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Insect mitochondrial genomics: the complete mitochondrial genome sequence of the meadow spittlebug Philaenus spumarius (Hemiptera: Auchenorrhyncha: Cercopoidae).

We present the complete mitochondrial genome sequence of the meadow spittlebug Philaenus spumarius (Auchenorrhyncha: Cercopoidae). This contribution represents the second mitochondrial genome from the Hemiptera and the second of the three hemipteran suborders sampled. The genome is a circular molecule of 16 324 bp with a total A+T content of 77.0% and 76.7% for coding regions only. The gene content, order, and structure are consistent with the Drosophila yakuba genome structure (Clary and Wolstenholme 1985) and the hypothesized ancestral arthropod genome arrangement (Crease 1999). Nucleotide composition and codon usage are near the means observed in other insect mitochondria sequenced to date but have a higher A+T richness compared with the other hemipteran example, the kissing bug Triatoma dimidiata (Dotson and Beard. 2001. Insect Mol. Biol. 10: 205-215). The major noncoding region (the A+T rich region or putative control region) between the small ribosomal subunit and the tRNAIle gene includes two extensive repeat regions. The first repeat region includes 19 tandem repeats of a 46-bp sequence, whereas the second contains a longer sequence (146 bp) tandemly repeated four times.

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B chromosomes in Sternorrhyncha (Hemiptera, Insecta).

In the hemipteroid insects of the suborder Sternorrhyncha, B chromosomes are relatively common in comparison with other suborders of Hemiptera. However, the occurrence of supernumerary chromosomes is restricted, in most cases, to several genera or closely related species. At least in some species of Psylloidea with the XY sex determination system, a mitotically stable B chromosome integrated into an achiasmatic segregation system with the X, and became fixed as a Y chromosome. In some Aphidoidea with a multiple X system of sex determination, B chromosomes appear to be in fact non-functional X chromosomes. Supernumerary chromosomes thus probably play an important role in the evolution of sex determination systems in Sternorrhyncha.

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Partial morphological and functional characterization of the corpus allatum-corpus cardiacum complex from the two-spotted stinkbug, Perillus bioculatus (Hemiptera: Pentatomidae).

Selected morphological and physiological properties of the corpus allatum (CA)-corpus cardiacum (CC) complex from the two-spotted stinkbug, Perillus bioculatus (Hemiptera: Pentatomidae), were studied. The CAs play an important role in insect physiology because of their production of the juvenile hormones (JHs), i.e., key hormones involved in development and reproduction. We found that the P. bioculatus CA-CC complex is present in two distinct morphological forms, the more frequently observed complex containing one "fused" CA between two CCs and the more rarely observed complex having one CA laterally attached to each CC. These complexes were tested for their ability to synthesize JH-like compounds. We found that the primary lipophilic compound synthesized by the CA-CCs migrated differently from JH III (a JH found in numerous insect species) when subjected to thin-layer chromatography. Furthermore, the synthesis of this compound is stimulated by 2E,6E-farnesol, a known precursor for JH III. These data indicate that the P. bioculatus CA- CC product has chemical properties similar to that of other (as of yet unidentified) hemipteran JHs. In addition, we found that the synthesis of this product is sensitive to pH and buffer type; minimally or not affected by the absence of the CC; expressed at similar levels in days 5-30 postemergent adults; and inhibited or decreased in adults reared under low temperature-short day conditions.

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Development of trypanosomatids in the phytophagous insect Dysdercus peruvianus (Hemiptera: Pyrrochoridae). A light and eletron microscopic study.

Experimental infections of the phytophagous Hemiptera Dysdercus peruvianus with different trypanosomatids were studied for up to 55 days by light microscopy while the course of infection with Leptomonas seymouri and the Leptomonas isolate 49/553G.O. was analyzed by electron microscopy. Rates of infection of D. peruvianus varied according to the infecting flagellate. The lower part of the midgut was found to be the preferential site of colonization where most flagellates were found isolated or arranged in clumps or rosettes. Specialized junctional structures with host cells were never observed. Flagellates could also be seen inside midgut cells within a parasitophorous vacuole. Infection of haemocoele and salivary glands was also observed.

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Soil temperature and diapause maintenance in eggs of the spittlebug, Deois flavopicta (Hemiptera: Cercopidae).

Diapausing eggs of the neotropical pasture pest, Deois flavopicta (Stal) (Hemiptera: Cercopidae), were exposed to low overnight temperatures that simulated field conditions during the dry season (23/12, 23/15 and 23/18 degrees C day/night), for different periods (0-60 days). After treatment, eggs were kept at 28 degrees C and contact water (100% humidity) until hatching. A group of diapausing eggs were kept all the time under this last condition as a control treatment. Time for hatching (in degree-days) was reduced with decrease in low overnight temperature and increase of exposure time to these cold shocks, although there was no interaction between the factors. Regression of exposure time to cold shock influencing the expected mean hatching time produced independent equations for temperatures below 18 degrees C and 15 degrees C. We constructed a model that simulates the expected proportion of the population hatching after the beginning of rainy season based on regression equations to mean hatching time and associated standard deviation. The simulation generated for the model correlated significantly with nymphal population observed in the field. These results showed that overnight soil temperatures below 18 degrees C, as occurs in Central and South-eastern Brazil between May and August, shorten the period of diapause, increase quiescent eggs in the soil, and may have synchronize population hatching.

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Movement pattern of Spartocera dentiventris (Berg) (Hemiptera: Coreidae) in an experimental crop of Nicotiana tabacum L. (Solanaceae).

In the present work we investigated the movement pattern of Spartocera (= Corecoris) dentiventris (Berg) (Hemiptera: Coreidae) adults through the mark-release-recapture method in an experimental tobacco crop (Nicotiana tabacum L., Solanaceae) in Porto Alegre, RS, Brazil. We marked 217 specimens from August 1999 to April 2000. Females moved 2.94 +/- 0.198 times and males 1.46 +/- 0.171 times among host plants. The number of movements per capture was 0.53 +/- 0.03 for females and 0.54 +/- 0.06 for males. Newly recruited adults took 12.75 +/- 0.919 days to leave their original plant, females moved 9.87 +/- 0.711 m and males 9.16 +/- 1.29 m in this first movement. We estimate that females move an average of 26.22 m and males 13.89 m during their permanence in the area.

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Morphogenesis of galls induced by Baccharopelma dracunculifoliae (Hemiptera: Psyllidae) on Baccharis dracunculifolia (Asteraceae) leaves.

The commonest insect gall on Baccharis dracunculifolia (Asteraceae) leaves is induced by Baccharopelma dracunculifoliae (Hemiptera, Psyllidae). The gall-inducing insect attacks young leaves in both the unfolded and the fully expanded stages. Four developmental phases were observed in this type of gall: 1) A folding phase, during which the leaf lamina folded upward alongside the midrib and the edges of the upper portion of the leaf approached each other, forming a longitudinal slit. A single chamber was formed on the adaxial surface of the leaf; 2) A swelling phase, in which the folded leaf tissues thickened and the edges of the leaf drew closer together, narrowing the slit. In this phase the gall matured, turning succulent, fusiform and pale green. The single nymphal chamber was lined with white wax and was able to house from one to several nymphs; 3) A dehiscence phase, characterized by the opening of the slit to release inducers; and 4) A senescence phase, when the gall turned dark and dry. The dermal system of the mature gall was composed of a single-layered epidermis. The mesophyll was swollen, and the swelling was due mainly to hyperplasia of the parenchyma. The vascular tissues along the midrib vein were conspicuous and the perivascular fibers resembled parenchymal cells. The hypertrophied secretory cavities contained low lipophylic content. This gall does not form nutritive tissue, but salivary sheaths left by the inducers were observed near the parenchyma, vascular bundles and secretory cavities. This study complements our current knowledge of gall biology and sheds further light on the plasticity of plant tissues stimulated by biotic factors.

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Beet leafhopper (Hemiptera: Cicadellidae) transmits the Columbia Basin potato purple top phytoplasma to potatoes, beets, and weeds.

Experiments were conducted to determine whether the beet leafhopper, Circulifer tenellus (Baker) (Hemiptera: Cicadellidae), transmits the purple top phytoplasma to potato, Solanum tuberosum L.; beets, Beta vulgaris L.; and selected weed hosts. The beet leafhopper-transmitted virescence agent (BLTVA) phytoplasma was identified as the causal agent of the potato purple top disease outbreaks that recently occurred in the Columbia Basin of Washington and Oregon. The phytoplasma previously was found to be associated almost exclusively with the beet leafhopper, suggesting that this insect is the probable vector of BLTVA in this important potato-growing region. Eight potato cultivars, including 'Russet Burbank', 'Ranger Russet', 'Shepody', 'Umatilla Russet', 'Atlantic', 'FL-1879', 'FL-1867', and 'FL-1833', were exposed for a week to BLTVA-infected beet leafhoppers. After exposure, the plants were maintained outdoors in large cages and then tested for BLTVA by using polymerase chain reaction after 6 to 7 wk. The leafhoppers transmitted BLTVA to seven of the eight exposed potato cultivars. Sixty-four percent of the exposed plants tested positive for the phytoplasma. In addition, 81% of the BLTVA-infected potato plants developed distinct potato purple top disease symptoms. Beet leafhoppers also transmitted BLTVA to beets and several weeds, including groundsel, Senecio vulgaris L.; shepherd's purse, Capsella bursa-pastoris (L.) Medik); kochia, Kochia scoparia (L.) Schrad; and Russian thistle, Salsola kali L. This is the first report of transmission of BLTVA to potatoes, beets, and the above-mentioned four weed species. Results of the current study prove that the beet leafhopper is a vector of the potato purple top disease.

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Mortality factors of eggs of Leptocorisa chinensis (Hemiptera: Alydidae) in rice fields.

To clarify factors causing mortality of Leptocorisa chinensis Dallas (Hemiptera: Alydidae) eggs in rice fields, sentinel egg masses were exposed for seven days in two rice, Oryza sativa L. (Poaceae), fields. An insecticide was sprayed to remove natural enemies in one-half of each field before exposing egg masses to predation. An egg mass containing 14 eggs was glued to a plastic label, and 21 egg masses each were placed in the sprayed and unsprayed control plots. During exposure, the number of hatched and missing eggs was observed daily. Egg predators were sampled in the fields before and after insecticide application. After the egg masses were retrieved from the fields, mortality factors of the unhatched eggs were assessed in the laboratory. The mean number of hatched and missing eggs was not significantly different between the sprayed and control plots in field A. In field B, however, the numbers were significantly different. The percentage of missing eggs in damaged egg masses ranged from 80 to 100%. In the laboratory, we observed that feeding marks caused by the grasshopper Conocephalus chinensis (Redtenbacher) were similar to those on the eggs exposed in the rice fields. The density of C. chinensis was low in control plots of field A. In contrast, the density was high in those of field B. These observations suggest that the density of egg predators, e.g., C. chinensis, is a mortality factor of L. chinensis eggs in rice fields.

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Feasibility of tracking within-field movements of Homalodisca coagulata (Hemiptera: Cicadellidae) and estimating its densities using fluorescent dusts in mark-release-recapture experiments.

A mark-release-recapture technique was developed and tested for use in tracking the field movements of adult glassy-winged sharpshooters, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae), in various agricultural and urban plantings. Greenhouse experiments in which adult H. coagulata were marked with one of five colored fluorescent dusts (Aurora Pink-All, Horizon Blue-A19, Blaze Orange-A15N, Saturn Yellow-A17, and Corona Magenta-A21) and released into cages with citrus seedlings showed that their mortality rates during a 30-d period were statistically similar to that of the undusted controls. Adults marked with a sixth dust color (Signal Green-A18N) suffered higher rates of mortality than did the undusted controls and thus were eliminated from further consideration. Adult H. coagulata marked with one of the five accepted colors of fluorescent dust were able to fly beyond 100 m in a field devoid of vegetation within minutes of their release, and the marking did not affect overall flight behavior significantly compared with that of the undusted controls. However, at wind speeds above 5 m/s, percentage recapture was significantly reduced, which indicates that both dusted and undusted adults were unable to orient their flight. In total, 41,124 marked and unmarked adults were released in the three field experiments in southern California (Riverside and Kern counties) during 2000 and 2001 to evaluate flight dispersal and estimate densities of H. coagulata. The mark-release-recapture and feral data obtained during the June, July, and August 2001 studies, when coupled with the Lincoln index, yielded estimates of adult H. coagulata of 1.2 and 2.2 million per ha, respectively, at a San Joaquin Valley (Kern Co.) and a southern California (Riverside Co.) citrus grove. The use of colored dusts to mark H. coagulata proved to be reliable, cost-effective, and time-efficient for mark-release-recapture studies with this insect within a citrus grove, but they are less likely to be useful for studies of adult H. coagulata movements among plantings.

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Detection and differentiation of Xylella fastidiosa strains acquired and retained by glassy-winged sharpshooters (Hemiptera: Cicadellidae) using a mixture of strain-specific primer sets.

Xylella fastidiosa Wells is a bacterial pathogen that causes a variety of plant diseases, including Pierce's disease (PD) of grapevine, almond leaf scorch, alfalfa dwarf, citrus variegated chlorosis, and oleander leaf scorch (OLS). Numerous strains of this pathogen have been genetically characterized, and several different strains occur in the United States. The dominant vector in southern California is the glassy-winged sharpshooter, Homalodisca coagulata (Say) (Hemiptera: Cicadellidae). The high mobility of this insect, and its use of large numbers of host plant species, provides this vector with ample exposure to multiple strains of X. fastidiosa during its lifetime. To learn more about the ability of this vector to acquire, retain, and transmit multiple strains of the pathogen, we developed a polymerase chain reaction (PCR)-based method to detect and differentiate strains of X. fastidiosa present in individual glassy-winged sharpshooter adults. Insects were sequentially exposed to plants infected with a PD strain in grapevine and an OLS strain in oleander. After sequential exposure, a few insects tested positive for both strains (7%); however, in most cases individuals tested positive for only one strain (29% PD, 41% OLS). In transmission studies, individual adults transmitted either the PD or OLS strain of the pathogen at a rate (39%) similar to that previously reported after exposure to a single strain, but no single individual transmitted both strains of the pathogen. PD and OLS strains of X. fastidiosa remained detectable in glassy-winged sharpshooter, even when insects were fed on a plant species that was not a host of the strain for 1 wk.

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Yield and forage quality of glandular-haired alfalfa under alfalfa weevil (Coleoptera: Curculionidae) and potato leafhopper (Hemiptera: Cicadellidae) pest pressure in Virginia.

Cultivars of glandular-haired alfalfa, Medicago sativa L., such as '54H69', are currently available and marketed as being resistant to potato leafhopper, Empoasca fabae (Harris) (Hemiptera: Cicadellidae). 54H69 and a standard, nonglandular-haired alfalfa 'Choice' were evaluated at two locations in Virginia over a 3-yr period. Dry matter yields and concentrations of crude protein and acid detergent fiber were compared at the first, second, and third harvests. Overall, the two cultivars produced similar dry matter yields of comparable forage quality in the absence of insecticides at both locations in each year. Untreated 54H69 did not produce greater dry matter yields than untreated Choice under either light or heavier potato leafhopper pest pressure. Concentrations of crude protein did not vary between the two cultivars at any harvest. Some differences in concentrations of acid detergent fiber were detected between cultivars, but these differences were not consistent among years, harvests, or between locations. Further comparisons between untreated 54H69 and treated Choice were made, but few significant differences were detected in dry matter yields or forage quality. An economic analysis for the study indicated that a grower planting 54H69 would realize less net revenue than a grower planting Choice, largely because of the seed premium for the glandular-haired cultivar and the evident need to treat 54H69 with insecticide for control of alfalfa weevil, Hypera postica (Gyllenhal) (Coleoptera: Curculionidae), and potato leafhopper.

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Activity and residues of imidacloprid applied to soil and tree trunks to control hemlock woolly adelgid (Hemiptera: Adelgidae) in forests.

We studied imidacloprid application methods and timing to control the hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), in forests. The methods compared were 1) soil injection near the trunk; 2) soil injection dispersed throughout the area under the canopy; 3) soil drench near the base of the trunk; and trunk injection with the 4) Arborjet, 5) Wedgle, and 6) Mauget systems. The applications were made in the fall and the following spring. Adelgid populations on the hemlocks (Tsuga spp.) were assessed in the fall of two successive years after the treatments. Relative to the untreated control trees, all the soil applications resulted in population reductions, but none of the trunk injections resulted in reductions. Fall and spring treatment efficacy did not differ. Reductions by the soil treatments were between 50 and 100% (avg 80%) by the first fall and 83-100% (avg 98.5%) by the second fall. Analysis of imidacloprid residues using enzyme-linked immunosorbent assay found residues in sap, needles, and twigs 1 mo to 3-yr after application. A laboratory dose-response bioassay using excised, adelgid-infested hemlock branches with cut ends immersed in serial dilutions of imidacloprid determined the LC50 value to be 300 ppb, based on an exposure of 20 d. A high degree of suppression of the adelgid on forest trees was associated with residues in hemlock tissue > 120 ppb 2 yr after soil treatment. Although precise relationships between residues and efficacy are elusive, it is clear that soil application of imidacloprid resulted in chronic residues of imidacloprid in tissues and suppression of adelgid populations for > 2 yr.

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Pheromone-based mating disruption of Planococcus ficus (Hemiptera: Pseudococcidae) in California vineyards.

Experiments were conducted to test a mating disruption program for the mealybug Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) in California vineyards. The sprayable, microencapsulated formulation of the racemic sex pheromone lavandulyl senecioate was applied with an air-blast sprayer, using three and four applications in 2003 and 2004, respectively. Mating disruption was combined with an application of buprofezin (2004) in June. Compared with a no-pheromone control, there were significantly lower season-long trap catches of adult males, season-long mealybug densities (2003 only), and crop damage in mating disruption plots. The amount of mealybug reduction and mechanisms that resulted in lower crop damage in mating disruption plots is discussed. In samples taken during the growing season (April to September), mealybug density was only 12.0 +/- 15.6 and 31.1 +/- 11.6% lower in the mating disruption plots than in control plots in 2003 and 2004, respectively. In the mating disruption treatment, mealybug egg production was significantly lower (2003 only), as were the proportion of ovisacs and crawlers produced. There was no treatment impact on percentage of parasitism. Mealybug density influenced treatment impact. In 2004, vines were categorized as having low, medium, or high mealybug densities during a preapplication survey. After treatment application, mealybug density was reduced by 86.3 +/- 6.3% on vines in the low mealybug density category, but it was unchanged on vines in the high density category. Another factor that reduced treatment impact was the relatively short effective lifetime of the sprayable formulation.

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Binomial sequential sampling plan for hemlock woolly adelgid (Hemiptera: Adelgidae) sistens infesting individual eastern hemlock trees.

The hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), is an exotic insect pest that is killing eastern hemlock, Tsuga canadensis (L.) Carrière, and Carolina hemlock, Tsuga caroliniana Engelmann, in the eastern United States. We used the sequential interval procedure to develop a binomial sequential sampling plan for A. tsugae sistens on individual eastern hemlock trees that uses nondestructive sampling of new shoots. The actual a (type I) and beta (type II) error rates were essentially 5 and 10%, respectively. Tallies of new shoots infested by at least one A. tsugae sistens were compared with stop values for thresholds of 10 and 30% of new hemlock shoots infested. Twenty to 80 new shoots had to be examined per tree to render a low, high, or indeterminate classification, which took < 2 min per tree regardless of the threshold used. This plan should be an efficient and cost-effective tool in the management of A. tsugae infestations on individual, high-value eastern hemlock trees.

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Functional response of the tiger beetle Megacephala carolina carolina (Coleoptera: Carabidae) on twolined spittlebug (Hemiptera: Cercopidae) and fall armyworm (Lepidoptera: Noctuidae).

The functional response of the tiger beetle Megacephala carolina carolina L. (Coleoptera: Carabidae) was determined on adult twolined spittlebug, Prosapia bicincta (Say) (Hemiptera: Cercopidae), and fourth instars of fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), in single-prey and two-prey systems. In the laboratory, M. carolina carolina demonstrated a type II functional response for P. bicincta and S. frugiperda in both single- and two-prey systems. Search efficiency of M. carolina declined for both prey as the initial number of prey increased. Of the total prey consumed, M. carolina carolina killed significantly more S. frugiperda than P. bicincta in the single-prey system (8.0 and 4.5, respectively) and the two-prey system (5.0 and 2.0, respectively). Estimates of attack coefficient, a, were not significantly different for P. bicincta and S. frugiperda in the single-prey (0.07 and 0.02) and two-prey systems (0.04 and 0.06), respectively. The handling time, T(h), was significantly greater for P. bicincta (5.02 and 10.64 h) than for S. frugiperda (2.66 and 4.41 h) in single- and two-prey systems, respectively. Estimations of attack coefficient and handling time in the single-prey system were used to predict prey preference of M. carolina carolina. No strong prey switching response was observed. M. carolina carolina showed no preference for either prey. However, in the presence of S. frugiperda, the functional response of the predator for P. bicincta was reduced. M. carolina carolina is a potential predator of one or more turfgrass pests and should be considered in conservation efforts.

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Susceptibility of twolined spittlebug (Hemiptera: Cercopidae) life stages to entomophagous arthropods in turfgrass.

Prosapia bicincta (Say) (Hemiptera: Cercopidae), the twolined spittlebug, is an economic pest of turfgrass in the southeastern United States. No data concerning natural enemies of P. bicincta in turfgrass have been reported previously. We compared predation of spittlebug eggs, nymphs, and adults in the laboratory by potential generalist predators commonly found in turfgrass: bigeyed bugs Geocoris uliginosus Say and Geocoris punctipes Say; red imported fire ant, Solenopsis invicta Buren; wolf spiders (Lycosa sp. Walckenaer); carabid beetles Harpalaus pensylvanicus DeGeer and Calosoma sayi Dejean; and tiger beetles Megacephala carolina carolina L. Eggs were readily consumed by generalist predators. S. invicta consumed 100% of the eggs offered. H. pensylvanicus and C. sayi were also significant predators of P. bicincta eggs. Nymphs live in spittlemasses that protect them from attack by predators, but exposed nymphs were susceptible to attack when mechanically removed from their spittlemasses. S. invicta and M. carolina carolina caused significant mortality of exposed nymphs. P. bicincta adults are aposematic and have the ability to reflex bleed; however, reflex bleeding did not prevent attack by predators. S. invicta and M. carolina carolina killed 100% of the adult spittlebugs offered in laboratory bioassays. Lycosa sp. are less voracious predators of adults. Sound background knowledge about P. bicincta and its potential natural enemy complex is important for the development and implementation of a detailed, site-specific, biologically based pest management program in turfgrass.

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