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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]

The present study aimed to compare some commercial traps baited with ammonium carbonate for their efficacy in monitoring Bactrocera oleae (Rossi) and their selectivity toward beneficial insects, under the conditions of olive groves located in five Mediterranean countries (Greece, Italy, Lebanon, Spain, Tunisia). The selectivity of the devices was evaluated toward several groups of non-target insects, namely lacewings (Chrysopidae), hoverflies (Syrphidae), ladybirds (Coccinellidae), bees (Apoidea) and hymenopteran parasitoids. The following devices were compared: yellow sticky panel, green sticky panel, Jackson trap with different combinations of yellow and white colours of the device and/or the panel, and McPhail trap. In most cases, the McPhail trap and the yellow panel showed the highest efficacy in monitoring male and female B. oleae flies; however, the yellow panel was most attractive for some groups of non-target insects, particularly the hymenopteran parasitoids, whereas the comparative non-target effects were sitespecific for the other arthropod groups. In the case of Chrysopidae, McPhail caught more individuals than the other traps in Italy and Spain, whereas in Lebanon and Greece, the highest number of individuals was captured in the two sticky panels. Coccinellidae were found in very low numbers only in Lebanon on yellow panels. Syrphidae were captured mainly on sticky panels in Greece and Tunisia. Apoidea were found only on yellow panels in Lebanon. The choice between the McPhail trap and the yellow panel should be made on the basis of various evaluations carried out at the site, including operational ones. [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 genus Metarhizium has an increasingly important role in the development of Integrated Pest Control against Tephritid fruit flies in aerial sprays targeting adults and soil treatments targeting preimaginals. Indeed, the soil is considered the main habitat and reservoir of Metarhizium spp., which may be a plant-beneficial microorganism due to its lifestyle as an endophyte and/or rhizosphere-competent fungus. This key role of Metarhizium spp. for eco-sustainable agriculture highlights the priority of developing proper monitoring tools not only to follow the presence of the fungus in the soil and to correlate it with its performance against Tephritid preimaginals but also for risk assessment studies for patenting and registering biocontrol strains. The present study aimed at understanding the population dynamics of M. brunneum strain EAMb 09/01-Su, which is a candidate strain for olive fruit fly Bactrocera oleae (Rossi, 1790) preimaginal control in the soil, when applied to the soil at the field using different formulations and propagules. For this, strain-specific DNA markers were developed and used to track the levels of EAMb 09/01-Su in the soil of 4 field trials. The fungus persists over 250 days in the soil, and the levels of the fungus remained higher when applied as an oil-dispersion formulation than when applied as a wettable powder or encapsulated microsclerotia. Peak concentrations of EAMb 09/01-Su depend on the exogenous input and weakly on environmental conditions. These results will help us to optimize the application patterns and perform accurate risk assessments during further development of this and other entomopathogenic fungus-based bioinsecticides. [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]

Microsclerotia (MS) produced by some species of Metarhizium can be used as active ingredients in mycoinsecticides for the control of soil-dwelling stages of geophilic pests. In this study, the MS production potential of two Metarhizium brunneum strains and one M. robertsii strain was evaluated. The three strains were able to produce MS in liquid fermentation, yielding between 4.0 × 106 (M. robertsii EAMa 01/158-Su strain) and 1.0 × 107 (M. brunneum EAMa 01/58-Su strain) infective propagules (CFU) per gram of MS. The EAMa 01/58-Su strain was selected for further investigation into the effects of key abiotic factors on their survival and conidial yield. The MS were demonstrated to be stable at different storage temperatures (−80, −18, and 4 °C), with a shelf-life up to one year. The best temperature for MS storage was −80 °C, ensuring good viability of MS for up to one year (4.9 × 1010 CFU/g MS). Moreover, soil texture significantly affected CFU production by MS; sandy soils were the best driver of infective propagule production. Finally, the best combination of soil temperature and humidity for MS germination was 22.7 °C and 7.3% (wt./wt.), with no significant effect of UV-B exposure time on MS viability. These results provide key insights into the handling and storage of MS, and for decision making on MS dosage and timing of application. [ABSTRACT FROM AUTHOR]

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]

Abstract Background The entomopathogenic anamorphic fungus Beauveria bassiana is currently used as a biocontrol agent (BCA) of insects. Fifty-seven Beauveria bassiana isolates -53 from Spain- were characterized, integrating group I intron insertion patterns at the 3'-end of the nuclear large subunit ribosomal gene (LSU rDNA) and elongation factor 1-alpha (EF1-α) phylogenetic information, in order to assess the genetic structure and diversity of this Spanish collection of B. bassiana. Results Group I intron genotype analysis was based on the four highly conserved insertion sites of the LSU (Ec2653, Ec2449, Ec2066, Ec1921). Of the 16 possible combinations/genotypes, only four were detected, two of which were predominant, containing 44 and 9 members out of 57 isolates, respectively. Interestingly, the members of the latter two genotypes showed unique differences in their growth temperatures. In follow, EF1-α phylogeny served to classify most of the strains in the B. bassiana s.s. (sensu stricto) group and separate them into 5 molecular subgroups, all of which contained a group I intron belonging to the IC1 subtype at the Ec1921 position. A number of parameters such as thermal growth or origin (host, geographic location and climatic conditions) were also examined but in general no association could be found. Conclusion Most Spanish B. bassiana isolates (77.2%) are grouped into a major phylogenetic subgroup with word-wide distribution. However, high phylogenetic diversity was also detected among Spanish isolates from close geographic zones with low climatic variation. In general, no correlation was observed between the molecular distribution and geographic origin or climatic characteristics where the Spanish B. bassiana isolates were sampled.

Abstract Background The entomopathogenic anamorphic fungus Metarhizum anisopliae is currently used as a biocontrol agent (BCA) of insects. In the present work, we analyzed the sequence data obtained from group I introns in the large subunit (LSU) of rDNA genes with a view to determining the genetic diversity present in an autochthonous collection of twenty-six M. anisopliae isolates selected as BCAs. Results DNA fragments corresponding to the 3'-end of the nuclear LSU rDNA genes of 26 M. anisopliae isolates were amplified by PCR. The amplicon sizes ranged from 0.8 to 3.4-kb. Four intron insertion sites, according to Escherichia coli J01695 numbering, were detected- Ec1921, Ec2066, Ec2449 and Ec2563- after sequencing and analysis of the PCR products. The presence/absence of introns allowed the 26 isolates to be distributed into seven genotypes. Nine of the isolates tested showed no introns, 4 had only one, 3 two, and 10 displayed three introns. The most frequent insertion sites were Ec1921 and Ec2449. Of the 26 isolates, 11 showed insertions at Ec2563 and a 1754-bp sequence was observed in ten of them. The most-parsimonious (MP) tree obtained from parsimony analysis of the introns revealed a main set containing four-groups that corresponded to the four insertion sites. Conclusion Four insertion sites of group I introns in the LSU rDNA genes allowed the establishment of seven genotypes among the twenty-six biocontrol isolates of M. anisopliae. Intron insertions at the Ec2563 site were observed for first time in this species.

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]

BACKGROUND: The cotton leafworm, Spodoptera littoralis, is one of the most destructive pests in the Mediterranean basin, being predominantly controlled using synthetic chemical pesticides. Strain EAMa 01/58‐Su of the fungus Metarhizium brunneum and the parasitoid Hyposoter didymator are promising biological control agents for this pest. In this study, we assessed the compatibility between these two agents to control S. littoralis under joint attack scenarios. RESULTS: Firstly, the direct and indirect effects of the fungus towards parasitoid adults were studied. The fungus significantly decreased life expectancy of the parasitoid (mortality = 62.5%; mean lethal concentration = 1.85 × 106 conidia ml−1; average survival time = 92.2 h) when applied at high concentrations (108 conidia ml−1), whereas it did not affect the reproductive potential of the parasitoid females during the 3 days after treatment. Secondly, the combinations between the two agents to control S. littoralis under different simultaneous use scenarios (inoculation of S. littoralis larvae with the fungus before being exposed to parasitoid females and vice versa) were investigated, with additive effect in all cases. A significant effect on fitness (preimaginal development time and reproductive potential) of the F1 parasitoid generation were detected. Moreover, parasitization significantly reduced the total hemocytes in S. littoralis hemolymph compared with the control, promoting fungal infection. Finally, parasitoids showed a significant preference for non‐inoculated S. littoralis larvae. CONCLUSIONS: We demonstrated compatibility (additive effect) between fungus and parasitoid under different joint attack scenarios to control S. littoralis in laboratory conditions. However, this will be supported by our ongoing greenhouse and field studies. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: The susceptibility of 20 widely distributed mill and table olive varieties to Bactrocera oleae (Rossi) as affected by irrigation, and fruit diameter and oil content was evaluated in a 3‐year trial in Southern Spain. Bactrocera oleae was bivoltine life cycle in the experimental site, with significant differences among population size throughout the study. Even though the olive fruit fly damaged all varieties, significant differences in susceptibility were detected. Among the mill olive varieties “Nevadillo Blanco de Jaén” was the most susceptible, with average infestation levels ranging between 6.7% and 52.2% and between 10.3% and 69.2% under rainfed and irrigated conditions, respectively, and “Arbequina” was the least susceptible, with average infestation levels ranging between 0.6% and 12.7% and between 2.3% and 18.5% under rainfed and irrigated conditions, respectively. Among the table olive varieties, “Gordal Sevillana,” “Ascolana Tenera” and “Ocal” were the most susceptible (with average infestation levels reaching 39.7%, 36.5% and 33.3%, respectively), while “Callosina” was the least susceptible (with infestation levels of only 8.4%). Irrigation tended to promote both B. oleae infestation and its earlier occurrence compared to the rainfed condition. Even though the diameter and oil content were positively correlated with B. oleae fruit infestation (correlation coefficients ranged between 0.5 and 0.95), the present work reveals that other yet‐unknown factors may influence B. oleae oviposition preferences. The results of this study can be useful for breeding programmes to develop olive varieties resistant to B. oleae and provide key information for wide‐area olive fly pest management decisions. [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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Soil treatments with Metarhizium brunneum EAMa 01/58-Su strain conducted in both Northern and Southern Spain reduced the olive fly (Bactrocera oleae) population density emerging from the soil during spring up to 70% in treated plots compared with controls. A model to determine the influence of rainfall on the conidial wash into different soil types was developed, with most of the conidia retained at the first 5 cm, regardless of soil type, with relative percentages of conidia recovered ranging between 56 and 95%. Furthermore, the possible effect of UV-B exposure time on the pathogenicity of this strain against B. oleae adults coming from surviving preimaginals and carrying conidia from the soil at adult emergence was also evaluated. The UV-B irradiance has no significant effect on M. brunneum EAMa 01/58-Su pathogenicity with B. oleae adult mortalities of 93, 90, 79, and 77% after 0, 2, 4, and 6 of UV-B irradiance exposure, respectively. In a next step for the use of these M. brunneum EAMa 01/58-Sun soil treatments within a B. oleae IPM strategy, its possible effect of on the B. oleae cosmopolitan parasitoid Psyttalia concolor, its compatibility with the herbicide oxyfluorfen 24% commonly used in olive orchards and the possible presence of the fungus in the olive oil resulting from olives previously placed in contact with the fungus were investigated. Only the highest conidial concentration (1 x 108 conidia ml-) caused significant P. concolor adult mortality (22%) with enduing mycosis in 13% of the cadavers. There were no fungal propagules in olive oil samples resulting from olives previously contaminated by EAMa 01/58-Su conidia. Finally, the strain was demonstrated to be compatible with herbicide since the soil application of the fungus reduced the B. oleae population density up to 50% even when it was mixed with the herbicide in the same tank. The fungal inoculum reached basal levels 4 months after treatments (1.6 x 10³ conidia g soil -1). These results reveal both the efficacy and environmental and food safety of this B. oleae control method, protecting olive groves and improving olive oil quality without negative effects on the natural enemy P. concolor. [ABSTRACT FROM AUTHOR]

Although entomopathogenic fungi (EPF) are best known for their ability to protect crops against insect pests, they may have other beneficial effects on their host plants. These effects, which include promoting plant growth and conferring resistance against abiotic stresses, have been examined in recent years to acquire a better understanding of them. The primary purposes of the present study were (i) to ascertain in vitro whether three different strains of EPF (viz., Metarhizium, Beauveria and Isaria) would increase the Fe bioavailability in calcareous or non-calcareous media containing various Fe sources (ferrihydrite, hematite and goethite) and (ii) to assess the influence of the EPF inoculation method (seed dressing, soil treatment or leaf spraying) on the extent of the endophytic colonization of sorghum and the improvement in the Fe nutrition of pot-grown sorghum plants on an artificial calcareous substrate. All the EPFs studied were found to increase the Fe availability during the in vitro assay. The most efficient EPF was M. brunneum EAMa 01/58–Su, which lowered the pH of the calcareous medium, suggesting that it used a different strategy (organic acid release) than the other two fungi that raised the pH of the non-calcareous medium. The three methods used to inoculate sorghum plants with B. bassiana and M. brunneum in the pot experiment led to differences in re-isolation from plant tissues and in the plant height. These three inoculation methods increased the leaf chlorophyll content of young leaves when the Fe deficiency symptoms were most apparent in the control plants (without fungal inoculation) as well as the Fe content of the above-ground biomass in the plants at the end of the experiment. The total root lengths and fine roots were also increased in response to fungal applications with the three inoculation methods. However, the soil treatment was the most efficient method; thus, its effect on the leaf chlorophyll content was the most persistent, and the effects on the total root length and fine roots were the most apparent. In conclusion, EPF improved the Fe nutrition of the sorghum plants, but their effects depended on the inoculation method. [ABSTRACT FROM AUTHOR]

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]

Metarhizium anisopliae, a fungal pathogen of terrestrial arthropods, kills the aquatic larvae of Aedes aegypti, the vector of dengue and yellow fever. The fungus kills without adhering to the host cuticle. Ingested conidia also fail to germinate and are expelled in fecal pellets. This study investigates the mechanism by which this fungus adapted to terrestrial hosts kills aquatic mosquito larvae. Genes associated with the M. anisopliae early pathogenic response (proteinases Pr1 and Pr2, and adhesins, Mad1 and Mad2) are upregulated in the presence of larvae, but the established infection process observed in terrestrial hosts does not progress and insecticidal destruxins were not detected. Protease inhibitors reduce larval mortality indicating the importance of proteases in the host interaction. The Ae. aegypti immune response to M. anisopliae appears limited, whilst the oxidative stress response gene encoding for thiol peroxidase is upregulated. Cecropin and Hsp70 genes are downregulated as larval death occurs, and insect mortality appears to be linked to autolysis through caspase activity regulated by Hsp70 and inhibited, in infected larvae, by protease inhibitors. Evidence is presented that a traditional host-pathogen response does not occur as the species have not evolved to interact. M. anisopliae retains pre-formed pathogenic determinants which mediate host mortality, but unlike true aquatic fungal pathogens, does not recognise and colonise the larval host. [ABSTRACT FROM AUTHOR]

Conidial spores are often used as the infectious agent during insect biocontrol applications of entomopathogenic fungi. Here we show differential virulence of conidia derived fromMetarhizium anisopliaestrain EAMa 01/58-Su depending upon the solid substrata used for cultivation, where LC50values differed by up to ~10-fold (5.3×106−4.5×105conidia/ml) and LT50values by ~40% (9.8−7.1 d). This fungal strain is also known to secrete proteins that are toxic towards adult Mediterranean fruit flies,Ceratitis capitata, and the Greater wax moth,Galleria mellonella, larvae.In vitroproduction and intrahemoceol injection usingG. mellonellaas the host was used to test fractions during purification of the protein toxins, demonstrating that they elicited defence-related responses including melanisation and tissue necrosis. Production of these proteins/peptides along with a number of potential cuticle degrading enzymes was confirmed bothin vitroand during the infection process (in vivo). Two-dimensional gel electrophoresis, followed by gel elution and bioassay, was used to identify at least three proteins or peptides (molecular mass=11, 15 and 15 kDa) as mediating the observed insect toxicity. These data demonstrate thatin vitroscreening for insect toxins can mimicin vivo(i.e. during the infection process) secretion and applies the use of proteomics to invertebrate pathology. [ABSTRACT FROM AUTHOR]

Background: The entomopathogenic anamorphic fungus Beauveria bassiana is currently used as a biocontrol agent (BCA) of insects. Fifty-seven Beauveria bassiana isolates -53 from Spain- were characterized, integrating group I intron insertion patterns at the 3'-end of the nuclear large subunit ribosomal gene (LSU rDNA) and elongation factor 1-alpha (EF1-α) phylogenetic information, in order to assess the genetic structure and diversity of this Spanish collection of B. bassiana. Results: Group I intron genotype analysis was based on the four highly conserved insertion sites of the LSU (Ec2653, Ec2449, Ec2066, Ec1921). Of the 16 possible combinations/genotypes, only four were detected, two of which were predominant, containing 44 and 9 members out of 57 isolates, respectively. Interestingly, the members of the latter two genotypes showed unique differences in their growth temperatures. In follow, EF1-a phylogeny served to classify most of the strains in the B. bassiana s.s. (sensu stricto) group and separate them into 5 molecular subgroups, all of which contained a group I intron belonging to the IC1 subtype at the Ec1921 position. A number of parameters such as thermal growth or origin (host, geographic location and climatic conditions) were also examined but in general no association could be found. Conclusion: Most Spanish B. bassiana isolates (77.2%) are grouped into a major phylogenetic subgroup with wordwide distribution. However, high phylogenetic diversity was also detected among Spanish isolates from close geographic zones with low climatic variation. In general, no correlation was observed between the molecular distribution and geographic origin or climatic characteristics where the Spanish B. bassiana isolates were sampled. [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]

The article presents a study on the effect of blastophore yields, fungal biomass, and toxicity of fungal proteins of Metarhizium anisopliae against the insecticidal protein control of Ceratitis capitata adults. Based on results, concentration of nitrogen source influences the production of insecticidal protein and toxicity of the crude soluble protein extract (CSPE). In conclusion, insecticidal effect against C. capitata can be improved through optimization of culture media.

The article presents a study on the use of entomopathogenic fungus Metarhizium anisopliae in controlling the Ceratitis capitata. The researchers gathered Metarhizium anisopliae strain at the University of Cordoba in Cordoba, Spain, and the conodial culture of the insecticidal fungal proteins were prepared using the Adamek's liquid medium. Accordingly, the secreted chemical from the crude soluble protein extract (CSPE) were tested in a diet test. The results have revealed four fungal proteins.

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]

The aim of the current study was to delve into the causes of mortality of Spodoptera littoralis larvae feeding on Metarhizium-colonized plants in the absence of fungal outgrowth on the cadavers as previous studies reported and to elucidate the possible indirect effects of this fungus-colonized diet. The effect was evaluated in experiments conducted using leaf discs of colonized plants and in planta using fungus-colonized whole plants. The mortality rates of larvae fed on Metarhizium-colonized melon leaves were 45.0% and 87.5%, and the average survival times were 6.6 and 3.1 days in experiments performed with discs and in planta, respectively. Notably, these mortality levels were not associated with observed apoptosis mediated by caspases 1, 3-7 and 8; thus, further investigation into the possible immune system reaction of the insect after the ingestion of colonized plants is required. The leaf consumption of S. littoralis larvae fed on melon-colonized leaves was lower than that on control plants in the disc experiments but not in experiments conducted in planta. In this regard, in experiments performed in planta, plant damage increased larval mortality in both fungally challenged and control larvae. There was also a meaningful effect of exposure to Metarhizium-colonized melon leaf discs on S. littoralis fitness, with significant reductions in 39.0% and 22.0% in female fecundity and egg fertility, respectively, detected in females emerging from pupae developing from larvae surviving exposure to colonized plant discs; all larvae died in the in planta experiments. Hence, the present work presents new findings revealing the high potential of endophytic entomopathogenic fungi to improve the outcome of foliar applications against chewing insects in the short, mid- and long term, by the reduction of the reproductive potential of surviving adults and reveals new insights into the development of bioassays with whole plants for more detailed evaluation of the impact of these fungi as endophytes used for plant protection. [ABSTRACT FROM AUTHOR]