Isolation, identification and virulence of indigenous entomopathogenic fungal strains against the peach-potato aphid, Myzus persicae Sulzer (Hemiptera: Aphididae), and the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

Background As different biogeographic strains and isolates of entomopathogenic fungi vary in their genetic, enzymatic and pathogenic characteristics, this study assessed the virulence of 2 indigenous strains of Beauveria bassiana (Balsam) Vuillemin and Metarhizium anisopliae (Metschn.) Sorokin (Ascomycota, Hypocreales: Clavicipitaceae), isolated from naturally infected insect cadavers, against the 3rd instar nymphs of Myzus persicae (Sulzer) (Hemiptera: Aphididae) and 3rd instar larvae of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) using leaf-dip and larval-dip methods, respectively. Results Both fungal isolates exhibited considerable pathogenicity against M. persicae and S. frugiperda. Mortality in all bioassays was conidial concentration and exposure time dependent and increased significantly along with both factors (R2 = 0.86–0.99 for B. bassiana and 0.82–0.94 for M. anisopliae). Moreover, M. anisopliae isolate appeared more virulent to S. frugiperda larvae than B. bassiana isolate, while the later fungal isolate was more lethal to M. persicae nymphs than the former one. At the highest conidial concentration (1.0 × 109 conidia/ml), M. anisopliae caused maximum mean mortality of S. frugiperda (88%) and M. persicae (65%) and B. bassiana exhibited maximum mean mortality of S. frugiperda (76%) and M. persicae (94%). Moreover, probit regression analyses showed LT50 values for M. persicae of 4.57 and 6.86 days at 1.0 × 109 conidia/ml for the isolates of B. bassiana and M. anisopliae, respectively, while LC50 values were 7.75 × 106 and 8.70 × 107 conidia/ml after 10th day of application, for the isolates of B. bassiana and M. anisopliae, respectively, against M. persicae. Similarly, LT50 values for S. frugiperda were 7.75 and 7.03 days for 1.0 × 109 conidia/ml concentration and LC50 values were 2.84 × 107 and 8.84 × 105 conidia/ml at 10th day data for the isolates of B. bassiana and M. anisopliae, respectively. Conclusion Overall study results demonstrated the effectiveness of B. bassiana and M. anisopliae against M. persicae and S. frugiperda, respectively. However, field evaluations of these indigenously isolated promising fungal strains against these insect pests.

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice

Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104, 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Late effects of Beauveria bassiana on larval stages of Aedes aegypti Linneo, 1762 (Diptera: Culicidae)

Aedes aegypti is a culicide that has gained relevance over the years due to its ability to transmit various viruses that cause diseases in humans that all the years cause high mortality rates in the world population. The main problem is that Ae. aegypti has managed to establish and maintain a close relationship with humans and their habitat, which is why the search for alternatives to control vector populations becomes imperative. The objective of the present work was to study the effects of two Beauveria bassiana strains on Aedes aegypti. Third instar larvae of Ae. aegypti in 250 mL plastic containers were inoculated with the GHA and NB3 strains at different concentrations (1.5 × 104, 1.5× 105, 1.5 × 106 and 1.5 × 107 conidia/mL). The NB3 strain presented highest mortality values with 63% in the highest concentration i.e., 1.5 × 107, while for the GHA strain the highest mortality value was 30.7% at the same concentration. The results showed significant difference in mortality with respect to the strain and days post treatment (P = 0.0001), but not with respect to the conidial concentration (P = 0.634). The average mortality of larvae per day for the NB3 for different concentrations ranged from 20 to 25 larvae per day, while for the GHA daily mortality ranged from 5 to 12 larvae. In post-treatment mortality, the highest mortality was recorded in the third stage larvae for the NB3, while for GHA the highest percentage mortality was observed in individuals who managed to reach the adult state. The findings of the current research depicted the noteworthy role of B. bassiana for the management of an important vector of human disease.

Development of a Rapid Sporulation Method of Fusarium graminearum Using Liquid Cultivation

Fusarium graminearum is an important fungus causing a variety of maize diseases, including stalk rot, ear rot and sheath rot. However, conidia of F. graminearum are not easily obtained under normal culture conditions, which seriously affects the identification and pathogenicity assessment of the isolates and screening of resistance sources. This study was undertaken to develop and utilize a rapid sporulation technique of F. graminearum using liquid cultivation, which could meet the needs of various tests. The results show that the optimum conditions for sporulation of F. graminearum were as follows: culture medium, 0.154 mol/L saline; temperature, 28-30℃; incubation time, 96 h; initial pH, 9-10; illumination, continuous ultraviolet light; and shaking speed, 150 rpm. Using this culture method, conidial concentration of tested F. graminearum strains can reach more than 1.5×105 conidia/ml. Compared with the existing methods using mung bean (MB) and carboxylmethyl cellulose (CMC) as matrix, saline is relatively low expensive, and the culture process is relatively quick. Overall, this study provided a systematic, rapid, and simple method to obtain a large number of conidia of F. graminearum.

Virulence of Five Isolates of Indigenous Beauveria Bassiana Against Eggs and Nymphs of Bemisia Tabaci Gennadius (Hemiptera: Aleyrodidae)

This research aims to study the virulence of five isolates of Beauveria bassiana to eggs and nymphs of Bemisia tabaci on tomato. In the experiment eggs and second instar nymphs of B. Tabaci were used. Five isolates of the fungus, i.e., WS, TD312, PD114, PA221, PB211, were tested. Conidial concentration of B. bassiana used were 108 conidia/ml. Experimental parameters included mortality of eggs and nymph and percentage of adult emergence. The results showed that all B. bassiana isolates tested were able to kill B. tabaci eggs but with very low mortality (2-19%). Mortality of second instar B. tabaci nymphs was dependent on the fungal isolates. Isolate WS had the highest virulence, which caused 70% mortality of 2nd instar nymphs, with a LT50 of 4.87 days. Nymphs of B. tabaci were highly susceptible to B. bassiana infection compared with eggs. B. bassiana applicated to nymphs of B. tabaci can decrease the percentage of adult emergence.

Factors affecting the infection of sweet cherry (Prunus avium) fruit by Podosphaera cerasi

Powdery mildew caused by Podosphaera cerasi is the most important fungal disease of sweet cherries in the Pacific Northwest of the United States. In this study, several factors related to disease epidemiology were evaluated. The experiments were conducted to investigate flower susceptibility to P. cerasi infection by in planta and in vitro inoculation. The susceptibility of fruit at various developmental stages was investigated using defined concentrations of P. cerasi conidia. Furthermore, the threshold of conidial concentration required for fruit infection was determined. The pathogen activity during full bloom was limited and not related to fruit disease incidence and severity at harvest. Foliar infections always preceded fruit infections by an average of 42 days during the three years of the study. The onset of fruit infection followed, on average, 66 days after full bloom and appeared simultaneously on all susceptible cherry cultivars in the research orchard. Disease symptoms were only observed on fruit in Biologische Bundesanstalt, Bundessortenamt, and Chemical industry (BBCH) scale 8 (maturity) in all cultivars examined. During this stage, a concentration of 500 conidia/ ml was sufficient to cause fruit infection at harvest. Interaction between the inoculation dates and inoculum conidial concentration revealed a dependency of disease development on the host stage at the time of conidial inoculation; the younger the fruit, the more conidia are needed to cause disease at harvest. Molecular studies showed a rapid increase in conidia viability at the transition from asymptomatic to the symptomatic disease of fruit. No evidence of ontogenic resistance of fruit to powdery mildew infection was observed.

Hypothesis and Theory: Fungal Spores in Stemflow and Potential Bark Sources

The study of stemflow fungi began over 50 years ago. Past work has been performed in different climatic regions of the world, with different sampling methods, by mycologists focusing on different taxonomical groups. Therefore, we aim to synthesize this work to delineate major conclusions and emerging hypothesis. Here, we present: (1) a systematic compilation of observations on stemflow conidial concentration, flux, and species composition; (2) an evaluation of the methods underlying these observations; (3) a testable theory to understand spatiotemporal dynamics in stemflow (including honeydews) conidial assemblages, with a focus on their relationship to bark structure and microhabitats; and (4) a discussion of major hypotheses based on past observations and new data. This represents a knowledge gap in our understanding of fungal dispersal mechanisms in forests, in a spatially-concentrated hydrologic flux that interacts with habitats throughout the forest microbiome. The literature synthesis and new data represent observations for 228 fungal species’ conidia in stemflow collected from 58 tree species, 6 palm species, and 1 bamboo species. Hypothetical relationships were identified regarding stemflow production and conidial concentration, flux, and species composition. These relationships appear to be driven by bark physico-chemical properties, tree canopy setting, the diversity of in-canopy microenvironments (e.g., tree holes, bark fissures, and epiphytes), and several possible conidia exchange processes (teleomorph aerosols, epi-faunal exchanges, fungal colonization of canopy microhabitats, and droplet impacts, etc.). The review reveals a more complex function of stemflow fungi, having a role in self-cleaning tree surfaces (which play air quality-related ecoservices themselves), and, on the other hand, these fungi may have a role in the protection of the host plant.

Evaluation of the Potential of Fungal Biopesticides for the Biological Control of the Seed Bug, Elasmolomus pallens (Dallas) (Hemiptera: Rhyparochromidae)

Elasmolomus pallens is a post-harvest insect pest of groundnuts which causes severe yield loss to farmers, particularly in Africa and Asia. Resistance to synthetic chemicals has been on the rise among insects and is a constraint on insecticides regulations. In view of the drive for alternative approaches to synthetic insecticides, this study evaluated the potential of biopesticides based on entomopathogenic fungi against E. pallens under laboratory conditions. Fungal isolates from the bug cadaver including Fusarium proliferatum F1, Aspergillus tamarii F2, A. flavus F3, Trichoderma atroviride F4, A. niger F4, and Metarhizium anisopliae (Meschn.) Sorokin, originating from the cadaver of Zonocerus variegatus were screened for virulence against the bug. Adult bugs were dipped briefly in conidial concentration 1 × 108 conidial mL−1 and observed at 25 ± 2 °C, 80 ± 10 RH and 14: 10 L:D for 10 days. The fungal isolates caused mortality ranging from 48 to 100% based on their potential to infect and kill the bug. Five conidial concentrations (1 × 104 to 1 × 108 conidia mL−1) were evaluated against adult bugs in the multiple-dose virulence bioassay. Lethal concentrations (LC50 and LC90) values of 6.75 × 106 and 4.42 × 109 conidia mL−1 were obtained for A. flavus F3 while M. anisopliae had 8.0 × 106 and 6.14 × 108 conidia mL−1 respectively. Lethal time (LT50 and LT90) values were 3.3 and 6.2 days for A. flavus F3 compared to 3.6 and 5.6 days for M. anisopliae, respectively. Thus, A. flavus F3 showed potential against E. pallens; and can be considered as an ideal isolate for incorporation into formulations for field applications.

Biocontrol Potential of Some Entomopathogenic Fungal Strains Against Bean Aphid Megoura japonica (Matsumura)

This research reported the in vitro pathogenicity of Verticillium lecanii strains, L2 and L5, and Beauveria bassiana strains, B76 and B252, against Megoura japonica using leaf-dip method. The virulence potential of these four entomopathogenic fungal strains of V. lecanii and of B. bassiana were compared between fungi conidia (concentrations 1 × 106, 1 × 107, and 1 × 108 conidia mL−1) and culture filtrate. Moreover, binary combination of four different fungal strains (L2 + B76, B76 + L5, L2 + B252, and L2 + B76 + B252 + L5) were evaluated against M. japonica under control condition. Aphid mortality was recorded after two, four, six, and eight days of post-treatment. In the conidial concentration bioassay, strain B76 showed maximal mortality (85.3%) against bean aphid, and strain L5 showed the lowest effect (60.0%) at the highest concentration (1 × 108 conidia mL−1) at eight days post-treatment. Whereas, binary combinations of B76 and L2 strains showed the highest effect against M. japonica (90.5%) than other combinations. Moreover, in comparison with the effect of filtrate and conidia bioassay, 91.4% and 84.1% were achieved in strain B76, and the lowest effect (63.8% and 55.1%) was recorded in strain L5.