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BACKGROUND: UV‐B radiation represents a significant challenge for the widespread use of entomopathogenic fungi in pest management. This study focused on research of the asynchronous response between virulence and conidial viability against Ceratitis capitata adults using specific statistical models. Moreover, it was also investigated whether the observed differences in susceptibility to UV‐B radiation in in vitro assays among three selected isolates of Beauveria bassiana were reflected in the above‐mentioned asynchrony. RESULTS: While the irradiation of the three isolates of B. bassiana was associated with a significant loss of conidial viability, their virulence was not significantly affected compared to nonirradiated treatments when exposed to 1200 mW m−2 for 6 h before or after the inoculation of C. capitata. In fact, the irradiation time needed to reduce the mortality to 50% compared to the controls was 34.69 h for EABb 10/225‐Fil, 16.36 h for EABb 09/20‐Fil, and 24.59 h for EABb 09/28‐Fil. Meanwhile, the irradiation time necessary to reduce conidial viability to 50% was 9.89 h for EABb 10/225‐Fil, 8.74 h for EABb 09/20‐Fil, and 4.71 h for EABb 09/28‐Fil. CONCLUSION: These results highlight the importance of modeling the response of entomopathogenic fungi virulence and conidial susceptibility when exposed to UV‐B radiation for the selection of environmentally competent isolates, regardless of the results obtained in previous in vitro assays on conidial germination. This strategic approach is critical in overcoming the challenges posed by UV‐B radiation and holds the key to realizing the full potential of entomopathogenic fungi in pest management. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi have a high potential as microbial control agents of insect pests, both through direct application and through endophytic colonisation and the subsequent effects on the phytophagous insects. Recently, such symbiotic relationships with their host plants and compatibility with other natural enemies of insect pests have been characterized, including the modification of plant volatile emissions and the consequent effect on the insects. The present research aimed at investigating whether systemic Beauveria bassiana (Balsamo) Vuillemin colonisation of melon plants reached the floral tissues and affected flowering, pollen production and pollinator behaviour. Melon plants sprayed with B. bassiana conidial suspensions were endophytically colonised throughout the entire plant growth cycle and reached the floral tissues. In addition, the endophytically-colonized plants produced a significantly higher number of pollen grains, and flowered earlier than control plants. A foraging preference of honeybees and bumblebees for the control plants in choice assays was detected, likely due to the advanced maturation stage of colonised plants flowers. Noteworthy, changes in the emitted blend of volatiles were observed in the endophytically-colonised plants, which emitted more of the pollinator attractants D-limonene and benzeneacetaldehyde than control plants. Future research should consider the mechanisms for endophytemediated early flowering and variations in volatile profiles throughout the blooming process, together with their impact on pollination under greenhouse conditions. [ABSTRACT FROM AUTHOR]

Endophytic insect pathogenic fungi have a multifunctional lifestyle; in addition to its well-known function as biocontrol agents, it may also help plants respond to other biotic and abiotic stresses, such as iron (Fe) deficiency. This study explores M. brunneum EAMa 01/58-Su strain attributes for Fe acquisition. Firstly, direct attributes include siderophore exudation (in vitro assay) and Fe content in shoots and in the substrate (in vivo assay) were evaluated for three strains of Beauveria bassiana and Metarhizium bruneum. The M. brunneum EAMa 01/58-Su strain showed a great ability to exudate iron siderophores (58.4% surface siderophores exudation) and provided higher Fe content in both dry matter and substrate compared to the control and was therefore selected for further research to unravel the possible induction of Fe deficiency responses, Ferric Reductase Activity (FRA), and relative expression of Fe acquisition genes by qRT-PCR in melon and cucumber plants.. In addition, root priming by M. brunneum EAMa 01/58-Su strain elicited Fe deficiency responses at transcriptional level. Our results show an early up-regulation (24, 48 or 72 h post inoculation) of the Fe acquisition genes FRO1, FRO2, IRT1, HA1, and FIT as well as the FRA. These results highlight the mechanisms involved in the Fe acquisition as mediated by IPF M. brunneum EAMa 01/58-Su strain. [ABSTRACT FROM AUTHOR]

The influence of treatment on the number of larvae that produced a cocoon was analyzed with an ANCOVA, where the factors were the age of the larvae treated and the treatment, and the covariate was the number of larvae that survived the treatment. The effect of the I B. bassiana i treatment was detectable from the third day on the L2-L3 larvae (Figure 2A), from the fourth day on the L4-L5 larvae (Figure 2B) and from the fifth day on the L6 larvae (Figure 2C). The strain of I B. bassiana i obtained from the larvae collected in the forest was sequenced and aligned with the I B. bassiana i species and has shown a different virulence against I C. perspectalis i larvae depending on the age of the larvae. Several larvae from the bioassays with L2-L3 and L4-L5 larvae prepared a cocoon (Table 5), remaining inside it as diapausing larvae, ceasing feeding and development. [Extracted from the article]

Entomopathogenic fungi are gaining acceptance in Integrated Pest Management (IPM) systems as effective and environmental safety biological control agents to protect a great variety of crops against pest insects. Many of these insect-pathogenic fungi can establish themselves as endophytes and thereby may induce the plant immune system. The activation of plant defenses by the fungal endophytic colonization can have a direct impact on herbivores and plant pathogens. An integral component of many plant defense responses is also the release of volatile organic compounds, which may serve as an indirect defense by attracting the natural enemies of herbivores. Here we investigated the effect of endophytic colonization by the entomopathogenic fungus Beauveria bassiana on the volatile emission by melon and cotton plants, either unharmed or after being damaged by sap-sucking aphids or leaf chewing caterpillars. We found that when the plants are colonized by B. bassiana they emit a different blend of volatile compounds compared to uncolonized control plants. Some of the emitted compounds have been reported previously to be released in response to herbivory and have been implicated in natural enemy attraction. Several of the compounds are also known to have antimicrobial properties. Therefore, endophytic colonization by B. bassiana might help to not only direct control insect pests but also increase the resistance of plants against agronomically important pests and phytopathogens. [ABSTRACT FROM AUTHOR]

The false carmine cochineal Dactylopius opuntiae has been originally used as biological control agent of the prickly pear cactus Opuntia ficus-indica Miller, considered as an exotic invasive species in many countries including Spain. However, the extremely high level of infestation has resulted in great inconvenience to the population of local residents and a serious threat to wipe out a plant that is already part of the Andalusian landscape. In the present research, different control means were evaluated towards this cochineal at field condition in a site in the province of Cadiz (Spain). The populations of the insect in the study site were confirmed to belong to D. opuntiae. We detected this insect throughout the study area with a very high population density in terms of number of colonies per cladode and the extent of the infestation. We also evaluated the efficacy of four different methods (Chlorpyrifos-methyl, Potassium soap, Cryptolaemus montrouzieri larvae, the entomopathogenic fungus Beauveria bassiana) for the control of D. opuntiae. Our results demonstrated that the Potassium soap and Chlorpyrifos-methyl were the best treatments in reducing D. opuntiae populations, with relative population density values being negative only in these two treatments, and efficacies of 91.5 and 76.7% respectively. The results of this investigation showed that the current situation of false carmine cochineal is worrisome, but nevertheless, the use of Potassium soap for its control is recommended because of its high efficacy and low environmental impact. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi (EF) have so far aroused little interest as plant growth promoters or nutritional enhancers while protecting the plant against insect pests and mites. In this work, we examined the performance of Triticum durum and Triticum aestivum plants pot‐grown on two unsterilized Zn‐poor calcareous Vertisols and inoculated with Beauveria bassiana or Metarhizium brunneum by application to soil or seed dressing. The effects of fungal inoculation were found to depend on crop species, soil type, sampling time and fungus application method. While the height of the control plants tended to be significantly greater than the height of inoculated plants at early and intermediate growth stages, the trend changed at more advanced plant stage of development, 56 days after sowing (DAS). This was supported by the significant increase in aerial dry matter (ADM) in T. aestivum plants treated with B. bassiana by seed dressing (17.4%) or with M. brunneum with soil application and seed dressing (20.6 and 26.9%, respectively) 69 DAS. In T. durum, at harvest, seed dressing with B. bassiana or M. brunneum had an overall positive but not significant effect on ADM or grain yield, whereas direct application to the soil slightly decreased grain yield (6.6 and 5.5%, respectively) and ADM (5.3 and 6.1%, respectively). In T. aestivum, none of the fungus–application method combinations affected yield at harvest significantly. Fungal inoculation had little influence on nutrient uptake, whereas application to soil increased Mn uptake and grain Mn concentration in T. durum. Zinc uptake was significantly increased only in T. aestivum grown on the soil with the lowest Zn content treated with M. brunneum. Grain Zn concentration tended to be inversely related to grain yield. These results support seed dressing with EF as a promising green technology for sustainable crop protection and production with no adverse effects on plant performance. [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: A new Beauveria bassiana‐based attract and infect device (AID) to control Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) was developed. The virulence and persistence of the fungal formulation used in the AID were evaluated in the laboratory. Semi‐field and field trials were carried out to validate the results and establish the potential of this device as a control tool. RESULTS: In laboratory conditions, a 50% lethal time (LT50) of 4.33 days was obtained when adults (7–10 days old) were exposed to the inoculation tunnel (IT) containing 1 × 1010 conidia g–1 in an oil‐based fungal formulation. This formulation maintained conidium viability at 50% for up to 2 months. Moreover, when adults were exposed to 2.5‐month field‐aged ITs, mortality still reached 50% 40 days after exposure. In addition, no differences were observed between ITs aged in early spring and those aged in summer, suggesting that the fungal formulation is not strongly affected by environmental factors in Mediterranean basin conditions. Semi‐field assays showed that the device allowed an easy transit of weevils through the IT, which were effectively attracted and infected. Using the AIDs in 4‐ha plot field trials, a reduction of >50% in the percentage of infested sentinel palms was obtained. CONCLUSION: Based on the results obtained in terms of the efficacy and persistence of this new AID in the field and its potential in reducing R. ferrugineus populations and palm infestation, this device could become a key tool for the management of R. ferrugineus. © 2018 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

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Beauveria bassiana strain 04/01-Tip, obtained from a larva of the opium poppy stem gall wasp Iraella luteipes (Hymenoptera; Cynipidae), endophytically colonizes opium poppy (Papaver somniferum L.) plants and protects them against this pest. The goal of this study was to monitor the dynamics of endophytic colonization of opium poppy by B. bassiana after the fungus was applied to the seed and to ascertain whether the fungus is transmitted vertically via seeds. Using a species-specific nested PCR protocol and DNA extracted from surface-sterilised leaf pieces or seeds of B. bassiana-inoculated opium poppy plants, the fungus was detected within the plant beginning at the growth stage of rosette building and them throughout the entire plant growth cycle (about 120–140 days after sowing). The fungus was also detected in seeds from 50% of the capsules sampled. Seeds that showed positive amplification for B. bassiana were planted in sterile soil and the endophyte was again detected in more than 42% of the plants sampled during all plant growth stages. Beauveria bassiana was transmitted to seeds in 25% of the plants from the second generation that formed a mature capsule. These results demonstrate for the first time the vertical transmission of an entomopathogenic fungus from endophytically colonised maternal plants. This information is crucial to better understand the ecological role of entomopathogenic fungi as plant endophytes and may allow development of a sustainable and cost effective strategy for I. luteipes management in P. somniferum. [ABSTRACT FROM AUTHOR]

Abstract: The potential of a strain of Beauveria bassiana (Ascomycota: Clavicipitaceae) obtained from a naturally infected Rhynchophorus ferrugineus (Coleoptera: Curculionidae) pupa as a biological control agent against this weevil was evaluated both in the laboratory and in semi-field assays. Laboratory results indicate that this strain of B. bassiana can infect eggs, larvae and adults of R. ferrugineus (LC50 from 6.3×107 to 3.0×109 conidia per ml). However, mortality was not the only indicator of treatment efficacy because adults of either sex inoculated with the fungus efficiently transmitted the disease to untreated adults of the opposite sex, with male-to-female and female-to-male rates of transmission of 55% and 60%, respectively. In addition, treatment with B. bassiana significantly reduced fecundity (up to 62.6%) and egg hatching (32.8%) in pairing combinations with fungus-challenged males, females or both sexes. Likewise, 30–35% increase in larval mortality was observed in larvae obtained from eggs from fungus-challenged females or from untreated females coupled with inoculated males, resulting in an overall 78% progeny reduction. Semi-field preventive assays on potted 5-year old Phoenix canariensis palms, with efficacies up to 85.7%, confirmed the potential of this strain as a biological control agent against R. ferrugineus. [Copyright &y& Elsevier]

Abstract: Factors affecting the occurrence and distribution of entomopathogenic fungi in 244 soil samples collected from natural and cultivated areas in Spain were studied using an integrated approach based on univariate and multivariate analyses. Entomopathogenic fungi were isolated from 175 of the 244 (71.7%) soil samples, with only two species found, Beauveria bassiana and Metarhizium anisopliae. Of the 244 soil samples, 104 yielded B. bassiana (42.6%), 18 yielded M. anisopliae (7.3%), and 53 soil samples (21.7%) harboured both fungi. Log-linear models indicated no significant effect of habitat on the occurrence of B. bassiana, but a strong association between M. anisopliae and soils from cultivated habitats, particularly field crops. Also, irrespective of habitat type, B. bassiana predominated over M. anisopliae in soils with a higher clay content, higher pH, and lower organic matter content. Logistic regression analyses showed that pH and clay content were predictive variables for the occurrence of B. bassiana, whereas organic matter content was the predictive variable for M. anisopliae. Also, latitude and longitude predicted the occurrence of these same species, but in opposite directions. Altitude was found to be predictive for the occurrence of B. bassiana. Using principal component analysis, four factors (1 to 4) accounted for 86% of the total variance; 32.8, 22.9, 19.6 and 10.4% of the cumulative variance explained, respectively. Factor 1 was associated with high positive weights for soil clay and silt content and high negative weights for soil sand content. Factor 2 was associated with high positive weights for soil organic matter content and high negative weights for soil pH. Factor 3 was associated with high positive weights for latitude and longitude of the sampled localities and factor 4, had high positive weights only for the altitude. Bi-plot displays representing soil samples were developed for different factor combinations and indicated that, irrespective of geographical location, absence of both fungal species was determined by alkaline sandy soils with low organic matter content, whereas heaviness of soil texture, acidity and increasing organic matter content led to progressively higher percentages of samples harbouring entomopathogenic fungi. These results could aid decision-making as to whether or not a particular cultivated or natural soil is suitable for using entomopathogenic fungi as a pest control measure and for selecting the fungal species best suited to a particular soil. [Copyright &y& Elsevier]

Entomopathogenic fungi are sprayed commercially for aphid control in greenhouses. Recently, their ability to grow endophytically within plants was discovered, offering the opportunity for systemic biological control. Endophytic colonization of host plants could also influence life-table parameters and behavior of herbivores. We investigated lethal and pre-mortality effects of Beauveria bassiana and Metarhizium brunneum on Aphis gossypii; aphids either received inoculum while feeding on recently sprayed leaves (surface inoculum and endophytically-colonized) or while feeding on unsprayed but endophytically-colonized leaves. We used choice assays to identify any preferences for endophytically-colonized or control plants. Volatile emissions from endophytically-colonized plants and control plants were also compared. Aphid mortality rates ranged between 48.2 and 56.9% on sprayed leaves, and between 37.7 and 50.0 on endophytically-colonized leaves. There was a significant effect of endophytic colonization on the rate of nymph production, but this did not result in an overall increase in the aphid population. Endophytic colonization did not influence host-plant selection even though there were qualitative and quantitative differences in the blend of volatiles released by endophytically-colonized and control plants. Although endophytic colonization did not change herbivore behavior, plants still benefit via indirect defense, resistance to plant pathogens or abiotic stress tolerance. [ABSTRACT FROM AUTHOR]

The use of mycoviruses to manipulate the virulence of entomopathogenic fungi employed as biocontrol agents may lead to the development of novel methods to control attacks by insect pests. Such approaches are urgently required, as existing agrochemicals are being withdrawn from the market due to environmental and health concerns. The aim of this work is to investigate the presence and diversity of mycoviruses in large panels of entomopathogenic fungi, mostly from Spain and Denmark. In total, 151 isolates belonging to the genera Beauveria, Metarhizium, Lecanicillium, Purpureocillium, Isaria, and Paecilomyces were screened for the presence of dsRNA elements and 12 Spanish B. bassiana isolates were found to harbor mycoviruses. All identified mycoviruses belong to three previously characterised species, the officially recognised Beauveria bassiana victorivirus 1 (BbVV-1) and the proposed Beauveria bassiana partitivirus 2 (BbPV-2) and Beauveria bassiana polymycovirus 1 (BbPmV-1); individual B. bassiana isolates may harbor up to three of these mycoviruses. Notably, these mycovirus species are under distinct selection pressures, while recombination of viral genomes increases population diversity. Phylogenetic analysis of the RNA-dependent RNA polymerase gene sequences revealed that the current population structure in Spain is potentially a result of both vertical and horizontal mycovirus transmission. Finally, pathogenicity experiments using the Mediterranean fruit fly Ceratitis capitata showed no direct correlation between the presence of any particular mycovirus and the virulence of the B. bassiana isolates, but illustrated potentially interesting isolates that exhibit relatively high virulence, which will be used in more detailed virulence experimentation in the future. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi (EF) provide a potent biocontrol tool; also, their endophytic behavior has broadened their contribution to integrated pest management (IPM) and crop production. In this work, Beauveria bassiana and Metarhizium brunneum were applied to bread wheat (Triticum aestivum) seedlings to elucidate how fungal colonization influences plant growth and the relative expression of 24 genes involved in hormonal syntheses and plant immune mechanisms. A preliminary assay was used to determine the time needed for fungal colonization and assess its effect on wheat growth. Then, plant material collected at various times after inoculation (viz., 2, 8, 20, and 36 h and 9 and 15 days) was used to investigate gene expression by quantitative reverse transcription PCR (RT-qPCR). During the colonization time, B. bassiana and M. brunneum caused strong downregulation of most genes associated with plant immunity and the synthesis of hormones like auxin, cytokinin, and gibberellin. This effect was concomitant with a slowdown of endophytic-colonization-related plant growth until 19 days postinoculation (dpi). However, the wheat started to recover at 15 dpi, simultaneously with upregulation of auxin- and gibberellin-related genes. The results suggest that the EF trigger induced systemic resistance rather than acquired systemic resistance during early plant-microbe cross talk in wheat. Also, they confirm that the hormone and immune responses of wheat triggered by EF inoculation influenced plant growth, which can be useful with a view to optimizing management of these microorganisms for sustainable agriculture. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi are known due to their ability to kill insects and besides they can play other roles, such as promoting plant growth or improving nutrient uptake. The main aim of this work was to assess the potential of Beauveria bassiana strain EABb 04/01-Tip for improving iron (Fe) nutrition in plants grown on calcareous substrates. Plants of a dicot (tomato) and a monocot (wheat) were pot-grown on an artificial substrate consisting of 0–240 g kg −1 Fe oxide-coated sand (FOCS), 250 g kg −1 calcium carbonate sand and quartz sand. The plants were subjected to two different treatments, namely seed dressing, which involved inoculating seeds with a conidial suspension of B. bassiana before sowing, and a control without inoculation. Leaf chlorophyll concentrations (SPAD) and Fe uptake by plants were correlated with the FOCS content of the substrate, therefore the experimental design was appropriate to see different levels of Fe chlorosis symptoms. B. bassiana was able to colonise both tomato and wheat plants without a negative effect on plant height, plant dry weight, root development in tomato or grain production in wheat. In addition, B. bassiana alleviated Fe chlorosis symptoms (described for first time) in both crops during early growing stages (<50 days after sowing) but the intensity of the effect depended on the plant species and available Fe on substrate. Nutritional alterations in K due to fungal application were detected in tomato and wheat plants. [ABSTRACT FROM AUTHOR]

Highlights: [•] The tillage effect on entomopathogenic fungi (EF) associated to Prays oleae was evaluated. [•] No significant differences were found between the different soil management practices. [•] Seasonal variation on abundance and diversity of EF was detected. [•] Beauveria bassiana was the most abundant entomopathogenic fungi in both soil types. [•] Results provide new insights to the dynamics of EF in the olive grove ecosystem [Copyright &y& Elsevier]

Viruses have been discovered in numerous fungal species, but unlike most known animal or plant viruses, they are rarely associated with deleterious effects on their hosts. The knowledge about viruses among entomopathogenic fungi is very limited, although their existence is suspected because of the presence of virus-like double-stranded RNA (dsRNA) in isolates of several spedes. Beauveria bassiana is one of the most-studied species of entomopathogenic fungi; it has a cosmopolitan distribution and is used as a biological control agent against invertebrates in agriculture. We analyzed a collection of 73 isolates obtained at different locations and from different habitats in Spain and Portugal, searching for dsRNA elements indicative of viral infections. The results revealed that the prevalence of viral infections is high; 54.8% of the isolates contained dsRNA elements with viral characteristics. The dsRNA electropherotypes of infected isolates indicated that virus diversity was high in the collection analyzed and that mixed virus infections occurred in fungal isolates. However, a hybridization experiment indicated that dsRNA bands that are similar in size do not always have similar sequences. Particular virus species or dsRNA profiles were not associated with locations or types of habitats, probably because of the ubiquity and efficient dispersion of this fungus as an airborne species. The sequence of one of the most common dsRNA elements corresponded to the 5.2-kbp genome of a previously undescribed member of the Totiviridae family, termed B. bassiana RNA virus 1 (BbRV1). [ABSTRACT FROM AUTHOR]

• Direct exposure of Philaenus to Metarhizium brunneum reduces its survivorship. • M. brunneum successfully colonises the host plant and reduces survival of Philaenus. • Philaenus reaches the xylem faster on M. brunneum -colonised plants. • Xylem sap ingestion by Philaenus is similar on non-colonised and colonised plants. • Contact with M. brunneum does not alter the egestion pattern of Philaenus. The most effective way to contain spread of the plant pathogen Xylella fastidiosa in Europe is to manage populations of its main insect vector, Philaenus spumarius L. (Hemiptera: Aphrophoridae). The action of some entomopathogenic endophytic fungi (EEF) on insect pests, directly through contact or indirectly by internal colonization of host plant tissues, may offer an excellent control option within integrated pest management (IPM) programmes. This study evaluated the impact of two EEF strains as control agents of P. spumarius (Metarhizium brunneum strain EAMa 01/58-Su and Beauveria bassiana strain EABb 01/33-Su [Hypocreales: Clavicipitaceae and Cordycipitaceae, respectively]); both strains were known to be excellent control agents of many other serious chewing and sap-sucking agricultural pests. First, the pathogenicity of strains applied directly to P. spumarius adults was evaluated, resulting in 50.0% and 32.0% mortality and a mean survival time of 4.7 and 6.2 days for M. brunneum and B. bassiana , respectively. Furthermore, colonization of Sonchus oleraceus by EEFs was observed after both soil and foliar applications. Greatest colonization (91.0%) was observed in plants treated with foliar applications of M. brunneum. Based on these results, we evaluated the feeding behaviour of P. spumarius on S. oleraceus plants previously colonized by M. brunneum using the 'Electrical Penetration Graph' technique. P. spumarius fed in a similar way on EEF-colonized and non-colonized S. oleraceus plants, with the exception that insects feeding on colonized plants reached xylem vessels faster than insects feeding on non-colonized plants. In addition, significantly higher mortality of P. spumarius was observed when feeding on plants treated by M. brunneum compared with the control. The duration of xylem ingestion events indicates that P. spumarius preferred to feed on basal leaves of the plant compared with apical leaves. Our results indicate strong potential for strain EAMa 01/58-Su of M. brunneum to be used as part of an X. fastidiosa IPM programme. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • M. acridum IMI 330189 was more virulent than B. bassiana EABb 90/2-Dm at low dosages. • Most treatments reduced per capita feeding (32–51%) during the interval of no thermoregulation. • Only IMI 330189 reduced per capita feeding (50%) when insects were allowed to thermoregulate. • Insect mortality suggested additive interaction in the lethal effects between the two strains. • For sublethal effects (feeding and fecundity) a potential antagonistic interaction was detected. Abstract The effects of single and mixed infections with Beauveria bassiana (EABb 90/2-Dm) and Metarhizium acridum (IMI 330189) strains on survival, feeding and reproduction of thermoregulating Dociostaurus maroccanus were evaluated. Adult locusts (2–3 days post fledging) were treated with low dosages of both fungal pathogens alone and in mixture (total dosage for single treatments and combinations = 1 × 102 and 1 × 103 spores per insect). M. acridum IMI 330189 was more virulent than B. bassiana EABb 90/2-Dm at both dosages. In the mixed infections, in which half of the infective units of the more virulent pathogen were replaced by the less virulent pathogen, the analysis of the cumulative insect mortality after 30 days suggested additive interaction in the lethal effects between the two strains. All fungal treatments, except EABb 90/2-Dm at 1 × 102 spores per insect showed reduction in per capita feeding, as indicated by fecal production per insect per day when insects were maintained at 27 ± 2 °C (32–51% of reduction compared with the control); but only IMI 330189 caused significant reduction in per capita feeding (50%) when those insects were allowed to thermoregulate. Both strains and their mixtures caused a significant reduction of locust fecundity, with a 21–53% reduction in the number of egg-pods per female, and 30–65% reduction in the number of fertile eggs per female. In both sublethal effects (feeding and fecundity) a potential antagonistic interaction between the fungal strains was detected. Locust fecundity (egg-pods per female) and per capita feeding were positively correlated (r = 0.783). Implications of these findings on the potential use of both strains to control D. maroccanus populations are discussed. [ABSTRACT FROM AUTHOR]

The response of entomopathogenic mitosporic ascomycete (EMAs) to abiotic stresses might be adapted to the microhabitats in which they inhabit. In phylloplane, these organisms are more exposed to such stresses than they are in soil, which may have led to adaptation to this environment. In the present work, we investigate whether Beauveria bassiana genotype or isolation habitat, i.e., soil or phylloplane, within the same geographic area influences their responses to key environmental stresses, such as temperature, moisture and ultraviolet radiation (UV-B), which can affect their successful use in microbial control. Twenty isolates of B. bassiana obtained from the soil and phylloplane in two ecosystems from southern Spain (holm oak dehesa and a reforested area) were selected to study the population distribution of these isolates and evaluate their thermal, humidity and UV-B requirements. Molecular characterization was conducted by using elongation factor-1α (EF-1α), the intergenic nuclear region Bloc and 15 microsatellite primers. The cluster analysis based on concatenated EF-1α and Bloc sequences grouped the 20 isolates into five clades within B. basiana , with Clades a, b, d and e containing both soil and phylloplane isolates and Clade c including three phylloplane isolates. The dendrogram and the minimal spanning network generated from the genetic distances among multilocus genotypes showed four divergent groups corresponding to the five clades obtained based on the sequence data (Clades b and d were represented in the same group), with a high degree of shared alleles within groups and few alleles shared among groups. Although no relationship was found between MLG and the habitat (soil or phylloplane) of isolation, isolates grouped into Clade c, all of which were collected from phylloplane, formed a separate group of MLGs. To investigate our hypothesis, the responses to temperature (germination and colony growth evaluated in the range 15–35 °C), water activity (conidia germination evaluated against values of a w between 1 and 0.862) and UV-B exposure (conidia exposed to 920 or 1200 mW m −2 for 2, 4 or 6 h) of the soil and phylloplane isolates from the five clades were investigated. No associations of isolate-specific genetic or physiological characteristics with isolate habitat, i.e., soil or phylloplane, were found. These results provide no support for the hypothesis that EMAs strains from the phylloplane have evolved to resist unfavourable environmental conditions. [ABSTRACT FROM AUTHOR]

[Display omitted] • B. bassiana endophytically colonized melon, tomato and cotton plants. • Cotton was the the least suitable host plant for endophytic colonization. • B. bassiana re-isolation rate from cotton plants increased after the first passage. • This is the first report an of improvement of endophytic capacity by passaging. • Virulence stability after successive passage through a host plant was demonstrated. Systemic crop protection using endophytic isolates of entomopathogenic fungi is at the forefront of IPM. Nonetheless, any potential trade-offs between virulence and endophytic behaviour must be elucidated if they are to be effectively used in pest management strategies. Here we investigated endophytic adaptation in an isolate of Beauveria bassiana following successive passage through melon, tomato and cotton tissues. Plants were sprayed with a suspension of B. bassiana endophytic isolate EABb 04/01-Tip to initiate endophytic colonization. Once colonization was established, the fungus was re-isolated from the plant, applied to another plant and re-isolated again; this was repeated to achieve three passages. After each passage, a conidial suspension of each isolate was used in bioassays to evaluate both virulence against 4th instar larvae of the model insect Galleria mellonella and to quantify the extent of endophytic activity in each respective host plant species. When sprayed leaves were inspected for fungal colonization, differences in percentage tissue colonization amongst the plant species were detected after the first re-isolation. Endophytic colonization rates in melon and tomato, which varied from 70 to 100%, were higher than those observed in cotton, which ranged from 40 to 50%; endophytic colonization in cotton increased to 75–100% after the third passage. This improvement in endophytic behaviour in cotton, an apparently suboptimal plant for fungal colonization, suggests an evolutionary adaptation to localized or transient endophytic colonization, while further assays are needed. Meanwhile, when endophytic colonization of non-sprayed leaves distant from the sprayed ones was investigated, endophytic activity was evident in all three crop species suggesting that movement within plants after successive passage increased the extent of endophytic colonization from transient to systemic. The present research highlights the potential for adaptation to endophytic behaviour in crops that are less suitable for endophytic colonization. Furthermore, we demonstrated stability in virulence after multiple passages through host plants. This is a key result for the development of IPM strategies based on endophytic entomopathogenic fungi. [ABSTRACT FROM AUTHOR]

Abstract: We used a newly developed bioassay method to demonstrate for the first time the potential of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae to be used for the control of neonate larvae of Capnodis tenebrionis, a major threat to stone-fruit orchards in several countries. Four B. bassiana and four M. anisopliae isolates were all pathogenic for neonate larvae of C. tenebrionis; mortality rates 10 days after inoculation by dipping in a suspension with 108 conidia/ml varied from 23.5% to 100%. Three of the four M. anisopliae isolates caused 100% mortality. In most cases, postmortem hyphal growth and sporulation of M. anisopliae or B. bassiana was observed covering the larvae in their galleries. The eight isolates were also evaluated for pathogenicity to C. tenebrionis eggs at the same dosage. Only two B. bassiana isolates caused significant egg hatching reduction of 84.5% and 94.5%. Our results indicate that M. anisopliae and B. bassiana may be considered as promising for a new approach to prevent larval infestations by C. tenebrionis. [Copyright &y& Elsevier]