[Show abstract][Hide abstract] ABSTRACT: Pollen beetles Meligethes aeneus were collected in oilseed rape fields at different sites in Switzerland in spring 2004-2005 and 32 isolates of the fungal genus Beauveria occurring as latent infections in the beetles were obtained and molecularly characterized. Three major clades, Beauveria bassiana sensu stricto (Clade A: n=13), Beauveriabrongniartii (Clade B: n=1) and Beauveria Clade C (n=18) were identified among the isolates based on sequences of the ITS region and the 5' end of EF1-α. B. bassiana s.s. was further separated in the two clades, Eu_1 (n=10) and Eu_4 (n=3). The intergenic region Bloc provided best resolution of the individual clades B. bassiana s.s. Eu_1, Eu_4 and B. brongniartii. No specific clade of Beauveria appeared to be associated with adult M. aeneus populations. However, data suggested high relative abundance of Beauveria Clade C among the fungal entomopathogens infecting M. aeneus. Characterization of the isolates by simple sequence repeats (SSR) revealed further genotypic diversity within the clades except B. bassiana s.s. Eu_4 which appeared to be clonal. However, the individual SSR markers were differentially amplifiable from isolates of the different clades. It is therefore important to identify the underlying phylogenetic affinity of Beauveria isolates to interpret results based on SSR markers. The data suggest that not all available SSR markers are suitable for reliable characterization of diversity within Beauveria Clade C.
Journal of Invertebrate Pathology 01/2012; 109(1):76-82. · 2.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorokin (Hypocreales: Clavicipitaceae) was applied in maize fields to control the Western Corn Rootworm Diabrotica virgifera virgifera Le Conte (Coleoptera: Chrysomelidae). Establishment and persistence of two strains of M. anisopliae were investigated after application as ‘fungal colonized barley kernels’ (FCBK) into the soil and as a spore suspension on maize leaves and on the soil surface in 2006 and 2007 at two locations in Hungary.The applied fungal strains were able to establish at both locations and a long-term persistence of at least 15 months could be recorded in the soil. A positive correlation between density of colony forming units (CFU) in the soil and the soil inhabiting stages of the host insect D. v. virgifera could be found. M. anisopliae spores applied on maize leaves were able to survive for no longer than 3 days after application, whereas on the soil surface a noticeably increase of fungus densities were found after treatments. Molecular markers were used to identify the applied M. anisopliae strains before and after application of FCBK into the soil of the maize field.
[Show abstract][Hide abstract] ABSTRACT: Diabroticina is a speciose subtribe of New World Chrysomelidae (Subfamily Galerucinae: Tribe Luperini) that includes pests such as corn rootworms, cucumber beetles and bean leaf beetles (e.g. Diabrotica, Acalymma, Cerotoma species). The evolution and spread of pesticide resistance, the European invasion of Diabrotica v. virgifera LeConte, and possible development of resistance due to the large-scale deployment of Diabrotica-active Bt maize in North America have generated a sense of urgency in developing biological control options against Diabroticina pests. In the present study, we review available knowledge on biological control options, including 290 publications on natural enemy-Diabroticina associations in the New World. Several natural enemy species or groups appear to be promising candidates for control strategies with different ecological rationales. We propose that future research should pursue: (1) development of inundative biological control products, particularly mass-produced entomopathogenic nematodes and fungi, (2) understanding of specific natural enemies of Diabroticina larvae throughout the Americas and of adults particularly in higher altitudes of Central America or northern South America including potential classical biological control agents against D. v. virgifera; (3) enhancement of natural enemies through cultural practices, i.e., reduced tillage, reduced weed control, cover crops, diversified crop rotations or soil amendments. Research and action must be coordinated to accelerate the exploration of biological control options.
Biocontrol Science and Technology 01/2009; · 0.71 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: An important mechanism for insect pest control should be the use of fungal entomopathogens. Even though these organisms have been studied for more than 100 y, their effective use in the field remains elusive. Recently, however, it has been discovered that many of these entomopathogenic fungi play additional roles in nature. They are endophytes, antagonists of plant pathogens, associates with the rhizosphere, and possibly even plant growth promoting agents. These findings indicate that the ecological role of these fungi in the environment is not fully understood and limits our ability to employ them successfully for pest management. In this paper, we review the recently discovered roles played by many entomopathogenic fungi and propose new research strategies focused on alternate uses for these fungi. It seems likely that these agents can be used in multiple roles in protecting plants from pests and diseases and at the same time promoting plant growth.
[Show abstract][Hide abstract] ABSTRACT: All three larval instars of Diabrotica virgifera virgifera LeConte (western corn rootworm, Coleoptera: Chrysomelidae) feed on the roots of maize, Zea mays (L.). We assessed the efficacies of the following four agents in controlling these larvae: (1) the entomopathogenic fungus
Metarhizium anisopliae (Metsch.) Sorokin (Hypocreales: Clavicipitaceae), (2) the nematode Heterorhabditis bacteriophora Poinar (Nematoda: Rhabditida), (3) a tefluthrin-based soil insecticide and (4) clothianidin-coated seeds. The agents were
applied in field plot experiments in southern Hungary in 2006 and 2007. Efficacy was assessed by comparing the number of emerging
D. v. virgifera adults and corresponding root damage among treatments and untreated controls. All agents significantly reduced D. v. virgifera numbers and root damage, but the relative success of each treatment was variable. On average across fields and years, the
nematode and the two insecticides reduced D. v. virgifera by 65±34% SD, while the fungus reduced D. v. virgifera by 31±7%. According to the node injury scale, the agents prevented 23–95% of potential root damage. Large-scale commercialisation
of these biological agents could offer viable and practical control options against D. v. virgifera.
[Show abstract][Hide abstract] ABSTRACT: Pandora neoaphidis is one of the most important fungal pathogens of aphids and has a great potential for use in biocontrol. Little is known on how this fungus persists in an area and in particular on its overwintering strategies. It is hypothesized that natural areas play an important role for survival and that soil may serve as a source of inoculum for new aphid populations in spring. To test these hypotheses, a cultivation-independent PCR-based diagnostic tool was developed, that allows the detection of P. neoaphidis in the environment. Two P. neoaphidis specific PCR primer pairs were designed, targeting sequences in the ribosomal RNA gene cluster. Specificity of both primer pairs was demonstrated with P. neoaphidis and non-target close entomophthoralean relatives. Moreover, single amplicons of expected sizes were obtained with both primer pairs from various environmental sample types, including aphid cadavers, plant material, and soil. The PCR-based diagnostic tool was applied to investigate the persistence of P. neoaphidis in soil samples obtained in 2004/2005 from a nettle field harboring infected aphids in fall 2004. P. neoaphidis was detected in every sample collected in November 2004 and March 2005, suggesting an overwintering stage of P. neoaphidis in top soil layers. The developed cultivation-independent PCR-based tool will be valuable for further investigation of the ecology of P. neoaphidis and for the development and future implementation of management strategies against aphids involving conservation biocontrol.
Journal of Invertebrate Pathology 04/2008; 99(1):49-56. · 2.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The Western Corn Rootworm D. virgifera virgifera Le Conte (Coleoptera: Chrysomelidae), a serious pest of maize, has been recently introduced into Europe. Several approaches
for its control are presently under investigation including microbial agents. In order to get information on the role of naturally
occurring pathogens in the regulation of Diabrotica populations, we started an investigation in established populations in Hungary, Romania, Serbia, Austria, and Italy in 2005
and 2006. In infested maize fields in Hungary, plants and their root systems were grubbed out and larvae and pupae were collected.
Adult D. v. virgifera were collected in Hungary, Austria, Romania, Serbia and Italy. Additionally, the occurrence of entomopathogenic fungi in
soils of maize fields was determined using Galleria mellonella and Tenebrio molitor larvae as bait insects. The density of entomopathogenic fungi was obtained by plating soil suspension on selective medium.
Metarhizium anisopliae and Beauveria spp. infections were found in 1.4% of field collected larvae, 0.2% of field collected pupae and 0.05% of field collected
adults. Whereas natural infections of D. v. virgifera were rarely found, a high density of insect pathogenic fungi was recorded in Hungarian soils. M. anisopliae could be detected in every maize field either using the “bait method” or a “selective medium” method. This is the first report
of a natural occurrence of entomoparasitic nematodes (Heterorhabditis sp., Steinernema sp.) in Diabrotica v. virgifera in Europe.
[Show abstract][Hide abstract] ABSTRACT: The western corn rootworm Diabrotica virgifera virgifera Le Conte (Col., Chrysomelidae), a serious pest of maize, has been recently introduced into Europe. Several approaches for its control are presently under investigation including microbial agents. During a field survey in Hungary in 2005, naturally occurring entomopathogenic fungi were found to attack this pest. These novel isolates together with standard isolates were tested for virulence against D. v. virgifera larvae and adults. Twenty strains of Metarhizium anisopliae, Beauveria bassiana and Beauveria brongniartii were used in bioassays in the laboratory. Larvae and adults were dipped into a spore suspension with a concentration of 1 × 107 conidia (con.)/ml. They were kept for 14 days at 22°C (±2°C) and 70% relative humidity. The number of infected larvae and adults were counted and infection rates were calculated. Adults were significantly more susceptible to entomopathogenic fungi than larvae. The most virulent isolate infected about 47% of larvae (M. anisopliae Ma2277), whereas the infection rate in adults was up to 97% (M. anisopliae Ma2275). Isolates of M. anisopliae caused significantly higher mortalities than isolates of B. brongniartii and B. bassiana. Most of the adult beetles were killed within 12 days. Isolates from D. v. virgifera were more virulent than those from other hosts.
[Show abstract][Hide abstract] ABSTRACT: Fourteen polymorphic microsatellite markers were isolated from the entomopathogenic fungus, Metarhizium anisopliae, based on enriched genomic libraries. In order to assess allelic variability, the microsatellite loci were analysed in a collection of 34 isolates sampled from across Switzerland. The number of detected alleles in 14 loci ranged from two to eight and expected heterozygosity from 0.265 to 0.808. Because of the high expected heterozygosity, the 14 microsatellite loci are very useful for ecological studies and analysis of population diversity, and to identifying, monitoring, and tracking M. anisopliae strains applied as biological control agents.
[Show abstract][Hide abstract] ABSTRACT: In field tests between 1985 and 1992, various sites which were infested with Melolontha melolontha were treated with the biocontrol fungus Beauveria brongniartii. The fungus was applied either as blastospores or as commercially available fungus colonized barley kernels (FCBK). In 1998/1999, soil samples were collected from the test sites and B. brongniartii isolates recovered and maintained in a culture collection. Isolates from this collection were subjected to genetic analyses by use of specific microsatellite markers. The applied B. brongniartii strains were detected at all test sites up to 14 years after their application. At some sites, the B. brongniartii populations consisted of the applied strains exclusively, whereas at other sites, indigenous populations or isolates that may have arisen from mutations or interactions among the applied strains and indigenous isolates were present in addition to the applied strains. The results suggested that applied B. brongniartii strains and indigenous B. brongniartii populations could coexist in the same habitat. Demonstration of long-term persistence of the applied B. brongniartii strains in the fields supported the success of this approach for the biological control of M. melolontha.
[Show abstract][Hide abstract] ABSTRACT: We have identified ten microsatellite markers in the entomopathogenic fungus Beauveria brongniartii from three genomic libraries enriched for (AAG)n-, (TG)n-, or (TC)n-repeats. The levels of polymorphism of the identified microsatellite loci were assessed in a collection of Beauveria strains originating from different countries, areas, and host insects. Two geographically separated Swiss populations of B. brongniartii originating from European cockchafer (Melolontha melolontha) were also analysed. Microsatellites containing (AAG)n-repeats generally displayed high levels of polymorphism, whereas microsatellites containing either (TG)n- or (TC)n-repeats displayed lower levels of polymorphism. Cluster analysis revealed that strains isolated from M. melolontha larvae, and two strains isolated from Melolontha hippocastani or Amphimallon solstitiale larvae, formed one cluster which was separated from strains isolated from adult M. melolontha and other adult insects. A high degree of biodiversity was detected among B. brongniartii strains of the two separated Swiss populations. Distinct haplotypes were identified in 29 of 35 B. brongniartii strains from population A and in 12 of 28 B. brongniartii strains from population B. The high discrimination power of the identified microsatellites makes them a valuable tool, suited for the characterization and identification of B. brongniartii strains used as biocontrol agents. In addition, they may be applied to monitor biological control strains of B. brongniartii in the field and possibly to investigate their interactions with indigenous B. brongniartii isolates.
Mycological Research 01/2001; 105(9):1079-1087. · 2.81 Impact Factor