Bassiana Infection Research Articles

DECIPHERING THE RELATIONSHIPS AMONG ENZYMATIC SYSTEMS AND VIRULENCE OF Beauveria bassiana: A REVIEW

Intensive crop production and extensive use of harmful synthetic chemical pesticides create numerous socio-economic problems worldwide. Therefore, sustainable solutions are needed for insect pest control, such as biological control agents. The fungal insect pathogen Beauveria bassiana has shown considerable potential as a biological control agent against a broad range of insects. The insight into the virulence mechanism of B. bassiana is essential to show the robustness of its use. B. bassiana has several determinants of virulence, including the production of cuticle-degrading enzymes (CDEs), such as proteases, chitinases, and lipases. CDEs are essential in the infection process as they hydrolyze the significant components of the insect's cuticle. Moreover, B. bassiana has evolved effective antioxidant mechanisms that include enzyme families that act as reactive oxygen species (ROS) scavengers, e.g., superoxide dismutases, catalases, peroxidases, and thioredoxins. In B. bassiana, the number of CDEs and antioxidant enzymes are characterized in recent years. These enzymes are believed to be crucial player of evolutionary process in this organism and their role in various mechanism of biological control. Recent discoveries have significantly increased our potential understanding on several potentially wanted unknown mechanisms of B. bassiana infection. This review focuses on the progress detailed in the studies of these enzymes and provides an overview of enzymatic activities and their contributions to virulence.

Pathogen‐mediated modulation of host metabolism and trophic interactions in Spodoptera littoralis larvae

AbstractThe success and sustainability of entomopathogens in insect control depend on their stress potential and ability to modulate certain physiological aspects of their insect hosts. In the present study, newly moulted fourth instars of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) were incubated for 24 h with the median lethal dose (LD50) of Steinernema riobrave, Heterorhabditis bacteriophora, or Beauveria bassiana to determine their effects on certain metabolic activities and nutritional physiology compared to those of uninfected (control) larvae. Infection decreased total protein, lipid, and carbohydrate contents. Carbohydrase activity was found to be pathogen and enzyme dependent, with S. riobrave and H. bacteriophora infection increasing amylase, invertase, and trehalase activities. Beauveria bassiana infection decreased amylase and invertase activities but increased that of trehalase. Infection with the three pathogens also enhanced phosphatase activity. All pathogens reduced transaminase activity. Changes in the nutritional indices varied not only with the variation in the candidate entomopathogen and surviving instar but also with the variation in the age of the same instar, with a profound change in late last instars (3‐day‐old sixth instars). The link between the metabolic activities and nutritional indices was discussed. Overall, the entomopathogen‐host interaction appears to be primarily nutritional. The pleiotropic effects of the median lethal infections on the insect pest might eventually help in the biological control potential of the entomopathogens tested.

Immunotranscriptome analysis of Plutella xylostella reveals differences in innate immune responses to low- and high-virulence Beauveria bassiana strain challenges.

Entomopathogenic fungi have developed multiple strategies to overcome the immune defenses of their target insects, whereas insect pests have devised various defense mechanisms to combat fungal infection. However, differences in the molecular mechanisms of the innate immune defense strategies of insects upon infection with different fungal strains from the same species have not been reported. Two Beauveria bassiana strains were obtained that significantly varied in their pathogenicity but were comparable in terms of growth, conidial yield, and cuticle penetration. To investigate the molecular mechanisms underlying the immune response of Plutella xylostella infected with these two strains, RNA-Seq was performed 48 h after infection. A total of 1027 differentially expressed genes (DEGs) were identified, and more than 200 DEGs were enriched in Kyoto Encyclopedia of Genes and Genome (KEGG) pathways involved in disease response, revealing differences in the immune response of P. xylostella to different B. bassiana infections at 48 h. Twenty-eight of the DEGs were related to innate immune functions, such as pathogen recognition, immune system activation and antimicrobial reactions. RNA interference (RNAi)-mediated gene silencing assays showed that PxApoLIII and PxCSP played critical roles in the P. xylostella immune response. PxApoLIII was expressed at higher levels during infection with the high-virulence strain, whereas PxCSP showed the opposite expression pattern during infection with the low-virulence strain, indicating that PxApoLIII and PxCSP might participate in P. xylostella innate immune defense against high- and low-virulence B. bassiana strains. The present findings demonstrate that strains of a single species of pathogenic fungi that differ in virulence can induce the expression of different genes in P. xylostella. These results advance our knowledge of the molecular mechanisms underlying fungi-pest interactions.

Comparative Genomic and Metabolomic Analyses of Two Pseudomonas aeruginosa Strains With Different Antifungal Activities.

Pseudomonas aeruginosa isolated from the plant rhizosphere has been widely used as an effective strain in biological control against plant disease. This bacterium promotes plant growth and protect plants against various phytopathogens through the production of phenazine metabolites. In this study, the strain P. aeruginosa Y12 with anti-Beauveria bassiana activity was isolated from the gut of housefly larvae. It was comparatively analyzed with the strain P. aeruginosa P18, which showed no anti-B. bassiana activity. Genomic and metabolomic methods were used to obtain a comprehensive understanding of the antimicrobial mechanism of Y12. After whole-genome resequencing of the two strains, a total of 7,087 non-synonymous single-nucleotide polymorphisms (nsSNPs), 1079 insertions and deletions (InDels), 62 copy-number variations (CNVs) and 42 structural variations (SV) were found in both strains. We analyzed the differentially abundant metabolites between Y12 and P18, and identified six bioactive compounds that could be associated with the antimicrobial activity of Y12. Additionally, we found that, unlike other previously reported rhizospheric P. aeruginosa strains, Y12 could produce both phenazine-1,6-dicarboxylic acid (PDC) and pyocyanin (PYO) at significantly higher concentrations than P18. As B. bassiana is an effective biological insecticide that can cause high mortality in adult houseflies but has little effect on housefly larvae, we believe that P. aeruginosa Y12, identified in housefly larvae but not in adults, were beneficial for the development of housefly larvae and could protect them from B. bassiana infection through the production of toxic metabolites.

Immunological regulation by a β-adrenergic-like octopamine receptor gene in crowded larvae of the oriental Armyworm, Mythmina separata

Recent reports demonstrate that octopamine plays an important immunological role in crowded larvae of the Oriental Armyworm, Mythmina separata. We identified an octopamine receptor, the β-adrenergic-like gene (designated MsOctβ2R), with a 1191 bp open reading frame that encodes 396 amino acids and contains seven conserved hydrophobic transmembrane domains. Multiple sequence alignments and a phylogenetic analysis indicated that MsOctβ2R was orthologous to Octβ2R that is present in other lepidopterans. MsOctβ2R was expressed throughout all developmental stages with higher relative expression during the fourth instar and adult stages. MsOctβ2R was highly expressed in the ventral nerve cord and the fat body relative to other examined tissues. Elevated MsOctβ2R expression was observed in larvae that were under higher-density conditions (7 and 10 larvae per jar). Silencing MsOctβ2R expression via dsRNA injections in larvae from higher-density conditions significantly decreased phenoloxidase (PO) and lysozyme activity, total haemocyte counts, and survival rates against Beauveria bassiana infections (54.06%, 9.91%, 36.22%, and 23.53%, respectively) when compared with control larvae. These results suggest that high-density conditions might alter prophylactic immunity in larvae by regulating the MsOctβ2R gene in M. separara and provide new insights into density-dependent prophylaxis in insects.

Thermoregulatory response of Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) to infection by Beauveria bassiana, and its effect on survivorship and reproductive success

Behavioral thermoregulation is a defensive strategy employed by some insects to counter infections by parasites and pathogens. Most reported examples of this type of thermoregulatory response involve behavioral fevering. However depending upon the life history of a host-insect and that of the parasite or pathogen, the host may respond by cold-seeking behavior. Thermoregulation is not only ecologically important; it may affect the success of parasites and pathogens as biological control agents. We examined if Frankliniella occidentalis (Pergande) thermoregulates in response to infection by Beauveria bassiana, a fungal pathogen commonly used for biological control. Fungal-infected thrips preferentially moved to cooler areas (~12 °C) while healthy thrips sought out warmer temperatures (~24 °C). This cold-seeking behavior suppressed the growth of B. bassiana in infected thrips, and significantly improved survivorship of infected thrips. At 24 °C, males only survived up to 10 d after infection and females up to 20 d after infection, which was substantially poorer survivorship than that of corresponding healthy individuals. However, individuals of both sexes survived up to 48 d after infection at 12 °C, which was a much less severe reduction in survivorship compared with the effect of B. bassiana infection at 24 °C. The proportion of females among progeny from infected thrips at 12 °C was higher than at 24 °C. Therefore, cold-seeking behavior is beneficial to F. occidentalis when infected by B. bassiana, and its effects should be considered in the use of B. bassiana in biological control programs.

Invasion success of a widespread invasive predator may be explained by a high predatory efficacy but may be influenced by pathogen infection

Invasive alien species (IAS) can drive community change through ecological interactions. Parasites and pathogens can play an important role in community function including mitigating or enhancing IAS impacts. Despite this, the degree to which pathogen pressure influences IAS impacts remains poorly understood. We quantified the predatory behaviour of the highly invasive alien harlequin ladybird (Harmonia axyridis) and two UK native species, the 7-spot (Coccinella septempunctata) and 2-spot (Adalia bipunctata) ladybirds, using comparative functional response experiments. We investigated the impacts of pathogen infection on the predatory ability of the ladybirds by exposing individuals to Beauveria bassiana, a widespread entomopathogen. Invasive H. axyridis was a more efficient predator than both the native A. bipunctata and C. septempunctata, often having higher attack and/or lower prey handling time coefficients, whereas native A. bipunctata were the least efficient predators. These differences were found in both adult and larval life-stages. Beauveria bassiana infection significantly altered the predatory efficiency of adult and larval ladybird predators. The effects of pathogenic infection differed between species and life-stage but in many cases infection resulted in a reduced predatory ability. We suggest that the interactions between IAS and pathogens are integral to determining invasion success and impact.

Label-free LC-MS/MS proteomic analysis of the hemolymph of silkworm larvae infected with Beauveria bassiana

Beauveria bassiana, a pathogen of the economically important silkworm (Bombyx mori), causes serious losses in the sericulture industry; however, the mechanisms underlying B. bassiana infection and the silkworm response are not fully understood. To obtain new insights into the interaction between B. bassiana and its host, hemolymph samples from fifth instar silkworm larvae infected with B. bassiana were analyzed at 36-h post-inoculation using a label-free LC-MS/MS proteomic technique. In total, 671 proteins were identified in the hemolymph, including 87 differentially expressed proteins, 42 up-regulated and 45 down-regulated in infected larvae. Six were detected only in infected larvae, and five were detected only in uninfected larvae. Based on GO annotations, 48 of the differentially expressed proteins were involved in molecular functions, 42 were involved in biological processes, and 39 were involved in cell components. A KEGG pathway analysis indicated that these differentially expressed proteins participate in 85 signal transduction pathways, including the amoebiasis, MAPK signaling, Hippo signaling, Toll and Imd signaling, and lysosome pathways. The silkworm hemolymph is the main site for B. bassiana replication. We identified differentially expressed proteins involved in the regulation of the host response to B. bassiana infection, providing important experimental data for the identification of key factors contributing to the interaction between the pathogenic fungus and its host.

Effect of Bark Application With Beauveria bassiana and Permethrin Insecticide on the Walnut Twig Beetle (Coleoptera: Curculionidae) in Black Walnut Bolts.

Formulations of entomopathogenic (insect-killing) fungi represent alternatives to synthetic insecticides in the management of forest and shade tree insects. We evaluated bark spray applications of the entomopathogen Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) strain GHA (BotaniGardES), permethrin insecticide (Astro), and water (control) on colonization of black walnut (Juglans nigra L.) (Fagales: Juglandaceae) bolts by the walnut twig beetle (Pityophthorus juglandis Blackman) (Coleoptera: Curculionidae), vector of the fungus that causes thousand cankers disease. Treated bolts were baited with a P. juglandis aggregation pheromone lure and deployed in infested walnut trees. Bark application of permethrin prevented P. juglandis colonization of the phloem. Although treatment of bolts with the B. bassiana suspension did not reduce P. juglandis colonization or short-term emergence relative to the control treatment, it increased the B. bassiana infection rate from 25 to 62% of emerged adults. Results suggest that commercial applications of B. bassiana strain GHA may help augment natural levels of infection by this entomopathogen in the eastern United States, and support continued exploration of entomopathogens for biological control of the walnut twig beetle.

Dual application of entomopathogenic nematodes and fungi on immune and antioxidant enzymes of the greater wax moth, Galleria mellonella L.

Pathogenicity and immunity effects of both the entomopathogenic nematode, (Heterorhabditis zealandica) and the entomopathogenic fungus (Beauveria bassiana) on the last larval instar of the greater wax moth, Galleria mellonella L. (Lepidoptera: Pyralidae), were investigated. Both pathogens were either applied separately or in a combination. The fungus was inoculated first, followed by the nematode on time manner 0, 2, 4, and 6 days. LC50 values for both pathogens were calculated and then used for determination of the changes in immune response-mediated phenoloxidase (PO) and detoxifying enzymes glutathione S-transferase (GST) and non-specific esterase (EST) activities. The results indicated that a positive correlation was found between pathogen concentration and host mortality percentage. LC50 values were 6.49 IJs/larva for H. zealandica and 3.1 × 102 conidia/ml for B. bassiana. Synergistic interactions were found in all combined applications. The degree of synergism increased (reaching 100% mortality), when the nematode was applied 2 days post-fungal infection. PO activity increased significantly (p < 0.05) in a time-dependent manner post-B. bassiana infection. In contrary, the combination of H. zealandica + B. bassiana or H. zealandica alone produced a significant suppressive effect on PO activity over time. GST activity increased significantly (p < 0.05) in 36 h, then decreased at 48 h post-combined application, while the fungal infection enhanced significantly GST activity in time-dependent manner than the control and other treatments. EST activity increased significantly (p < 0.05) in both combined application and the single nematode infection than the single fungal infection, which increased during the initial period only. The increased mortality rates and suppression of phenoloxidase and glutathione S-transferase enzymes, following the combined application suggests a strong synergistic effect between both pathogens. It could be concluded that the tested combined pathogens are compatible element for integrated pest management.

The limpet transcription factors of Triatoma infestans regulate the response to fungal infection and modulate the expression pattern of defensin genes

As part of the innate humoral response to microbial attack, insects activate the expression of antimicrobial peptides (AMP). Understanding the regulatory mechanisms of this response in the Chagas disease vector Triatoma infestans is important since biological control strategies against pyrethroid-resistant insect populations were recently addressed by using the entomopathogenic fungus Beauveria bassiana. By bioinformatics, gene expression, and silencing techniques in T. infestans nymphs, we achieved sequence and functional characterization of two variants of the limpet transcription factor (Tilimpet) and studied their role as regulators of the AMP expression, particularly defensins, in fungus-infected insects. We found that Tilimpet variants may act differentially since they have divergent sequences and different relative expression ratios, suggesting that Tilimpet-2 could be the main regulator of the higher expressed defensins and Tilimpet-1 might play a complementary or more general role. Also, the six defensins (Tidef-1 to Tidef-6) exhibited different expression levels in fungus-infected nymphs, consistent with their phylogenetic clustering. This study aims to contribute to a better understanding of T. infestans immune response in which limpet is involved, after challenge by B. bassiana infection.

Beauveria bassiana infection reduces the vectorial capacity of Aedes albopictus for the Zika virus

Zika virus (ZIKV), a mosquito-borne flavivirus, poses a serious threat to public health worldwide, and Aedes albopictus is one of its vectors. To evaluate the potential of the entomopathogenic fungus Beauveria bassiana for ZIKV vector control, we compared the vectorial capacity of Ae. albopictus females blood-fed with ZIKV with or without exposure to Beauveria bassiana. We found that fungal infection significantly decreased the amount of ZIKV by 3.6-, 12.3- and 7.8-fold in mosquito midguts, heads and salivary glands, respectively. Similarly, fungal infection also reduced the rates of ZIKV dissemination, potential transmission and potential population transmission for mosquitoes by 26.8%, 38.4% and 35.2%, respectively. On the other hand, the median survival time and fecundity of fungus-infected mosquitoes were reduced by 84.2% and 39.8% in comparison with those of the non-fungus-infected mosquitoes. The first gonotrophic cycle length was increased by 15.3% because of fungal infection. This study revealed that B. bassiana infection significantly reduced the vectorial capacity of A. albopictus for ZIKV, which suggested that B. bassiana could be broadly and efficiently used in the field for the control of Zika vectors.

Evaluation of the compatibility of entomopathogenic fungi and two botanical insecticides tondexir and palizin for controlling Galleria mellonella L. (Lepidoptera: Pyralidae)

Interactions between insect pathogenic fungi and insecticides can be both positive and negative for pest control. In the first part of this study, the insecticidal efficacy of entomopathogenic fungi including Beauveria bassiana (Bals.) Vuill., Lecanicillium (=Verticillium) lecanii (Zimm.) Zare & Gams, and Metarhizium anisopliae (Metsch.) Sor. was tested against different larval instars of Galleria mellonella using a dipping bioassay method. In addition, the toxicity of two botanical insecticides, tondexir (TON) and palizin (PAL) was evaluated against 7th instar G. mellonella. In the second part of this study, the effect of TON and PAL on spore germination of B. bassiana and L. lecanii was determined. Subsequently, the effect of the application of a sub-lethal concentration of TON (414.24 ppm) or PAL (116.44 ppm) at 24 h prior to the application of an LC50 of B. bassiana or L. lecanii was evaluated in 7th instar G. mellonella. The lowest LC50 values were found against 7th instar G. mellonella which were 4.64 × 105 and 1.83 × 106 conidia ml−1 for B. bassiana and L. lecanii, respectively. The LC50 values for TON and PAL against 7th instar G. mellonella were 1868 and 735.68 ppm, respectively. TON and PAL showed significantly lower effects on the germination of B. bassiana than on the germination of L. lecanii. The highest inhibition of germination was caused by TON at concentrations of 2500 and 4000 ppm (20.8 and 24.3% inhibition, respectively). Both insecticides significantly reduced the susceptibility of larval G. mellonella to B. bassiana infection. PAL reduced the effectiveness of L. lecanii against larval G. mellonella. However, the infectivity of L. lecanii was not affected by the application of a sublethal concentration of TON. The use of these botanical insecticides with fungi in integrated control programs for G. mellonella is discussed.

Simultaneous effects of thermal stress and fungal infection on lipid peroxidation and antioxidant system of rice-striped stem borer, Chilo suppressalis Walker (Lepidoptera: Crambidae)

ABSTRACTThe effects of thermal stress, time of exposure, and infection by Beauveria bassiana were determined on the lipid peroxidation and antioxidant system of Chilo suppressalis Walker. The larvae were exposed to 24°C and 34°C for, respectively, 24 h and 72 h. Then, these two groups of larvae were divided into three subgroups: intact larvae, larvae injected with 80 μg/ml of Tween-80 (0.02%), and larvae injected with 104 spores/ml of Beauveria bassiana suspended in 80 μg/ml of Tween-80 (0.02%). The highest activities of superoxide dismutase and peroxidase were found in larvae injected with B. bassiana at both thermal regimes, while the activity of catalase was lowest in these treatments. Highest activities of glutathione S-transferase, glucose-6-phosphate dehydrogenase, and ascorbate peroxidase were observed in the larvae injected with B. bassiana at both thermal regimes. Finally, thermal stress and fungal injection led to the highest amounts of Malondialdehyde and the ratio of oxidized and reduced thiols at both thermal regimes. In our study, the simultaneous exposure of C. suppressalis to thermal stress and B. bassiana infection significantly increased the concentration of oxidative agents in the body of treated larvae, indicating that the antioxidant system may have a role in larval defense.Abbreviations: SOD: Superoxide dismutase; POX: Peroxidase; CAT: Catalase; APOX: Ascorbate peroxidase; GPDH: Glucose-6-Phosphate dehydrogenase; GSTs: Glutathione S-transferases; MDA: Malondialdehyde; RSSR/RSH: Oxidized and reduced thiols.

Repressed Beauveria bassiana infections in Delia antiqua due to associated microbiota.

Insects form both mutualistic and antagonistic relationships with microbes, and some antagonistic microbes have been used as biocontrol agents (BCAs) in pest management. Contextually, BCAs may be inhibited by beneficial insect symbionts, which can become potential barriers to entomopathogen-dependent pest biocontrol. Using the symbioses formed by one devastating dipteran pest, Delia antiqua, and its associated microbes as a model system, we sought to determine whether the antagonistic interaction between BCAs and microbial symbionts could affect the outcome of entomopathogen-dependent pest biocontrol. The result showed that in contrast to non-axenic D. antiqua larvae, i.e., onion maggots, axenic larvae lost resistance to the entomopathogenic Beauveria bassiana, and the re-inoculation of microbiota increased the resistance of axenic larvae to B. bassiana. Furthermore, bacteria frequently isolated from larvae, including Citrobacter freundii, Enterobacter ludwigii, Pseudomonas protegens, Serratia plymuthica, Sphingobacterium faecium and Stenotrophomonas maltophilia, suppressed B. bassiana conidia germination and hyphal growth, and the re-inoculation of specific individual bacteria enhanced the resistance of axenic larvae to B. bassiana. Bacteria associated with larvae, including C. freundii, E. ludwigii, P. protegens, S. plymuthica, S. faecium and S. maltophilia, can inhibit B. bassiana infection. Removing the microbiota can suppress larval resistance to fungal infection. © 2018 Society of Chemical Industry.

Mortality of Solenopsis invicta Workers (Hymenoptera: Formicidae) After Indirect Exposure to Spores of Three Entomopathogenic Fungi

Mortality caused by indirect exposure to Metarhizium brunneum and Beauveria bassiana (GHA and NI8) to the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae), workers was evaluated. Groups of 50 workers were placed in one side of dual-box arenas. The opposite side of the arenas was lined with filter paper squares previously sprayed with unformulated purified spores (106 spores/ml) suspended in 0.2% Ethal TDA 3, HLB 8 of the three fungal strains, or untreated filter paper squares as the control. Daily observations were done for 1 wk to determine mortality. Dead ants from each treatment and control were collected, surface cleaned, and placed in PDA media and incubated at 27°C, 60% RH for 7 d to detect fungal growth. The presence of fungal growth in the dead ants confirmed that fungal spores infected workers while walking on the treated paper. In the M. brunneum and B. bassiana GHA treatments, 51.35 and 56.68% of the workers died, respectively, during days 1 and 2. However, only 9.47 and 35.96% of the mortality could be explained by fungal infection by M. brunneum and B. bassiana GHA, respectively. Most of the mortality observed in the B. bassiana NI8 treatment (84.48%) occurred later (between days 4–6) and most of this mortality occurring during day 4 (89.06%) could be explained by B. bassiana infection. Overall mortality was significantly higher in the B. bassiana NI8 treatment than the other two fungi tested and control. Potential application of these fungal strains for fire ant control are discussed.

CTL14, a recognition receptor induced in late stage larvae, modulates anti-fungal immunity in cotton bollworm Helicoverpa armigera

C-type lectin (CTL) is usually considered as pattern recognition receptors in insect innate immunity. Here we found that CTL14 of Helicoverpa armigera was only activated in the fifth instar larvae not in the second instar by entomopathogen Beauveria bassiana infection. Recombinant CTL14 protein was found to form aggregates with zymosan and B. bassiana in vitro. Immunoprecipitation studies demonstrated that CTL14 interacted with serine proteinases (SP), serine proteinase inhibitor (serpin), prophenoloxidases (PPO) and vitellogenin (Vg) in the larval hemolymph. Furthermore, depletion of CTL14 using dsRNA led to dramatic decrease in the expression level of PPO1. Additionally, CTL14 depleted H. armigera decreased the resistance to fungal challenge. Taken together, our study showed the direct involvement of CTL14 in the anti-fungal immunity of H. armigera, which further explained the stronger immune responses in the fifth instar compared to the second instar larvae.

Protein deficiency lowers resistance of Mormon crickets to the pathogenic fungus Beauveria bassiana

Little is known about the effects of dietary macronutrients on the capacity of insects to ward off a fungal pathogen. Here we tested the hypothesis that Mormon crickets fed restricted protein diets have lower enzymatic assays of generalized immunity, slower rates of encapsulation of foreign bodies, and greater mortality from infection by Beauveria bassiana, a fungal pathogen. Beginning in the last nymphal instar, Mormon crickets were fed a high, intermediate, or low protein diet with correspondingly low, intermediate, or high carbohydrate proportions. After they eclosed to adult, we drew hemolymph, topically applied B. bassiana, maintained them on diet treatments, and measured mortality for 21 days. Mormon crickets fed high protein diets had higher prophenoloxidase titers, greater encapsulation response, and higher survivorship to Beauveria fungal infection than those on low protein diets. We replicated the study adding very high and very low protein diets to the treatments. A high protein diet increased phenoloxidase titers, and those fed the very high protein diet had more circulating prophenoloxidase. Mormon crickets fed the very low protein diet were the most susceptible to B. bassiana infection, but the more concentrated phenoloxidase and prophenoloxidase associated with the highest protein diets did not confer the greatest protection from the fungal pathogen as in the first replicate. We conclude that protein-restricted diets caused Mormon crickets to have lower phenoloxidase titers, slower encapsulation of foreign bodies, and greater mortality from B. bassiana infection than those fed high protein diets. These results support the nutrition-based dichotomy of migrating Mormon crickets, protein-deficient ones are more susceptible to pathogenic fungi whereas carbohydrate-deficient ones are more vulnerable to bacterial challenge.

Genes involved in Beauveria bassiana infection to Galleria mellonella.

The ascomycete fungus Beauveria bassiana is a natural pathogen of hundreds of insect species and is commercially produced as an environmentally friendly mycoinsecticide. Many genes involved in fungal insecticide infection have been identified but few have been further explored. In this study, we constructed three transcriptomes of B. bassiana at 24, 48 and 72h post infection of insect pests (BbI) or control (BbC). There were 3148, 3613 and 4922 genes differentially expressed at 24, 48 and 72h post BbI/BbC infection, respectively. A large number of genes and pathways involved in infection were identified. To further analyze those genes, expression patterns across different infection stages (0, 12, 24, 36, 48, 60, 72 and 84h) were studied using quantitative RT-PCR. This analysis showed that the infection-related genes could be divided into four patterns: highly expressed throughout the whole infection process (thioredoxin 1); highly expressed during early stages of infection but lowly expressed after the insect death (adhesin protein Mad1); lowly expressed during early infection but highly expressed after insect death (cation transporter, OpS13); or lowly expressed across the entire infection process (catalase protein). The data provide novel insights into the insect-pathogen interaction and help to uncover the molecular mechanisms involved in fungal infection of insect pests.

Evaluation of Beauveria bassiana infection in thehemolymph serum proteins of the housefly, Musca domestica L. (Diptera: Muscidae).

Beauveria bassiana plays a prominent role in biocontrol of houseflies, Musca domestica (L.). Thus, a deeper insight into immune response of M. domestica during B. bassiana infection was warranted to assist the production of more efficient mycoinsecticides. The present study investigates changes in protein profile of M. domestica hemolymph serum post B. bassiana infection using two-dimensional difference gel electrophoresis (2D-DIGE) followed by identification of selected proteins by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). The non-infected or control group of flies showed an expression of 54 proteins, while M. domestica infected with B. bassiana expressed a total of 68 hemolymph serum proteins. Thirty three proteins were expressed in both groups of houseflies, whereas 35 proteins were exclusively expressed in infected flies and 21 proteins were exclusively expressed in control flies. Among the 33 proteins which were expressed in both groups of houseflies, 17 proteins showed downregulation, while16 proteins were upregulated in the infected flies compared to the non-infected ones. The results from this study are expected to facilitate better understanding of insect's immune response mechanism.