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Aphids are associated with several facultative bacterial endosymbionts that may influence their interactions with other organisms. We show here that one of the three most common facultative symbionts of pea aphid (Acyrthosiphon pisum), the bacterium Regiella insecticola, has a major effect on host resistance to a fungal pathogen. Experimental establishment of the bacterium in uninfected aphid clones led to higher survival after fungal attack. The bacteria also increased the aphid's inclusive fitness, because the presence of the symbiont reduced the probability of fungal sporulation on aphid cadavers, hence lowering the rate of transmission of the disease to nearby related aphids.

Meconema meridionale Costa (Orthoptera, Tettigoniidae) infected with Zoophthora giardii Bałazy (Entomophthorales, Entomophthoraceae) were collected at five localities in the northern half of Switzerland. At one of these sites, the fungus caused epizootics in two subsequent years. Symptoms and morphological data coincide with those given by Bałazy who found the fungus on Meconema thalassinum De Geer. Conidiobolus gustafssonii Bałazy (Entomophthorales, Ancylistaceae) was found on a single Ectobius vittiventris Costa (Blattodea, Ectobiidae). Symptoms and dimensions of the primary conidia correspond with the original description given by Bałazy who found the fungus on Ectobius lapponicus L. The two fungi are new for Switzerland and Meconema meridionale and E. vittiventris represent new hosts for these pathogens.

The entomophthoralean fungus Zoophthora rhagonycharum (Bałazy) S. Keller, previously recorded in Europe from Poland and Switzerland, is now reported in North America from New York State, United States of America. On both continents, this obligate insect pathogen is known only from resting spores found within dead, adult native soldier beetles (Cantharidae) of the genus Rhagonycha Eschscholtz, 1830. Resting spores have undulating, light brown episporia. In New York, columnar rhizoids attach cadavers tightly to the undersides of leaves in the understory of hardwood forests in late June and early July.

Primary conidia of the entomopathogens Erynia (subgenus Neopandora) delphacis (1 isolate) and Erynia (Neopandora) neoaphidis (3 isolates) were stimulated to form germ-tubes with Tween 20 and with free, long-chain fatty acids, each incorporated into Entomophthora complete medium (ECM). When combined with other basal media (three tested), these compounds did not stimulate germ-tube formation. Triacylglycerols and vegetable oils, added to the same media, allowed almost complete resporulation in the fungi. In both species, Tween 20 (0.1%) encouraged greater germ-tube production (41-69%) than the fatty acids (0.1%) ( less than or equal to 36%). For E. delphacis, Tween 20 and the fatty acids differed significantly, but for E. neoaphidis the differences were almost always insignificant. Myristic and oleic acids stimulated germ-tube formation in both species. Palmitic acid allowed almost complete resporulation of the fungi, except for one isolate of E. neoaphidis that formed germ-tubes. Linoleic acid, tested only for E. delphacis, was fungistatic to most conidia. Higher concentrations of the fatty acids ( less than or equal to 1%) did not increase germ-tube formation, except 1% oleic acid which affected E. delphacis alone (>80% germination and germ-tubes). Linoleic acid, and sometimes also myristic and oleic, were fungistatic and/or toxic, depending on their concentration and on medium composition. Addition of fatty acids to ECM usually extended the lag period, and altered the morphology of the conidia and germ-tubes. These phenomena were not observed with Tween 20. Colonies were formed by E. delphacis alone, stimulated by ECM supplemented with Tween 20 or fatty acids. The results are discussed with respect to biological and physiological aspects of germination, and with respect to the mode of action of the fatty acids and the surfactant.

The alfalfa weevil (Hypera postica Gyllenhal, Coleoptera: Curculionidae) is a major pest of alfalfa crops. Chemical control measures are inefficient, but the larvae are often infested by parasitoid wasps of the genus Bathyplectes Forster (Hymenoptera: Ichneumonidae), which offer a potential biological control strategy. The development of effective biological control requires the identification of parasitoid species, but conventional methods involve the rearing of parasitoids to the puparium stage for morphological confirmation. Here we designed a PCR method in which two pairs of primers are used to detect and identify Bathyplectes curculionis Thomson and Bathyplectes anura Thomson larvae in a faster way. We compared conventional rearing to the new method as a means to determine the parasitism rates caused by each species in Spain during the 2019 and 2020 seasons. In 2019, the PCR method detected five times as many B. curculionis events and twice as many B. anura events. Similarly, in 2020, the PCR method detected seven times as many B. curculionis events and twice as many B. anura events. High mortality of H. postica larvae was recorded in 2020 due to an epizootic attack by the fungus Zoophthora phytonomi Arthur (Zygomycetes: Entomophthorales), explaining the lower overall rate of parasitism and the widespread death of the larvae. Statistical analysis confirmed that the PCR method is more sensitive than conventional rearing for the detection of target parasitoids. However, only rearing can confirm the effective parasitism or the presence of unexpected species. We therefore recommend the use of both methods in parallel when evaluating host-parasitoid systems.

The fungal order Entomophthorales in the Zoopagomycota includes many fungal pathogens of arthropods. This review explores six genera in the subfamily Erynioideae within the family Entomophthoraceae, namely, Erynia, Furia, Orthomyces, Pandora, Strongwellsea, and Zoophthora. This is the largest subfamily in the Entomophthorales, including 126 described species. The species diversity, global distribution, and host range of this subfamily are summarized. Relatively few taxa are geographically widespread, and few have broad host ranges, which contrasts with many species with single reports from one location and one host species. The insect orders infected by the greatest numbers of species are the Diptera and Hemiptera. Across the subfamily, relatively few species have been cultivated in vitro, and those that have require more specialized media than many other fungi. Given their potential to attack arthropods and their position in the fungal evolutionary tree, we discuss which species might be adopted for biological control purposes or biotechnological innovations. Current challenges in the implementation of these species in biotechnology include the limited ability or difficulty in culturing many in vitro, a correlated paucity of genomic resources, and considerations regarding the host ranges of different species.

Pandora neoaphidis and Entomophthora planchoniana (phylum Entomophthoromycota) are important fungal pathogens on cereal aphids, Sitobion avenae and Rhopalosiphum padi. Here, we evaluated and compared for the first time the virulence of these two fungi, both produced in S. avenae cadavers, against the two aphid species subjected to the same exposure. Two laboratory bioassays were carried out using a method imitating entomophthoralean transmission in the field. Healthy colonies of the two aphid species were exposed to the same conidial shower of P. neoaphidis or E. planchoniana, in both cases from a cadaver of S. avenae. The experiments were performed under LD 18:6 h at 21 °C and a successful transmission was monitored for a period of nine days after initial exposure. Susceptibility of both S. avenae and R. padi to fungal infection showed a sigmoid trend. The fitted nonlinear model showed that the conspecific host, S. avenae, was more susceptible to E. planchoniana infection than the heterospecific host R. padi, was. In the case of P. neoaphidis, LT50 for S. avenae was 5.0 days compared to 5.9 days for R. padi. For E. planchoniana, the LT50 for S. avenae was 4.9 days, while the measured infection level in R. padi was always below 50 percent. Our results suggest that transmission from conspecific aphid host to heterospecific aphid host can occur in the field, but with expected highest transmission success to the conspecific host.

Aphids commonly harbor bacterial facultative symbionts that have a variety of effects upon their aphid hosts, including defense against hymenopteran parasitoids and fungal pathogens. The soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is infected with the symbiont Arsenophonus sp., which has an unknown role in its aphid host. Our research goals were to document the infection frequency and diversity of the symbiont in field-collected soybean aphids, and to determine whether Arsenophonus is defending soybean aphid against natural enemies. We performed diagnostic PCR and sequenced four Arsenophonus genes in soybean aphids from their native and introduced range to estimate infection frequency and genetic diversity, and found that Arsenophonus infection is highly prevalent and genetically uniform. To evaluate the defensive role of Arsenophonus , we cured two aphid genotypes of their natural Arsenophonus infection through ampicillin microinjection, resulting in infected and uninfected isolines within the same genetic background. These isolines were subjected to parasitoid assays using a recently introduced biological control agent, Binodoxys communis [Braconidae], a naturally recruited parasitoid, Aphelinus certus [Aphelinidae], and a commercially available biological control agent, Aphidius colemani [Braconidae]. We also assayed the effect of the common aphid fungal pathogen, Pandora neoaphidis (Remaudiere & Hennebert) Humber (Entomophthorales: Entomophthoraceae), on the same aphid isolines. We did not find differences in successful parasitism for any of the parasitoid species, nor did we find differences in P. neoaphidis infection between our treatments. Our conclusion is that Arsenophonus does not defend its soybean aphid host against these major parasitoid and fungal natural enemies.

The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.

A three-year survey of entomophthoralean pathogenic fungi of aphids from horticultural crops in La Plata, Argentina, was conducted. Nine species of aphids, including Aphis fabae Scopoli, Aphis gossypii Glover, Brevicoryne brassicae (L.), Lipaphis erysimi (Kaltenbach), Macrosiphum euphorbiae (Thomas), Myzus sp., Myzus persicae (Sulzer), Nasonovia ribisnigri (Mosley) and Capitophorus elaeagni (del Guercio) were recorded as hosts of entomopathogenic fungi. Six species of Entomophthorales that infected and killed aphids were found in vegetable crops. The fungal species identified were Conidiobolus obscurus (Hall & Dunn) Remaudière & Keller, Entomophthora planchoniana Cornu, Neozygites fresenii (Nowakowski) Remaudière & Keller, Pandora neoaphidis (Remaudière & Hennebert) Humber, Zoophthoraradicans (Brefeld) Batko and Zoophthora sp. Pandora neoaphidis was the most predominant pathogen of aphids and was found throughout the summer (December–March) 2004. The recovery of C. obscurus, N. fresenii and P. neoaphidis represent first records of these fungi for South America.