Compatibility of the entomopathogenic fungus Beauveria bassiana with flufenoxuron and azadirachtin against Tetranychus urticae

Laboratory studies were developed to evaluate the compatibility of flufenoxuron and azadirachtin with Beauveria bassiana against Tetranychus urticae larvae along with the required Probit analysis of the involved chemicals on all of the life stages of this mite. Flufenoxuron displayed parallel regression lines for the mortality of eggs, deutonymphs and adults. Larvae and protonymphs were the most susceptible life stages. Protonymphs were 35 times more sensitive than eggs and adults. Azadirachtin gave equal mortality on proto- and deutonymphs. The response of eggs and adults was equivalent when treated with azadirachtin. The regression lines for proto- and deutonymphs were parallel to those of adults and eggs yet three times more sensitive. The effects of separate combinations of the entomopathogenic fungus Beauveria bassiana at its LC₂₀ with flufenoxuron and azadirachtin at their corresponding LC₄₀ were evaluated on mite larvae. The application of flufenoxuron with B. bassiana revealed a clear synergy. While the combination of azadirachtin and B. bassiana had an additive effect. These combinations with B. bassiana could improve mite control by contributing to a decline in the likelihood of resistance so often described in the literature.     

Demography of the twospotted spider mite,Tetranychus urticae Koch

Summary A simple life table model was constructed for Tetranychus urticae in which daily survivorship of eggs and motil stages, fecundity, and development time was altered to assess the impact of each parameter on the intrinsic rate of increase. r. Interpretation of the trade-offs focused on management considerations.A second aspect of the study concerned age and stage structure in mite populations including the time path of convergence to a stable age distribution and the effect of changes in birth and death rates on the age profile. The stable stage distributions of 7 tetranychid mite species were computed using 25 separate life tables. In spite of the wide range of r-values induced by different experimental conditions, all of the stage distributions were quite similar averaging roughly 66% eggs, 26% immatures, and 8% adults. Several population studies were cited which reported stage distributions of growing mite populations. The empirical evidence suggested that natural mite populations are often quite near this stable distribution.A practical problem involving the extent to which hormoligosis (insecticide stimulation) affects mite population growth rate was addressed using the life table model and laboratory data from controlled studies. The findings suggested that mite populations treated with insecticide may attain a 1.4- to a 4.2-fold difference in population size relative to an untreated population after 2 generations and over a 1,300-fold potential difference after 10 generations.     

Behavioral aspects of dicofol resistance in the twospotted spider mite Tetranychus urticae

Behavioral analysis of near-isogenic dicofol-resistant and dicofol-susceptible spider mites did not uncover behavior patterns, present in resistant individuals yet absent in susceptible individuals, that would have reduced exposure to dicofol. On the contrary, homozygous resistant individuals were more likely than homozygous susceptible individuals to remain in contact with treated leaf areas. In the case of dicofol resistance in twospotted spider mites, resistance appears to have resulted in the loss, rather than the acquisition, of behavioral traits that lessen exposure to pesticide. Therefore, we conclude that behavioral resistance does not play a major role in twospotted spider mite resistance to dicofol.Deux génotypes de T. urticae Koch (un sensible et un résistant) ont servi à étudier les aspects comportementaux de la résistance aux pesticides: la durée et la fréquence des comportements de nutrition, de marche et d'attente ont été mesurés chez cet acarien. Dans toutes les expériences, des femelles de mêmes âges avaient été placées sur des feuilles jeunes de haricot (Phaseolus vulgaris) portant des résidus de dicofol.3 séries d'expériences ont été réalisées pour examiner:

Resistance status of the carmine spider mite,Tetranychus cinnabarinus and the twospotted spider mite,Tetranychus urticae to selected acaricides on strawberries

The carmine spider mite, Tetranychus cinnabarinus (Boisduval) and the twospotted spider mite, Tetranychus urticae Koch, are serious pests of strawberries and many other horticultural crops. Control of these pests has been heavily dependent upon chemical acaricides. Objectives of this study were to determine the resistance status of these two pest species to commonly used acaricides on strawberries in a year‐round intensive horticultural production region. LC₉₀ of abamectin for adult carmine spider mites was 4% whereas that for adult twospotted spider mites was 24% of the top label rate. LC_90s of spiromesifen, etoxazole, hexythiazox and bifenazate were 0.5%, 0.5%, 1.4% and 83% of their respective highest label rates for carmine spider mite eggs, 0.7%, 2.7%, 12.1% and 347% of their respective highest label rates for the nymphs. LC_90s of spiromesifen, etoxazole, hexythiazox and bifenazate were 4.6%, 11.1%, 310% and 62% of their respective highest label rates for twospotted spider mite eggs, 3%, 13%, 432,214% and 15% of their respective highest label rates for the nymphs. Our results suggest that T. cinnabarinus have developed resistance to bifenazate and that the T. urticae have developed resistance to hexythiazox. These results strongly emphasize the need to develop resistance management strategies in the region.     

Effects of triflumuron on the two-spotted spider mite,Tetranychus urticae (Acari: Tetranichydae)

The effects of triflumuron on the mortality, fecundity, and fertility of the two-spotted spider mite, Tetranychus urticae, were evaluated in the laboratory. No differences in toxicity for larvae, protonymphs and deutonymphs were observed, but immature stages were 3.8-times more susceptible than adults at the LC₅₀. The compound exhibited a direct contact ovicidal activity influenced by eggs age. 48–72 hold eggs were significantly more sensitive than eggs of the other age classes. No hatch inhibition was observed in eggs laid by treated adult females using a sublethal dose at two physiological times (<12 and 48–72h old). However, fecundity decreased in younger treated females, but it increased in the older-ones. The toxicity for immatures and eggs, and the sublethal effects described suggest that triflumuron could be an interesting incorporation in integrated pest programs of T. urticae. This revised version was published online in July 2006 with corrections to the Cover Date.     

Sublethal effects of Beauveria bassiana on life table parameters of two–spotted spider mite,Tetranychus urticae (Acari: Tetranychidae)

Effects of the entomopathogenic fungus Beauveria bassiana were studied on life table parameters of two-spotted spider mite, Tetranychus urticae feeding on bean and cucumber under laboratory conditions. The developmental periods for all immature stages were not affected by fungal infection on each host plant but the duration of larval stage was significantly longer on bean. The female and male longevity, oviposition period and fecundity were significantly lower on fungus-treated mites but were not different between two host plants. Significant reductions were found on the intrinsic rate of increase (r _m ), the net reproductive rate (R ₀), the finite rate of increase (λ), the mean generation time (T _c ) and the population doubling time (D _t ) as a result of mycosis. Only the mean generation time (T _c ) was influenced considering the effect of host plant, which was shorter on cucumber.     

Comparative toxicity of some acaricides to the predatory mite, Phytoseiulus persimilis and the twospotted spider mite, Tetranychus urticae

The relative toxicity of someacaricides to the predatory mite, Phytoseiulus persimilis and the twospottedspider mite, Tetranychus urticae (Acari: Phytoseiidae, Tetranychidae) wasevaluated in laboratory. Five of theacaricides tested, including bifenazate,acequinocyl, chlorfenapyr, flufenoxuron andfenbutatin oxide, were much less toxic to adultfemales and immatures of P. persimilisthan to those of T. urticae, and adultfemale predators treated with these fiveacaricides produced 84±96% as many eggs as didcontrol females. Etoxazole did not seriouslyaffect the survival and reproduction of adultfemale predators but caused high mortalityrates in eggs and larvae of P.persimilis. Milbemectin and fenazaquin werevery toxic to adult females and immatures ofP. persimilis. Adult female predatorssurvived on a diet of spider mites treated withbifenazate, acequinocyl, chlorfenapyr,flufenoxuron and fenbutatin oxide, and theirfecundity, prey consumption and the sex ratioof the progeny were not substantially affected. Based on the results, bifenazate, acequinocyl,chlorfenapyr, flufenoxuron and fenbutatin oxideappeared to be the promising candidates for usein integrated mite management programs whereP. persimilis is the major naturalenemy.     

Role of the entomogenous fungus, Neozygites floridana, in population declines of the twospotted spider mite, Tetranychus urticae, on field corn

In field plots in which populations of predaceous species were suppressed using carbaryl, populations of the twospotted spider mite, Tetranychus urticae Koch, on field corn declined in association with aerial dispersal of the mites or with epizootics of the entomogenous fungus, Neozygites floridana Weiser & Muma. The relative importance of these two factors varied from year to year depending on environmental conditions. When moist weather conditions induced epizootics of N. floridana before corn plants became heavily infested with mites, the pathogen appeared to be the major factor causing mite population declines. When dry weather conditions allowed mite populations to expand unchecked until corn plants became entirely infested, mite aerial dispersal was the major factor associated with mite population declines. In every field in which there was an epizootic of N. floridana, routine applications of the fungicide maneb delayed epizootics and reduced or delayed the impact of the pathogen on mite populations. The total number of Neozygites-infected mites produced in maneb-treated plots throughout the test period was significantly greater than that in untreated plots when epizootics were initiated during periods of mite population growth. Fewer total infected mites were produced in maneb-treated plots compared to untreated plots when epizootics were initiated during periods of mite population decline.

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Résumé Dans des parcelles de maïs dont les populations d'espèces prédatrices avaient été éliminées par du carbaryl, des populations de T. urticae ont régressé en fonction de la dispersion aérienne des acariens ou d'épizooties provoquées par N. floridana. L'importance relative de ces deux causes varie d'année suivant les conditions écologiques. Le pathogène paraissait la principale cause de déclin des populations quand le temps humide a induit les épizooties par N. floridana avant que le maïs n'ait été largement contaminé par les acariens. Quand le temps sec a permis aux populations d'acariens de se développer sans frein jusqu'à contamination totale du maïs, la dispersion aérienne des acariens était la principale cause de déclin des populations d'acariens. Dans tous les champs où il y avait eu épizootie de N. floridana, des traitements de routine avec le fongicide meneb ont retardé les épizooties et réduit ou retardé l'impact du pathogène sur les populations d'acariens. Pendant toute la période d'étude, le nombre total d'acariens infestés par Neozygites était significativement plus élevé dans les parcelles traitées au maneb que dans les parcelles non traitées, lorsque les épizooties étaient provoquées pendant la période de croissance des populations d'acariens. Quand les épizooties ont été déclenchées pendant la période de déclin des populations d'acariens, un nombre plus faible d'acariens infestés était observé dans les parcelles traitées au maneb que dans les parcelles sans traitement.

     

Effects of cultivar and irrigation management on population growth of the twospotted spider mite Tetranychus urticae on greenhouse ivy geranium

Greenhouse experiments were conducted to determine whether different ivy geranium cultivars and different irrigation frequencies influenced population growth of the twospotted spider mite (TSM), Tetranychus urticae. Results showed that mite numbers at harvest (about 4 weeks after plants were artificially infested) were very low on 'Sybil Holmes', but significantly higher on 'Amethyst 96'. Irrigation frequency of ivy geraniums had no measurable effect on TSM populations over a range in which water was applied when pot weight dropped 15, 30 or 45% from container capacity. However, the most frequently irrigated plants (15% treatment) produced as much or more growth than plants that received less water. These plants also had the lowest incidence of oedema. There was no significant interaction between water management and host plant resistance, implying the difference in host plant resistance to TSM between the two cultivars should remain constant, at least within the range of the irrigation frequencies studied. Our findings suggest that growers can take advantage of plant resistance to TSM on ivy geranium cultivars of current commercial importance, and that watering practices can be adjusted to optimize plant production without affecting pest populations.     

Side effects of the entomopathogenic fungus Beauveria bassiana on the predator Coccinella undecimpunctata

The effect on predators of the biopesticide based on Beauveria bassiana spores was studied. A high concentration of the pathogen showed adverse effects on the predator (Coccinella undecimpunctata) and its biological aspects: 2, 4 and 8 g/L eliminated egg deposition in treated females. Spores at 4 g/L affected significantly the longevity of adults, duration of larvae and pupal period.     

Postharvest disinfestation of diapausing and non-diapausing twospotted spider mite (Tetranychus urticae) on persimmons: hot water immersion and coolstorage

The mortality response of diapausing and non-diapausing twospotted spider mite (Tetranychus urticae Koch) on persimmons to hot water immersion treatments between 44 and 54 °C was examined, for potential as a quarantine treatment. The mean immersion time for mean 99% mortality (LT₉₉) of diapausing mites at 44 °C was 211 min, and this time decreased with increasing temperature to 3.6 min at 54 °C. Non-diapausing mites were found to be less tolerant to temperatures below 48 °C, with an estimated LT₉₉ of 102 min at 44 °C, but had similar thermotolerance above 48 °C. In 47 °C water the immersion time required to kill 99% of diapausing mites was estimated at 67 min. This time was not reduced by subsequent coolstorage at 0 °C for up to eight weeks. Rather, coolstorage had the effect of keeping mites alive, relative to LT₉₉ estimates calculated for mites stored at 20 °C. Similarly the thermotolerance of mites did not change with increased time in diapause, even though mites in diapause for 12 weeks had high control mortality. Hot water immersion appears to be a potentially useful disinfestation method for persimmons.     

Light stimulates conidiation of the entomopathogenic fungus Beauveria bassiana

Beauveria bassiana is a commercially important entomopathogenic fungus. Like other insect fungal pathogens, B. bassiana usually produces asexual reproductive bodies, conidia, for dispersal, transmission and infection of insects. Adequate mass-production of high quality conidia is crucial to development of an efficient B. bassiana insecticide. However, little is known about details of conidiation in this fungus in response to environmental signals, which limits understanding of the mechanism of conidiation and improvement in conidia production. Here, morphologenetic changes of B. bassiana under different light conditions are reported. When cultured in total darkness, B. bassiana hyphae can grow continuously with few reproductive structures differentiated, while illumination with white light resulted in prolific formation of conidiophores bearing abundant conidia, indicating that light could stimulate conidiation of B. bassiana. Among the single colour lights tested, blue light was the most effective to stimulating sporulation. Colonies became adapted for blue light stimulus only after hyphae had grown in total darkness for at least 96 h, whereas the photoadaptation obviously declined after 144 h. For the exposure time, 3 min of blue light pulse was enough to stimulate conidiation in the photoadapted mycelia, while prolonged light exposure over 3 min resulted in a decrease in conidia yield. Our results provided useful clues for understanding the mechanism of conidiation mediated by light in B. bassiana.     

Ultrahistology of oogenesis and vitellogenesis in the spider mite Tetranychus urticae

Histology of the ovary of the spider mite Tetranychus urticae is described light and electron microscopically with special reference to oogenesis and vitellogenesis of this mite. Morphology of the ovary is comparable to the typical sac-like chelicerate ovary with oocytes protruding from the ovarian surface, thus resulting in a grape-like appearance. According to different oogenetic stages, a germ, pre-vitellogenic and vitellogenic region can be observed. Oogonia and primary oocytes characterized by extranuclear material or 'yolk nuclei' are situated in the germ region. Primary oocytes develop into three-nucleated nurse cells situated in the periphery of the pre-vitellogenic region, and into pre-vitellogenic oocytes protruding from the ovarian surface. Growth of oocytes is performed while they are in ovarian pouches by uptake of nurse cell cytoplasm and following extraovarian yolk precursors. Intraoocyte yolk synthesis interpreted from altered cytoplasmic organelles also occurs. Processes taking place during oogenesis and vitellogenesis in T. urticae are compared to published information on yolk synthesis of other animal species.     

Fertilization in the two-spotted spider mite (Tetranychus urticae: Acari)

Different aspects of the fertilization of the two-spotted spider mite (Tetranychus urticae Koch) were studied. It appeared that after the mating act of virgin females, haploid eggs were produced first before the appearance of diploid eggs. From this and other facts, it was concluded that fertilization of the oocytes occurs in a very early stage of egg development. The fertilization is assumed to take place in the ovary and not in the oviduct.By using genetic markers, it could be proved that in most cases the first mating is effective. Later matings usually are ineffective. It is suggested that the sperm supply in the first mating determines the success of later matings.

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Zusammenfassung Die Befruchtung der Bohnenspinnmilbe wurde unter verschiedenen Gesichtspunkten untersucht. Nach der Begattung jungfräulicher Weibchen wurden zunächst haploide Eier abgelegt, bevor diploide Eier produziert wurden. Daraus und aus anderen Grüunden wird geschlossen, daß die Befruchtung der Oocyten in einem sehr frühen Stadium der Entwicklung erfolgt. Die Befruchtung findet vermutlicht im Ovar und nicht im Ovidukt statt.Unter Verwendung genetischer Markierung wurde nachgewiesen, daß in den meisten Fällen die erste Begattung ausschlaggebend ist. Spätere Kopulationen bleiben gewöhnlich wirkungslos. Es wird vermutet, daß die Spermafüllung bei der ersten Begattung den Erfolg späterer Begattungen bestimmt.

     

Development of the entomopathogenic fungus Beauveria bassiana in liquid cultures

Growth and development of B. bassiana was followed in four liquid media: peptone, peptone-glucose, glucose and glucose-peptone-yeast extract. Six developmental stages were defined: (I) the unswollen conidium, (II) the swollen conidium, (III) emergence of the germ tube, (IV) elongation of the germ tube and formation of the first septum, (V) polar and bipolar elongation (growth) of the resulting mycelium and initiation of a blastospore and, (VI) seccession of that blastospore. Conidia of B. bassiana produced germ tubes in all liquid media. Blastospores were produced in all liquid media except glucose. In peptone-glucose, the yield of blastospores was four-fold higher than in glucose-peptone-yeast extract. However, biomass production was highest in peptone-glucose-yeast extract.     

Volatile organic compounds released by the entomopathogenic fungus Beauveria bassiana

The composition of volatile organic compounds (VOC) released by the entomopathogenic fungus Beauveria bassiana (Hyphomycete: Deuteromycotina) utilizing two different carbon sources was investigated. Analyses were performed by solid-phase microextraction (SPME) coupled to capillary gas chromatography (CGC) and CGC-mass spectrometry (MS). Major components in glucose-grown cultures were diisopropyl naphthalenes, ethanol, and sesquiterpenes. Alkane-grown fungal VOC switched to a fingerprint with prevalence of n-decane. This is the first report on the volatiles released by entomopathogenic fungi.     

Agrobacterium tumefaciens-mediated genetic transformation of the entomopathogenic fungus Beauveria bassiana

Agrobacterium tumefaciens-mediated transformation (agro-transformation) was successfully applied to the entomogenous fungus Beauveria bassiana. Conidia of B. bassiana were transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selective trait, under the control of a heterologous fungal promoter and the Aspergillus nidulans trpC terminator. The efficiency of transformation was up to 28 and 96 transformants per 10(4) and 10(5) target conidia, respectively, using three distinct vectors. High mitotic stability of the transformants (80-100%) was demonstrated after five successive transfers on a nonselective medium. Abortive transformants were observed for all the hph(r) vectors used. Putative transformants were analysed for the presence of the hph gene by PCR and Southern analysis. The latter analysis revealed the integration of two or more copies of the hph gene in the genome. The agro-transformation method was found to be effective for the isolation of B. bassiana hygromycin resistant transformants and may represent a useful tool for insertional mutagenesis studies in this fungus.