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PubMed2003

Molecular cloning of trypsin-like cDNAs and comparison of proteinase activities in the salivary glands and gut of the tarnished plant bug Lygus lineolaris (Heteroptera: Miridae).

Using specific proteinase inhibitors, we demonstrated that serine proteinases in the tarnished plant bug, Lygus lineolaris, are major proteinases in both salivary glands and gut tissues. Gut proteinases were less sensitive to inhibition than proteinases from the salivary glands. Up to 80% azocaseinase and 90% of BApNAse activities in the salivary glands were inhibited by aprotinin, benzamidine, and PMSF, whereas only 46% azocaseinase and 60% BApNAse activities in the gut were suppressed by benzamidine, leupeptin, and TLCK. The pH optima for azocaseinase activity in salivary glands ranged from 6.2 to 10.6, whereas the pH optima for gut proteinases was acidic for general and alkaline for tryptic proteinases. Zymogram analysis demonstrated that approximately 26-kDa proteinases from salivary glands were active against both gelatin and casein substrates. Three trypsin-like cDNAs, LlSgP2-4, and one trypsin-like cDNA, L1GtP1, were cloned from salivary glands and gut, respectively. Putative trypsin precursors from all cloned cDNAs contained a signal peptide, activation peptide, and conserved N-termini (IVGG). Other structural features included His, Asp, and Ser residues for the catalytic amino acid triad of serine proteinase active sites, residues for the binding pocket, and four pairs of cysteine residues for disulfide bridges. Deduced trypsin-like proteins from LlSgP2, LlSgP3, and LlGtP1 cDNAs shared 98-99% sequence identity with a previously reported trypsin-like precursor, whereas the trypsin-like protein of LlSgP4 shared only 44% sequence identity with all other trypsin-like proteins, indicating multi-trypsin forms are present in L. lineolaris.

Amino Acid Sequence↗
PubMed2000

Comparison of alpha-amylase and protease activities of a zoophytophagous and two phytozoophagous Heteroptera.

To better understand the nature of facultative phytophagy in the zoophytophagous Geocoris punctipes (Say), and facultative zoophagy in phytozoophagous Lygus hesperus (Knight) and Lygus lineolaris (Palisot de Beauvois), we compared the activities of both the starch digesting enzyme alpha-amylase and of general proteases in these species. The alpha-amylases and proteases were demonstrated in L. hesperus, L. lineolaris and G. punctipes. The presence of alpha-amylase in the salivary gland complexes of G. punctipes indicates a disposition of this species toward utilization of nutrients that can be derived only from plants, either directly from ingestion of plant macromolecules or from second-hand ingestion of plant material from the digestive system of their prey. The alpha-amylase activity in G. punctipes was much less than those of phytozoophagous L. hesperus and L. lineolaris. The relative importance of amylolytic activity and proteolytic activity is also discussed.

Adaptation, Physiological↗
PubMed1998

Wing morph-related physiological differences in adults of temperate population of Pyrrhocoris apterus (L.) (Heteroptera: Pyrrhocoridae).

The reproductive and diapausing adult females of brachypterous morph and macropterous females with reproductive arrest of non-diapause type, originating from the laboratory cultures of Pyrrhocoris apterus, were studied for their feeding and drinking behaviour, digestive enzyme activities, and carbohydrate and lipid contents. The highest feeding and drinking activities were observed in reproductive brachypters, the lowest in macropters. Macropters also differed from brachypters by lower activities of gut lipase, peptidase and protease, lower concentration of haemolymph sugars, and lower weight of fat body, which probably reflects their low feeding activity. The total content of fat body lipids was also lower in macropters (0.6 mg) than in reproductive and diapausing brachypters (4.6 and 7.5 mg, respectively) on day 14. A very high amount of glycogen was found in the fat body of diapausing brachypters, 363 micrograms on day 14, as opposed to 15 and 80 micrograms in macropterous and reproductive brachypterous females, respectively. The obtained data indicate that the most important difference between macropterous and brachypterous females with different types of reproductive arrest consists of an enhanced mobilization of lipids for dispersal in macropters and accumulation of energetic reserves for hibernation in brachypters.

Amylases↗
PubMed2001

Cryoprotective role of polyols independent of the increase in supercooling capacity in diapausing adults of Pyrrhocoris apterus (Heteroptera: Insecta).

Diapausing cold-acclimated adults of the bug Pyrrhocoris apterus accumulate four 'winter' polyols, ribitol, sorbitol, mannitol and arabinitol, in total concentrations of up to 100 mM. The accumulation started only when the temperatures dropped below a threshold of +5 degrees C in laboratory acclimated insects. The supercooling capacity (SCP) was not affected by polyol accumulation and remained constant at approximately -17 degrees C. Cold hardiness, measured as survival time (Lt50) at -15 degrees C, increased from approximately 1 day to approximately 1 week in parallel with polyol accumulation. There was a tight correlation (r=0.98) between the concentration of 'winter' polyols in haemolymph and Lt50(-15). When a mixture of ribitol and sorbitol was injected into the haemolymph of the bugs acclimated to +5 degrees C, the concentration of polyols increased from 2.5 to 83.1 mM in haemolymph, or from 0.07 to 6.61 microg/mg of fresh weight in the whole body, the SCP remained unchanged and survival after exposure to -14 degrees C for 3 days increased approximately three-fold in comparison to untreated controls. Such results were interpreted as evidence for the cryoprotective role of accumulated polyols independent of the depression of SCP. Acclimation protocol using thermoperiod, mimicking daily temperature oscillations, resulted in moderately lower SCP, higher sum of polyols accumulated and significantly longer Lt50(-15) than at acclimation protocol with constant temperatures.

Adaptation, Physiological↗
PubMed2002

Phospholipid and triacylglycerol fatty acid composition of major life stages of sunn pest, Eurygaster integriceps (Heteroptera: Scutelleridae).

Phospholipid and triacylglycerol fatty acid compositions of whole animals from all life stages of Eurygaster integriceps, including eggs, nymphs, pre-diapausing adults and diapausing adults, were determined. The fatty acid composition of total lipids of their food, wheat, was also determined. The major components of the insects and their food were the expected C16 and C18 saturated and unsaturated fatty acids. Since fatty acid compositions of third-stadium nymphs were not similar to the profiles of their food, most likely, dietary fatty acids are modified by the insect. The fact is that the food does not provide C20 polyunsaturated fatty acids, but the insect tissue lipids include these components. We suggest biosynthesis of the C20 components by elongation/desaturation of C18:2n-6, an abundant component of the diets. We also show differences in fatty acid profiles from each of the life stages.

Aging↗
PubMed2003

Pheromones of milkweed bugs (Heteroptera: Lygaeidae) attract wayward plant bugs: Phytocoris mirid sex pheromone.

The synthetic aggregation pheromone of the large milkweed bug, Oncopeltus fasciatus (Dallas) (Lygaeinae), also attracted males of the plant bug, Phytocoris difficilis Knight (Miridae). Field testing partial blends against the six-component blend comprising the Oncopeltus pheromone showed that cross-attraction of P. difficilis males was due to synergism between (E)-2-octenyl acetate and (E,E)-2,4-hexadienyl acetate. Hexyl acetate was abundant in the metathoracic scent gland (MSG) secretion of P. difficilis males, but because female P. difficilis could not initially be found in the field, further combinatorial tests were guided by prior research on the pheromones of two Phytocoris species in the western United States. The combination of hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates was as attractive to P. difficilis males as the milkweed bug pheromone, yet no milkweed bugs were drawn to this blend. Gas chromatographic (GC)-electroantennographic detection (EAD) and GC-mass spectrometric (MS) analyses of female P. difficilis MSGs determined that their secretion contained predominantly hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates (all strongly EAD-active)-the latter two compounds found only in trace amounts from males-plus five minor female-specific compounds, three of which were EAD-active. (E,E)-2,4-Hexadienyl acetate was not detected from P. difficilis females or males. The blend of the three major components, hexyl, (E)-2-hexenyl, and (E)-2-octenyl acetates (2:1.5:1 by volume), was as attractive as the blend of all six EAD-active compounds identified from females, indicating that this ternary blend constitutes the sex pheromone of P. difficilis. Hexyl acetate with (E)-2-octenyl acetate also attracted males of another species, P. breviusculus Reuter, but addition of (E)-2-hexenyl acetate and/or (E,E)-2,4-hexadienyl acetate inhibited attraction of P. breviusculus males. Attraction of P. difficilis males occurred mainly during the first half of scotophase. The possible neurophysiological basis for this asymmetrical cross-attraction is discussed.

Adaptation, Physiological↗
PubMed2003

Behaviour of ring bivalents in holokinetic systems: alternative sites of spindle attachment in Pachylis argentinus and Nezara viridula (Heteroptera).

Heteropteran chromosomes are holokinetic; during mitosis, sister chromatids segregate parallel to each other but, during meiosis, kinetic activity is restricted to one pair of telomeric regions. This meiotic behaviour has been corroborated for all rod bivalents. For ring bivalents, we have previously proposed that one of the two chiasmata releases first, and a telokinetic activity is also achieved. In the present work we analyse the meiotic behaviour of ring bivalents in Pachylis argentinus (Coreidae) and Nezara viridula (Pentatomidae) and we describe for the first time the chromosome complement and male meiosis of the former (2n = 12 + 2m + X0, pre-reduction of the X). Both species possess a large chromosome pair with a secondary constriction which is a nucleolus organizer region as revealed by in-situ hybridization. Here we propose a new mode of segregation for ring bivalents: when the chromosome pair bears a secondary constriction, it is not essential that one of the chiasmata releases first since these regions or repetitive DNA sequences adjacent to them become functional as alternative sites for microtubule attachment and they undertake chromosome segregation to the poles during anaphase I.

Anaphase↗
PubMed2004

The evolution of a neo-XY1Y2 sex chromosome system by autosome-sex chromosome fusion in Dundocoris nodulicarinus Jacobs (Heteroptera: Aradidae: Carventinae).

Sibling subspecies of Dundocoris nodulicarinus, inhabiting different isolated indigenous evergreen forests in South Africa, have chromosome numbers of 2n(male) = 14XY, 9XY1Y2 and 7XY1Y2. The ancestral chromosome number of Dundocoris is probably 2n(male) = 28XY and several chromosome fusions were involved in the karyotype evolution of these taxa. The XY1Y2 sex chromosome system of the 9XY1Y2 D. nodulicarinus novenus originated by the fusion of a large autosome with the X-chromosome, forming a neo-X with the homologue of the fused autosome forming the neo-Y (=Y1) and the original Y-chromosome, the Y2. While the original X- and Y-chromosomes are heterochromatic and heteropycnotic during prophase I, the autosomal part of the neo-X and the neo-Y stay euchromatic and behave like a normal autosomal pair, forming synapsis and chiasmata. The XY1Y2 sex chromosome system of the 7XY1Y2 D. nodulicarinus septeni probably originated from the 9XY1Y2 karyotype when the homologous chromosomes of a small autosomal pair fused with the original X- and Y-chromosomes, respectively. In both the subspecies with the neo-XY1Y2 systems, the original sex chromosomes still undergo chromatid segregation at anaphase I (= post-reductional). The evolution and behaviour of the karyotypes and sex chromosome systems during the course of meiosis in the subspecies of D. nodulicarinus are described, discussed and illustrated.

Animals↗
PubMed2004

Achiasmatic male meiosis in Tenagobia (Fuscagobia) fuscata (Stål) (Heteroptera, Corixoidea, Micronectidae).

Male karyotype and meiosis of Tenagobia fuscata (Corixoidea, Micronectidae) are studied. The species possesses a male diploid chromosome number 2n = 28 + XY, holokinetic chromosomes, absence of m chromosomes and an achiasmatic male meiosis. Autosomes divide pre-reductionally while the sex chromosomes do so post-reductionally. Banding techniques (C, DAPI and CMA) show that large heterochromatic AT-GC rich bands are generally terminally located, although some interstitial bands are also detected. Many bivalents are heteromorphic for heterochromatin amount and location. This is the first report of a species with achiasmatic male meiosis within the Nepomorpha. These cytogenetic features markedly differ from all previous reports for 26 species of the superfamily Corixoidea. T. fuscata occurs in permanent shallow water bodies, and most known individuals are brachypterous. Their dispersion depends on occasional floodings of the water bodies they occupy. Since achiasmatic meiosis maintains groups of co-adapted genes, this feature could be an adaptive strategy of the species to the particular type of habitat and ecological niche it occupies.

Animals↗
PubMed2002

Meiotic studies in Dysdercus Guérin Meneville, 1831 (Heteroptera: Pyrrhocoridae). II. Evidence on variations of the diffuse stage between wild and laboratory-inbred populations of Dysdercus chaquensis Freiberg, 1948.

Dysdercus Guerin Méneville, 1831 comprises insect species that are often serious pests of cotton both in the Old and New World, representing the only taxon from Pyrrhocoridae in the Neotropical Region. The genus is cytologically characterized by possession of holokinetic chromosomes and a prereductional type of meiosis. So far, only seven species from the Old World and five species from the Neotropical Region have been cytogenetically described. In the present report we compare the male meiosis from both natural and inbred populations of Dysdercus chaquensis Freiberg, 1948. Our results demonstrated that even though both populations share the same diploid chromosome number, the presence of a diffuse stage was found to be committed to the wild population of the species. Furthermore, the diffuse stage was found in a high frequency in all analysed wild specimens. indicating the long duration of this period among the regular meiosis of D. chaquensis. Taking into account that the diffuse stage is connected with an intense and long period of cellular growth, and with an important transcriptional activity, the absence of this stage in all the inbred specimens of D. chaquensis could be related with the lack of unfavourable physiological conditions due to the environmental uniformity along seven years of inbreeding.

Heteroptera↗
PubMed2003

Cytochrome P450 CYP6X1 cDNAs and mRNA expression levels in three strains of the tarnished plant bug Lygus lineolaris (Heteroptera: Miridae) having different susceptibilities to pyrethroid insecticide.

Three cDNAs, cloned from both pyrethroid-susceptible and -resistant strains of Lygus lineolaris, contained a 1548 nucleotide open reading frame encoding a 516 amino acid residue protein. Predicted cytochrome P450s from cDNAs were classified as the first three new members of subfamily CYP6X, CYP6X1v1 for a susceptible strain and CYP6X1v2 and CYP6X1v3 for two resistant strains. Putative cytochrome P450 CYP6X1s from L. lineolaris were highly similar (up to 42% amino acid sequence identity) to several insect CYP6s that are responsible for reduced sensitivity to pyrethroid insecticides. A total of twenty-six nucleotide substitutions were revealed between cDNAs of susceptible and resistant strains. Two nucleotide substitutions resulted in amino acid changes, Asp373 to Ala373 and Ser487 to Ala487, between susceptible and resistant strains. The resistant laboratory strain contained 2.1-fold higher cytochrome P450 mRNA per microgram total RNA than the susceptible laboratory strain. Topical treatment with 10 ng permethrin elevated cytochrome P450 mRNA levels by approximately 2-fold. The results of this study indicated that cytochrome P450 gene mutation, coupled with up-regulation, was present only in the pyrethroid resistant strains, and was possibly related to resistance development in the tarnished plant bug.

Alleles↗
PubMed2006

Controlling Lygus plant bugs (Heteroptera: Miridae) with European Peristenus relictus (Hymenoptera: Braconidae) in Canada--risky or not?

The European Peristenus relictus Loan (syn. P. stygicus) has been considered for biological control of Lygus plant bugs native to Canada. Laboratory and field studies were conducted in the area of origin to evaluate the host specificity of P. relictus. Laboratory choice and no-choice tests demonstrated that P. relictus attacked all non-target species offered (fundamental host range). However, closely related non-target mirids (tribe Mirini) were generally well accepted by P. relictus, while hosts from the tribe Stenodemini were less frequently attacked and less suitable for parasitoid development. To validate the laboratory results, a thorough examination of the parasitoid complex of common mirids in Europe was conducted to determine which non-target species may serve as alternative hosts for P. relictus in a natural situation (ecological host range). When comparing both approaches, the fundamental host range of P. relictus matched its ecological host range. In addition to three Lygus species, the ecological host range of P. relictus in the area of Schleswig-Holstein, northern Germany, contains at least 16 non-target species, including hosts belonging to the subfamilies Mirinae, Phylinae and Bryocorinae. A broad ecological and fundamental host range suggests that P. relictus is a generalist; however, P. relictus was not the primary contributor to parasitism of most non-target hosts studied. Although P. relictus is assumed to be of minor importance for regulating non-target populations in the area of investigation, the results of the present study indicate that P. relictus has the potential to use non-target host populations for reproduction.

Animals↗
PubMed2006

Morphology and preliminary enzyme characterization of the salivary glands from the predatory bug Podisus nigrispinus (Heteroptera: Pentatomidae).

Podisus nigrispinus (Dallas) is a common predator in agricultural and natural systems in Neotropical America. Its feeding strategy involves extra-oral digestion and to better understand this process its salivary glands were extracted and subjected to morphological and preliminary enzyme characterization. The salivary glands of P. nigrispinus are formed by a pair of main and accessory gland complexes. The main salivary glands are further divided into an anterior and a posterior lobe. The compartmentalization of the salivary gland complex is likely to be important for the production, activation and release of the digestive enzymes used in the extra-oral digestion of prey items. Proteases and lipase, important digestive enzymes involved in zoophagy, were detected in the salivary glands of P. nigrispinus. The prevailing trypsin-like protease activity was characterized by using the serine-protease substrate N-alpha-benzoyl-L-Arg-p-nitroanilidine (L-BApNA) and the trypsin inhibitors tosyl-L-lysine chloromethyl ketone (TLCK) and benzamidine. The KM value obtained for trypsin-like activity was 1.57 mm and the different peaks of optimum pH and temperature activity suggest the presence of multiple forms of this enzyme in P. nigrispinus. Detection of amylase activity in the salivary glands of this predator suggests its ability to digest starch and obtain nutrients from plants, which may have adaptative value under prey scarcity.

Amylases↗
PubMed2003

Development of a bioassay system for the predator, Xylocoris flavipes (Heteroptera: Anthocoridae), and its use in subchronic toxicity/pathogenicity studies of Beauveria bassiana strain GHA.

Microbial biocontrol agents are useful commercially only if they do not harm other natural biocontrol organisms, at recommended use rate in the environment where the microorganism is being used. To test the hypothesis that the predatory warehouse pirate bug, Xylocoris flavipes (Reuter), is not adversely affected by the entomopathogen, Beauveria bassiana (Balsamo) Vuillemin strain GHA, we developed a method using individually confined bugs during a 10-d feeding regime. Three concentrations of a conidial suspension were applied to assay surfaces (filter paper disks) to achieve 2.7 x 10(7), 2.6 x 10(6), and 2.6 x 10(5) conidia per cm2, representing 100X, 10X, and a field rate of 2.6 x 10(13) conidia per ha (+/-10%), respectively. Fifth instar X. flavipes suffered 0% infection at the field rate when confined to treated filter paper for 10 d (16% and 42% infection, respectively, at 10X and 100X the field rate). Second instar migratory grasshoppers, Melanoplus sanguinipes (F.), exposed to the same doses suffered 97, 92, and 100% mortality at the three respective doses 10 d after exposure. These data indicate that B. bassiana can be used safely at recommended application levels without significant effect on fifth instar populations of X. flavipes.

Animals↗
PubMed2003

Thermal requirements for the development and reproduction of Nysius huttoni White (Heteroptera: Lygaeidae).

Nysius huttoni White is an economically important pest of wheat and brassica crops in New Zealand. Because of its frequent presence in export fruit packages, it is also considered an important quarantine pest to countries that trade with New Zealand. To provide critical information for the pest risk analysis, forecast and management of N. huttoni, we investigated the effect of five consistent temperatures (10, 15, 20, 25, and 30 degrees C) on its development, survival and reproduction. At 10 degrees C both eggs and nymphs did not develop but the latter grew. Nymphs could survive 10 degrees C for >1.5 mo, with the fifth instar nymphs surviving for up to 145 d. Adults could live for at least 100 d at this temperature. This species could not complete its lifecycle at or below 15 degrees C. Between 15 and 30 degrees C, fifth instar stage was significantly longer than other nymphal stages. Egg hatch rate and total survival rate for all stages were significantly higher at 20 degrees C than at other test temperatures. The developmental rate of different life stages increased linearly with the increase of temperatures from 15 to 30 degrees C. The estimated low temperature threshold for the completion of lifecycle was 11.9 degrees C, and that for mating and oviposition was 12.3 and 16.8 degrees C, respectively. The thermal requirement for completing a life cycle of N. huttoni was 625 DD. The time needed for completing a life cycle was similar for both sexes. Temperature had little effect on adult body weight and sex ratio. Implications of the above findings are discussed.

Animals↗
PubMed2003

Resistance to greenbug (Heteroptera: Aphididae) biotype I in Aegilops tauschii synthetic wheats.

The greenbug, Schizaphis graminum (Rondani), is a major pest of wheat in North America, reducing U.S. wheat production by 60 to 100 million dollars each year. In this research, 149 wheat lines containing genes from Aegilops tauschii (Coss.) Schmal. were evaluated for resistance to greenbug biotype I. More than 50% of the lines sustained moderate foliar chlorosis from greenbug feeding, and approximately one third of all the lines were highly resistant. All lines with chlorosis scores similar to the resistant control 'Largo' expressed high levels of antibiosis, producing greenbug populations with mean weights ranging from 0.05 to 11.8 mg. There was no significant difference between greenbug weights on these lines and those reared on 'Largo', but the mean weight of individuals reared on the susceptible control 'Thunderbird' was significantly greater than those reared on 'Largo' or any of the test lines. The mean population size of greenbugs produced on plants of each line was significantly correlated with mean greenbug weight. Tolerance was not evident in any of the lines examined, but was unexpectedly apparent in 'Thunderbird' at a level similar to that in the tolerant control cultivar 'Largo'.

Heteroptera↗