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Developing SCAR markers to study predation on Trialeurodes vaporariorum.

DNA markers of Trialeurodes vaporariorum were developed to detect remains of these whitefly in the gut of the predator Dicyphus tamaninii. A 2400-bp DNA fragment of T. vaporariorum, absent in other closely related prey species and in the predator banding pattern, was identified by random amplified polymorphic DNA (RAPD) analysis. After cloning and sequencing this fragment, two pairs of sequence-characterized amplified region (SCAR) primers were developed, amplifying single bands of 2100 bp and 310 bp, respectively. Detection of T. vaporariorum DNA in the predator gut was only possible using the primers that amplified the shortest fragment. Specificity tests performed with this pair of primers showed the presence of the 310-bp band for T. vaporariorum in all stages.

Animals↗

The molecular elasticity of the insect flight muscle proteins projectin and kettin.

Projectin and kettin are titin-like proteins mainly responsible for the high passive stiffness of insect indirect flight muscles, which is needed to generate oscillatory work during flight. Here we report the mechanical properties of kettin and projectin by single-molecule force spectroscopy. Force-extension and force-clamp curves obtained from Lethocerus projectin and Drosophila recombinant projectin or kettin fragments revealed that fibronectin type III domains in projectin are mechanically weaker (unfolding force, F(u) approximately 50-150 pN) than Ig-domains (F(u) approximately 150-250 pN). Among Ig domains in Sls/kettin, the domains near the N terminus are less stable than those near the C terminus. Projectin domains refolded very fast [85% at 15 s(-1) (25 degrees C)] and even under high forces (15-30 pN). Temperature affected the unfolding forces with a Q(10) of 1.3, whereas the refolding speed had a Q(10) of 2-3, probably reflecting the cooperative nature of the folding mechanism. High bending rigidities of projectin and kettin indicated that straightening the proteins requires low forces. Our results suggest that titin-like proteins in indirect flight muscles could function according to a folding-based-spring mechanism.

Animals↗

The pupal specifier broad directs progressive morphogenesis in a direct-developing insect.

A key regulatory gene in metamorphosing (holometabolous) insect life histories is the transcription factor broad (br), which specifies pupal development. To determine the role of br in a direct-developing (hemimetabolous) insect that lacks a pupal stage, we cloned br from the milkweed bug, Oncopeltus fasciatus (Of'br). We find that, unlike metamorphosing insects, in which br expression is restricted to the larval-pupal transition, Of'br mRNA is expressed during embryonic development and is maintained at each nymphal molt but then disappears at the molt to the adult. Induction of a supernumerary nymphal stage with a juvenile hormone (JH) mimic prevented the disappearance of br mRNA. In contrast, induction of a precocious adult molt by application of precocene II to third-stage nymphs caused a loss of br mRNA at the precocious adult molt. Thus, JH is necessary to maintain br expression during the nymphal stages. Injection of Of'br dsRNA into either early third- or fourth-stage nymphs caused a repetition of stage-specific pigmentation patterns and prevented the normal anisometric growth of the wing pads without affecting isometric growth or molting. Therefore, br is necessary for the mutable (heteromorphic) changes that occur during hemimetabolous development. Our results suggest that metamorphosis in insects arose as expression of br, which conveys competence for change, became restricted to one postembryonic instar. After this shift in br expression, the progressive changes that occur within the nymphal series in basal insects became compressed to the one short period of morphogenesis seen in the larva-to-pupa transition of holometabolous insects.

Animals↗

Binding mode of the first aminoacyl-tRNA in translation initiation mediated by Plautia stali intestine virus internal ribosome entry site.

Eukaryotic ribosomes directly bind to the intergenic region-internal ribosome entry site (IGR-IRES) of Plautia stali intestine virus (PSIV) and initiate translation without either initiation factors or initiator Met-tRNA. We have investigated the mode of binding of the first aminoacyl-tRNA in translation initiation mediated by the IGR-IRES. Binding ability of aminoacyl-tRNA to the first codon within the IGR-IRES/80 S ribosome complex was very low in the presence of eukaryotic elongation factor 1A (eEF1A) alone but markedly enhanced by the translocase eEF2. Moreover, eEF2-dependent GTPase activity of the IRES/80 S ribosome complex was 3-fold higher than that of the free 80 S ribosome. This activation was suppressed by addition of the antibiotics pactamycin and hygromycin B, which are inhibitors of translocation. The results suggest that translocation by the action of eEF2 is essential for stable tRNA binding to the first codon of the PSIV-IRES in the ribosome. Chemical probing analysis showed that IRES binding causes a conformational change in helix 18 of 18 S rRNA at the A site such that IRES destabilizes the conserved pseudoknot within the helix. This conformational change caused by the PSIV-IRES may be responsible for the activation of eEF2 action and stimulation of the first tRNA binding to the P site without initiation factors.

Animals↗

Biological characteristics of the mirids Macrolophus costalis and Macrolophus pygmaeus preying on the tobacco form of Myzus persicae (Hemiptera: Aphididae).

Biological and population parameters of the predatory bugs Macrolophus costalis Fieber and Macrolophus pygmaeus Rambur preying on the tobacco aphid Myzus persicae (Sulzer) were examined. Tobacco was used as host plant and all experiments were carried out at 23 degrees C and L16:D8. In M. costalis, the developmental time for eggs and the total duration of nymphal instars was 13.1 and 22.2 days and in M. pygmaeus 13.0 and 20.2 days, respectively. Both species completed their nymphal development feeding only on plant juices although they required significantly more time to attain adulthood than when feeding on prey. The longevity of M. costalis females and the preoviposition period were 49.9 and 5.6 days and those of M. pygmaeus were 50.3 and 6.3 days, respectively. Adult males showed a higher longevity than females in both species. The intrinsic rate of increase and the mean total number of eggs laid were 0.0644 and 121 in M. costalis and 0.0615 and 104 in M. pygmaeus, respectively. Mean total aphid consumption by nymphs of M. costalis was 61 aphids, about twice that of M. pygmaeus (37 aphids). Total aphid consumption by a male and female pair of M. costalis and M. pygmaeus until the death of the female was 244 and 285 aphids, respectively. The results of the study are discussed in relation to the impact of the two predatory bugs on tobacco aphid populations.

Animals↗

Genetic differentiation of geographically separated populations of the southern green stink bug Nezara viridula (Hemiptera: Pentatomidae).

Genetic variation in the southern green stink bug Nezara viridula (Linnaeus) from 11 geographically separated sampling locations (Slovenia, France, Greece, Italy, Madeira, Japan, Guadeloupe, Galapagos, California, Brazil and Botswana) was studied by sequencing 16S and 28S rDNA, cytochrome b and cytochrome c oxidase subunit I gene fragments and random amplified polymorphic DNA (RAPD) analysis. Sequencing revealed 11 distinct haplotypes clustering into lineages A, B and C. Lineage C was characteristic for a single analysed specimen from Botswana. Lineage B was detected in Japan, and it probably arose in Asia. Haplotypes of European and American specimens belonged to lineage A; specimens from France, Slovenia, Madeira and Brazil shared highly similar haplotypes (>99%) from subgroup A1, while all the specimens from Greece, California, Galapagos and Guadeloupe shared a haplotype from subgroup A2. RAPD data were more variable but consistent with mtDNA sequences, revealing the same clustering. They separated the Botswanian specimen from Japanese specimens and from a group of more closely related specimens from Europe and America. Sequence and RAPD results both support the African origin of N. viridula, followed by dispersal to Asia (lineage B) and, more recently, by expansion to Europe and America (lineage A). RAPD analysis revealed two highly supported subgroups in Japan, congruent with mtDNA lineages A2 and B, suggesting multiple colonization of Japan. Invariant sequences at the 28S rDNA combined with other results do not support the hypothesis that cryptic (sibling) species exist within the populations investigated in this study.

Animals↗

Occupational asthma due to grain pests Eurygaster and Ephestia.

BACKGROUND: Workers occupationally exposed to grain dust have a high prevalence of asthma. The pathogenesis of their respiratory symptoms remains obscure when sensitization to cereal allergens has not been proved. Given the ubiquity of arthropods in stored vegetable products, we have studied the allergenic potential of two very prevalent grain pests, Eurygaster and Ephestia, as a cause of occupational asthma. We have also studied the allergenic relationship between Anisakis simplex (AS) and these pests. METHODS: We selected 15 asthmatic workers exposed to cereal dust, in whom sensitization to cereal allergens was not clear. As controls, we selected a patient who suffered from anaphylaxis after the ingestion of cereals, 6 patients sensitized to different arthropods, 1 patient who suffered from asthma after inhaling fish flour contaminated with AS, and a pool of 40 asthmatic patients with different ethiologies not due to arthropods or cereals. We performed prick tests with these pests, AS, and pure and parasitized flours, bronchial challenges, specific IgE determination, and RAST inhibition. RESULTS: All of the 15 cereal-exposed workers were sensitized to Eurygaster and Ephestia. Only 2 had detectable levels of serum IgE to these pests, but 13 presented a positive prick test result and IgE to AS without problems after eating fish. Bronchial challenges were positive to Eurygaster in 7 patients and to Ephestia in 2 patients. Twelve patients had positive prick tests to parasitized flour but not to pure flour. The patient with cereal anaplylaxis presented sensitization to both flours but not to AS. In the control group, the patient who suffered from asthma after the inhalation of AS-parasitized fish flour presented a positive prick test result and high levels of specific IgE to these pests. The RAST inhibition showed cross-reactivity between Eurygaster and AS in this patient, but the inhibition was not as evident with the sera of other workers. The same tests in the group of 40 asthmatic patients were negative. CONCLUSIONS: The clinical importance of pests as a cause of baker's asthma should not be underestimated. Sensitization to Eurygaster and Ephestia could be important in the asthma episodes suffered by our patients, and the parasitized wheat was found to have a higher allergenic potential than pure cereal flour.

Adolescent↗

Allometry for sexual size dimorphism: testing two hypotheses for Rensch's rule in the water strider Aquarius remigis.

Within any given clade, male size and female size typically covary, but male size often varies more than female size. This generates a pattern of allometry for sexual size dimorphism (SSD) known as Rensch's rule. I use allometry for SSD among populations of the water strider Aquarius remigis (Hemiptera, Gerridae) to test the hypothesis that Rensch's rule evolves in response to sexual selection on male secondary sexual traits and an alternative hypothesis that it is caused by greater phenotypic plasticity of body size in males. Comparisons of three populations reared under two temperature regimes are combined with an analysis of allometry for genital and somatic components of body size among 25 field populations. Contrary to the sexual-selection hypothesis, genital length, the target of sexual selection, shows the lowest allometric slope of all the assayed traits. Instead, the results support a novel interpretation of the differential-plasticity hypothesis: that the traits most closely associated with reproductive fitness (abdomen length in females and genital length in males) are "adaptively canalized." While this hypothesis is unlikely to explain Rensch's rule among species or higher clades, it may explain widespread patterns of intraspecific variation in SSD recently documented for many insect species.

Animals↗

Within-field manipulation of potato leafhopper (Homoptera: Cicadellidae) and insect predator populations using an uncut alfalfa strip.

The effects of a 3-m wide uncut alfalfa strip on the within field distribution of Empoasca fabae (Harris) (Homoptera: Cicadellidae) and several species of insect predators were examined for 3 yr (1998-2000). The objectives were to determine whether this uncut strip would serve as a trap crop for E. fabae and a refuge for insect predators at first harvest. Empoasca fabae and insect predators in the families Coccinellidae, Nabidae, Anthocoridae, Chrysopidae, and Hemerobiidae were collected weekly using sweep samples and sticky traps from the uncut strips and up to 40 m into the alfalfa regrowth. For 2 yr, both small- (0.34 ha) and large-scale (approximately 11.3 ha) field trials showed higher numbers of E. fabae in 73% of the uncut strips for 2-3 wk after harvest. Similarly, the number of insect predators found within <50% of the uncut strips was also higher during the first or second week after harvest. In 1999, however, we did not observe higher numbers of E. fabae in the uncut strips. Differences may be because of higher E. fabae population numbers in 1999 compared with 1998 and 2000. This research provides alfalfa growers a potential cultural management technique for E. fabae while conserving predatory insects.

Animals↗

Survival of stored-product insect natural enemies in spinosad-treated wheat.

The survival of stored product insect natural enemies in wheat treated with spinosad was investigated in laboratory and pilot scale experiments. The predator Xylocoris flavipes (Reuter), the warehouse pirate bug, and the parasitoids Habrobracon hebetor (Say), Theocolax elegans (Westwood), and Anisopteromalus calandrae (Howard) were exposed to wheat treated with aliquots of water or spinosad at 0.05-1 mg ([AI])/kg. X. flavipes was the only species that survived (92% survival) in spinosad-treated wheat at 1 mg/kg. X. flavipes suppressed populations of immature Tribolium castaneum (Herbst), the red flour beetle, by nearly 90% compared with a water-treated control, but 100% suppression of immatures was achieved in wheat receiving spinosad or spinosad + X. flavipes treatments. A 3-mo pilot scale experiment to evaluate T. castaneum suppression in drums holding 163.3 kg of wheat showed that the pest populations increased throughout the study in the control treatment, but peaked after 1 mo in the X. flavipes-treated drums. By comparison, better T. castaneum population suppression was achieved in spinosad or spinosad + X. flavipes treatments. Although X. flavipes can survive and reproduce in spinosad-treated wheat, under our test conditions spinosad alone provided adequate suppression of T. castaneum populations in stored wheat.

Animals↗

Injury to preflowering and flowering cotton by brown stink bug and southern green stink bug.

The impact of brown stink bug, Euschistus servus (Say), and southern green stink bug, Nezara viridula (L.), injury was evaluated on preflowering and flowering cotton, Gossypium hirsutum L., plants in no-choice tests. Vegetative stage cotton seedlings and reproductive structures, including flower buds (square) and bolls, were infested with adults and/or nymphs of both species. There were no significant differences in height, height to node ratio, square retention, and flower initiation for cotton seedlings or plants with a match-head square between southern green stink bug adult- or brown stink bug adult-infested and noninfested treatments. Abscission for individual large squares (precandle) and multiple squares (medium and small square on the same sympodial branch) was not significantly different among infested and noninfested treatments for the following species and developmental stages: brown stink bug adults, southern green stink bug adults, and third and fourth to fifth instar southern green stink bug nymphs. In boll infestation studies, the relationship between boll maturity, expressed as heat units beyond anthesis, and boll growth, abscission, hard locked carpels, seedcotton yield, and seed germination was measured. Brown stink bug induced abscission in bolls that had accumulated > 0-350 heat units beyond anthesis. Boll growth and seedcotton yield was significantly lower for bolls infested with brown stink bug through 266.5 and 550 heat units beyond anthesis, respectively. The proportion of hard locked carpels per boll was significantly greater for the infested treatment in a cohort of bolls that accumulated from 51 to 400 heat units beyond anthesis. Seed germination in bolls infested with brown stink bug was significantly lower in bolls aged 101-600 heat units beyond anthesis.

Animals↗

Toxicity of three acaricides to Tetranychus urticae (Tetranychidae: Acari) and Orius insidiosus (Anthocoridae: Hemiptera).

Management for twospotted spider mite, Tetranychus urticae Koch, populations in peanut, Arachis hypogaea L., relies on acaricides. The outcomes of acaricide applications are most predictable when complete information on their toxicity and specificity is available. Specifically, the degrees to which acaricides impact different stages of T. urticae and natural enemies combined determine the overall efficacy of an acaricide application. The objectives of this study were to determine stage-specific direct and residual efficacies of three acaricides (fenpropathrin, etoxazole, and propargite) against T. urticae, and the direct and residual toxicity of the acaricides to Orius insidiosus (Say) adults. Direct toxicity of acaricides to T. urticae was measured on peanut cuttings. All acaricide treatments caused significant mortality to a mixed stage population of T. urticae, and mortality did not differ among the acaricides 7 d after treatment. When toxicity to eggs was tested, the proportion of eggs that hatched for all acaricide treatments was significantly lower than the control, with etoxazole and propargite causing 100% mortality. Exposure to acaricide residues caused < 30% mortality of T. urticae adults 1 and 2 d after treatment and was not significantly different from the control. Fenpropathrin and propargite caused 100% mortality and etoxazole caused > 50% mortality of O. insidious adults after direct exposure to the acaricides. Residual toxicity of acaricides to O. insidiosus adults varied but remained toxic to O. insidiosus longer than to T. urticae. Fenpropathrin had the longest residual effect on O. insidiosus adults, causing > 95% mortality after 14 d; etoxazole and propargite caused < 30% mortality after 14 d.

Acari↗

Molecular distinction between populations of Gonatocerus morrilli, egg parasitoids of the glassy-winged sharpshooter from Texas and California: do cryptic species exist?

Two molecular methods were utilized to distinguish geographic populations of Gonatocerus morrilli (Howard) from Texas and California and to test the possibility that this species could exist as a species-complex. Inter-Simple Sequence Repeat-Polymerase Chain Reactions (ISSR-PCR) were performed with a 5'-anchored ISSR primer. Twenty-five markers were generated with four populations (40 individuals) of G. morrilli. Twenty-three were polymorphic and the percentage of polymorphic loci was 92%. Most markers could be considered diagnostic since there was no band sharing between the Texas and California populations. Such differences typically are not found unless the populations are reproductively isolated. Exact tests for population differentiation indicated significant differences in marker frequencies among the populations. Comparison of other genetic differentiation estimates, which evaluate the degree of genetic subdivision, demonstrated excellent agreement between GST and theta values, 0.92 and 0.94, respectively, indicating that about 92 to 94% of the variance was distributed among populations. The average genetic divergence (D), as measured by genetic distance, was extremely high (Nei = 0.82 and Reynolds = 2.79). A dendrogram based on Nei's genetic distance separated the Texas and California populations into two clusters, respectively. Amplification of the Internal Transcribed Spacer-1 (ITS-1) region showed no size differences, whereas the ITS-2 DNA fragment varied in size between the two geographic populations. The ITS-2 fragment sizes were about 865 and 1099 base pairs for the California and Texas populations, respectively. The present study using the two molecular methods provides novel data critical to the glassy-winged sharpshooter/Pierce's disease biological control program in California.

Animals↗

Antagonistic pre- and post-copulatory sexual selection on male body size in a water strider (Gerris lacustris).

A crucial question in sexual selection theory is whether post-copulatory sexual selection reinforces or counteracts conventional pre-copulatory sexual selection. Male body size is one of the traits most generally favoured by pre-copulatory sexual selection; and recent studies of sperm competition often suggest that large male size is also favoured by post-copulatory sexual selection. In contrast to this general pattern, this study shows that pre- and post-copulatory sexual selection act antagonistically on male body size in Gerris lacustris. One large and one small male were kept together with two females in this experiment. Large males had a significant mating advantage, but small males copulated longer and gained higher fertilization success from each mating. Large and small males, however, gained similar reproductive success, and there was no overall correlation between mating success and reproductive success. These results suggest that estimates of male fitness based solely on mating success should be viewed with caution, because of potentially counteracting post-copulatory selection.

Animals↗

Fruit or aposematic insect? Context-dependent colour preferences in domestic chicks.

Colours are common stimuli in signalling systems. Requirements to function well as a signal sometimes conflict between different signallers, and the same colour stimulus is used to convey completely different messages to the same receiver. Fruits and aposematic insects both use red coloration as a signal, in the former case to signal profitability and in the latter case as a warning signal. In two experiments, we investigated whether the domestic chick, an omnivorous predator, differed in its unconditioned preference or avoidance of red and green stimuli depending on whether or not the stimulus was an insect. The experiments were designed as preference tests between red and green painted prey. The prey were live insects and artificial fruits (experiment 1), and, to investigate the effect of movement, live and dead insects (experiment 2). The chicks did not show any difference in pecking preference between red and green when fruit-like stimuli were used, but when the prey were insects, green prey were strongly preferred to red prey, and prey movement did not affect this bias. Thus, young chicks may recognize prey as insects and then discriminate between different prey colorations, or one type of food may elicit an unlearned colour preference-avoidance response that is absent with another type of food.

Animals↗

Why do multiple traits determine mating success? Differential use in female choice and male competition in a water boatman.

Mating success is often determined by multiple traits, but why this occurs is largely unknown. Much attention has been paid to female preferences for multiple traits, but surprisingly few researchers have addressed the possibility that multiple traits are important because they serve different functions in female choice and male-male competition. Differential trait function could result from a conflict of interest between the sexes or from constraints forcing the sexes to pay attention to different traits. I show that traits determined at distinct life-history stages differ in their importance in female choice and male-male competition in a water boatman Sigara falleni. Juvenile conditions determined body and foreleg pala size and were the main determinants of mating success under female choice, whereas adult conditions determined body mass and influenced mating success when male competition was included. This differential use of condition-dependent traits under the two selection regimes appeared to arise partly from a conflict between the sexes, since the two selection forces (female choice and male competition) conflict for selection on pala size, and partly from constraints, as females appeared unable to assess adult condition.

Age Factors↗

Maternal food provisioning in relation to condition-dependent offspring odours in burrower bugs (Sehirus cinctus).

The sensory modalities used for communication among family members have at least partly evolved within an organism's pre-existing sensory context. Given the well-known general importance of chemical communication in insects, we hypothesized in sub-social insects with parental care that chemical signals emitted by larvae to influence parental care (i.e. solicitation pheromones) would have evolved. To test this hypothesis, we performed an experiment in the burrower bug Sehirus cinctus (Hemiptera: Cydnidae) where nymphs were hand-reared under high- or low-food conditions. These hand-reared clutches were used as a source of volatiles. The volatiles were collected for chemical analysis and delivered to caring mothers to quantify their behavioural response. As predicted, mothers exposed to volatiles from nymphs in poor condition provisioned significantly more food than those exposed to air (controls) or volatiles from high-condition nymphs. Chemical analysis revealed that nymphs emitted a blend of eight compounds of which alpha-pinene and camphene showed the strongest relationship with food treatment. Exposure to pure synthetic alpha-pinene and camphene did not affect maternal provisioning, however, suggesting that the functional significance of alpha-pinene and/or camphene may occur in a blend with other compounds. This study shows a clear effect of condition-dependent offspring odours on maternal food provisioning and identifies, for the first time, candidate compounds for a potential chemical offspring begging signal.

Animal Communication↗

Manipulation of endogenous trypsin proteinase inhibitor production in Nicotiana attenuata demonstrates their function as antiherbivore defenses.

Evidence for the in planta defensive function of trypsin protease inhibitors (TPIs) comes from observations of enhanced herbivore resistance after heterologous TPI expression or the manipulation of signal cascades that activate numerous defense responses, including TPI production; no studies have altered the expression of an endogenous pi gene to examine defensive function. We isolated two genes with seven- and six-repeat TPI domains from Nicotiana attenuata from the potato (Solanum tuberosum) PI-II family. To determine whether endogenous TPIs in N. attenuata function defensively against the native herbivores, hornworm (Manduca sexta) and mirids (Tupiocoris notatus), we expressed 175 bp of the seven-domain pi from N. attenuata in an antisense orientation in a TPI-producing genotype to reduce TPI expression and expressed the full-length seven-domain pi in a sense orientation under control of a constitutive promoter to restore TPI activity in a natural genotype from Arizona unable to produce TPIs. Constitutive and inducible TPI production in two antisense lines were diminished by 80% to 90% and 33% to 52%, respectively, and sense expression restored 67% of the activity found in the TPI-producing genotype after caterpillar attack in the TPI-deficient A genotype. Hornworm larvae fed on genotypes with low or no TPI activity grew faster, had higher survivorship, and produced heavier pupae than those that fed on genotypes with high TPI activity. T. notatus showed higher preference for genotypes with low or no TPI activity than for genotypes with high TPI levels. We conclude that endogenous TPIs are an effective defense against these native herbivores.

Amino Acid Sequence↗