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In this study, the entomopathogenic fungi; Beauveria bassiana, Isaria fumosorosea and Metarhizium anisopliae were evaluated against fourth nymphs and adults of the green shield bug, P. prasina. The experiment was conducted both under field and laboratory conditions. Isolates included six Beauveria bassiana, one Metarhizium anisopliae and one Isaria fumosorosea. Isolates were bioassayed against nymphs and adults in both under field and laboratory conditions. Four replications of the bioassays were analyzed and evaluated daily for 14 days. LT50 and LT95 values for the experimented isolates ranged from 4.19 to 7.11 days and from 6.68 to 11.30 days, respectively in laboratory for nymphs, ranged from 4.98 to 7.18 days and from 8.03 to 12.22 days, respectively in laboratory for adults. LT50 and LT95 values for the experimented isolates ranged from 6.69 to 10.70 days and from 10.29 to 17.91 days, respectively in field for nymphs, ranged from 7.29 to 10.70 days and from 11.23 to 17.91 days, respectively in field for adults. It was obvious that BB1/21b (Beauveria bassiana) was the most virulent on adults of P. prasina. As a result, while the effect of the isolates used in the study was high due to controlled conditions in the laboratory, it was low in field conditions.

In the natural ecosystem, fungal entomopathogens are the most efficient biocontrol agents against insect pests. In this study we offer an alternative for conventional fungal diagnostic, Surface-enhanced Raman spectroscopy (SERS) technique combine with principal component analysis (PCA) for detection and identification three entomopathogenic fungi, namely, IBCB 66 Beauveria bassiana, IBCB 130 Isaria fumosorosea, and IBCB 425 Metarhizium anisopliae. Using a simple preparation approach, highly active silver nanoparticles suitable for detecting complex biomolecules were produced for application in the SERS technique. Entomopathogens fungi produced highly enhanced and reproducible Raman signals based on their biochemical composition due to the high density of hot spots at the confluence of silver nano-aggregates, allowing the three entomopathogens species to be differentiated in the SERS spectrum fingerprint region, 550-1700 cm-1. The SERS method, along with PCA analysis, accounted for over 99 % of total variance and allowed for very high probability discrimination between the three entomopathogens, allowing taxonomic affiliation to be determined in a short period of time. These findings suggest that the SERS methodology can be used to develop a new, fast, accurate, and cost-effective diagnostic method for fungal entomopathogens.

Made available in DSpace on 2022-04-29T08:33:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-04-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) In the Neotropics, leaf-cutting ants are pests of great importance to various crops. The natural defenses and complexity of these ants reduce the impact of potentially useful pathogens for biological control. Furthermore, several types of research with entomopathogenic fungi usually focus on only a few isolates of well-known species, like Beauveria bassiana and Metarhizium anisopliae, in addition to not always use isolates obtained from leaf-cutting ants. Here, we evaluated the pathogenicity of several filamentous fungi to workers of the leaf-cutting ant Atta sexdens rubropilosa. For that, we used fungi previously isolated in an extensive isolation study in Atta spp. reproductive females. Conidia suspensions of Aspergillus flavus, B. bassiana, Isaria fumosorosea and Purpureocillium lilacinum were prepared to a final concentration of 108 conidia mL− 1. In addition, we included sterilized distilled water as control and commercial B. bassiana as standard. Ant workers were immersed in these treatments for eight seconds. The number of dead workers was counted daily. Several fungal isolates killed more, or faster than the standard B. bassiana, showing that they may have a better potential for use in leaf-cutting ants´ biological control. Additionally, the present study reports for the first time the pathogenicity of P. lilacinum and I. fumosorosea towards Atta workers. Thus, we showed that an extensive and specific prospection is of importance for the development of biopesticides for leaf-cutting ants control. Departamento de Proteção Vegetal Faculdade de Ciências Agronômicas Universidade Estadual Paulista (UNESP), SP Universidade Federal do Piauí (UFPI), Piauí Departamento de Biologia Geral e Aplicada Instituto de Biociências de Rio Claro Universidade Estadual Paulista (UNESP), SP Departamento de Proteção Vegetal Faculdade de Ciências Agronômicas Universidade Estadual Paulista (UNESP), SP Departamento de Biologia Geral e Aplicada Instituto de Biociências de Rio Claro Universidade Estadual Paulista (UNESP), SP FAPESP: 2008/07032-7

Exotic invasive whiteflies (Hemiptera: Aleyrodidae) in India cause direct and indirect yield losses in agriculture, horticulture and forestry crop plants. Around 25 years ago, the spiralling whitefly, Aleurodicus dispersus Russell invaded and established on many host plants including economically important crops in India. Recently, within a span of five years, seven whiteflies invaded India viz., solanum whitefly, Aleurothrixus trachoides (Back) reported to breed on 37 plant species; rugose spiraling whitefly (RSW), Aleurodicus rugioperculatus Martin found breeding on 40 host plants; nesting whiteflies, Paraleyrodes bondari Peracchi on 34 host plants and P. minei Ιaccarino infest about 25 host plants; legume feeding whitefly, Tetraleurodes acaciae (Quaintance) infesting 5 host plants; palm infesting whitefly, Aleurotrachelus atratus Hempel on 4 host plants and woolly whitefly, Aleurothrixus floccosus (Maskell) infesting guava. These invasive species are native to the Neotropical region, mostly from Central America and the Caribbean. Extensive spread along the coastal regions and gardens near the backwater of India is predicted owing to the favorable weather factors and availability of host plants. Species of exotic whiteflies with similar habits co-exist in more or less the same niche and have a similar pattern of growth and development. The intensity of infestation of RSW on coconut, banana and oil palm, the woolly whitefly on guava and the palm infesting whitefly and nesting whiteflies on coconut was severe. The exotic aphelinid parasitoid, Encarsia guadeloupae Viggiani (Hymenoptera: Aphelinidae), a predator Pseudomallada astur (Banks) (Neuroptera: Chrysopidae) and the entomopathogenic fungus, Isaria fumosorosea Wize (Hypocreales: Clavicipitaceae) play a major role in reducing the population of these invasives. The most insidious spread of these species in India is likely mediated by humans through the movement of infested seedlings and plant materials. Extensive surveys revealed that these species spread rapidly in the large geographical region of India mostly through transportation of infested seedlings. This study reports a major expansion of the geographic and host range and the patterns of co-occurrence, damage and economic impact of these exotic species in India and their natural enemies.

Galleria mellonella is a pest of the honey bee (Apis mellifera L.) and causes significant losses to the beekeeping industry; therefore, experiments are needed to decode the effects of entomopathogenic fungi on insect physiology. The gas chromatography–mass spectrometry (GC-MS) method was successful for the determination of the organic compounds of Galleria mellonella larvae, noninfected and infected by three fungal species: M. robertsii, M. flavoviride, and I. fumosorosea. A total of 46 compounds were identified in G. mellonella, including fatty acids, other acids, fatty acid methyl esters, monoacylglycerols, amino acids, sterols, and several other organic compounds. The lipids of G. mellonella larvae after M. robertsii, M. flavoviride, and I. fumosorosea exposure contained 40, 35, and 33 organic compounds, respectively. The following organic compounds, present in the noninfected larvae, were absent from the infected larvae: fatty acids C22:0 and C24:0, glutaric acid, urocanic acid, hydroxycinnamic acid, dihydroxycinnamic acid, 10-oxodecanoic acid, glycine, aspartic acid, glutamic acid, lysine, tyrosine, tryptophan, 2-aminobutyric acid, and tyramine. These compounds can be used by fungi as an exogenous source of carbon. The following organic compounds, present in the infected larvae, were absent from the noninfected larvae: fatty acids C10:0, C11:0, C13:0, and C20:0, suberic acid, phenylacetic acid, fatty acid methyl ester (FAME) C16:0, FAME C18:2, FAME C18:1, glycerol 2-monopalmitate, norvaline, proline, sitosterol, and 2-dekanal. These compounds can be synthesized as an insect response to fungal infection.

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is considered as a highly invasive and destructive agricultural pest due to its strong dispersal and adaptive capacity. We evaluated the susceptibility of B. dorsalis pupae to six entomopathogenic fungi (Isaria fumosorosea IFCF01, Beauveria bassiana Bb02, B. bassiana Bb04, Metarhizium anisopliae Ma01, M. anisopliae Ma04, and M. anisopliae Ma09) in the laboratory through impregnation method (immersing fly pupae into conidial suspension). Results indicated that fungus treatment during the pupal stage of B. dorsalis, not only caused significant mortality of target pupae (4.4–14.4% mortality), but also influence the survival of the emerged adults (38.9–72.8% mortality on the 10 days post-emergence). Among the six entomopathogenic fungi, M. anisopliae Ma04 presented the highest virulence against B. dorsalis. Results of virulence bioassay indicated that the LC50 values of M. anisopliae Ma04 against B. dorsalis declined from 5.2 × 1028 to 5.2 × 107 conidia/ml over a 1–10-day period post adult emergence, and the LT50 values decreased from 5.25 to 2.78 days with the concentrations of conidial suspension increasing from 1.0 × 108 to 1.0 × 1010 conidia/ml. Therefore, M. anisopliae Ma04 have a greater potential for B. dorsalis control.

The compatibility of Isaria fumosorosea IF-1106 with 12 commercial pesticide adjuvants was evaluated using the predictive parameter SC90 (the adjuvant concentration at which 90% of conidia survived...

Implementing a predator-mediated pathogen dissemination system can meet the goal of simultaneous biocontrol of harmful mites and other pests. But for the system to be effective, it must satisfy the requirement of high lethality of the pathogen to the target pest(s) with low lethality to predatory mites in a suitable environment. Prior testing showed that the fungal entomopathogen Isaria fumosorosea (Wize) Brown & Smith exhibited high toxicity to aphids yet low toxicity to the predatory mite Neoseiulus cucumeris (Oudemans). In practical application, the humidity conditions of conidia carried by predatory mites to plants are not necessarily constant. Therefore, we set two different humidity conditions to observe the changes of spore attachment quantity and the differences of germination and infection between predatory mites and aphids after their bodies were stained with spores. Under 50±1% relative humidity (RH), predatory mites cleaned most spores in a short time; 24 hours after dusting, the number of spores (mean ± SD) on body was (0.64±0.10)×103, just (4.43±0.35)% of that at 0h, but spores on aphids were kept (16.58±1.48)% ((6.46±0.91)×103 spores) of that at beginning. There was a significant difference in the percentage of spores retained on aphids and predatory mites (p=0.000

Some botanical extracts affect the cellular immune system of insects by interfering with normal hemocyte function. This gives rise to diminution in cellular immunity and causes a decrease in insect...

The objectives of this study were to quantify the virulence of four entomopathogenic fungal species to pupae of Rhagoletis pomonella (Walsh) (Diptera: Tephritidae) and to determine the potential to combine entomopathogenic fungi (EPFs) and entomopathogenic nematodes (EPNs) for biological control of this pest. The four species of EPFs included Beauveria bassiana (strain GHA), Metarhizium brunneum (strain F52), Isaria javanica (wf GA17), and Isaria fumosorosea (Apopka 97 strain). In laboratory assays, all fungi reduced adult emergence but there were no differences between fungal species. Isaria javanica and M. brunneum were examined further in a EPFs and EPNs bioassay that also included the EPNs Steinernema carpocapsae (ALL strain) and S. riobrave (355 strain). All nematodes and fungi were applied either alone or in combination (fungus + nematode). There were no differences between species within the same entomopathogen group (fungi and nematodes). However, the treatment with S. riobrave resulted in lower R. pomonella emergence than either fungal species. The combination of S. riobrave and I. javanica resulted in the lowest R. pomonella emergence (3%) at fourth-week interval, which was significantly lower than any of the single-agent applications, yet virulence of the other three combination treatments was not different from their respective nematode treatments applied alone. Additive interactions were detected for all fungus–nematode combinations. This study suggests that application of entomopathogenic nematodes and fungi could be an effective option to suppress R. pomonella populations.

12 páginas, 4 figuras y 2 tablas., The pine engraver beetle, Ips acuminatus Gyll, is a bark beetle that causes important damages in Scots pine (Pinus sylvestris) forests and plantations. As almost all higher organisms, Ips acuminatus harbours a microbiome, although the role of most members of its microbiome is not well understood. As part of a work in which we analysed the bacterial diversity associated to Ips acuminatus, we isolated the strain Arthrobacter sp. IA7. In order to study its potential role within the bark beetle holobiont, we sequenced and explored its genome and performed a pan-genome analysis of the genus Arthrobacter, showing specific genes of strain IA7 that might be related with its particular role in its niche. Based on these investigations, we suggest several potential roles of the bacterium within the beetle. Analysis of genes related to secondary metabolism indicated potential antifungal capability, confirmed by the inhibition of several entomopathogenic fungal strains (Metarhizium anisopliae CCF0966, Lecanicillium muscarium CCF6041, L. muscarium CCF3297, Isaria fumosorosea CCF4401, I. farinosa CCF4808, Beauveria bassiana CCF4422 and B. brongniartii CCF1547). Phylogenetic analyses of the 16S rRNA gene, six concatenated housekeeping genes (tuf-secY-rpoB-recA-fusA-atpD) and genome sequences indicated that strain IA7 is closely related to A. globiformis NBRC 12137T but forms a new species within the genus Arthrobacter; this was confirmed by digital DNA-DNA hybridization (37.10%) and average nucleotide identity (ANIb) (88.9%). Based on phenotypic and genotypic features, we propose strain IA7T as the novel species Arthrobacter ipsi sp. nov. (type strain IA7T = CECT 30100T = LMG 31782T) and suggest its protective role for its host., The project was funded by Czech Science Foundation project GACR 19-09072S. ZSS received a grant from the Junta de Castilla y León, Spanish Regional Government, and LIGD received a grant supported by Junta de Castilla y León (Escalera de Excelencia CLU-2018-04), co-funded by the P.O. FEDER of Castilla y León 2014-2020.

Highly polyphagous rugose spiralling whitefly (RSW), Aleurodicus rugioperculatus Martin (Hemiptera: Aleyrodidae) invaded India during 2016. Pathogenicity of two strains (ICAR-NBAIR Pfu-1 & Pfu-5) o...

Spotted lanternfly, Lycorma delicatula (White), is an invasive Asian insect that was initially found in Berks County, Pennsylvania, in 2014. As of early 2020, this pest had been found in five more eastern states and it is expected to continue to expand its geographical range. Lycorma delicatula is highly polyphagous but seems to prefer tree-of-heaven, Ailanthus altissima. However, grape growers in Pennsylvania have reported significant damage and loss of vines caused by L. delicatula adults. In fall 2018, two fungal entomopathogens (Beauveria bassiana and Batkoa major) drove localized collapses in L. delicatula populations in Berks County, Pennsylvania. In 2019, we tested applications of a commercialized mycoinsecticide based on B. bassiana strain GHA on L. delicatula populations in a public park in southeastern Pennsylvania. A single application of B. bassiana reduced fourth instar nymphs by 48% after 14 d. Applications of B. bassiana to L. delicatula adults in the same park resulted in 43% mortality after 14 d. Beauveria bassiana spores remained viable on foliage for 5–7 d after spraying. We also conducted semi-field bioassays with B. bassiana GHA (formulated as BoteGHA and Aprehend) and another mycoinsecticide containing Isaria fumosorosea Apopka Strain 97 against L. delicatula adults feeding on potted grapes. All the mycoinsecticides killed ≥90% of adults after 9 d using direct applications. Aprehend killed 99% of adults after 9 d with exposure to residues on sprayed grapes. These data show that fungal entomopathogens can help to suppress populations of L. delicatula in agroecosystems and natural areas.

The red flour beetle is a very important primary insect pest of wheat worldwide. The experiments were performed to check the virulence of Beauveria bassiana and Isaria fumosorosea by insect dipping method on the adults and 2nd instar of Tribolium castaneum. Conidia were taken from 15 days old fungi and subsequently four concentrations i.e., 2×108, 3×108, 4×108 and 5×108 spores/ml of both fungi were prepared in 0.05% Tween 80 solution. Minimum 12.5% and maximum 32.5% mortality of adult insects was recorded on 7th day after the treatment at 2×108 and 5×108 spores/ml concentrations of B. bassiana, respectively while on larval stages, minimum 2.5% on 5th day and maximum 80.0% mortality was observed on 7th day post treatment of B. bassiana, respectively. On the other side, minimum 7.5 and maximum 22.5 mortality percentage was noted on 7th day post application of I. fumosorosea, respectively while on immatures minimum 5% on 6th day and highest 70% mortality was noted on 7th day post infection with 2×108 and 5×108 spores/ml of I. fumosorosea, correspondingly. This study showed the effectiveness of insect pathogenic fungi against the important stored grain insect pest and proved to be a positive management strategy.

Entomopathogenic fungi and the egg parasitoid Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) might be used together in biological control. However, the effects of these fungi on T. pretiosum are not known. Thus, this study aimed to determine the effect of the entomopathogenic fungus Isaria fumosorosea, on the biological parameters of T. pretiosum. Two isolates of I. fumosorosea (IBCB 367 and IBCB 394) were used for this purpose. (1) In a free choice test: cards (1.0 × 5.0 cm) with non-parasitized eggs of Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) were either sprayed with 0.2 mL of the fungus suspension (1.0 × 109 conidia.mL-1) or with sterile distilled water containing Tween® 80 (0.01%), which were then offered to females of T. pretiosum. (2) No choice test: the isolates were sprayed at a concentration of 1.0 × 109 conidia.mL-1 on cards (1.0 × 5.0 cm) with A. kuehniella eggs. The control consisted of spraying sterile distilled water containing Tween® 80 (0.01%). Individual females of T. pretiosum were confined for 24 h with the cards. The number of eggs parasitized, percentage of emergence, longevity, duration of the egg-adult period and sex ratio were evaluated, as well as the longevity of the females that parasitized the eggs and the mortality of the emerging adults evaluated. IBCB 367 isolate repelled T. pretiosum. The pre-parasitism and post-parasitism sprays did not affect the number of eggs parasitized or the sex ratio, however, the pre-parasitism IBCB 394 treatment the females and males survived for longer, whereas the survival of females in post-parasitism treatment with the same isolate was reduced.The presence of conidia on and mycelium of the fungus in T. pretiosum was confirmed using Scanning Electron Microscopy and a histological analysis. Isolates IBCB 367 and IBCB 394 from I. fumosorosea are selective to T. pretiosum in the laboratory.

This study was conducted to evaluate the insecticidal activity of the endophytic fungus Isaria fumosorosea Wize isolated from Ricinus communis L. against Locusta migratoria L. and Acanthoscelides obtectus Say. To evaluate the insecticidal activity, a mixture of fungal extract and spore suspensions (1x107, 1x106 and 1x105 spores/ml of dissolved extract) were applied against the insects. It was found that I. fumosorosea had a good insecticidal activity, positively proportional to concentration and time after exposure (24, 48, 72 hours). A maximum mortality rate of 80% for A. obtectus and 70% for L. migratoria at 1x107 spores/ml of dissolved extract was recorded. Preliminary phytochemical analyses of the fungal extract revealed the presence of alkaloids, flavonoids and phenols. Additionally, a fungal development on the insects bodies has also been noticed; that is undoubtedly explained the recorded insecticidal activity.

Fusarium spp. are dangerous pathogens of several commercially important crops around the world. Among non-chemical alternatives, beneficial microorganisms offer a potential control option for Fusarium and other soilborne pathogens. A greenhouse study was conducted using cotton as a model plant and F. oxysporum f. sp. vasinfectum Race 4 (FOV Race 4) as the pathogen to evaluate the efficacy of commercial formulations of microbial and botanical biopesticides. Streptomyces lydicus (Actinovate® AG), Bacillus amyloliquefaciens (Stargus®), giant knotweed extract (Regalia®), entomopathogenic fungi - Beauveria bassiana (BotaniGard® ES), Isaria fumosorosea (PFR-97™ 20% WDG), and Metarhizium brunneum (M. anisopliae; Met52® EC) were applied to FOV Race 4 infested soil in three treatment regimens, and the growth of susceptible Pima cotton seedling growth and health was monitored for 5 weeks. Treatments were generally effective against antagonizing FOV Race 4 especially when applied multiple times at a lower rate or once at a higher rate. Entomopathogenic fungi were as effective as other products that are exclusively used for disease management. Beauveria bassiana provided superior control for a long period compared to other treatments. This study reports the efficacy of non-chemical alternatives against FOV Race 4 and the potential of using entomopathogenic fungi for disease management in addition to their normal role of arthropod pest management.

The present study investigates the effects of the fungal entomopathogens Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea, following their endophytic colonization of the Solanum tuberosum L plants. Our results showed the studied fungal species isolates had no effects in promoting or suppressing the growth of S. tuberosum potato but affect the infestation of the serious pests.

CONGRATULATIONS to YIA Senior winner Isabella Daza (above) from Port St. Lucie, Florida. Her project examines the use of the entomopathogenic fungus Isaria fumosorosea Wize as a method for biological control of the invasive yellowmargined leaf beetle, Microtheca ochroloma Stal. The project’s main objective is to evaluate in the laboratory the repellency of the fungus to reduce feeding by the beetle on crucifer crops. Her results were presented at the Treasure Coast Science and Engineering Fair in February 2014.

Flavonoids, including chalcones, are more stable and bioavailable in the form of glycosylated and methylated derivatives. The combined chemical and biotechnological methods can be applied to obtain such compounds. In the present study, 2′-hydroxy-2-methylchalcone was synthesized and biotransformed in the cultures of entomopathogenic filamentous fungi Beauveria bassiana KCH J1.5, Isaria fumosorosea KCH J2 and Isaria farinosa KCH J2.6, which have been known for their extensive enzymatic system and ability to perform glycosylation of flavonoids. As a result, five new glycosylated dihydrochalcones were obtained. Biotransformation of 2′-hydroxy-2-methylchalcone by B. bassiana KCH J1.5 resulted in four glycosylated dihydrochalcones: 2′-hydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside, 2′,3-dihydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside, 2′-hydroxy-2-hydroxymethyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside, and 2′,4-dihydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside. In the culture of I. fumosorosea KCH J2 only one product was formed—3-hydroxy-2-methyldihydrochalcone 2′-O-β-d-(4″-O-methyl)-glucopyranoside. Biotransformation performed by I. farinosa KCH J2.6 resulted in the formation of two products: 2′-hydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside and 2′,3-dihydroxy-2-methyldihydrochalcone 3′-O-β-d-(4″-O-methyl)-glucopyranoside. The structures of all obtained products were established based on the NMR spectroscopy. All products mentioned above may be used in further studies as potentially bioactive compounds with improved stability and bioavailability. These compounds can be considered as flavor enhancers and potential sweeteners.

Simple Summary Citrus greening is devastating the citrus industry in Florida, and conventional synthetic pesticide applications are rapidly becoming unsustainable for the control of the vector, the Asian citrus psyllid (AsCP), Diaphorina citri. Laboratory experiments indicate that the entomopathogenic fungus Cordyceps javanica, alone and in combination with horticultural oils, may offer a more sustainable strategy to manage AsCP. The field study in this paper indicated that C. javanica alone, C. javanica mixed with white oil, and spinetoram (the chemical standard) mixed with white oil significantly suppressed AsCP populations by 60–90% and 61–83% 7 and 14 days after treatment, respectively, in 2018, compared to white oil only and the untreated control treatments. Only spinetoram + oil suppressed AsCP 100% up to 7 days after treatment in 2019. AsCP’s natural enemies, mostly lady beetles, were observed on fungal-treated citrus trees and the untreated control. Overall, C. javanica was as effective in suppressing AsCP adults as spinetoram for up to 14 days in field conditions and was compatible with the psyllids’ natural enemies. The use of C. javanica in citrus-integrated pest management is suggested. Abstract Citrus greening disease is devastating the citrus industry in Florida, and the conventional synthetic pesticide applications used to control the vector, the Asian citrus psyllid (AsCP), Diaphorina citri, are rapidly becoming unsustainable. Various laboratory experiments indicate that the entomopathogenic fungus Cordyceps javanica, alone and in combination with horticultural oils, may offer a more sustainable strategy for the management of AsCP. Field studies conducted in 2018 and 2019 in mature citrus indicated that C. javanica alone, C. javanica mixed with white oil, and the chemical standard spinetoram mixed with white oil significantly suppressed AsCP adult populations by 61–83% up to 14 days after treatment in 2018, although colony-forming units of C. javanica were still present on the leaves 21 days after treatment (DAT). Only spinetoram + oil significantly suppressed AsCP, by 100%, up to 7 DAT in 2019. Natural enemies of AsCP, including lady beetles, lacewing larvae and the parasitoid Tamarixia radiata, were observed in the fungal treatments and the untreated control. The AsCP suppression by C. javanica and its compatibility with beneficial organisms suggest the potential use of this entomopathogenic fungus in citrus-integrated pest management.

Yellow mite is a serious pest of many field and horticultural crops including jute. As repeated use of acaricides possess problem of resistance, alternatively, the infectivity of talc based formulation of three entomopathogens, i.e., Lecanicillium lecanii (Ll), Isaria fumosorosea (= Paecilomyces fumosoroseus ) (Pf) and Beauveria bassiana (Bb) at 4x10 8 cfu/ml, and 6x10 8 cfu/ml concentrations were evaluated against yellow mite under laboratory condition. In both the concentrations, Pf and Bb recorded significantly higher mortality of yellow mite than Ll. At 3-DPT (days post treatment) significantly highest mortality was observed in Pf (30.35%) followed by Bb (21.59%) and Ll (4.87%). Later on at 4 and 5-DPT period the level of mortality in Pf and Bb treated population was at par but significantly higher than Ll. At 5-DPT, Pf recorded maximum cumulative mortality (40.92%) at par with Bb (35.99%) and significantly higher than Ll (12.92%). Significantly higher cumulative mortality at 6x10 8 CFU/l concentration during 5 days after treatment was 34.35%. The infectivity of talc based formulation of Ll at both the concentrations was significantly less than Pf and Bb. Higher infectivity of I. fumosorosea indicates it to be ideal for yellow mite control in jute.

Understanding the interaction between pathogen virulence and host responses is very important for selecting virulent isolates of entomopathogenic fungi that control an economically important pest, such as the date palm dust mite, Oligonychus afrasiaticus (McGregor). In this study, the virulence of Metarhizium anisopliae (ARSEF 8453 and ARSEF 9374), Beauveria bassiana (ARSEF 6884 and ARSEF 8037), and Isaria fumosorosea (ARSEF 9602 and ARSEF 7284) was evaluated by virulence-regulating traits and host detoxification-related enzymatic analysis. The results revealed significant differences in virulence among all tested isolates of entomopathogenic fungi. The most virulent isolate of B. bassiana ARSEF 6884 (LT50 = 6.3 days; LC50 = 2.7 × 106 conidia ml−1) observed in the current study revealed the highest activity of cuticle-degrading protease Pr1 (2.12 ± 0.10 μmol NA ml−1 min−1) compared with the least virulent isolate of I. fumosorosea ARSEF 7284 (LT50 = 13.1 days; LC50 = 7.9 × 107 conidia ml−1). Virulent isolates successfully infected the target host and revealed lower activities of antioxidant (catalase and superoxide dismutase) and detoxification (glutathione S-transferases) enzymes at late stage infection. In conclusion, isolates ARSEF 6884 and ARSEF 9374 were found to be promising for the development of an environmentally friendly management strategy to control date palm dust mites.

The European large raspberry aphid, Amphorophora idaei (Borner) (Hemiptera: Aphididae), is one of the most important pests of the raspberry, causing significant economic damage due to its high mobility and its role as a vector of various viral plant pathogens. In the absence of effective control solutions, 16 strains of entomopathogenic fungi comprising six genera, i.e. Isaria fumosorosea, Lecanicillium longisporum, Lecanicillium muscarium, Beauveria bassiana, Metarhizium anisopliae and Metarhizium brunneum (formally known as Metarhizium anisopliae var. anisopliae) were screened for their comparative virulence against A. idaei. Three different concentrations of fungal conidial suspensions (1 × 105, 1 × 106 and 1 × 108 ml−1) of M. brunneum, I. fumosorosea and L. longisporum were sprayed on detached raspberry leaves infested with A. idaei under laboratory conditions, and further trials were conducted in the glasshouse. The results showed that these entomopathogenic fungal strains were able to reduce the number of A. idaei. However, virulence varied significantly between different fungal strains/species. The most virulent species were L. longisporum and I. fumosorosea, followed by M. brunneum. All species of B. bassiana showed least virulence. The virulence of three selected strains M. brunneum BNL102, I. fumosorosea FE 9901 and L. longisporum Vertalec were dependent on dose and insect life stage. LT50 values of A. idaei exposed to pre-treated leaves with entomopathogenic fungi were relatively lower than those of aphids that received direct inocula by spraying. Similarly, significant reduction in A. idaei populations were observed in the glasshouse. Overall, this study demonstrates the effectiveness of fungal strains that could be commercially exploited against A. idaei.

En el estado de Veracruz, el control de mosca blanca con insecticidas sintéticos en las hortalizas ha provocado problemas de contaminación. El objetivo de la presente investigación consistió en evaluar la eficiencia de productos biorracionales en el control de mosca blanca en el cultivo de jitomate en condiciones de invernadero y en los cultivos de chile y calabacita en condiciones de cielo abierto. Para jitomate los tratamientos fueron Nim aceite 4% IA, Nim aceite + Metarhizium anisopliae 1x107 mL-1, Nim aceite + Beauveria bassiana 1x107 mL-1, Nim aceite + Isaria javanica 1x 107 mL-1 y un testigo (sin aplicación), y para chile y calabacita Metarhizium anisopliae 1x107 mL-1, Isaria fumosorosea 1x107 mL-1, Beauveria bassiana 1x107 mL-1 y un testigo (sin aplicación). Los tratamientos de Nim aceite + M. anisopliae y Nim aceite + I. javanica fueron los más eficientes en jitomate y los insecticidas a base de B. bassiana y M. anisopliae en chile y calabacita, respectivamente. Existe un potencial para el uso de hongos entomopatógenos para control de mosca blanca en los cultivos de jitomate en invernadero, chile y calabacitas a cielo abierto, potencial aplicable a condiciones de alta tecnología y de producción tradicional.ABSTRACTIn the state of Veracruz, the control of whitefly with synthetic insecticides in vegetables has caused contamination problems. The objective of the present investigation was to evaluate the efficiency of biorational products for whitefly control in tomato cultivation under greenhouse conditions, and in chili and squash under open sky conditions. In tomato, treatments were Nim oil 4% IA, Nim oil + Metarhizium anisopliae 1x107 mL-1, Nim oil + Beauveria bassiana 1x107 mL-1, Nim oil + Isaria javanica 1x107 mL-1 and Blank (control). In chili and squash, treatments were Metarhizium anisopliae 1x107 mL-1, Isaria fumosorosea 1x107 mL-1, Beauveria bassiana 1x107 mL-1 and Blank (control). The Nim oil + M. anisopliae and Nim oil + I. javanica treatments were the most efficient in tomato and the B. bassiana and M. anisopliae based insecticides in chili and zucchini, respectively. There is a potential for the use of entomopathogenic fungi for whitefly control in tomato under greenhouse conditions, and chili and squash under open sky conditions, a potential applicable to high technology and traditional production conditions.

In this investigation, the impact of the predatory mite, Phytoseiulus persimilis (PM) and entomopathogenic fungus, Isaria fumosorosea (F) at three levels (F1=104, F2=106 and F3=108 conidia/ml) alone or in grouping state as compared with Ortus (acaricide) was evaluated on eggs and motile stages of two-spotted spider mites (TSSM). Nine treatments (CK=control, F1, F2, F3, PM, F1+PM, F2+PM, F3+PM and Ortus) were performed. On the other hand, the effect of the used entomopathogenic fungus on numbers of eggs and motile stages of the predatory mite was detected only at four treatments (PM, PM+F1, PM+F2, and PM+F3). Applications of I. fumosorosea at all chosen doses caused significant decreases in the mean population density of TSSM eggs (50.15-58.81% in 2016 and 59.63-68.37% in 2017) and motile stages (62.61-74-66% in 2016 and 68.47-75.32%) in comparison to the control. The effect of PM addition on TSSM eggs was higher than that of I. fumosorosea using but on TSSM motile stages, the pattern differed. The combination of both I. fumosorosea and P. persimilis showed greater influence on TSSM eggs and motile stages than the use of I. fumosorosea or P. persimilis individually. Spraying I. fumosorosea with releasing P. persimilis led to small reductions in eggs (11.88-22.42% in 2016 and 7.86-21.73% in 2017), and motile stages (9.84-21.33% in 2016 and 6.75-17%) of P. persimilis. Results of this study concluded that the combination of entomopathogenic fungus, I. fumosorosea and predatory mite, P. persimilis was strongly succeeded in the biological control process of TSSM on soybean plants under field conditions.

Estrone, estradiol, ethynylestradiol and estrone 3-methyl ether underwent a biotransformation process in the submerged culture of Isaria fumosorosea KCh J2. Estrone was transformed into seven metabolites, four of which were glycosylated. Estradiol was selectively glycosylated at C-3 and then transformed to D-ring lactone. Ethynylestradiol was coupled with methylglucoside and 6β-hydroxyderivative was obtained. Estrone 3-methyl ether was not transformed indicating that a free hydroxyl group at C-3 is necessary for glycosylation. Baeyer–Villiger oxidation combined with hydroxylation and glycosylation was observed. All glycosides obtained in this study are 3-O-β-methylglucosides.

Background Nanoparticles can be used for effective pest management as a combined formulation of metal and some other material that has proven efficacy against a given pest. This study reports the synthesis, characterization and efficacy of Isaria fumosorosea-based zero-valent iron (ZVI) nanoparticles against sweet potato whitefly Bemisia tabaci (Gennadius). Results The I. fumosorosea-ZVI nanoparticles showed a characteristic surface plasmon absorption band at 470 nm during UV-visible spectroscopy. The scanning electron micrographs of nanoparticles showed spherical shaped nanoparticles with sizes ranging between 1.71 and 3.0 µm. The EDX analysis showed the characteristic peak of iron at 0.6 and 6.8 KeV. The XRD analysis showed characteristic peaks at 44.72°, 65.070°, 82.339° and 82.65°. The bioassay results indicated that the percentage of larval mortality of B. tabaci challenged with I. fumosorosea ZVI nanoparticles was both concentration and age dependent. Isaria fumosorosea ZVI nanoparticles showed high pathogenicity against second and third instar nymphs, and pupae with LC50 values of 19.17, 26.10 and 37.71 ppm, respectively. The LT50 was lowest for second instar nymphs (3.15 days) and highest for pupae (4.22 days) when inoculated with a concentration of 50 ppm. Conclusion Isaria fumosorosea ZVI nanoparticles can be an eco-friendly tool for effective B. tabaci management. © 2019 Society of Chemical Industry.

Entomopathogenic fungi based microbial insecticides are considered as safe alternatives to chemical pesticides, which secretes several bioactive compounds to kill the host insects. In this study, we report a new approach for the synthesis and characterization of insecticide toxic protein IF8 produced by the Isaria fumosorosea 08, and to evaluate the mycotoxin level against the vector of Huanglongbing (HLB) or citrus greening disease, the Asian citrus psyllid, Diaphorina citri. Soluble toxic metabolites extracted from I. fumosorosea 08 through submerged liquid state culture had a molecular weight of 43 kDa when subjected by to sodium dodecyl sulfate-poly-acrylamide (SDS-PAGE) gel electrophoresis. The most abundant of toxic protein IF8 was determined by High-performance liquid chromatography (HPLC) and liquid chromatography electrospray ionization-mass spectroscopy (LC-ESI-MS) for the analysis of its molecular mass weight and purity. Further Matrix-assisted laser desorption ionization-time of flight (MALDI-TOFF) analysis confirmed the presence of toxic metabolites in liquid culture. Subsequently, mycotoxic effect of toxic protein IF8 was tested against D. citri at three different concentrations (1%, 2%, and 3%). The results showed the insecticidal activity of >80% when administered at three different concentrations at 48–120 hour post-application. Additionally, we also investigated the physicochemical properties and stability of IF8 by using computational biological tools. This is the first study to report the characterization of fungal mediated synthesis of the protein IF8 toxic to the insect D. citri. These results suggest the mycotoxin control of D. citri and prevention of HLB transmission by using a natural toxic compound which is eco-friendly and can be potentially used for the integrated management of D. citri.

Entomopathogenic fungi are essential participants in ecosystems that help regulate arthropod communities. Many of them have a symbiotic-endophytic relationship with plants, endowing them with insecticidal or insect-repellent traits, which could be exploited for insect control in agriculture. Among insects, aphids are one of the most harmful taxa; they damage plants directly by sucking plant sap or indirectly by transmitting economically important viruses. Plant infestation by aphids depends on prefeeding activities related to host identification, particularly leaf walking and short probing. Although several aspects of aphid prefeeding activities have been studied in different hosts, this behavior has not been examined on plants colonized by entomopathogenic fungi. Here, we studied the effects of three strawberry endophytic entomopathogenic fungi, namely, Beauveria bassiana, Isaria fumosorosea and Metarhizium anisopliae var. robertsii, on the prefeeding activity of the aphid Myzus persicae by monitoring aphids feeding on leaves of fungus colonized and uncolonized plants. Our analysis revealed that the endophytes significantly affected aphid prefeeding behavior, which was typified by a reduction of the expected mean number of short probes, an avoidance to make a second probe, and a high propensity for abandoning the leaf. We discuss possible mechanisms of fungal interference with aphid activity.

Fourier Transform Infrared Spectroscopy (FTIR) methods are the most commonly used spectroscopic techniques for differentiation of fungi species, however reflectance spectroscopy as a non-invasive technique can also be used. The aim of the study was to develop a method to rapidly differentiate fungi by means of reflectance spectroscopy using visible-infrared spectrum. Spectral measurements were conducted on six entomopathogenic fungi: Beauveria bassiana, Isaria fumosorosea, I. farinosa, I. tenuipes, Lecanicillium lecanii, L. muscarium cultured on Petri-dishes. The FieldSpec3 ASD spectroradiometer. Recording reflected radiance in the range 350–2500 nm was used. Measurements were performed in two modes: contact and proximal and obtained spectra were transformed using two methods: Savitzky-Golay (SG) and baseline alignment (BA) smoothing and derivative. The success rate of 100% in differentiate between fungi species was achieved with spectra recorded in visible-near infrared range with contact and proximal measurement and after SG transformation. Two wavelengths (411 nm and 520 nm) were needed to differentiate fungi using SG and proximal measurement while seven wavelengths were necessary to get full separation with contact measurement. BA spectra transformation method gave separation accuracy of 84, and 90% with four to five wavelengths for contact and proximal measurements, respectively, however, BA do not require full spectrum of wavelengths to fungi discrimination. Proposed reflectance spectroscopy method could discriminate between fungi species very similar macroscopically e.g. L. lecanii and L. muscarium until recently recognized as one species.

The bulb mite, Rhizoglyphus robini, is a serious pest of garlic, onion and other crops. The mite is usually found in association with dangerous fungal pathogens such as Fusarium spp. Control of this pest has relied upon the use of synthetic acaricides but chemical control of the bulb mite is difficult because it is able to develop resistance quickly. Thus, alternative control methods, e.g. biological control, need to be developed and implemented. The aim of this study was to assess efficacy of selected strains of entomopathogenic fungi (EPF) against adult females of R. robini under laboratory conditions. New EPF strains were isolated from soil samples collected in onion and garlic fields in the Czech Republic and Israel using soil elution and cultivation on selective media. Fungal species were determined using macroscopic, microscopic and molecular markers. The efficacy against R. robini females was tested in 17 isolated and 3 reference strains of EPF. Results revealed high variability among species and strains. The highest efficacy against R. robini mites was found in strains of Metarhizium anisopliae isolated from soil samples collected in the Czech Republic which caused mortality up to 99.3%, and a Metarhizium indigoticum strain from Israel causing 98.3% mortality after four days of bioassay. Isaria fumosorosea strains did not caused mortality higher than 40%. The lowest virulence was found in Beauveria spp. strains causing mortality of mites between 5 and 25%. Median lethal time (LT50) and median lethal concentration (LC50) in the three most virulent strains ranged between 2 and 4 days and between 1.01×104 and 2.36×105 spores/ml, respectively. The concentration-response models indicated that the M. indigoticum strain is more lethal than M. anisopliae strains. The present study showed that some strains of entomopathogenic fungi, especially from the genus Metarhizium, could be perspective biocontrol agents against R. robini.

Gnomoniopsis castaneae is the cause of the chestnut brown rot but has been also regarded as an important mortality factor for the chestnut gall wasp Dryocosmus kuriphilus. The question to whether G. castaneae could serve as a natural biocontrol agent against insect pests is investigated in the present study. We used three serious insect pests as experimental model insects: Plodia interpuctella and Trogoderma granarium, which are important pests of stored products, and Myzus persicae, a cosmopolitan, serious pest of annual and perennial crop plants. Although chemical pesticides represent effective control means, they are also related to several environmental and health risks. In search for alternative pest management methods, scientific interest has been focused, inter alia, on the use of entomopathogenic fungi. While Isaria&nbsp, fumosorosea has long been recognized as an effective control agent against several pests, G.castaneae has been very little studied. The present study examined whether and to what extent G. castaneae and I. fumosorosea exhibit insecticidal activity against fourth-instar larvae of P. interpunctella and T. granarium and adults of M. persicae. Mortality was examined in interrelation with dosage and time exposure intervals. Both fungi exhibited pesticidal action. However, G. castaneae induced noteworthy mortality only at very high doses. In general, we concluded that G. castaneae failed to cause high insect pathogenicity at normal doses and may not be an efficient biocontrol agent compared with other entomopathogens. On the other hand, our study reiterates the pathogenic potential of I. fumosorosea. More studies are needed to further our insight into the potential of EF species as a component of IPM.

The effect of two entomopathogenic agents, nematodes and fungi, was tested at three concentrations and three incubation temperatures in a laboratory against fourth-instar larvae of plum curculio, Conotrachelus nenuphar (Herbst). After inoculation by immersion, larvae were statistically more susceptible to Isaria fumosorosea fungus at an incubation temperature of 5°C and to Beauveria bassiana at 25°C. Fungal concentration had no effect on mortality. More larvae were killed by nematodes at warmer temperatures and higher concentrations of either species, Steneirnema feltiae or S. riobrave. Based on preliminary laboratory results, all entomopathogens suppressed plum curculio larvae, even at low concentrations, with 35% killed by fungi and 22% by nematodes. The results are promising for further studies on control of C. nenuphar by biological agents at field conditions.

Entomopathogenic nematodes and fungi are globally distributed soil organisms that are frequently used as bioagents in biological control and integrated pest management. Many studies have demonstrated that the combination of biocontrol agents can increase their efficacy against target hosts. In our study, we focused on another potential benefit of the synergy of two species of nematodes, Steinernema feltiae and Heterorhabditis bacteriophora, and the fungus Isaria fumosorosea. According to our hypothesis, these nematodes may be able to disseminate this fungus into the environment. To test this hypothesis, we studied fungal dispersal by the nematodes in different arenas, including potato dextrose agar (PDA) plates, sand heaps, sand barriers, and glass tubes filled with soil. The results of our study showed, for the first time, that the spreading of both conidia and blastospores of I. fumosorosea is significantly enhanced by the presence of entomopathogenic nematodes, but the efficacy of dissemination is negatively influenced by the heterogeneity of the testing arena. We also found that H. bacteriophora spread fungi more effectively than S. feltiae. This phenomenon could be explained by the differences in the presence and persistence of second-stage cuticles or by different foraging behavior. Finally, we observed that blastospores are disseminated more effectively than conidia, which might be due to the different adherence of these spores (conidia are hydrophobic, while blastospores are hydrophilic). The obtained results showed that entomopathogenic nematodes (EPNs) can enhance the efficiency of fungal dispersal.

Buxus sp. is an important native and ornamental tree in Europe threatened by a serious invasive pest Cydalima perspectalis. The larvae of this moth are able to defoliate box trees and cause their death. The development of novel biopesticides targeting this pest might help protect Buxus trees grown wildly or in city parks. Laboratory experiments were conducted to assess the efficacy of entomopathogenic fungus Isaria fumosorosea strain CCM 8367 against C. perspectalis. The last-instar larvae of the box tree moth were treated by the suspension of fungus conidia at concentrations ranging from 1 × 104 to 1 × 108 spores per 1 mL. Fungus infection was observed mostly in pupae, but the maximum mortality did not exceed 60%, indicating a very low susceptibility of C. perspectalis to I. fumosorosea. Furthermore, a number of ungerminated fungal conidia were found on larval cuticles using a low-temperature scanning electron microscopy. Our data also reveal that the hydroalcoholic extract from B. sempervirens leaves significantly inhibits both the germination of I. fumosorosea conidia and fungus growth. It can be speculated that the strain CCM 8367 of I. fumosorosea is not a potent biocontrol agent against C. perspectalis and low virulence of the fungus might be due to the accumulation of host plant phytochemicals having antimicrobial activity in larval cuticle of the pest.

Flavonoids are known for their numerous biological activities, however, their pharmacological application is limited by poor bioavailability. Glycosides are usually more stable and more soluble in water and in this form, flavonoids are present in nature. Likewise, the presence of the methyl group in the flavonoid skeleton results in facilitated absorption and greater bioavailability. Entomopathogenic filamentous fungi are effective in the biotransformation of flavonoids, they are known especially for efficient glycosylation. In the current study we used strains of Beauveria bassiana KCH J1.5 and Isaria fumosorosea KCH J2 to biotransform flavonoids with a single methyl group. 2&prime, Hydroxy-5&prime, methylchalcone was biotransformed by both strains into 2&prime, hydroxy-5&prime, methylchalcone 3-O-&beta, D-(4&Prime, O-methyl)-glucopyranoside. In the culture of B. bassiana KCH J1.5 four products were obtained from 6-methylflavanone: 4&prime, hydroxy-6-methylflavanone 3&prime, O-&beta, O-methyl)-glucopyranoside, 4&prime, hydroxyflavanone 6-methylene-O-&beta, 6-hydroxymethylflavanone 3&prime, O-methyl)-glucopyranoside and 4&prime, hydroxy-6-hydroxymethylflavanone 3&prime, O-methyl)-glucopyranoside. Biotransformation with I. fumosorosea KCH J2 as a biocatalyst resulted in the formation of 6-methylflavanone 4&prime, O-methyl)-glucopyranoside and 2-phenyl-6-methylchromane 4-O-&beta, O-methyl)-glucopyranoside. All of these flavonoids can be used in biological activity tests and can be useful in studies concerning structure&mdash, bioactivity relationships.

Simple Summary The solanum whitefly, Aleurothrixus trachoides, is a polyphagous pest known to attack > 70 crops worldwide. Endemic to the Neotropical region, in the past few years, it has emerged as a significant pest of several horticultural crops including pepper in the United States. To develop an eco-friendly sustainable management strategy for this pest, in this study, we evaluated the efficacy and compatibility of two commercially available biological control agents, predatory beetle Delphastus catalinae (Dc) and entomopathogenic fungi Cordyceps fumosorosea (Cfr) under laboratory conditions. Results showed both Cfr, treatments (alone or in combination with Dc) were efficacious, and Cfr did not negatively impact the beetle longevity. This study’s outcome is important not only for organic horticultural growers, but also for conventional growers as it offers a low-risk alternative tool for solanum whitefly integrated pest management in Florida and other affected regions. Abstract Aleurothrixus (formerly known as Aleurotrachelus) trachoides Back (Hemiptera: Aleyrodidae), commonly known as pepper or solanum whitefly, is a new emerging whitefly pest of several horticultural crops in the United States. During the preliminary survey for pepper whitefly infestation in Florida, a whitefly-specific predatory beetle Delphastus catalinae Horn (Coleoptera: Coccinellidae) was observed associated with this pest in the natural ecosystem. The current study was undertaken to determine the efficacy of this naturally occurring predator, D. catalinae, and an entomopathogenic fungus, Cordyceps (formerly known as Isaria) fumosorosea (Cfr) (Wize) (Hypocreales: Cordycipitaceae), alone or in combination, under controlled laboratory conditions. Whitefly mortality for Cfr (88%), beetle (100%) and Cfr + beetle (100%) treatments were similar and significantly higher compared to control. In the combination treatment, Cfr did not impact D. catalinae longevity and daily food intake, indicating a neutral interaction between the two treatments. Results showed that both of these natural biological control agents could potentially offer an effective alternative in the battle against invasive whiteflies such as pepper whitefly in U.S. horticulture production, either as a stand-alone strategy or in an integrated approach. Although these findings are promising, compatibility of the two treatments needs to be evaluated further under greenhouse and semi-field conditions before recommending to commercial growers.

The synthesis and biotransformation of five flavones containing methoxy substituents in the B ring: 2&prime, 3&prime, 4&prime, methoxyflavones, 2&prime, 5&prime, dimethoxyflavone and 3&prime, trimethoxyflavone are described. Strains of entomopathogenic filamentous fungi were used as biocatalysts. Five strains of the species Beauveria bassiana (KCh J1.5, J2.1, J3.2, J1, BBT), two of the species Beauveria caledonica (KCh J3.3, J3.4), one of Isaria fumosorosea (KCh J2) and one of Isaria farinosa (KCh KW 1.1) were investigated. Both the number and the place of attachment of the methoxy groups in the flavonoid structure influenced the biotransformation rate and the amount of nascent products. Based on the structures of products and semi-products, it can be concluded that their formation is the result of a cascading process. As a result of enzymes produced in the cells of the tested strains, the test compounds undergo progressive demethylation and/or hydroxylation and 4-O-methylglucosylation. Thirteen novel flavonoid 4-O-methylglucosides and five hydroxy flavones were isolated and identified.

Systena frontalis (F.) is a major insect pest of nursery production systems in the Midwest and Northeastern regions of the United States with the adults feeding on plant leaves, which reduces salability. However, there is conflicting information on overwintering, and no information on host-plant selection or insecticide susceptibility of S. frontalis adults. Therefore, we conducted a series of experiments under greenhouse, laboratory, and field conditions from 2015 to 2019. The overwintering experiment was isolated in a greenhouse to assess adult emergence from growing medium of containerized plants collected from a wholesale nursery. We found that S. frontalis overwinters in growing medium based on adults captured on yellow sticky cards and the presence of adults on new plant growth. Host-plant selection experiments were conducted at the wholesale nursery and under laboratory conditions to determine feeding selection based on S. frontalis adult feeding damage on whole plants using a foliar damage ranking scale for different cultivars of Itea virginica L., Hydrangea paniculata Siebold, Weigela florida (Bunge), and Cornus sericea L., plants. We found that S. frontalis adults exhibited no preference for the leaves of the plant species or cultivars tested in the field or laboratory. Insecticide efficacy experiments were conducted under field and laboratory conditions. In the field experiments, the insecticides acetamiprid, dinotefuran, and Isaria fumosorosea (Wize) (Hypocreales: Clavicipitaceae) provided better protection of plants from S. frontalis adult feeding than the untreated check. In the laboratory experiments, the acetamiprid and pyrethrins with canola oil treatments provided the highest percent S. frontalis adult mortality.

The sugarcane aphid [Melanaphis sacchari (Zehntner)] has become a persistent problem to sorghum [Sorghum bicolor (L.) Moench] producers in the United States since its first identification on grain sorghum near Beaumont, TX, in 2013. Since then, this aphid “super‐clone” has spread to almost all sorghum‐growing areas of the country. Growers have managed the sugarcane aphid using Insecticide Resistance Action Committee (IRAC) Group 4 insecticides and the use of resistant hybrids. In order to provide additional methods of control especially for organic sorghum production, two strains of the fungal entomopathogens Beauveria bassiana and one strain of Isaria fumosorosea as well as water and Sivanto (flupyradifurone) were applied to the sugarcane aphid susceptible sorghum hybrid DKS53‐53 in a randomized complete block design (RCBD) with four replications at Tifton and Fort Valley, GA. Aphid number, plant damage, grain yield, plant dry biomass, and plant height were measured for each treatment. As compared to the treatment with water, only the plots treated with Sivanto had significantly less aphid number and plant damage and significantly more grain yield and increased plant height. Thus, the applied entomopathogens did not reduce sugarcane aphid number, did not reduce plant damage, and did not increase grain yield in either location as compared to the treatment with water (the negative control). Thus, these three strains of entomopathogens were ineffective for controlling the sugarcane aphid under these field conditions.

Entomopathogenic fungi (EPF) have primarily been applied as an inundative approach to manage pests. However, in recent decade multifunctional role of EPF have been documented which provide multiple benefits to host plants when colonized as an endophyte. In this study five fungal isolates from the genus Beauveria (three), Isaria (one) and Lecanicillium (one) were evaluated for their ability to colonize common bean, Phaseolus vulgaris and to assess their effects in planta on plant growth promotion and possible negative effects on the two-spotted spider mites, Tetranychus urticae. All the tested isolates in this study were able to endophytically colonize root, stem and even leaves of inoculated plants examined at 7 and 14 days post inoculation, indicating the systemic colonization of EPF. Colonized plants showed increased plant heights, fresh shoot and root weights compared to plants without inoculation. Survivorship of T. urticae significantly differed among the treatments with higher survival probability in control plants. Significant reduction in larval development, adult longevity and female fecundity of spider mites were observed when fed on treated plants compared to control plants. The negative effects were found to be carried over the second generation fed on fresh plants. Overall, our results show (i) the positive effects of fungal endophytes on plant growth, (ii) reduction in population growth rate and (iii) negative effects of endophytes on growth and reproduction of spider mites in successive generations. The study presents reports on the endophytic management of plant-feeding mites and highlights the possibility of utilizing entomopathogenic fungal endophytes in the integrated pest management program.

Isaria fumosorosea is an entomopathogenic fungus that is used as a control alternative for nymphs and adults of Bemisia tabaci. Currently there are some commercial products, however, in greenhouse ...

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae), is the vector of the pathogen, Candidatus Liberibacter asiaticus that causes Huanglongbing (HLB) or citrus greening disease, the serious threats to citrus industry worldwide. The objective of this study was to evaluate the role of the entomopathogenic fungi (EPF) Beauveria bassiana (Hyphomycetes: Moniliales) and Isaria fumosorosea (Hypocreales: Cordycipitaceae) in controlling the adult citrus psyllid of D. citri under laboratory and semi-field conditions. Bioassays were performed by exposing the adults to the EPF at the concentration of (1 × 108 conidia/ml). The results showed that both EPF had the potential to control the adult citrus psyllid, giving 64–74% mortality rate in the laboratory and 61–72% under semi-filed conditions. These results support the use of both EPF as effective biopesticides for integrated management of the Asian citrus psyllid, D. citri.

A field experiment was carried out to evaluate the effectiveness of synthetic and natural preparations in protecting potato against Leptinotarsa decemlineata (Colorado Potato Beetle). Seven substances and microorganisms with proved or potential toxic effect on the pest were used: Neem extract (Azadirachta indica), suspensions of Beauveria bassiana Balsamo (Vuillemin) (Hypocreales: Cordycipitaceae) and Isaria fumosorosea Wize (Hypocreales: Cordycipitaceae) entomopathogenic fungal spores, natural pyrethrin and three chemical insecticides (lambda-cyhalothrin, thiamethoxam and imidacloprid). Plots where no protection measures were applied served as control. Chemical insecticides were more effective in controlling L. decemlineata larvae than natural preparations. It was found that chemical insecticides protected potato yield more effectively than the preparations containing natural pyrethrin or spores of entomopathogenic fungi. Tuber yield from the plots protected with entomopathogenic fungi and Neem extract was significantly higher than that on unprotected plots but still lower than the yield obtained from the plots protected with a chemical preparation.

The lady beetle Thalassa montezumae and the entomopathogenic fungus Isaria fumosorosea (Ifr) were assessed alone and in combination to suppress green croton scale, Phalacrococcus howertoni, populations on croton plants using laboratory bioassays and greenhouse cage studies. The acquisition of Ifr blastospores by beetle larvae (3rd instar) and adults during contamination in well plates was used to simulate exposure to direct spraying and subsequent possible fungal infection was assessed. Spore dispersal by the insects was determined after the blastospore-contaminated T. montezumae life stages roamed on agar plates for 24 h by counting the number of colony-forming units (CFUs) produced in the plates. There were no significant differences in survival times at 14 days post-treatment between beetle larvae and adults exposed to Ifr and those exposed to water only. Mean survival time of larvae exposed to Ifr was 14 days and water 12 days, whereas for adults it was 13 days compared to 13 days, respectively. Plates with Ifr blastospore-contaminated T. montezumae adults roaming on the agar surface displayed significantly more fungal trails as CFUs compared to plates with larvae. In greenhouse cage studies, the mean mortality rates of the scale exposed to beetle larvae, either alone (80.8%) or in combination with Ifr (89.1%), were not significantly different. Scale mortality rates in the fungus-only (60.5%) and beetle larvae-only treatments were statistically similar. The treatment with both biocontrol agents had a significantly higher scale mortality rate compared to the treatment with Ifr only. Therefore, spraying Ifr prior to releasing T. montezumae is an effective and compatible biological control strategy for management of the green croton scale on croton plants.

The low survival of insect-pathogenic fungi when used for insect control in agriculture is mainly due to the deleterious effects of ultraviolet radiation and heat from solar irradiation. In this study, conidia of 15 species of entomopathogenic fungi were exposed to simulated full-spectrum solar radiation emitted by a Xenon Test Chamber Q-SUN XE-3-HC 340S (Q-LAB® Corporation, Westlake, OH, USA), which very closely simulates full-spectrum solar radiation. A dendrogram obtained from cluster analyses, based on lethal time 50 % and 90 % calculated by Probit analyses, separated the fungi into three clusters: cluster 3 contains species with highest tolerance to simulated full-spectrum solar radiation, included Metarhizium acridum, Cladosporium herbarum, and Trichothecium roseum with LT50 > 200 min irradiation. Cluster 2 contains eight species with moderate UV tolerance: Aschersonia aleyrodis, Isaria fumosorosea, Mariannaea pruinosa, Metarhizium anisopliae, Metarhizium brunneum, Metarhizium robertsii, Simplicillium lanosoniveum, and Torrubiella homopterorum with LT50 between 120 and 150 min irradiation. The four species in cluster 1 had the lowest UV tolerance: Lecanicillium aphanocladii, Beauveria bassiana, Tolypocladium cylindrosporum, and Tolypocladium inflatum with LT50

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) cryptic species complex represents a major threat to several important crops including vegetables, cereals, fruits, and ornamentals worldwide. One important mode of its dispersal is through the trans-boundary (domestic, international) movement of infested plant materials. To prevent the spread of this invasive insect pest on inter-state ornamental shipments, the efficacy of biopesticides was tested through dip treatments of plant material pre-shipping. In several experiments, clean Mentha sp. (Lamiaceae), mint plants were exposed to adults of B. tabaci for 1 or 6 days. Adults were removed from plants and eggs, and immature stages were treated with one or a combination of the following: entomopathogenic fungi Beauveria bassiana (BotaniGard®), Isaria fumosorosea (Preferal®), the surfactant Natur'l Oil™, and soap (Publix®). After treatment application, plants were placed in commercial cardboard boxes and shipped from Apopka, Florida (FL) to Fort Pierce, FL. Assessments for the treatment efficacies were conducted at 3, 7, and 14 days post arrival of the shipment. Fungal entomopathogens; B. bassiana, I. fumosorosea, and the surfactant Natur'l Oil™, showed significant reduction in the whitefly population compared to the untreated control (P