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Abstract Chalcones are intermediate products in the biosynthesis of flavonoids, which possess a wide range of biological properties, including antimicrobial and anticancer activities. The introduction of a chlorine atom and the glucosyl moiety into their structure may increase their bioavailability, bioactivity, and pharmacological use. The combined chemical and biotechnological methods can be applied to obtain such compounds. Therefore, 2-chloro-2′-hydroxychalcone and 3-chloro-2′-hydroxychalcone were synthesized and biotransformed in cultures of two strains of filamentous fungi, i.e. Isaria fumosorosea KCH J2 and Beauveria bassiana KCH J1.5 to obtain their novel glycosylated derivatives. Pharmacokinetics, drug-likeness, and biological activity of them were predicted using cheminformatics tools. 2-Chloro-2′-hydroxychalcone, 3-chloro-2′-hydroxychalcone, their main glycosylation products, and 2′-hydrochychalcone were screened for antimicrobial activity against several microbial strains. The growth of Escherichia coli 10,536 was completely inhibited by chalcones with a chlorine atom and 3-chlorodihydrochalcone 2′-O-β-d-(4″-O-methyl)-glucopyranoside. The strain Pseudomonas aeruginosa DSM 939 was the most resistant to the action of the tested compounds. However, chalcone aglycones and glycosides with a chlorine atom almost completely inhibited the growth of bacteria Staphylococcus aureus DSM 799 and yeast Candida albicans DSM 1386. The tested compounds had different effects on lactic acid bacteria depending on the tested species. In general, chlorinated chalcones were more effective in the inhibition of the tested microbial strains than their unchlorinated counterparts and aglycones were a little more effective than their glycosides.

Chalcones, secondary plant metabolites, exhibit various biological properties. The introduction of a chlorine and a glucosyl substituent to the chalcone could enhance its bioactivity and bioavailability. Such compounds can be obtained through a combination of chemical and biotechnological methods. Therefore, 4-chloro-2′-hydroxychalcone and 5′-chloro-2′-hydroxychalcone were obtained by synthesis and then glycosylated in two filamentous fungi strains cultures, i.e., Isaria fumosorosea KCH J2 and Beauveria bassiana KCH J1.5. The main site of the glycosylation of both compounds by I. fumosorosea KCH J2 was C-2′ and C-3 when the second strain was utilized. The pharmacokinetics of these compounds were predicted using chemoinformatics tools. Furthermore, antimicrobial activity tests were performed. Compounds significantly inhibited the growth of the bacteria strains Escherichia coli 10536, Staphylococcus aureus DSM 799, and yeast Candida albicans DSM 1386. Nevertheless, the bacterial strain Pseudomonas aeruginosa DSM 939 exhibited significant resistance to their effects. The growth of lactic acid bacteria strain Lactococcus acidophilus KBiMZ 01 bacteria was moderately inhibited, but strains Lactococcus rhamnosus GG and Streptococcus thermophilus KBM-1 were completely inhibited. In summary, chalcones substituted with a chlorine demonstrated greater efficacy in inhibiting the microbial strains under examination compared to 2′-hydroxychalcone, while aglycones and their glycosides exhibited similar effectiveness. [ABSTRACT FROM AUTHOR]

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Entomopathogenic fungi are important component for regulation of pests in ecosystem. Isaria fumosorosea, as one of the entomopathogenic fungi, was reported to successfully controlled the outbreaks of forest defoliators attacked larch (Larix kaempferi) plantation in Furano, Japan and beech (Fagus crenata) forest in Hachimantai, Japan. Instead of semi-cultured method, in this paper, a culture-independent method based on DNA using qPCR was developed for specific detection and quantification of I. fumosorosea directly from soil DNA extract using specific primer. The primer IFU5821F/IFU6061R was designed and found to be the best primer pair for I. fumosorosea. Standard soil DNA was obtained with strong relationship and good fitting with five levels (R2= 0.989, E = 0.58). I. fumosorosea could not be detected from all soil samples which was possibility caused by low density of the fungi. The qPCR was likely a rapid and specific method to detect the fungus from soil.

Biological control nowadays is rapidly growing to reduce the incessant use of chemical insecticides for control of various insect pests. In the present study, entomopathogenic fungi are used to determine insecticidal activity against walnut blue butterfly under laboratory conditions. The experimental setup was completely randomized design (CRD) with two treatments along with control with different concentrations of entomopathogenic fungi. The bioassay was carried out by spraying second larval instar of Chaetoprocta odata [Hewitson 1865] with 1, 2, 3 & 4 % conidial concentration of Beauveria bassiana (Balsamo) Vuill. (1912) and Isaria fumosorosea Wize (1904). The results of this study showed that all the concentrations showed remarkable pathogenic activity but I. fumosorosea was highly pathogenic and recorded the highest mortality rate of 93.33 % after 144 hours compared to B. bassiana where 73.33 % mortality was reported. LC50 values for B. bassiana (4.15) was higher than that of I. fumosorosea (3.34) which indicates that I. fumosorosea was more effective against C. odata population. Among different concentrations of I. fumosorosea, 4 % concentration was the most effective with lowest LC50 values.

The citrus mealybug, Planococcus citri, is an insect pest of greenhousegrown horticultural crops. Citrus mealybug causes plant damage when feeding on plant leaves, stems, flowers, and fruits, resulting in a substantial economic loss. Insecticides are applied to manage citrus mealybug populations in greenhouse production systems. Anecdotal information suggests that mixing entomopathogenic fungal-based insecticides with insect growth regulators may be effective for managing citrus mealybug populations under greenhouse conditions. Consequently, we conducted two experiments in a research greenhouse at Kansas State University (Manhattan, KS, USA) in 2023. The experiments were designed to determine the efficacy of three commercially available entomopathogenic fungal-based insecticides [Beauveria bassiana Strain GHA (BotaniGard®), B. bassiana strain PPRI 5339 (Velifer™) and Isaria fumosorosea Apopka Strain 97 (Ancora®)] when mixed with three insect growth regulators [azadirachtin (Azatin®O), novaluron (Pedestal®), and pyriproxyfen (Distance®)] on citrus mealybug feeding on coleus, Solenostemon scutellarioides, plants. The entomopathogenic fungal-based insecticides alone or when mixed with the insect growth regulators were not effective in managing citrus mealybug populations, with <20% mortality during each experiment. In addition, all coleus plants treated with the entomopathogenic fungal-based insecticides had a white, powdery residue on the leaves. Our study demonstrates that entomopathogenic fungal-based insecticides, even when mixed with insect growth regulators, are not effective in managing citrus mealybug populations in greenhouses, which is likely because the environmental conditions (temperature and relative humidity) are not optimal for conidial germination and hyphal infection to occur. Therefore, entomopathogenic fungal-based insecticides have limited use for managing insect pests in greenhouse production systems. [ABSTRACT FROM AUTHOR]

Abstract Background Eublemma amabilis Moore (Lepidoptera: Noctuidae) is an important pest of lac insect, Kerria spp. (Hemiptera: Tachardiidae: Coccoidea) causing up to 20–25% damage of lac and its encrustation, which has immense industrial utilities. Extensive field monitoring in Regional Lac Insect Field Gene Bank (RLIFGB) resulted in collection of a large number of naturally occurring entomopathogenic fungi (EPF) infected insect cadavers on the lac encrustation of host plant, Flemingia semialata. Pathogenicity test under laboratory conditions showed its efficacy on E. amabilis larvae. Molecular characterization of this EPF by 18S rDNA identified it as Isaria fumosorosea (Wize) (accession number, MH414514.1). Results Being a potent EPF, biological parameters viz., conidial density and sporulation were determined and its pathogenicity were evaluated against eggs, larvae and pupae of E. amabilis at different spore dilution under laboratory conditions. The highest mortality rates of larvae and pupae (96 & 72%) and (88 & 72%) were recorded at 107spore/ml both in dipping and spraying methods, respectively. The egg mortality, however, was recorded as 40 and 35% for both methods, respectively, at 107spore/ml, which was significantly lower than other stages of E. amabilis. Field study at (107 spore/ml) recorded 81.18, 59.41 and 76.36% mean population reductions over control during monsoon 2019, winter 2019–2020 and summer 2020, respectively. Biosafety analysis of the native EPF on productivity linked parameters of lac insect was found to be safe suggesting that the native Isaria fumosorosea (MH414514.1) strain, reported first from Assam. Conclusions In this study, it was confirmed that the EPF I. fumosorosea can be offered as an effective microbial agent, alternate to insecticide against E. amabilis, either as a stand-alone strategy or in an integrated approach.

Combining chemical and microbiological methods using entomopathogenic filamentous fungi makes obtaining flavonoid glycosides possible. In the presented study, biotransformations were carried out in cultures of Beauveria bassiana KCH J1.5, Isaria fumosorosea KCH J2, and Isaria farinosa KCH J2.6 strains on six flavonoid compounds obtained in chemical synthesis. As a result of the biotransformation of 6-methyl-8-nitroflavanone using the strain I. fumosorosea KCH J2, two products were obtained: 6-methyl-8-nitro-2-phenylchromane 4-O-β-D-(4″-O-methyl)-glucopyranoside and 8-nitroflavan-4-ol 6-methylene-O-β-D-(4″-O-methyl)-glucopyranoside. 8-Bromo-6-chloroflavanone was transformed by this strain to 8-bromo-6-chloroflavan-4-ol 4′-O-β-D-(4″-O-methyl)-glucopyranoside. As a result of microbial transformation by I. farinosa KCH J2.6 effectively biotransformed only 8-bromo-6-chloroflavone into 8-bromo-6-chloroflavone 4′-O-β-D-(4″-O-methyl)-glucopyranoside. B. bassiana KCH J1.5 was able to transform 6-methyl-8-nitroflavone to 6-methyl-8-nitroflavone 4′-O-β-D-(4″-O-methyl)-glucopyranoside, and 3′-bromo-5′-chloro-2′-hydroxychalcone to 8-bromo-6-chloroflavanone 3′-O-β-D-(4″-O-methyl)-glucopyranoside. None of the filamentous fungi used transformed 2′-hydroxy-5′-methyl-3′-nitrochalcone effectively. Obtained flavonoid derivatives could be used to fight against antibiotic-resistant bacteria. To the best of our knowledge, all the substrates and products presented in this work are new compounds and are described for the first time. [ABSTRACT FROM AUTHOR]

The citrus mealybug, Planococcus citri, is an insect pest of greenhouse-grown horticultural crops. Citrus mealybug causes plant damage when feeding on plant leaves, stems, flowers, and fruits, resulting in a substantial economic loss. Insecticides are applied to manage citrus mealybug populations in greenhouse production systems. Anecdotal information suggests that mixing entomopathogenic fungal-based insecticides with insect growth regulators may be effective for managing citrus mealybug populations under greenhouse conditions. Consequently, we conducted two experiments in a research greenhouse at Kansas State University (Manhattan, KS, USA) in 2023. The experiments were designed to determine the efficacy of three commercially available entomopathogenic fungal-based insecticides [Beauveria bassiana Strain GHA (BotaniGard®), B. bassiana strain PPRI 5339 (Velifer™) and Isaria fumosorosea Apopka Strain 97 (Ancora®)] when mixed with three insect growth regulators [azadirachtin (Azatin® O), novaluron (Pedestal®), and pyriproxyfen (Distance®)] on citrus mealybug feeding on coleus, Solenostemon scutellarioides, plants. The entomopathogenic fungal-based insecticides alone or when mixed with the insect growth regulators were not effective in managing citrus mealybug populations, with

Entomopathogenic fungus are important component in pests regulation in ecosystem. Isaria fumosorosea, as one of the entomopathogenic fungi, was reported to successfully control outbreaks of forest defoliators known to attack larch (Larix kaempferi) plantation in Furano, Japan and beech (Fagus crenata) forest in Hachimantai, Japan. Instead of semi-cultured methods, this paper developed and used a culture-independent method based on DNA using qPCR for specific I. fumosorosea detection and quantification directly from soil DNA extraction using specific primer. The primer IFU5821F/IFU6061R was designed and the best primer pair for I. fumosorosea. Strong relationship and good fitting with five levels (R2 = 0.989, E = 0.58) were obtained standard soil DNA extraction. I. fumosorosea could not be detected from all soil samples possibly caused by low density of the fungi. The qPCR was likely a rapid and specific method to detect the fungus from soil. [ABSTRACT FROM AUTHOR]

Background: Eublemma amabilis Moore (Lepidoptera: Noctuidae) is an important pest of lac insect, Kerria spp. (Hemiptera: Tachardiidae: Coccoidea) causing up to 20–25% damage of lac and its encrustation, which has immense industrial utilities. Extensive field monitoring in Regional Lac Insect Field Gene Bank (RLIFGB) resulted in collection of a large number of naturally occurring entomopathogenic fungi (EPF) infected insect cadavers on the lac encrustation of host plant, Flemingia semialata. Pathogenicity test under laboratory conditions showed its efficacy on E. amabilis larvae. Molecular characterization of this EPF by 18S rDNA identified it as Isaria fumosorosea (Wize) (accession number, MH414514.1). Results: Being a potent EPF, biological parameters viz., conidial density and sporulation were determined and its pathogenicity were evaluated against eggs, larvae and pupae of E. amabilis at different spore dilution under laboratory conditions. The highest mortality rates of larvae and pupae (96 & 72%) and (88 & 72%) were recorded at 107spore/ml both in dipping and spraying methods, respectively. The egg mortality, however, was recorded as 40 and 35% for both methods, respectively, at 107spore/ml, which was significantly lower than other stages of E. amabilis. Field study at (107 spore/ml) recorded 81.18, 59.41 and 76.36% mean population reductions over control during monsoon 2019, winter 2019–2020 and summer 2020, respectively. Biosafety analysis of the native EPF on productivity linked parameters of lac insect was found to be safe suggesting that the native Isaria fumosorosea (MH414514.1) strain, reported first from Assam. Conclusions: In this study, it was confirmed that the EPF I. fumosorosea can be offered as an effective microbial agent, alternate to insecticide against E. amabilis, either as a stand-alone strategy or in an integrated approach. [ABSTRACT FROM AUTHOR]

Aquaponics has the potential to produce sustainable and accessible quality food through the integration of hydroponics and aquaculture. Plants take up dissolved nutrients in fish wastewater, allowing water reuse for fish. However, the simultaneous presence of fish and plants in the same water loop has made phytosanitary treatments of diseases such as powdery mildew problematic due to risks of toxicity for fish and beneficial bacteria, limiting its commercialization. Entomopathogenic and mycoparasitic fungi have been identified as safe biological control agents for a broad range of pests. This study aimed to investigate the efficacy of entomopathogenic fungi, Lecanicillium attenuatum (LLA), Isaria fumosorosea (IFR), and mycoparasitic fungus Trichoderma virens (TVI) against Podosphaera xanthii. Also, we investigated the possible harmful effects of the three fungal biocontrol agents in aquaponics by inoculating them in aquaponics water and monitoring their survival and growth. The findings showed that the three biocontrol agents significantly suppressed the powdery mildew at 107 CFU/ml concentration. Under greenhouse conditions (65-73% relative humidity (RH)), a significant disease reduction percentage of 85% was recorded in L. attenuatum-pretreated leaves. IFR-treated leaves had the least AUDPC (area under disease progress curve) of ~434.2 and disease severity of 32% under 65-73% RH. In addition, L. attenuatum spores were the most persistent on the leaves, the spores population increased to 9.54 × 103 CFUmm-2 from the initial 7.3 CFUmm-2 under 65-73%. In contrast, in hydroponics water, the LLA, IFR, and TVI spores significantly reduced by more than 99% after 96 hrs. Initial spore concentrations of LLA of 107 CFU/ml spores were reduced to 4 x 103 CFU after 96 hrs. Though the results from this study were intended for aquaponics systems, relevance of the results to other cultivation systems are discussed.

Contextualization: The andean blackberry (Rubus glaucus Benth) is one of the fruits of greater commercialization and importance in Colombia and in the entire Andean zone. This crop is affected by E. colombianus, an insect pest present in blackberry and other crops such as tree tomato, lulo, grape among others. Knowledge gap: The effectiveness of biocontrol agents in controlling E. colombianus under field conditions is not known. Purpose: Evaluate the effectiveness of the native biocontrol agents Metarhizium robertsii and Isaria fumosorosea using commercial products with two application methods. Methodology: The research was carried out at the Altamira Farm, in the municipality of Guacarí (Valle del Cauca), located at 2.990 meters above sea level. 120 plants were selected, and sample of individuals of E. colombianus was taken for reactivation of the fungi in the laboratory. Additionally, the initial population was evaluated, before applying the treatments, in an experimental design of subdivided plots with a 4 x 2 x 3 factorial arrangement. Later, evaluations were carried out every eight days from the month of the first application, and over a period of a month and a half. Results and conclusions: The average initial population of E. colombianus was about 31 individuals, at different stages, in the 120 plants. The most frequently found stages were the first and second, from the root neck to a depth of 80 to 120 cm. The treatment and method that had the greatest effectiveness on E. colombianus was Metarhizium robertsii applied with an injector, which caused an average of 78 % of dead individuals, followed by I. fumosorosea with an average of 75 %. In contrast, the M. robertsii treatment applied. With a pump was the one that presented the lowest average number of individuals, with a total of 17% of individuals killed by the fungi after 45 days. [ABSTRACT FROM AUTHOR]

Flavonoid compounds are secondary plant metabolites with numerous biological activities; they naturally occur mainly in the form of glycosides. The glucosyl moiety attached to the flavonoid core makes them more stable and water-soluble. The methyl derivatives of flavonoids also show increased stability and intestinal absorption. Our study showed that such flavonoids can be obtained by combined chemical and biotechnological methods with entomopathogenic filamentous fungi as glycosylation biocatalysts. In the current paper, two flavonoids, i.e., 2′-hydroxy-4-methylchalcone and 4′-methylflavone, have been synthesized and biotransformed in the cultures of two strains of entomopathogenic filamentous fungi Isaria fumosorosea KCH J2 and Beauveria bassiana KCH J1.5. Biotransformation of 2′-hydroxy-4-methylchalcone resulted in the formation of two dihydrochalcone glucopyranoside derivatives in the culture of I. fumosorosea KCH J2 and chalcone glucopyranoside derivative in the case of B. bassiana KCH J1.5. 4′-Methylflavone was transformed in the culture of I. fumosorosea KCH J2 into four products, i.e., 4′-hydroxymethylflavone, flavone 4′-methylene-O-β-d-(4″-O-methyl)-glucopyranoside, flavone 4′-carboxylic acid, and 4′-methylflavone 3-O-β-d-(4″-O-methyl)-glucopyranoside. 4′-Methylflavone was not efficiently biotransformed in the culture of B. bassiana KCH J1.5. The computer-aided simulations based on the chemical structures of the obtained compounds showed their improved physicochemical properties and antimicrobial, anticarcinogenic, hepatoprotective, and cardioprotective potential. [ABSTRACT FROM AUTHOR]

Flavonoid compounds exhibit numerous biological activities and significantly impact human health. The presence of methyl or glucosyl moieties attached to the flavonoid core remarkably modifies their physicochemical properties and improves intestinal absorption. Combined chemical and biotechnological methods can be applied to obtain such derivatives. In the presented study, 4′-methylflavanone was synthesized and biotransformed in the cultures of three strains of entomopathogenic filamentous fungi, i.e., Isaria fumosorosea KCH J2, Beauveria bassiana KCH J1.5, and Isaria farinosa KCH J2.1. The microbial transformation products in the culture of I. fumosorosea KCH J2, flavanone 4′-methylene-O-β-D-(4″-O-methyl)-glucopyranoside, 2-phenyl-(4′-hydroxymethyl)-4-hydroxychromane, and flavanone 4′-carboxylic acid were obtained. Biotransformation of 4′-methylflavanone in the culture of B. bassiana KCH J1.5 resulted in the formation of one main product, i.e., flavanone 4′-methylene-O-β-D-(4″-O-methyl)-glucopyranoside. In the case of I. farinosa KCH J2.6 as a biocatalyst, three products, i.e., flavanone 4′-methylene-O-β-D-(4″-O-methyl)-glucopyranoside, flavanone 4′-carboxylic acid, and 4′-hydroxymethylflavanone 4-O-β-D-(4″-O-methyl)-glucopyranoside were obtained. The Swiss-ADME online simulations confirmed the increase in water solubility of 4′-methylflavanone glycosides and analyses performed using the Way2Drug Pass Online prediction tool indicated that flavanone 4′-methylene-O-β-D-(4″-O-methyl)-glucopyranoside and 4′-hydroxymethylflavanone 4-O-β-D-(4″-O-methyl)-glucopyranoside, which had not been previously reported in the literature, are promising anticarcinogenic, antimicrobial, and hepatoprotective agents. [ABSTRACT FROM AUTHOR]

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.

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 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. [ABSTRACT FROM AUTHOR]

Abstract The efficacy of the native entomopathogenic fungus, Isaria fumosorosea TR-78-3, was evaluated against females of the bark and ambrosia beetles, Anisandrus dispar Fabricius and Xylosandrus germanus Blandford (Coleoptera: Curculionidae: Scolytinae), under laboratory conditions by two different methods as direct and indirect treatments. In the first method, conidial suspensions (1 × 106 and 1 × 108 conidia ml−1) of the fungus were directly applied to the beetles in Petri dishes (2 ml per dish), using a Potter spray tower. In the second method, the same conidial suspensions were applied on a sterile hazelnut branch placed in the Petri dishes. The LT50 and LT90 values of 1 × 108 conidia ml−1 were 4.78 and 5.94/days, for A. dispar in the direct application method, while they were 4.76 and 6.49/days in the branch application method. Similarly, LT50 and LT90 values of 1 × 108 conidia ml−1 for X. germanus were 4.18 and 5.62/days, and 5.11 and 7.89/days, for the direct and branch application methods, respectively. The efficiency of 1 × 106 conidia ml− 1 was lower than that of 1 × 108 against the beetles in both application methods. This study indicates that I. fumosorosea TR-78-3 had a significant potential as a biological control agent against A. dispar and X. germanus. Further studies are necessary to evaluate the efficacy of the isolate on the pests under field conditions.

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 x 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 x 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 x 109 conidia.mL-1 on cards (1.0 x 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. [ABSTRACT FROM AUTHOR]

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.

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 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 citrus rust mite, Phyllocoptruta oleivora Ashmead (Acari: Eriophyidae), is a major pest of citrus, affecting both quality and yield. Although entomopathogenic fungi such as Beauveria bassiana (Bals.-Criv.) Vuill. (Cordycipitaceae), Metarhizium anisopliae (Metschn.) Sorokīn (Clavicipitaceae), and Isaria fumosorosea Wize (Cordycipitaceae) have been used for insect and mite control, the last 2 have never been applied to control citrus rust mite. The aim of this study was to evaluate the effect of 3 concentrations of B. bassiana, M. anisopliae, and I. fumosorosea on mortality and growth rate (r) of P. oleivora populations under greenhouse conditions. The fungi were isolated from agricultural soil samples. Mobile stages of P. oleivora were collected from leaves of commercial Valencia orange groves. At a concentration of 1 × 108 conidia mL-1, B. bassiana and M. anisopliae caused > 60% mortality in P. oleivora populations, whereas mortality due to I. fumosorosea never exceeded 50%. There was a significant inverse relationship between the percent mortality and the LT50. Application of B. bassiana resulted in the slowest growth rate of P. oleivora populations, which was 10.5 × slower than the control population receiving no fungus application. Our results demonstrate that B. bassiana and M. anisopliae are efficient in control of P. oleivora populations under greenhouse conditions, whereas I. fumosorosea produces moderate control. [ABSTRACT FROM AUTHOR]

Western flower thrips, Frankliniella occidentalis, is one of the most destructive insect pests of greenhouse-grown horticultural crops. The primary method of managing western flower thrips populations involves applications of insecticides; however, there is no information associated with the effect of the insect growth regulator, pyriproxyfen, or the entomopathogenic fungus, Isaria fumosorosea, on western flower thrips pupae in growing media. Therefore, four laboratory experiments were conducted to determine the effect of pyriproxyfen and I. fumosorosea applied as a drench to growing media on western flower thrips pupae. Expt. 1 evaluated the efficacy of pyriproxyfen and I. fumosorosea on western flower thrips pupae. Based on the results from Expt. 1, Expt. 2 assessed the effect of pyriproxyfen in two growing media (LC1 and BM1) on western flower thrips pupae. Expts. 3 and 4 determined the residual activity of pyriproxyfen in growing media on western flower thrips pupae 3, 5, 7, and 14 days after treatments were applied. The pyriproxyfen treatment resulted in a significantly lower estimated mean probability of western flower thrips adults captured on yellow sticky cards (17%) compared with the water control (59%), untreated check (88%), and two I. fumosorosea treatments (46% for 1.0 g and 41% for 2.0 g of Ancora) in Expt. 1. However, for the two growing media in Expt. 2, the estimated mean probability of western flower thrips adults captured on yellow sticky cards was not significantly different between the pyriproxyfen treatment (LC1 = 15%; BM1 = 12%) and the water control (LC1 = 41%; BM1 = 24%). For either the pyriproxyfen treatment or the untreated check, there was no evidence of a significant difference between the two growing media on the estimated mean probability of western flower thrips adults captured on yellow sticky cards. Furthermore, there was no evidence of any residual activity 3 days after drench applications of pyriproxyfen. The results of the study have demonstrated that drench applications of pyriproxyfen are not affecting survival of western flower thrips pupae. [ABSTRACT FROM AUTHOR]

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 study involved six test fungi previously recorded in the literature as being endophytes (Beauveria bassiana, Metarhizium anisopliae, Isaria fumosorosea, Trichoderma harzianum, Fusarium proliferatum, Chaetomium globosum), two plant pathogenic fungi (Ascochyta fabae, Plenodomus lingam) and four host plants (Vicia faba, Bras­sica napus, Phaseolus vulgaris, Zea mays). Aerial conidia, blastospores, or ascospores, respectively were applied to leaf surfaces by spraying or by infiltrating spore suspensions through stomata directly into the leaves. Obser­vations using light microscopy showed that the test fungi germinated on the leaf surface but did not enter actively into the leaves. Within the leaves, germination of spores and growth of hyphae appeared to depend on the presence of damaged plant tissue. Various host reactions such as browning of epidermal cells and formation of papillae were observed. Colonization of healthy leaves by the test fungi in a manner similar to the pathogens A. fabae (on Faba bean) and P. lingam (on oilseed rape) was not observed. Spore germination and hyphal growth commenced when inoculated leaves were placed on agar medium. The results indicate that the test fungi possessed a saprotrophic rather than an endo­phytic life style when associated with leaf tissue of the studied hosts.

Entomopathogenic fungi can produce a series of chitinases, some of which function synergistically with proteases and other hydrolytic enzymes to degrade the insect cuticle. In the present study, the chitinase gene Ifu-chit2 from Isaria fumosorosea was investigated. The Ifu-chit2 gene is 1,435-bp long, interrupted by three short introns, and encodes a predicted protein of 423 amino acids with a 22 residue signal peptide. The predicted Ifu-Chit2 protein is highly homologous to Beauveria bassiana chitinase Bbchit2 and belongs to the glycohydrolase family 18. Ifu-Chit2 was expressed in Escherichia coli to verify chitinase activity, and the recombinant enzyme exhibited activity with a colloidal chitin substrate. Furthermore, the expression profiles of Ifu-chit2 were analyzed at different induction times under in vivo conditions. Quantitative real-time PCR analysis revealed that Ifu-chit2 expression peaked at two days post-induction. The expression of chitinase Ifu-chit2 in vivo suggests that the chitinase may play a role in the early stage of pathogenesis.

Entomopathogenic fungi are fungal species which are used as a potential source of biopesticides. These fungi produce secondary metabolites which in insects can cause disruption in the normal functioning of their bodies, disease or even death. In order to fully characterize the physiology of entomopathogenic fungi we should identify the volatile organic compounds which are involved in this process. Therefore, we conducted a qualitative and quantitative analysis of volatile compounds produced by entomopathogenic fungi. Seven different species of fungi were analyzed: Metarhizium anisopliae , Metarhizium flavoviride , Pandora sp., Isaria fumosorosea , Hirsutella danubiensis , Batkoa sp. and Beauveria bassiana . The analyses were performed using the HS-SPME/GC MS technique. In the analyzed fungi, 63 volatile compounds were identified and classified into the following groups: aldehydes, ketones, alcohols, esters, acids, terpenes and others. The results show that entomopathogenic fungi produce a wide profile of secondary metabolites. Principal Components Analysis was used to determine whether separate classes of fungi can be distinguished from one another based on their metabolite profiles. [ABSTRACT FROM AUTHOR]

The housefly, Musca domestica L. (Diptera: Muscidae), a major insect pest in poultry and dairy farms, has developed resistance against a variety of insecticides worldwide. In order to avoid resistance development and negative impacts of insecticides, the use of entomopathogenic fungi can be an effective option for the management of M. domestica. The present study was performed to derive toxic crude proteins from entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae var. anisopliae and Isaria fumosorosea and test their effect on survival of M. domestica. The crude proteins produced in Czapek medium by the six different isolates (two each) of the entomopathogenic fungi were tested against adults of M. domestica. A significant effect was observed on the survival of M. domestica, ranging from 52.0 to 91.0% mortality. Isolates Bb-01, Ma-4.1, and If-03 showed maximum percent mortalities. These three isolates also exhibited concentration and exposure time based response toward survival of M. domestica. The crude protein concentrations i.e., 8 and 10 mg/ml caused the maximum mortality (100.0%) of M. domestica population in 2.77 to 3.77 days. In addition, the lowest exposure of duration (96 hrs) of houseflies to B. bassiana (Bb-01) crude protein (10 mg/ml) caused 100.0% mortality of tested population as compared to other isolates. In conclusion, crude proteins of entomopathogenic fungi showed good potential for the eco-friendly management of M. domestica. However, further purification of the anti-insect proteins and their evaluation under field conditions is required. [ABSTRACT FROM AUTHOR]

The development of resistance by Plutella xylostella to almost all insecticides is of significant concern all over the world. Entomopathogenic fungi such as Isaria fumosorosea have been used as an alternative to insecticides. However, the knowledge of miRNA-regulated reactions against entomopathogenic fungi is still in its infant stage. In the present study, P. xylostella was challenged with I. fumosorosea at four different time points (12, 18, 24, and 36 h) including a control, to build miRNA libraries by Illumina sequencing. The results of differential expression analysis exhibited that 23 miRNAs were differentially expressed, compared to control, in all treatments. It is worth mentioning, of these, some conserved miRNAs such as miR-2, miR-9a, miR-745, miR-7b, and miR-2767, known to play critical roles in host-pathogen interaction, were also identified. Furthermore, differentially expressed miRNAs were validated by RT-qPCR. Our results provide an essential information for further functional studies of the interaction between I. fumosorosea and P. xylostella at the post-transcriptional level.

Most, if not all, entomopathogenic fungi have been used as alternative control agents to decrease the insect resistance and harmful effects of the insecticides on the environment. Among them, Isaria fumosorosea has also shown great potential to control different insect pests. In the present study, we explored the immune response of P. xylostella to the infection of I. fumosorosea at different time points by using RNA-Sequencing and differential gene expression technology at the genomic level. To gain insight into the host-pathogen interaction at the genomic level, five libraries of P. xylostella larvae at 12, 18, 24, and 36 h post-infection and a control were constructed. In total, 161 immunity-related genes were identified and grouped into four categories; immune recognition families, toll and Imd pathway, melanization, and antimicrobial peptides (AMPs). The results of differentially expressed immunity-related genes depicted that 15, 13, 53, and 14 up-regulated and 38, 51, 56, and 49 were down-regulated in P. xylostella at 12, 18, 24, and 36 h post-treatment, respectively. RNA-Seq results of immunity-related genes revealed that the expression of AMPs was reduced after treatment with I. fumosorosea. To validate RNA-Seq results by RT-qPCR, 22 immunity-related genes were randomly selected. In conclusion, our results demonstrate that I. fumosorosea has the potential to suppress the immune response of P. xylostella and can become a potential biopesticide for controlling P. xylostella.

Los agricultores colombianos utilizan productos químicos con categorías toxicológicas I, II y III para el control de insectos como la mosca blanca Bemisia tabaci (Gennadius, 1889) (Hemiptera: Aleyrodidae); pero su uso indiscriminado genera un riesgo para la salud y el medio ambiente. El control microbiológico con hongos entomopatógenos surge como alternativa de control ambientalmente sostenible. No obstante, su empleo exitoso dentro de una estrategia de manejo integrado de plagas (MIP) depende, en gran parte, del efecto de los productos químicos sobre el bioplaguicida. El objetivo del presente trabajo fue determinar la compatibilidad in vitro de un bioplaguicida con base en el hongo Isaria fumosorosea formulado como polvo mojable con cuatro fungicidas y cinco insecticidas comerciales. Para esto, se evaluó el efecto de tres concentraciones de los productos químicos: FR = dosis recomendada en campo, 0.5 x FR y 0.25 x FR sobre la germinación de los conidios y el número de unidades formadoras de colonia UFC/g de I. fumosorosea. Los cuatro fungicidas (Benlate, Carboxin Captan, Metalaxil-Mancozeb y Mancozeb) en las tres dosis evaluadas fueron incompatibles con el bioplaguicida, mientras que el insecticida Thiametoxam fue compatible cuando se utilizó la dosis de 0.25 x FR.

The interactions between the entomopathogenic fungus Beauveria bassiana Balsamo (Vuillemin) (Hypocreales: Cordycipitaceae) and the entomopathogenic fungus Isaria fumosorosea (Wize) Brown and Smith (Hypocreales: Clavicipitaceae) were examined on young adults of Sitophilus granarius (L.) (Coleoptera: Curculionidae) and S. oryzae (L.) (Coleoptera: Curculionidae). Conidial suspensions of these entomopathogenic fungi were applied both separately and in combination, at three dosages, 104, 106, and 108 conidia/mL. Mortality of experimental adults was recorded daily for 15 days. An overall positive interaction between the pathogenic microorganisms was observed. Mean weevil mortality caused by the separate acting fungi, B. bassiana, ranged from 26.7% to 53.3% and from 36.6% to 63.3% for S. granarius and S. oryzae, respectively. The respective values for I. fumosorosea were 20.0%−53.3% and 46.7%−66.7%. The combined treatments showed a distinct interaction between the pathogens; for S. granarius, the interaction between the pathogens was additive in all combinations, whereas, for S. oryzae, the interaction was additive in seven and competitive in two of the combinations. Applying both entomopathogenic microorganisms may offer a method for weevil control that could be more effective than using each pathogen alone.

Glycosylation is an efficient strategy to modulate the solubility, stability, bioavailability and bioactivity of drug-like natural products. Biological methods, such as whole-cell biocatalyst, promise a simple but highly effective approach to glycosylate biologically active small molecules with remarkable regio- and stereo-selectivity. Herein, we use the entomopathogenic filamentous fungus Isaria fumosorosea ACCC 37814 to biotransform a panel of phenolic natural products, including flavonoids and anthraquinone, into their glycosides. Six new flavonoid (4-O-methyl)glucopyranosides are obtained and structurally characterized using high resolution mass and nuclear magnetic resonance spectroscopic techniques. These compounds further expand the structural diversity of flavonoid glycosides and may be used in biological study.

Isaria fumosorosea and Isaria farinosa are important entomopathogenic fungi with a worldwide distribution and multiple host insects. However, the concerns about the safety risks of myco-pesticides have been attracting the attention of researchers and consumers. Secondary metabolites (SMs), especially the mycotoxins, closely affect the biosafety of Isaria myco-insecticides. In the last forty years, more than seventy SMs were identified and isolated from I. fumosorosea and I. farinose. The SMs of I. fumosorosea include the mycotoxins of non-ribosomal peptides (NRPs) (beauvericin and beauverolides), terpenes (trichocaranes and fumosorinone), lactone compounds (cepharosporolides), acids (dipicolinic acid and oxalic acid), etc. Meanwhile, the NRP mycotoxins (cycloaspeptides) and the terpene compounds (farinosones and militarinones) are the main SMs in I. farinosa. Although several researches reported the two Isaria have promised biosafety, the bioactivities and the safety risks of their SMs have not been studied in detail so far. However, based on existing knowledge, most SMs (i.e., mycotoxins) do not come from Isaria myco-insecticide itself, but are from the host insects infected by Isaria fungi, because only the hosts can provide the conditions for fungal proliferation. Furthermore, the SMs from Isaria fungi have a very limited possibility of entering into environments because many SMs are decomposed in insect cadavers. The biosafety of Isaria myco-insecticides and their SMs/mycotoxins are being monitored. Of course, SMs safety risks of Isaria myco-insecticides need further research.

Protein tyrosine phosphatase 1B (PTP1B) is implicated as a negative regulator of insulin receptor (IR) signaling and a potential drug target for the treatment of type II diabetes and other associated metabolic syndromes. Thus, small molecule inhibitors of PTP1B can be considered as an attractive approach for the design of new therapeutic agents of type II diabetes and cancer diseases. In a continuing search for new PTP1B inhibitors, a new tetramic acid possessing a rare pyrrolidinedione skeleton named fumosorinone A (1), together with five known ones 2-6 were isolated from the entomogenous fungus Isaria fumosorosea. The structures of 2-6 were elucidated by extensive spectroscopic analysis. Fumosorinone A (1) and beauvericin (6) showed significant PTP1B inhibitory activity with IC50 value of 3.24 μM and 0.59 μM. [ABSTRACT FROM AUTHOR]

The catalytic activity of enzymes produced by an entomopathogenic filamentous fungus (Isaria fumosorosea KCh J2) towards selected steroid compounds (androstenedione, adrenosterone, progesterone, 17α-methyltestosterone and dehydroepiandrosterone) was investigated. All tested substrates were efficiently transformed. The structure of the substrate has a crucial impact on regio- and stereoselectivity of hydroxylation since it affects binding to the active site of the enzyme. Androstenedione was hydroxylated in the 7α-position to give a key intermediate in the synthesis of the diuretic-7α-hydroxyandrost-4-ene-3,17-dione with 82% conversion. Adrenosterone and 17α-methyltestosterone were hydroxylated in the 6β-position. Hydroxylated derivatives such as 15β-hydroxy-17α-methyltestosterone and 6β,12β-dihydroxy-17α-methyltestosterone were also observed. In the culture of Isaria fumosorosea KCh J2, DHEA was effectively hydroxylated in the C-7 position and then oxidized to give 7-oxo-DHEA, 3β,7α- and 3β,7β-dihydroxy-17a-oxa-d-homo-androst-5-ene-17-one. We obtained 7β-OH-DHEA lactone with 82% yield during 3 days transformation of highly concentrated (5 g/L) DHEA. [ABSTRACT FROM AUTHOR]

This work investigated the production of the fungi Isaria fumosorosea and Isaria farinosa in biphasic fermentation using agro-industrial products and residues. Combinations of natural liquid substrates, alternative to the complete medium and potato dextrose medium, were evaluated. The best liquid media were sugarcane molasses + rice broth, rice broth + yeast and sugarcane molasses + yeast + rice broth, which resulted in the highest viable propagule concentration. The molasses + rice broth medium was selected for the next phase of the study in which the production of both fungal isolates was evaluated in solid grain substrates. In solid-state fermentation, the best conidia production was achieved with the soybean meal and broken corn for I. farinosa, and whole rice and broken rice for I. fumosorosea. Results demonstrated that the two fungal species could be rapidly produced with higher yield of conidia on agro-industrial resources by using biphasic fermentation techniques.Estudou-se a produção dos fungos Isaria fumosorosea (ESALQ1296) e Isaria farinosa (ESALQ1205) em fermentação líquida e sólida utilizando produtos e resíduos agroindustriais. Inicialmente, avaliaram-se combinações de substratos líquidos naturais e alternativos aos meios completo e batata-dextrose. Os meios líquidos mais promissores foram: melaço + caldo de arroz, caldo de arroz + levedura e melaço + levedura + caldo de arroz, pois resultaram nas maiores produções de propágulos viáveis. O meio melaço + caldo de arroz foi selecionado para etapa seguinte do trabalho, na qual se avaliou a produção dos fungos em grãos. A fermentação sólida foi mais eficiente quando se utilizaram os meios de farelo de soja e milho quirela para I. farinosa,e arroz inteiro e arroz quirela para I. fumosorosea. A fermentação bifásica para os fungos estudados a partir desses produtos agroindustriais selecionados proporcionou elevada produção de conídios em curto intervalo de tempo, podendo ser implantada em escala industrial.