Recovery of endophytic Beauveria bassiana on a culture medium based on cetyltrimethylammonium bromide

Selective media (potato dextrose agar plus yeast extract, different concentrations of cetyltrimethylammonium bromide (CTAB) and different antibacterials) were evaluated for the isolation of endophytic Beauveria cf. bassiana from dry bean (Phaseolus vulgaris) plants. CTAB amounts of 0.15 and 0.3?g/L plus either dihydrostreptomycin, oxytetracycline or doxycycline resulted in the lowest numbers of common saprophytic fungi and the highest proportional recovery of Beauveria colonies from internal plant tissues.     

Efficacy of Trichogramma wasps for controlling tomato leaf miner Tuta absoluta

Tomato is considered as one of the most essential vegetables after potato in Egypt. Tomato is infested by many insects; one of these serious insects is tomato leaf miner Tuta absoluta, which causes yield losses up to 100%. T absoluta is attacking leaves, flowers, stems and fruits. Trichogramma species is playing an important role in reducing this insect infestation. Efficiency of Trichogramma evanescens alone and in combination with insecticides were assessed against T. absoluta in tomato fields. Three release rates 500, 1000 and 1500 parasitized egg/card as well as 2 insecticides (Dimilin and Corgen) were applied. Number of normal and parasitized eggs, live and dead larvae and tunnel numbers were recorded. The results revealed that T. absoluta eggs were higher in control plots than those counted in Trichogramma plots. Among Trichogramma release rates, higher release rates, obtained higher parasitism rates of Tuta eggs (reached to 50%). Companying Trichogramma with IGR Dimilin had a little side effect on parasitism rate which it was smaller compared to plots of Trichogramma alone. Dimilin (IGR) caused reducing in Tuta larval population, which it differed significantly compared to control plots. Another IGR, Coragen resulted increasing the mortality of Tuta larvae than other treatments. In Trichogramma release rates, higher release rates, lower number of tunnels were found. In plots of companying Trichogramma with Coragen, the lowest tunnel numbers were recorded. Field release of T. evancences would be a promise biocontrol agent to suppress T. absoluta. These findings may encourage using T. evancences as a favorable strategy for integrated management of T. absoluta to produce safe tomato.     

Interrelated responses of tomato plants and the leaf miner Tuta absoluta to nitrogen supply

Plant–insect interactions are strongly modified by environmental factors. This study evaluates the influence of nitrogen fertilisation on the tomato (Solanum lycopersicum L.) cv. Santa clara and the leafminer (Tuta absoluta (Meyrick), Lepidoptera: Gelechiidae). Greenhouse‐grown tomato plants were fed hydroponically on a complete nutrient solution containing either a high nitrogen concentration (HN) sustaining maximum growth or a low nitrogen concentration (LN) limiting plant growth. Insect‐free plants were compared with plants attacked by T. absoluta. Seven and 14 days after artificial oviposition leading to efficacious hatching and larvae development, we measured total carbon, nitrogen and soluble protein as well as defence compounds (phenolics, glycoalkaloids, polyphenol oxidase activity) in the HN versus LN plants. Only in the HN treatment did T. absoluta infestation slightly impair leaf growth and induce polyphenol oxidase (PPO) activity in the foliage. Neither the concentration of phenolic compounds and proteins nor the distribution of nitrogen within the plant was affected by T. absoluta infestation. In contrast, LN nutrition impaired T. absoluta‐induced PPO activity. It decreased protein and total nitrogen concentration of plant organs and enhanced the accumulation of constitutive phenolics and tomatine. Moreover, LN nutrition impaired T. absoluta development by notably decreasing pupal weight and lengthening the development period from egg to adult. Adjusting the level of nitrogen nutrition may thus be a means of altering the life cycle of T. absoluta. This study provides a comprehensive dataset concerning interrelated responses of tomato plants and T. absoluta to nitrogen nutrition.     

First record of an entomopathogenic fungus of tomato leafminer,Tuta absoluta (Meyrick) in Tanzania

Tuta absoluta (Meyrick) is a destructive pest causing adverse environmental and economic effects globally. Management of T. absoluta by synthetic chemicals is of diminutive achievement due to resistance trick of the pest. Harnessing of biocontrols is currently fascinating and efficient for management of the tremendous pests. This study isolated an entomopathogenic fungus from dead larvae of T. absoluta on tomato leaves from farmer’s field Tanzania. Two Aspergillus fungal isolates were isolated and tested against larvae and adults of T. absoluta under conditions of 19.5°C and 50%RH, 30.4°C and 70% RH, respectively, in the laboratory. Sequence analysis of Aspergillus isolates that we referred as A-Tz1 and A-Tz2 were analysed with the BLAST program available at the National Center for Biological Information (NCBI). The two isolates (A-Tz1 and A-Tz2) showed 100% similarity with Aspergillus oryzae of GeneBank accession numbers EF121337.1 and MG519722.1. Such similarity implied that both A-Tz1 and A-Tz2 isolates were A. oryzae. To confirm pathogenicity, bioassays were conducted against the larvae, pupa and adults of T. absoluta. Results showed that at 1.0?×?10⁸?conidia/mL, fungal isolates caused up to 70% larval mortality 3 days post inoculation and consequently inhibited pupation by 84.5% and adult emergence by 74.4%. The identified A. oryzae reduced the life span of adult T. absoluta up to 5 days post inoculation at 1.0?×?10⁸?conidia/mL whereas in control T. absoluta survived up to 25 days. This study found that isolates of Aspegillus species have the potential to be used as active ingrediennt for the formulation of a new bioinjections for application by farmers to control the pest in fields.     

Virulence of Metarhizium anisopliae and Beauveria bassiana isolates and the effects of fungal infection on the reproduction potential of Rhiphicephalus microplus engorged females

The aim of the study was to assess the virulence of five Metarhizium anisopliae (Ma) and three of Beauveria bassiana (Bb) isolates, and the effect of the fungal infection to the reproduction of engorged females from two colonies of Rhiphicephalus microplus; one colony was collected from naturally infested cattle (Native) and the other one from a laboratory colony (Media Joya). Virulence was evaluated using the immersion technique at a concentration of 1?×?10⁸ conidia/ml; control groups received a water suspension with Tween 80 (0.1%). The Reproductive Efficiency Index ‘REI’ (eggs laid/engorged female weight) and the Reproductive Aptitude Index ‘RAI’ (eggs hatched as larvae/engorged female weight) were calculated for both groups. This experiment shows that two entomopathogenic fungal isolates, Bb115 and Ma136, caused high mortality from 5 days post-treatment (PT), reaching mortality rates of 99–100% at 15 days PT in both R. microplus colonies. The Bb115 isolate caused 98 and 79% reduction in egg oviposition in the field and laboratory colonies, respectively, while the reduction in egg hatchability was 98 and 89% in the field and laboratories colonies, respectively. In the case of Ma136, the egg oviposition was reduced in 73% in the field colony and 64% in laboratory colony, while in the field and laboratory colonies, with a reduction in egg hatchability of 73% and 86%, respectively. These results indicate the potential of Bb115 and Ma136 isolates as possible biological control agents of R. microplus.     

Introduction of Beauveria bassiana as a biological control agent against the Ommatissus lybicus in Kerman province

Dubas bug, Ommatissus lybicus is one of the major sucking pests of date palm. Data palm is the most important economic crop in Iran. In this research, we isolated the pathogenic strain KB 201, KB 206, KB 211, KB 215, KB 220 and KB 512, of entomopathogenic fungi Beauveria bassiana from soil and insects, which produced aerial and submerged conidia and blastospores in the laboratory conditions. We investigated the best conditions for the production and utilisation of the spore suspension to spray the nymph of O. lybicus, which is one of the important pests in Kerman province (Iran). A total of 180 isolates that naturally infected by O. lybicus were collected from date palm in Kerman province during spring and summer seasons 2010–2011. The pathogen city test was carried out with direct spray. To bioassay the isolates, three concentrations of the spore suspension were prepared as follows: 1?×?10⁶, 1?×?10⁷ and 1?×?10⁸?conidia/ml. The pests were sprayed by aerial conidial suspension, which was prepared by 0.01% Tween 80 in distilled water, and the controls were sprayed by 0.01% Tween 80in distilled water. After spraying the pests, the plates were incubated at 25?±?1?°C and 80% of relative humidity. Then, the treated pests were monitored every day for the fungal growth and mortality.     

Bioassay method for assessing the virulence of Beauveria bassiana against tarnished plant bug, Lygus lineolaris (Hem., Miridae)

A standard bioassay method for assessing the pathogenicity of Beauveria bassiana (Balsamo) Vuillemin (GHA strain) against second instar tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hem., Miridae) was developed. Several types of inoculation methods, assay containers and incubation times were tested. Our goal was to minimize control mortality and maximize treatment mortality. Five inoculation methods (immersing broccoli florets or bean pods, spraying broccoli florets or bean pods, and immersing insects) and four types of plastic containers (114‐, 171‐, 228‐ and 455‐ml) were tested. Immersing insects directly in a fungal suspension was the most effective inoculation method, which resulted in a treatment mortality of 70–81.3% at a concentration of 1 × 10⁷ conidia/ml. The 114‐ml plastic container was the most suitable assay container when 10 tarnished plant bug nymphs were treated together, resulting in a control mortality of only 6% 12 days after treatment. Within the first 6 days after treatment, 71.1% of the insects were killed, compared with a total mortality of 81.3% after 12 days. Nymphs infected with the test fungus changed colour from green to black. Mycelial outgrowth and sporulation on the cadavers demonstrated that most nymphs died of fungal infection. A total of 61.1 and 80.5% of the cadavers showed signs of mycelial outgrowth 9 days after death among those that were surface sterilized and those that were not, respectively.     

Efficacy of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against Thaumetopoea pityocampa Shiff. (Lepidoptera: Thaumatopoeidae)

Abstract

In this study, Metarhizium anisopliae TR 106 and Beauveria bassiana TR 217 was tested against fourth instar larvae of Thaumetopoea pityocampa. The LT₅₀ and LT₉₀ of 1?×?10⁶ concentration of M. anisopliae against T. pityocampa were 3.60 and 4.11 for direct application, while these were 2.87 and 3.60?days, respectively in leaves application. The LT₅₀ and LT₉₀ of the 1?×?10⁸ concentration of the same isolate were 2.50 and 2.95?days for direct application, and 2.98 and 3.74?days for leaves application. The LT₅₀ of insect and leaves application for 1?×?10⁶ of B. bassiana were 3.75 and 3.49?days, respectively. The LT₉₀ of same concentration for insect application was 4.48?days, while LT₉₀ for leaves application was 4.63?days. Similarly, LT₅₀ of insect and leaves application for 1?×?10⁸ of B. bassiana were 3.03 and 3.31?days, while LT₉₀ were 3.68 and 4.29?days, respectively. Approximate 100% mycosis was observed in all treatments.     

Introduction of Beauveria bassiana as a biological control agent for Tribolium castaneum in Kerman province

Barley, Hordeum vulgare, one of the important crops in the word, is used in malting, feed and food industries. The red flour beetle, Tribolium castaneum, was found wherever grains or other dried foods are stored. Disinfestations of barley using chemical methods to kill insects, in this research, for the first time we isolated the pathogenic KB512 of entomopathogenic fungi Beauveria bassiana from soil and insects, which produced aerial and submerged conidia and blastospores in laboratory conditions. We investigated the best conditions for the production and utilisation of spore suspension to spray the larvae of T. castaneum, which is one of the important pests in Kerman province (Iran). One hundred and eighty isolates that naturally infected by T. castaneum were reared during spring and summer seasons 2010–2011. The pathogenicity test was carried out with direct spray. To bioassay the isolates, three concentrations of the spore suspension were prepared as follows: 1?×?10⁶ and 1?×?10⁸ conidia/ml. The pests were sprayed by aerial conidial suspension, which was prepared by 0.01% Tween 80 in distilled water, and the controls were sprayed by 0.01% Tween 80 in distilled water. After spraying the pests, the plates were incubated at 25?±?1?°C and 80% of relative humidity. Then, the treated pests were monitored every day for the fungal growth and mortality.     

Interactions between Beauveria bassiana and Trichogramma pretiosum under laboratory conditions

Knowledge about interactions among biological control agents, used in sequence or simultaneously, may help to increase their efficiency, or in the case of conservation biological control, to avoid the impairment of efficiency. The aim of this study was to evaluate the interaction between two isolates of Beauveria bassiana (Bals.) Vuill. (Hypocreales) (Unioeste 47 and Unioeste 57) and Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae), varying the timing of sprays of the fungus. Isolates were sprayed at 10⁹ conidia ml^?1 upon cards (1 × 5 cm; i.e., 2 × 10⁵ conidia mm^?2) with eggs of Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) and the control group was sprayed with sterile distilled water with Tween^® 80 (0.01%). First, the percentage of parasitized eggs was assessed in a choice test: females of T. pretiosum were confined with two cards, one of them treated with either of the fungus isolates, the other a water‐treated control. Second, parasitism was compared on cards sprayed at various time intervals pre‐ vs. post‐parasitism: the number of parasitized eggs was assessed, as well as the parasitoid's emergence percentage, egg‐adult period, offspring sex ratio, and the longevity of parasitizing females. Trichogramma pretiosum preferred to parasitize control eggs (65 and 72%) and parasitization of the treated eggs did not differ between fungus isolates: 35% (Unioeste 47) vs. 28% (Unioeste 57). Spraying before or after parasitizing did not affect the number of parasitized eggs, nor offspring sex ratio; however, both treatments affected the percentage of emerged adults, egg‐adult period, and longevity. These findings suggested that isolates did not interfere in the longevity of parasitizing females. When the fungus was sprayed on the various development stages of the parasitoid within the host egg, or when it was offered to the parasitoid at different times after spraying, differences in adults' longevity, parasitism, emergency period from egg to adult, and sex ratio were observed. Beauveria bassiana may be used in conjunction with this parasitoid in a program of pest control.     

Lethal effects of Gliocladium virens,Beauveria bassiana and Metarhizium anisopliae on the malaria vector Anopheles albimanus (Diptera: Culicidae)

Control of Anopheles albimanus, the main vector of malaria on the coast of the State of Chiapas, is based mainly on application of chemical insecticides, which has resulted in resistance to most registered insecticides. Strategies for biological control may provide sustainable alternatives. We report on the lethal effects of a native isolate of Gliocladium virens on An. albimanus larvae and adults, compared to that of strains of Beauveria bassiana and Metarhizium anisopliae. Conidial suspensions of G. virens, B. bassiana and M. anisopliae cultured on Sabouraud agar were tested in bioassays with An. albimanus larvae and adults. Mosquito larvae were more susceptible to all fungi, compared to adults. On early and late instar larvae, M. anisopliae showed the most pathogenic effect (LC₅₀ of 1.4×10⁵ conidia/mL in early instars; 1.1×10⁵ conidia/mL in late instars), followed by G. virens (LC₅₀ of 3.3×10⁵ conidia/mL in early instars and 3.5×10⁶ conidia/mL in late instars). Metarhizium anisopliae sensu lato and the native G. virens could be considered good choices for An. albimanus control in southern Mexico.     

Spatial and temporal variation in the control failure likelihood of the tomato leaf miner,Tuta absoluta

Insecticide resistance is frequently associated with field control failure, but such an assessment of its likelihood seldom occurs. This phenomenon is a potential cause of the control failure of the tomato leaf miner Tuta absoluta (Lepidoptera: Gelechiidae); therefore we surveyed insecticide resistance as the control failure likelihood for a duration of 7 years in 20 field populations of this species. The insecticide label rate was used as the discriminating concentration, and the minimum efficacy threshold required for insecticides in Brazil (i.e. 80% efficacy) was the targeted efficacy. The spatial and temporal variations of the control failure likelihood were also assessed, as was the potential influence of land topography for the area‐wide pattern observed. Most populations of T. absoluta were susceptible to abamectin, chlorfenapyr and spinosad and not to bifenthrin, triflumuron and teflubenzuron. The indoxacarb susceptibility varied in space and time. Spatial dependence was observed for abamectin and indoxacarb. The control failure likelihood of T. absoluta was higher for bifenthrin, triflumuron and teflubenzuron. A flat topography and the wind direction may favour the spread of the control failure likelihood and should be considered in designing pest management programs for T. absoluta.     

Life table parameters of the tomato leaf miner Tuta absoluta (Lepidoptera: Gelechiidae) on different tomato cultivars

The tomato leaf miner Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating pest of tomato plants originating from South America. In this study, the effect of three tomato cultivars on the life table parameters of T. absoluta was investigated. Data were analysed based on age‐stage, two‐sex life table theory. The net reproduction rate (R₀) of T. absoluta on Falkato, Isabella and Grandella cultivars was determined as 15.73 ± 3.35, 17.66 ± 3.33 and 10.03 ± 4.10 eggs, respectively. The intrinsic rate of increase (r) value on those cultivars was 0.09 ± 0.007, 0.095 ± 0.006 and 0.074 ± 0.017 day^?1, respectively. Finite population rates of increase (λ) were 1.095 ± 0.008, 1.099 ± 0.007 and 1.076 ± 0.018 day^?1, and mean generation times (T) on the three cultivars were 30.09 ± 0.29, 30.22 ± 0.27 and 31.12 ± 0.65 days, respectively. The egg incubation period, pre‐adult duration and fecundity on these tomato cultivars was also calculated. The results showed no significant differences between r, R₀, and T on the three tested tomato cultivars. These findings can be useful for developing an integrated pest management strategy against this noxious pest.     

Insecticide toxicity and walking response of three pirate bug predators of the tomato leaf miner Tuta absoluta

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Organophosphate and pyrethroid resistances in the tomato leaf miner Tuta absoluta (Lepidoptera: Gelechiidae) from Iran

The tomato leafminer, Tuta absoluta (Meyrich) (Lepidoptera: Gelechiidae), is a serious pest of tomato crops worldwide. The intensive use of chemical pesticides to control it has led to the selection of resistant populations. This study investigated the resistance of T. absoluta populations to pyrethroid and the organophosphate insecticides from ten regions of Iran. The resistance ratios at LC₅₀ for chlorpyrifos and diazinon varied among populations from 4.3 to 12 and from 1.4 to 9.0, respectively. The resistance ratios of the pyrethroids cypermethrin, deltamethrin and permethrin varied from 1.3 to 3.7, 2.7 to 13 and 1.2 to 4.3, respectively. Inclusion of synergists in toxicological bioassays and the variation observed in the activity of esterases, glutathione S‐transferase and cytochrome P450‐dependent monooxygenase suggest the existence of metabolically based resistance. Esterase and P450 biochemical assays were positively correlated with deltamethrin, and cypermethrin tolerance and diazinon tolerance correlated with esterase activity. The genes encoding the organophosphate and pyrethroid target sites acetylcholinesterase (ace1) and sodium channel (kdr) were partly sequenced. The genotyping revealed mutations in high frequencies in all populations leading to an A201S substitution in ace1 and three substitutions in the sodium channel gene L1014F, M918T, T929I. In summary, our results indicate the presence of organophosphate and pyrethroid resistance in Iranian T. absoluta populations with involvement of both detoxification enzymes and target site alterations. Most likely the populations of T. absoluta imported to Iran were resistant upon arrival.     

Suitability of the tomato borer Tuta absoluta as prey for Macrolophus pygmaeus and Nesidiocoris tenuis

The tomato borer, Tuta absoluta (Meyrick) (Lep.: Gelechiidae), is an important tomato pest native to South America, which appeared in eastern Spain at the end of 2006. Prey suitability of T. absoluta eggs and larval instars was examined under laboratory conditions to evaluate whether two indigenous predators, Macrolophus pygmaeus (Rambur) and Nesidiocoris tenuis Reuter (Hem.: Miridae), can adapt to this invasive pest. Both predators preyed actively on T. absoluta eggs and all larval stages, although they preferred first‐instar larvae. Our results demonstrate that both mirids can adapt to this invasive pest, contributing to their value as biological control agents in tomato crops.     

Isolation and selection of virulent isolates of Beauveria bassiana for biological control of Ommatissus lybicus in Kerman province

Date palm (Phoenix dactylifera) is the most important economic crop in Kerman province. The palm date leafhopper (Ommatissus lybicus) is one of the most important and key pests that causes high level damage to date crop. Date is an organic fruit in this area, and biological control has been introduced as the best long-term strategy in this region. To investigate appropriate biological control agents in Kerman province during 2005–2007, 178 isolates were collected from naturally infected O. lybicus for the first time recorded in the world, and Beauveria bassiana was introduced as a suitable biological control agent for control of this pest.     

Soil application of Beauveria bassiana GHA against apple sawfly,Hoplocampa testudinea (Hymenoptera: Tenthredinidae): Field mortality and fungal persistence

Low impact alternatives to synthetic insecticides for the control of apple sawfly (Hoplocampa testudinea Klug) are scarce encumbering pest management in organic apple orchards. We investigated the soil persistence and field efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (BotaniGard) against apple sawfly under common organic orchard practices. We also assessed the efficacy of B. bassiana GHA and Metarhizium brunneum Petch (indigenous strain) against sawfly in the laboratory. Larvae treated with either fungus in the laboratory died faster than control larvae and displayed 49.4%–68.4% mycosis. In the field, B. bassiana density remained high in the week after application, during larval descent to the soil. Fungal density decreased to 25% at 49 d after application and to 0.4% after 55 weeks. Molecular markers revealed that the majority of fungal isolates recovered comprised the applied B. bassiana strain GHA. Larvae pupating in soil cages in the orchard for 49 d displayed 17% mycosis. The high efficacy under laboratory conditions was not seen in the field. B. bassiana application resulted in densities above the upper natural background level during the growing season, but reversion to background levels occurred within a year. It remains to be investigated whether this has a detrimental effect on nontarget organisms. Additional work is needed to bridge the knowledge gap between laboratory and field efficacy in orchards.     

Different host plant species modifies the susceptibility of Bactericera cockerelli to the entomopathogenic fungus Beauveria bassiana

Bactericera cockerelli (Sulc.) is a serious pest of solanaceous crops and a vector of the plant pathogen Candidatus Liberibacter psyllaurous. Entomopathogenic fungi are the most important biological control alternatives for this pest. Host plant species, however, can modify the outcomes of insect–pathogen interactions. We conducted laboratory experiments to quantify the virulence of two isolates of the entomopathogenic fungus Beauveria bassiana (Bals. [Vuill.]), BB40 and BB42, against third instar B. cockerelli nymphs maintained on chilli pepper plants. Owing to the lack of difference in virulence against B. cockerelli nymphs on chilli pepper between the two B. bassiana isolates, only BB42 was used to: compare virulence against nymphs maintained on either chilli pepper, potato or tomato; and in vivo conidia production from nymphs maintained on different host plants. Virulence of the two B. bassiana isolates against B. cockerelli nymphs was similar. Bactericera cockerelli nymphs maintained on tomato were more susceptible to B. bassiana than nymphs maintained on potato or chilli peppers. Infected nymphs maintained on chilli peppers produced the greatest number of conidia followed by infected nymphs maintained on tomato and potato. Host plant affected the susceptibility of B. cockerelli to B. bassiana isolate BB42 and subsequent conidia production. The implications of our results for microbial control of B. cockerelli by B. bassiana are discussed.     

Susceptibility of apple clearwing moth larvae,Synanthedon myopaeformis (Lepidoptera: Sesiidae) to Beauveria bassiana and Metarhizium brunneum

Apple clearwing moth larvae, Synanthedon myopaeformis (Lepidoptera: Sesiidae) were found to be susceptible to infection by two entomopathogenic fungi: an indigenous fungus isolated from S. myopaeformis cadavers and identified as Metarhizium brunneum (Petch); and Beauveria bassiana isolate GHA. In laboratory bioassays, larvae exhibited dose related mortality after exposure to both the M. brunneum and Beauveria bassiana with 7 day LC₅₀'s of 2.9×10⁵ and 3.4×10⁵ spores/mL, respectively. Larval mortalities caused by the two isolates at 1×10⁶ spores/mL were not significantly different and 73% of the M. brunneum-treated, and 76% of the B. bassiana-treated larvae were dead 7 days post treatment, with LT₅₀'s of 5.5 and 5.1 days, respectively.