, las cuatro cepas se sujetaron a prueba en tres diferentes medios de producción masiva (arroz, bagazo de caña y bagazo de uva). Para estas tres pruebas se evaluó crecimiento de micelio, viabilidad y esporulación. De manera conjunta se observó la adhesión de conidias de cada una de las cepas a ninfas de B. cockerelli bajo condiciones de laboratorio. Para concluir las evaluaciones, se realizaron dos experimentos en invernaderos para valorar las mortalidades generadas en dos cultivos diferentes (chile y tomate) a dos diferentes concentraciones (1x107conidias por mL y 1x108conidias por mL). De las evaluaciones se concluyó que para éstas cepas el medio de cultivo más adecuado es aquél enriquecido con levadura y que dichas cepas se desarrollan mejor a 25°C, a excepción de las cepas de B. bassiana, las cuales generan mayor cantidad de esporas a 30°C; en cuanto a la reproducción masiva, la búsqueda de sustratos alternativos debe continuar, ya que el bagazo de uva y el de caña no fueron capaces de dar las condiciones adecuadas para generar la misma cantidad de esporas que en el medio usual, el arroz. Finalmente, utilizar hongos entomopatógenos para el manejo de B. cockerelli bajo condiciones de invernadero promete ser una estrategia confiable, ya que dependiendo de la concentración y la cepa utilizada, pueden lograrse niveles de control que van desde un 50 a un 82%.

Compuestos cuticulares de Euschisthus heros (F.) (Heteroptera: pentatomidae) y su papel en las etapas inciales de la infección por hongos entomopatógenos.

2015

Efeito de diferentes concentrações do óleo de nim no crescimento micelial de fungos entomopatogênicos e Trichoderma harzianum.

A associação de extratos de origem vegetal com fungos entomopatogênicos pode aumentar a eficiência do controle biológico de pragas e doenças de plantas e ainda reduzir custos e impactos ambientais. No presente trabalho foi avaliado o efeito do óleo de nim, incorporado ao meio BDA (Batata-dextrose-ágar) nas concentrações de 0, 1, 10, 100, 1.000, 10.000 e 100.000 μ/L, sobre o crescimento micelial de Metarhizium anisopliae, Beauveria bassiana, Trichoderma harzianum e Lecanicillium lecanii. O óleo de nim foi adicionado ao meio antes e após a esterilização em autoclave por 20 min. e 1 atm. Na parte central das placas de Petri contendo os meios foi transferido um disco de micélio dos agentes de biocontrole e mantidas em sala de incubação a 27 ± 2°C. O crescimento da colônia foi avaliado diariamente por seis dias, considerando dois sentidos do diâmetro. Cada placa correspondeu a uma repetição sendo cinco placas por tratamento. O óleo de nim adicionado antes e após autoclavagem em todas as concentrações testadas, estimularam o crescimento micelial de todos os agentes de biocontrole. Esta associação é de grande valia para o controle biológico, pode aumentar a eficiência destes fungos e reduzir impactos ambientais.

Uso de Metarhizium spp. na produção de mudas pré-brotadas de cana-de-açúcar e seus efeitos na planta, em pragas e doenças

Os fungos do gênero Metarhizium são entomopatogênicos e ainda apresentam relações endofíticas e podem viver saprofiticamente no solo. A associação desses microrganismos com insetos é bem conhecida, mas as interações diretas com as plantas ainda são incipientes. Objetivou-se com este estudo, determinar a capacidade de colonização endofítica em raízes de cana-de-açúcar (Saccharum spp.) de M. robertsii, M. anisopliae e três linhagens brasileiras recém-descobertas, bem como revelar o potencial destes fungos como antagonistas a dois fungos fitopatogênicos responsáveis pela podridão-vermelha (Fusarium moniliforme e Colletotrichum falcatum) e no controle de duas importantes pragas, a broca da cana-de-açúcar, Diatraea saccharalis e o nematóide-das-galhas, Meloidogyne javanica. Foram utilizados 24 isolados de Metarhizium spp. nos experimentos de promoção de crescimento de plantas após a inoculação em gemas de cana-de-açúcar em condições de casa-de-vegetação e campo. O efeito da inoculação do fungo em mudas de cana-de-açúcar foi investigado quanto a mortalidade e redução no desenvolvimento de D. saccharalis e redução nas populações de M. javanica. Experimentos de inibição in vitro em cultivo pareado foram conduzidos com os fungos F. moniliforme e C. falcatum e Trichoderma harzianum. Além disto, foram realizados testes qualitativos \"in vitro\" para avaliar a capacidade desses fungos em produzir β-1,3-Glucanase e Sideróforos. A inoculação de Metarhizium de todas as espécies testadas promoveu o crescimento de parte aérea, peso úmido e seco de raízes em relação ao controle (água). Enquanto o tratamento de gemas rotineiramente usado na usina onde se realizou a produção de mudas pré-brotadas com aplicação do fungicida Comet® + o fertilizante Biozyme TF resultou em pegamento de 32% e 59,6%, apenas com o uso de isolados de Metarhizium spp. estes valores foram de até 50% e 86,4%, nos dois experimentos em campo, respectivamente. As mortalidades observadas em lagartas de D. saccharalis que se alimentaram das plantas inoculadas com isolados de Metarhizium spp. atingiram valores de até 80%, sendo a mortalidade confirmada pela esporulação do fungo de até 55%. As lagartas sobreviventes apresentaram menor peso do que aquelas do controle não tratado. A inoculação de Metarhizium spp. em mudas de cana-de-açúcar conferiu proteção a M. javanica, resultando em uma densidade de massa de ovos por grama de raiz nas mudas inoculadas com o fungo até 59,8% menor no primeiro experimento e 64,1% menor no segundo em relação as mudas sem inoculação. Os ensaios de antagonismo mostraram que, todos os isolados de Metarhizium spp. apresentaram alguma forma de inibição tanto para F. moniliforme como para C. falcatum. Dos 24 isolados testados, 20,8% produziram a enzima hidrolítica, β-1,3-glucanase, o que pode estar associado a capacidade de inibição dos fungos fitopatogênicos. Constatou-se que 8 (33,3%) dos isolados produziram sideróforos e sugere que esses fungos apresentam mecanismos de disponibilização do ferro. O conhecimento gerado com este estudo poderá subsidiar novas estratégias de utilização de Metarhizium spp. em campo na produção de MPB de cana-de-açúcar.

Starvation Induces Expression of the Plant-Adhesin Gene, Mad2, of the Entomopathogenic Fungus Metarhizium robertsii.

Metarhizium robertsii is an entomopathogenic fungus that is additionally plant rhizosphere competent. Two adhesin-encoding gens, Mad1 and Mad2, are involved in insect pathogenesis or plant root colonization, respectively. This study examined differential expression of the Mad genes for M robertsii grown on a variety of insectand plant-related substrates. Mad1 was up regulated in response to insect cuticles and up regulation of Mad2 resulted from root exudates, tomato stems and non-preferred carbohydrates. A time course analysis that compared water, minimal media, and nutrient rich broth revealed Mad2 gene expression increased as nutrient availability decreased. The regulation of Mad2 compared to known stress-related genes (Hsp30, Hsp70 and ssgA) under various stresses (nutrient, pH, osmotic, oxidative, temperature) revealed Mad2 to be generally up regulated by nutrient starvation only. Examination of the Mad2 promoter region revealed two copies of a stress-response element (S TRE) known to be regulated under the general stress response pathway.

The analysis of Metarhizium robertsii potential as endophytic plant root coloniser, plant growth enhancer and antagonist to bean root pathogen Fusarium solani f. sp. phaseolis.

The soil-inhabiting insect-pathogenic fungus Metarhizium robertsii also colonizes plant roots endophytically, thus showing potential as a plant symbiont. M robertsii is not randomly distributed in soils but preferentially associates with the plant rhizosphere when applied in agricultural settings. Root surface and endophytic colonization of switchgrass (Panicum virgatum) and haricot beans (Phaseolus vulgaris) by M robertsii were examined after inoculation with fungal conidia. Light and confocal microscopies were used to ascertain this rhizosphere association. Root lengths, root hair density and emergence of lateral roots were also measured. Initially, M robertsii conidia adhered to, germinated on, and colonized, roots. Furthermore, plant roots treated with Metarhizium grew faster and the density of plant root hairs increased when compared with control plants. The onset of plant root hair proliferation was initiated before germination of M robertsii on the root (within 1-2 days). Plants inoculated with M robertsii AMAD2 (plant adhesin gene) took significantly longer to show root hair proliferation than the wild type. Cell free extracts of M robertsii did not stimulate root hair proliferation. Longer term (60 days) associations showed that M robertsii endophytically colonized individual cortical cells within bean roots. Metarhizium appeared as an amorphous mycelial aggregate within root cortical cells as well as between the intercellular spaces with no apparent damage to the plant. These results suggested that not only is M robertsii rhizosphere competent but displays a beneficial endophytic association with plant roots that results in the proliferation of root hairs. The biocontrol of bean (Phaseolis vulgaris) root rot fungus Fusarium solani f. sp. phaseolis by Metarhizium robertsii was investigated in vitro and in vivo. Dual cultures on Petri dishes showed antagonism of M robertsii against F. solani. A relative inhibition of ca. 60% of F. solani growth was observed in these assays. Cell free culture filtrates of M robertsii inhibited the germination of F. solani conidia by 83% and the inhibitory metabolite was heat stable. Beans plants colonized by M robertsii then exposed to F. solani showed healthier plant profiles and lower disease indices compared to plants not colonized by M robertsii. These results suggested that the insect pathogenic/endophytic fungus M robertsii could also be utilized as a biocontrol agent against certain plant pathogens occurring in the rhizosphere.

Plant rhizosphere specificity and variability in the insect and plant adhesins, Madl and Mad2, within the genus Metarhizium suggest plant adaptation as an evolutionary force

Metarhizium is a soil-inhabiting fungus currently used as a biological control agent against various insect species, and research efforts are typically focused on its ability to kill insects. In section 1, we tested the hypothesis that species of Metarhizium are not randomly distributed in soils but show plant rhizosphere-specific associations. Results indicated an association of three Metarhizium species (Metarhizium robertsii, M. brunneum and M. guizhouense) with the rhizosphere of certain types of plant species. M. robertsii was the only species that was found associated with grass roots, suggesting a possible exclusion of M. brunneum and M. guizhouense, which was supported by in vitro experiments with grass root exudate. M. guizhouense and M. brunneum only associated with wildflower rhizosphere when co-occurring with M. robertsii. With the exception of these co-occurrences, M. guizhouense was found to associate exclusively with the rhizosphere of tree species, while M. brunneum was found to associate exclusively with the rhizosphere of shrubs and trees. These associations demonstrate that different species of Metarhizium associate with specific plant types. In section 2, we explored the variation in the insect adhesin, Madl, and the plant adhesin, Mad2, in fourteen isolates of Metarhizium representing seven different species. Analysis of the transcriptional elements within the Mad2 promoter region revealed variable STRE, PDS, degenerative TATA box, and TATA box-like regions. Phylogenetic analysis of 5' EF-Ia, which is used for species identification, as well as Madl and Mad2 sequences demonstrated that the Mad2 phylogeny is more congruent with 5' EF-1a than Madl. This suggests Mad2 has diverged among Metarhizium lineages, contributing to clade- and species-specific variation. While other abiotic and biotic factors cannot be excluded in contributing to divergence, it appears that plant associations have been the driving factor causing divergence among Metarhizium species.

Variability in the Insect and Plant Adhesins, Mad1 and Mad2, within the Fungal Genus Metarhizium Suggest Plant Adaptation as an Evolutionary Force

Several species of the insect pathogenic fungus Metarhizium are associated with certain plant types and genome analyses suggested a bifunctional lifestyle; as an insect pathogen and as a plant symbiont. Here we wanted to explore whether there was more variation in genes devoted to plant association (Mad2) or to insect association (Mad1) overall in the genus Metarhizium. Greater divergence within the genus Metarhizium in one of these genes may provide evidence for whether host insect or plant is a driving force in adaptation and evolution in the genus Metarhizium. We compared differences in variation in the insect adhesin gene, Mad1, which enables attachment to insect cuticle, and the plant adhesin gene, Mad2, which enables attachment to plants. Overall variation for the Mad1 promoter region (7.1%), Mad1 open reading frame (6.7%), and Mad2 open reading frame (7.4%) were similar, while it was higher in the Mad2 promoter region (9.9%). Analysis of the transcriptional elements within the Mad2 promoter region revealed variable STRE, PDS, degenerative TATA box, and TATA box-like regions, while this level of variation was not found for Mad1. Sequences were also phylogenetically compared to EF-1a, which is used for species identification, in 14 isolates representing 7 different species in the genus Metarhizium. Phylogenetic analysis demonstrated that the Mad2 phylogeny is more congruent with 59 EF-1a than Mad1. This would suggest that Mad2 has diverged among Metarhizium lineages, contributing to clade- and species-specific variation, while it appears that Mad1 has been largely conserved. While other abiotic and biotic factors cannot be excluded in contributing to divergence, these results suggest that plant relationships, rather than insect host, have been a major driving factor in the divergence of the genus Metarhizium.

Variability in the Insect and Plant Adhesins, Mad1 and Mad2, within the Fungal Genus Metarhizium Suggest Plant Adaptation as an Evolutionary Force

Several species of the insect pathogenic fungus Metarhizium are associated with certain plant types and genome analyses suggested a bifunctional lifestyle; as an insect pathogen and as a plant symbiont. Here we wanted to explore whether there was more variation in genes devoted to plant association (Mad2) or to insect association (Mad1) overall in the genus Metarhizium. Greater divergence within the genus Metarhizium in one of these genes may provide evidence for whether host insect or plant is a driving force in adaptation and evolution in the genus Metarhizium. We compared differences in variation in the insect adhesin gene, Mad1, which enables attachment to insect cuticle, and the plant adhesin gene, Mad2, which enables attachment to plants. Overall variation for the Mad1 promoter region (7.1%), Mad1 open reading frame (6.7%), and Mad2 open reading frame (7.4%) were similar, while it was higher in the Mad2 promoter region (9.9%). Analysis of the transcriptional elements within the Mad2 promoter region revealed variable STRE, PDS, degenerative TATA box, and TATA box-like regions, while this level of variation was not found for Mad1. Sequences were also phylogenetically compared to EF-1a, which is used for species identification, in 14 isolates representing 7 different species in the genus Metarhizium. Phylogenetic analysis demonstrated that the Mad2 phylogeny is more congruent with 59 EF-1a than Mad1. This would suggest that Mad2 has diverged among Metarhizium lineages, contributing to clade- and species-specific variation, while it appears that Mad1 has been largely conserved. While other abiotic and biotic factors cannot be excluded in contributing to divergence, these results suggest that plant relationships, rather than insect host, have been a major driving factor in the divergence of the genus Metarhizium.

Trading nitrogen for carbon: Nitrogen and carbon translocation in a plant/fungal (Metarhizium spp.) symbiosis

While nitrogen is critical for all plants, they are unable to utilize organically bound nitrogen in soils. Therefore, the majority of plants obtain useable nitrogen through nitrogen fixing bacteria and the microbial decomposition of organic matter. In the majority of cases, symbiotic microorganisms directly furnish plant roots with inorganic forms of nitrogen. More than 80% of all land plants form intimate symbiotic relationships with root colonizing fungi. These common plant/fungal interactions have been defined largely through nutrient exchange, where the plant receives limiting soil nutrients, such as nitrogen, in exchange for plant derived carbon. Fungal endophytes are common plant colonizers. A number of these fungal species have a dual life cycle, meaning that they are not solely plant colonizers, but also saprophytes, insect pathogens, or plant pathogens. By using 15N labeled, Metarhizium infected, wax moth larvae (Galleria mellonella) in soil microcosms, I demonstrated that the common endophytic, insect pathogenic fungi Metarhizium spp. are able to infect living soil borne insects, and subsequently colonize plant roots and furnish ts plant host with useable, insect-derived nitrogen. In addition, I showed that another ecologically important, endophytic, insect pathogenic fungi, Beauveria bassiana, is able to transfer insect-derived nitrogen to its plant host. I demonstrated that these relationships between various plant species and endophytic, insect pathogenic fungi help to improve overall plant health. By using 13C-labeled CO2, added to airtight plant growth chambers, coupled with nuclear magnetic resosnance spectroscopy, I was able to track the movement of carbon from the atmosphere, into the plant, and finally into the root colonized fungal biomass. This indicates that Metarhizium exists in a symbiotic partnership with plants, where insect nitrogen is exchanged for plant carbon. Overall these studies provide the first evidence of nutrient exchange between an insect pathogenic fungus and plants, a relationship that has potentially useful implications on plant primary production, soil health, and overall ecosystem stability.

Combinación de diferentes estrategias de aplicación de hongos entomopatógenos para el control de Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) y Bemisia tabaci (Gennadius) (Hemiptera:Aleyrodidae)

Los ascomicetos mitospóricos entomopatógenos, que actúan por vía tegumentaria, han sido utilizados para el control microbiano de plagas mediante distintas estrategias que permiten el contacto de sus conidios con los insectos diana. Sin embargo, se conocen nuevos aspectos sobre su función ecológica tales como su capacidad secretora de compuestos con actividad insecticida, así como su sorprendente comportamiento como endófitos, que complementan su empleo clásico y que pueden pantag ermitir el desarrollo de nuevas estrategias de control de plagas. El empleo conjunto de estos hongos y sus extractos para el control de un insecto polífago de importancia mundial Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) es abordado en el capítulo II, donde se pone de manifiesto la existencia de variación intra- e interespecífica en la virulencia y estrategias patogénicas de las cepas de Beauveria bassiana y Metarhizium brunneum evaluadas frente a larvas del noctuido. La aplicación conjunta de cepas con distintas estrategias patogénicas junto con sus extractos a larvas del lepidóptero tuvo efecto aditivo, y alguno antagónico, de lo que depende su empleo conjunto. Los capítulos III y IV revelan la existencia de colonización endofítica transitoria tras la aplicación foliar de suspensiones de conidios de cepas de las especies mencionadas en alfalfa, tomate y melón, así como el impacto de esta colonización sobre dos fitófagos con diferentes hábitos de alimentación, uno masticador S. littoralis y otro picador-suctor Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). El capítulo III muestra la contribución aditiva de la mortalidad causada por la alimentación de larvas de S. littoralis a expensas de material vegetal colonizado endofíticamente por cepas de B. bassiana y M. anisopliae con la debida al tratamiento tópico de las mismas con las suspensiones fúngicas. El origen de la mortalidad iniciada a través de la vía digestiva en larvas de este lepidóptero permanece incierto para las cepas de B. bassiana, pero podría estar asociado a la presencia en la planta de metabolitos fúngicos en las cepas de M. brunneum como denota la presencia de destruxina (dtx) A en el 11,0% de los cadáveres. El capítulo IV revela que cepas de ambas especies fúngicas pueden iniciar ciclos de infección en insectos picadores-suctores cuando se alimentan a expensas de sustrato vegetal colonizado endofíticamente, si bien, ambas especies presentan estrategias diferentes. Así, B. bassiana muestra una gran capacidad para colonizar el melón, incluso con efecto traslaminar, que causa la infección de ninfas de B. tabaci por contacto con el tegumento. Sin embargo, la mortalidad de las mismas causada por M. brunneum, de crecimiento mucho más localizado en la hoja, está relacionada una vez mas con la presencia de dtx A en el 43,0% de los cadáveres. Estos resultados deben ser considerados para la evaluación del impacto real de los tratamientos con hongos entomopatógenos, y abren nuevas vías en el control de plagas.