Impact of culture age on conidial germination,desiccation and UV tolerance of entomopathogenic fungi

The impact of culture age on conidial yields, germination and tolerance to UV exposure of freshly harvested and dry conidia produced by five entomopathogenic fungal (EPF) isolates was studied. Beauveria bassiana, Metarhizium anisopliae, Lecanicillium lecanii and Lecanicillium muscarium were grown on potato dextrose agar (PDA) medium for 7 or 14 days at 25°C. While the age of cultures had a significant impact on the germination rate of conidia produced by isolates L. lecanii CBS 122.175 and B. bassiana LMSA 1.01.093, other EPF isolates germinated at the same rate regardless of the culture age. When exposed to UV radiation, conidia produced by all isolates germinated at a lower rate compared to the non-irradiated conidia, although this decrease in germination (20–80% decrease) was unaffected by the culture age. Air-drying had only a slight impact on conidial germination (0–60% decrease). Under the conditions of this study, the stability of irradiated conidia produced by M. anisopliae LMSA 1.01.197 and B. bassiana CBS 110.25 was significantly increased when conidia were dried prior to UV exposure. This increase in tolerance to stress of dried conidia might be caused, at least partially, by the low metabolic activity associated with dehydration.     

Influence of two formulation types and moisture levels on the storage stability and insecticidal activity of Beauveria bassiana

The formulation of mycopesticides may require a physical separation of conidia from the substrate and subsequent drying. In the present study, Beauveria bassiana conidia produced by solid-state fermentation were harvested either through a dry or washing protocol. Washed conidia were used to design a water-dispersible granule (WG) formulation, whereas sieved conidia were mixed with an emulsifiable oil to achieve an oil-based formulation (OD). Potential harmful effects caused by the formulation type on the storage stability and insecticidal activity against Hypothenemus hampei were assessed. As expected, the time for initial conidial germination to drop 50% (GT₅₀) in all treatments was deeply influenced by storage temperatures, which varied from over 180 days at 4 °C to less than 90 days at 35°C. In all four tested temperatures, GT₅₀s for unformulated dry conidia were significantly higher than for those formulated as WG, and the latter was similar to conidia formulated as OD in the two highest temperatures. Residual water content in the OD formulation (1,600 vs. 340?ppm) had a negative influence on conidial survival under storage, whereas WG granules immediately dried after the washing protocol showed conidial germination similar to granules exposed to a slower dehydration regime. Mortality of H. hampei adults exposed to different concentrations of B. bassiana formulated as WG was slightly lower (10–15%) than either the OD or the unformulated conidia. In brief, we have demonstrated that formulation type and their moisture level can affect the storage stability and insecticidal activity of B. bassiana conidia toward the coffee berry borer. Of particular importance, we have shown that drying oils prior to formulation could improve the storage of mycopesticides, an approach that may find industrial applications.     

In vitro compatibility of Beauveria bassiana strain ATCC 74040 with copper fungicides

Copper fungicides and mycoinsecticides based on entomopathogenic fungi Beauveria spp. are the most common pesticides used in organic crop production systems. The in vitro effects of the copper fungicides copper oxide, copper hydroxide, copper oxychloride, copper sulphate, dicopper chloride trihydroxide and tribasic copper sulphate were investigated for mycelial growth, sporulation and conidial germination of the ATCC 74040 commercial strain of Beauveria bassiana. Mycelial growth was evaluated on potato dextrose agar plates with 100%, 75%, 50%, 25%, 12.5%, 6.25% and 0% of the recommended application rates of each fungicide at 15 and 25°C. Sporulation and conidial germination were determined at the recommended field doses of each fungicide at 25°C. All copper fungicides had fungistatic or fungicidal effects on B. bassiana that varied according to the dose. Only in two cases, copper oxide at 15°C and copper hydroxide at 25°C, at the lowest concentration of 6.5%, was mycelial growth not statistically significantly inhibited. Inhibition of mycelial growth depended both on the fungicide and its concentration, and partly on temperature. Both sporulation and conidial germination of B. bassiana were significantly inhibited by all fungicides. All fungicides inhibited the sporulation in a similar way (99.8%–100%). With the exception of copper oxychloride (inhibition, 13.6%), the other fungicides showed high detrimental effects on conidial germination (inhibition, 91.7%–100%). The fungus was strongly affected by some fungicides even at the lowest doses. The biological index used for the B. bassiana with copper fungicides ranged from 0.6 (copper sulphate) to 18.1 (copper oxychloride). Therefore, the tested fungicides were classified into the upper half of the highly toxic (T) category and are considered incompatible with the entomopathogenic fungus B. bassiana strain ATCC 74040 under in vivo experimental conditions. These results need to be further verified in vitro under both greenhouse and open-field conditions.     

Influence of commercial fungicides on the germination,growth and virulence of four species of entomopathogenic fungi

The influence of 15 commercially available fungicides on the germination, growth and virulence of Metarhizium anisopliae, Beauveria bassiana, Isaria fumosorosea, and Lecanillium longisporum was evaluated. The influence of the fungicides on conidial germination was dependant on the fungicide type and dose. Most fungicides retarded conidial germination of all the fungi tested at 10× and at the recommended rate of application, however, their toxicity declined at lower concentrations. Most of the fungicides inhibited mycelial growth of B. bassiana, whereas L. longisporum growth was unaffected. Only two and eight fungicides influenced mycelial growth of I. fumosorosea and M. anisopliae, respectively. None of the fungicides influenced the virulence of B. bassiana and L. longisporum, however, tolylfluanid and azoxystrobin reduced the virulence of M. anisopliae and I. fumosorosea, respectively. These studies clearly show that certain fungicides have the potential to inhibit germination of entomopathogenic fungi in vitro but appear to have little or no effect on their virulence against target insects.     

Selection of surfactant compounds to enhance the dispersion of Beauveria bassiana

This study aimed to select surfactant compounds to improve the dispersion of emulsifiable conidial suspensions of Beauveria bassiana for field application while maintaining high conidia viability. Surfactant compatibility was initially evaluated by colony growth in culture media amended with increasing doses of surfactants. In a second experiment, the conidia germination rate was evaluated after temporary exposure to the compounds under laboratory conditions. The effect of the hydrophilic–lipophilic balance (HLB) of the surfactants on conidia dispersion and viability was assessed using mixtures of the most compatible products and concentration initially selected. Unitol L/20 and Ultranex NP/100 did not show deleterious effect on colony growth on culture media, while Surfom 3409 strongly inhibited conidial germination. Unitol L/20, Ultranex NP/100 and Ultratan D did not interfere in the germination of conidia during the first 10 minutes of exposure. After 120 minutes, germination was greatly reduced at a 0.5% concentration, with the exception of Ultranex NP/100. The higher conidial dispersion and germination were obtained with HLB values between 8 and 12 and lower remaining clusters at HLB 10. Moreover, there was no significant effect of HLB values on insect mortality. Unitol L/20 and Ultranex NP/100 were the most compatible surfactants in concentrations lower than 0.5%. The HLB value selected for dispersion of B. bassiana on water based formulations was 10.     

Growth and insect assays of Beauveria bassiana with neem to test their compatibility and synergism

Beauveria bassiana is being used as a biopesticide for many insect pests. Neem oil (azadirachtin) is an eco-safe popular botanical pesticide. A biopesticde with a neem compatible isolate of B. bassiana will enable their simultaneous use in pest management. A sample of 30 isolates of B. bassiana from culture collections was screened for compatibility with a commercial formulation of neem oil (Margoside®) at the field recommended dose (0.3%, v/v). Compatibility was tested in vitro through germination and growth assays. In all isolates, conidial germination was delayed but not significantly decreased by neem. In the growth assays, 23 isolates were found compatible with neem. In the neem sensitive isolates, growth was decreased but not totally inhibited. The effect of combined treatment with B. bassiana and neem in comparison to single treatments with either of them on Spodoptera litura Fabricius was tested in laboratory bioassays. The combined treatment was found to have synergistic effect on insect mortality when a B. bassiana isolate compatible with neem was used, while, with an isolate sensitive to neem, an antagonistic effect was observed.     

Evaluation of Beauveria bassiana (Hyphomycetes) isolates as potential agents for control of Dendroctonus valens

Abstract The red turpentine beetle (RTB), Dendroctonus valens LeConte, as a destructive invasive pest, has become one of the most economically important forest pest in China. Effective control measures are desperately needed. Entomopathogenic fungi, such as Beauveria bassiana, have shown great potential for the management of some bark beetle species. In this study, 12 isolates of B. bassiana from bark beetle were examined for biological characteristics and virulence, to assess their potential as biocontrol agents for RTB. There were significant differences (at P= 0.05) in colony growth rate, conidial yield, conidial germination, tolerance to UV light and extracellular proteases activity among the tested B. bassiana isolates. Isolates, including Bb1801, Bb1906, Bb789 and Bb773, exhibited the best characteristics, because they have faster hyphal growth rate, higher spore production and faster spore germination, higher UV tolerance and protease (Pr1) production. The results of a pathogenicity test of B. bassiana on RTB larvae showed that most isolates of B. bassiana have demonstrated high efficacy and the highest virulent isolate was Bb1801, which killed 100% of the treated insects and had a median lethal time (LT₅₀) of 4.60 days at a concentration of 1×10⁷ conidia/mL. Therefore, isolate Bb1801 has a great potential for sustainable control of RTB in the forest. The correlation between biological characteristics and virulence of the fungal isolates is discussed and the possibility of combination of entomopathogenic fungi with semiochemicals, as one of the promising strategy for RTB control, is considered.     

Use of Beauveria bassiana in combination with commercial insecticides to manage Phauda flammans (Walker) (Lepidoptera: Phaudidae): Testing for compatibility and synergy

An entomopathogenic fungal strain, Beauveria bassiana PfBb, was identified from Phauda flammans (Lepidoptera: Phaudidae) larvae. The compatibility and synergy of B. bassiana PfBb employed in combination with three concentrations (i.e., recommended concentration, 20% and 10% of the recommended concentration) of five commercial insecticides were determined. Beta cypermethrin at 10% of recommended concentration had the lowest inhibitory effect on the mycelial growth of B. bassiana PfBb compared with other insecticides. Insecticides utilized at recommended concentration had no significant effect on the sporulation of B. bassiana PfBb, while the extent of their effect at 20% and 10% of recommended concentration differed among insecticides. Insecticides at 10% of recommended concentration had the lowest inhibition of sporulation and conidial germination compared with other concentrations. The conidial germination of B. bassiana PfBb was the highest after treatment with beta cypermethrin at 10% of recommended concentration. The cumulative mortality for 1 × 10⁷ spores/mL B. bassiana PfBb combined with each insecticide at 10% of recommended concentration was higher than that observed with the application of insecticides alone. The percent cadavers of Phauda flammans larvae observed after treatment with B. bassiana PfBb combined with beta cypermethrin at 10% of recommended concentration were not significantly different from those observed after infection with B. bassiana PfBb alone. Our findings demonstrate that B. bassiana PfBb combined with beta cypermethrin at 10% of recommended concentration could increase the efficiency of this insecticide.     

草菇VvLaeA调控球孢白僵菌的生长发育

食用真菌草菇[Volvariella volvacea(Bull.ex.Fr.)Sing.]VvLaeA在调控真菌发育方面的功能并不清楚。本研究在对草菇VvLaeA进行生物信息学分析基础上,通过聚合酶链式反应(polymerase chain reaction,PCR)的方法将超量表达启动子Vvgpd和草菇VvlaeA基因的开放阅读框(open reading frame,ORF)进行融合,并将融合片段克隆入pK2(bar)载体。采用农杆菌介导的转化方法将重组载体pK2(bar)-OEVvlaeA转入球孢白僵菌中进行表达,测定转化子的菌株发育情况。结果表明,草菇VvLaeA与其他真菌的蛋白同源性较低;和野生型相比,转化菌株的菌落直径显著增大,色素减少,分生孢子产量和萌发率均显著下降,对胁迫反应的敏感性增强。进一步研究发现,转化菌株分生孢子的细胞壁结构成分发生改变,和分生孢子发育相关基因的表达被显著下调。提示草菇VvLaeA能提高球孢白僵菌菌株的生长速率,而负调控色素沉积和分生孢子发育,这为草菇基因功能鉴定提供了启示。     

<Emphasis Type="Italic">Beauveria bassiana</Emphasis> pathogenicity to the cherry slugworm, <Emphasis Type="Italic">Caliroa cerasi</Emphasis> (Hymenoptera: Tenthredinidae) larvae

The cherry slugworm Caliroa cerasi is a significant destructive pest of sour cherries (Prunus cerasus) in Turkey. The potential of entomopathogenic fungi for controlling C. cerasi was evaluated. The effects of exposure methods and conidial concentrations (1 × 10⁶, 1.5 × 10⁶, 1 × 10⁷ and 1.5 × 10⁷ conidia/ml) on mature larvae of C. cerasi infected by Beauveria bassiana were investigated under laboratory conditions. Larvae sprayed directly with B. bassiana conidial suspensions and larvae exposed to B. bassiana-treated leaves resulted in 100% mortality within 2.90 and 2.77 days, respectively. The median lethal time (LT₅₀) and days to mortality were highest in the 1.0 × 10⁷ concentrations of conidia for both direct spray and leaf exposure. The present study suggests that B. bassiana has good potential for control of the cherry slugworm, C. cerasi.     

Isolation of Beauveria species from Lebanon and evaluation of its efficacy against the cedar web-spinning sawfly, Cephalcia tannourinensis

Larvae of the cedar web-spinning sawfly, Cephalcia tannourinensis Chevin (Hymenoptera: Pamphiliidae), infected with a white fungus were collected from the Tannourine-Hadath El-Jebbeh cedar forest. Macro- and micro-morphological data based on the examination of colonies, conidiophores, and conidial shape of the fungus suggested a Beauveria species. Sequence analysis of the internal transcribed spacer regions of the isolated fungus showed that it is most closely related to isolates of B. bassiana Clade C. The present study showed that the isolated B. bassiana is a naturally occurring entomopathogenic fungus parasitizing the larvae of C. tannourinensis in Lebanon. Laboratory bioassays showed that B. bassiana caused high mortality of eggs and larvae. The infected eggs turned brownish in color, while larvae of the first instar ceased feeding and showed immobility and rigidity within 5 days and before sporulating conidial mat appeared on their cuticle. Second and third larval instars took longer time to show fungal sporulation: mortalities ranged between 85 and 100% within 7 days when treated with different conidial concentrations. The efficacy of control of C. tannourinensis using B. bassiana was higher or equal to the reference insect growth regulator, diflubenzuron, suggesting the possibility of its success as a biological control agent.     

A conidial protein (CP15) of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> contributes to the conidial tolerance of the entomopathogenic fungus to thermal and oxidative stresses

Aerial conidia are central dispersing structures for most fungi and represent the infectious propagule for entomopathogenic fungus Beauveria bassiana, thus the active ingredients of commercial mycoinsecticides. Although a number of formic-acid-extractable (FAE) cell wall proteins from conidia have been characterized, the functions of many such proteins remain obscure. We report that a conidial FAE protein, termed CP15, isolated from B. bassiana is related to fungal tolerance to thermal and oxidative stresses. The full-length genomic sequence of CP15 was shown to lack introns, encoding for a 131 amino acid protein (15.0 kDa) with no sequence identity to any known proteins in the NCBI database. The function of this new gene with two genomic copies was examined using the antisense-RNA method. Five transgenic strains displayed various degrees of silenced CP15 expression, resulting in significantly reduced conidial FAE protein profiles. The FAE protein contents of the strains were linearly correlated to the survival indices of their conidia when exposed to 30-min wet stress at 48°C (r ² = 0.93). Under prolonged 75-min heat stress, the median lethal times (LT₅₀s) of their conidia were significantly reduced by 13.6–29.5%. The CP15 silenced strains were also 20–50% less resistant to oxidative stress but were not affected with respect to UV-B or hyperosmotic stress. Our data indicate that discrete conidial proteins may mediate resistance to some abiotic stresses, and that manipulation of such proteins may be a viable approach to enhancing the environmental fitness of B. bassiana for more persisting control of insect pests in warmer climates.     

Conidial water affinity is an important characteristic for thermotolerance in entomopathogenic fungi

The interspecific thermotolerance of several species of entomopathogenic fungi was evaluated based on the conidial water affinity. The species were divided between hydrophilic and hydrophobic conidia. The species with hydrophobic conidia were Beauveria bassiana (ARSEF 252), Metarhizium brunneum (ARSEF 1187), Metarhizium robertsii (ARSEF 2575), Isaria fumosorosea (ARSEF 3889) and Metarhizium anisopliae s.l. (ARSEF 5749). The species with hydrophilic conidia were Tolypocladium cylindrosporum (ARSEF 3392), Tolypocladium inflatum (ARSEF 4877), Simplicillium lanosoniveum (ARSEF 6430), Lecanicillium aphanocladii (ARSEF 6433), S. lanosoniveum (ARSEF 6651), Aschersonia placenta (ARSEF 7637) and Aschersonia aleyrodis (ARSEF 10276). The conidial surface tension of each isolate was also studied. Conidial suspensions were exposed to 38, 41 or 45 °C. After exposure, the suspensions were inoculated on media and conidial germination was evaluated. Considerable differences in thermotolerance were found among the 12 entomopathogenic fungal species. Species with hydrophobic conidia were generally more thermotolerant than species with hydrophilic conidia. All isolates with hydrophobic conidia showed higher conidial surface tension than the isolates with hydrophilic conidia.     

腺苷酸核糖基化因子BbarfA介导白僵菌孢子萌发和毒力

【目的】探究腺苷酸糖基化因子ARF在球孢白僵菌(Beauveria bassiana)中存在种类及生物学功能。【方法】利用BLASTp搜索球孢白僵菌非冗余蛋白数据库,鉴定ARF并进行聚类分析,结合表达分析、反义抑制、超量表达野生型基因和GTP解离位点与结合位点突变的基因,解析其中1个ARF与白僵菌发育分化、逆境胁迫反应和毒力的关系。【结果】球孢白僵菌中存在至少6个ARF或类似蛋白,分别聚类于酵母、人类ARF及其类似蛋白的不同类群。其中BBA_01574与人类的ARF3、ARF4和ARF5聚为一类,命名为BbarfA。BbarfA在成熟的分生孢子和球形膨大时期表达明显高于芽管伸长期。反义抑制BbarfA加速了孢子萌发,提高了菌株毒力,而超量表达BbarfA和点突变GTP解离区域的BbarfA则延迟了孢子萌发速度,降低了菌株毒力。尽管BbarfA转录受高盐、髙渗、氧化和高温胁迫的诱导,但遗传修饰的转化子与野生菌株对上述胁迫反应的敏感性无明显差异。【结论】BbarfA介导分生孢子萌发和毒力。     

Application of hydrophilic–lipophilic balance (HLB) number to optimize a compatible non-ionic surfactant for dried aerial conidia of Beauveria bassiana

The hydrophilic–lipophilic balance (HLB) number system was used to optimize a compatible non-ionic surfactant, TDA (polyoxyethylene tridecyl ether) in formulations for two Beauveria bassiana strains, NI8 and GHA. The optimal HLB number for TDA was determined on the basis of wetting times for conidial powders. The results indicated that optimal HLB number of TDA for B. bassiana strain NI8 was 8, while the optimum HLB number for strain GHA was 10. The optimized TDA surfactants required significantly less wetting times than the commonly used laboratory surfactants, Triton X-100, Span 80, and Tween 80. These optimized TDA surfactants were further characterized on their ability to produce conidial suspensions of the two strains after 5 min of mixing, TDA HLB 8 and TDA HLB 10 produced suspensions of 1.8 × 10⁸ and 1.6 × 10⁸ conidia/ml for NI8 and GHA, respectively. These conidial levels were significantly higher than those in Triton X-100, Span 80, and Tween 80 suspensions after the same mixing time. Germination assays showed that TDA HLB 8 promoted significantly higher germination rates of strain NI8 than those observed in other commonly used laboratory surfactants. However, the germination rates of the GHA strain were unaffected by any of the surfactants tested. The efficacy of the conidial suspensions was confirmed with assays against Lygus lineolaris. Bioassay results indicated that there were no significant differences in mortalities because of surfactants. These results suggest optimization based upon HLB number will not negatively impact parameters associated with efficacy, while providing desirable physical properties.     

Keimungsprozente und Keimungskurven der Konidien und Submers Gebildeten Blastosporen eines Stammes vonBeauveria bassiana (Bals.) Vuill.

Zusammenfassung Für Infektionsversuche und biologische Bek?mpfung k?nnen Massenkulturen vonBeauveria bassiana entweder Konidien oder Blastosporen liefern. Die qualitative Bewertung solcher Sporen und Sporenpr?parate und ebenso deren Standardisierung setzen voraus, dass auch das Keimverhalten der Sporen bekannt ist. Deshalb wurden die zeitabh?ngigen Keimungsprozente und Keimungskurven der Blastosporen und Konidien eines Stammes vonB. bassiana auf Malzextrakt-Pepton-Agar bei 24 und 18°C ermittelt und damit auch die Unterschiede, die durch Temperatur und Sporenart bedingt sind. Für die Bewertung und Standardisierung von Sporen und Sporenpr?paraten k?nnen die Keimprozente nach 24 und 36 Stunden oder die Zeit für 50% Keimung Kriterien sein und mit den hier wiedergegebenen Standardwerten verglichen werden.

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Summary Mass cultures ofBeauveria bassiana provide either conidia or blastospores for laboratory experiments and biological control measures. For qualitative evaluation of spores and spore preparations it is of primary importance to know the germination behaviour of spores. Therefore, the percentage germination of blastospores and conidia in relation to time (0–36 hours) and temperature (24 and 18°C) have been determined on malt-extract peptone agar (Table 1) and demonstrated by graphs (Fig. 1). The periods of time necessary for 50% germination of blastospores and conidia show differences depending on temperature and type of spore (Table 2). Pronounced differences of percentage germination as caused by the two temperatures become obvious with blastospores after 6–10, with conidia after 15–20 hours; after 24 hours, such differences are relatively low (Table 3). For evaluation and standardization of spores and spore preparations ofB. bassiana the percentage germination at 24°C as obtained after 24 and 36 hours or the period of time for 50% germination can be used as criteria and compared with the corresponding standard values presented.

     

Optimised fermentation conditions and improved collection efficiency using dual cyclone equipment to enhance fungal conidia production

Methods for enhancing conidial yield and for harvesting pure fungal conidia of entomopathogenic fungi were investigated. Fermentation conditions (liquid-to-solid ratio, MgSO₄·7H₂O, incubation temperature, inoculum sizes, KNO₃ and relative humidity) of Beauveria bassiana s.l. and Metarhizium anisopliae s.l. were optimised to increase the conidial yields that reached 11.2 mg/g and 24.5 mg/g, increases of 72% and 52% compared to the unoptimised yields of 6.5 mg/g and 16.1 mg/g, respectively. Three methods were compared for harvesting pure conidia of B. bassiana: dual cyclone equipment (DCE), sieving 200 and elution with 0.02% Tween-80 suspension. DCE performed the best, giving a conidial yield of 12.6 mg/g and 1.8 × 10¹⁰ conidia·g^–1. To further enhance the harvest efficiency, response surface methodology combined with a Box–Behnken design was employed, and the conidial yield of B. bassiana reached 20.9 mg/g, a total increase of 221% compared to the original conditions. Under these optimised harvest parameters, the conidial yield of M. anisopliae rose to 42.2 mg/g, an increase of 162%. The conidia of B. bassiana and M. anisopliae harvested in this way were pure, with no mycelial fragments or substrate visible in microscopic images.     

Nutrient-mediated germination of Beauveria bassiana conidia on the integument of the bark beetle Dendroctonus ponderosae (Coleoptera:Scolytidae)

Field-collected adult mountain pine beetles, Dendroctonus ponderosae, were inoculated with the white muscardine fungus, Beauveria bassiana, and thoroughly examined externally with a scanning electron microscope. Germinating conidia were found at a very low incidence, and only on antennal clubs. However, mortality of inoculated controls was high, and B. bassiana was confirmed as the causative agent. Germination on the cuticle was greatly increased by sonication of adult beetles. The hypothesis that hemolymph released through sonication-damaged membranes provided a nutrient stimulus that enhanced conidial germination was tested by placing hemolymph or yeast-malt extract broth on the cuticle of otherwise nontreated beetles, which were then inoculated with conidia. Significant germination occurred on the cuticle of these beetles. Therefore, a limiting factor for conidial germination on the sclerotized cuticle of D. ponderosae is probably a lack of sufficient nutrients.     

Influence of humidity on the infection of western flower thrips, <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis> (Thysanoptera: Thripidae), by <Emphasis Type="Italic">Beauveria bassiana</Emphasis>

The insecticidal activity of Beauveria bassiana GHA derived from a commercial mycoinsecticide BotaniGard ES against Frankliniella occidentalis was determined in a bioassay by dipping the female adults into a conidial suspension. The 90% lethal concentration of B. bassiana GHA was estimated to be 9.7 × 10⁶ conidia/ml. The lethal times for achieving 90% mortality of thrips inoculated with a 1/500-diluted solution of BotaniGard ES and a 10^7.5 (3.16 × 10⁷) conidia/ml suspension of B. bassiana GHA were estimated to be five and six days, respectively. When the treated thrips were exposed to a high relative humidity (RH) of over 99% for various periods and then transferred to 60% RH, the requisite lengths of the high-humidity period to achieve 90% mortality of the thrips at six days after inoculation were estimated to be 46 and 47 h in BotaniGard ES and B. bassiana GHA, respectively. Fungal multiplication in the thrips was detected between 48 to 60 h after inoculation by measuring Beauveria-specific DNA in the host following inoculation with a B. bassiana GHA suspension of 10^7.5 conidia/ml using a real-time quantitative PCR. The mycelial growth in the host hemocoel was not influenced by the low-humidity condition.