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PubMed2006

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.

Animals↗

PubMed2006

Boll weevil (Coleoptera: Curculionidae) damage to cotton bolls under standard and proactive spraying.

In some parts of the boll weevil's, Anthonomus grandis grandis (Boheman) (Coleoptera: Curculionidae), distribution from the United States to Argentina, insecticides are applied after cut-out (end of square production) when bolls are the predominant stage of fruiting body. This study demonstrates that the standard spray regime in southern Texas, which involves insecticide applications after cut-out, did not result in more bolls than a nonsprayed control. An alternative "proactive" spray regime focusing on protecting large squares before cut-out resulted in 1.9- to 2.5-fold more bolls in the lower half of the canopies than the control. At one of two experimental field locations, the percentage of damaged boll carpels was 3-fold greater in the standard spray regime's lower canopy than in the proactive spray regime, and the percentage in the control was 1.6-fold greater than in the standard regime. At both experimental field locations, the upper canopy control had 2.1- to 2.3-fold greater percentages of carpel damage than the proactive spray regime. The standard spray regime resulted in 2.3-fold greater percentage of carpel damage than the proactive regime. In the control and the standard spray regime, percentages of upper canopy nondamaged bolls were mostly lower than or not different from percentages of bolls with one, two, three, or all four carpels damaged, but in the proactive regime, percentage of nondamaged bolls in the upper canopy was greater than percentages of bolls with one or more damaged carpels. Reasons for the ineffectiveness of the standard spray regime and the benefits observed in the proactive approach are discussed.

Animals↗

PubMed2007

Oogenesis in summer females of the rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), in southern Zhejiang, China.

The rice water weevil, Lissorhoptrus oryzophilus Kuschel, has two generations in southern Zhejiang, China. To determine oogenesis in first-generation females (summer females) and its relations to temperature, females were collected from a rice field in early and mid-July and reared on young rice plants at 28, 31 and 34 degrees C in the laboratory. Percentage of females having oocytes, number of oocytes of different stages (stage-I, from early previtellogenesis to middle vitellogenesis; stage-II, late vitellogenesis; and mature-oocyte stage), and length of ovarioles were determined every 10 d of feeding. At each temperature, oogenesis took place in over 40% of females after 20~40 d of feeding, but only 0.0~3.3 stage-I, 0.0~0.8 stage-II and 0.0~1.1 mature oocytes were observed at each observation date. Temperature had significant effect on number of stage-I oocytes but not on number of stage-II and mature oocytes in early July females; temperature had no significant effect on number of oocytes of either stage in mid-July females. Conclusively, in southern Zhejiang, summer L. oryzophilus females have great potential to become reproductive on rice, but their oogenesis activity is very low, with the overall procedures little affected by temperature.

Animals↗

PubMed2005

Purification of a 6.5 kDa protease inhibitor from Amazon Inga umbratica seeds effective against serine proteases of the boll weevil Anthonomus grandis.

A 6.5 kDa serine protease inhibitor was purified by anion-exchange chromatography from the crude extract of the Inga umbratica seeds, containing inhibitor isoforms ranging from 6.3 to 6.7 kDa and protease inhibitors of approximately 19 kDa. The purified protein was characterized as a potent inhibitor against trypsin and chymotrypsin and it was named I. umbratica trypsin and chymotrypsin inhibitor (IUTCI). MALDI-TOF spectra of the IUTCI, in the presence of DTT, showed six disulfide bonds content, suggesting that this inhibitor belongs to Bowman-Birk family. The circular dichroism spectroscopy indicates that IUTCI is predominantly formed by unordered and beta-sheet secondary structure. It was also characterized, by fluorescence spectroscopy, as a stable protein at range of pH from 5.0 to 7.0. Moreover, this inhibitor at concentration of 75 microM presented a remarkable inhibitory activity (60%) against digestive serine proteases from boll weevil Anthonomus grandis, an important economical cotton pest.

Chromatography, Ion Exchange↗

PubMedDate not supplied

[Phylogenetic reconstruction of weevil superfamily Curculionoidea (Coleoptera) using the SYNAP method].

Phylogenetic relationships within the superfamily Curculionoidea were reconstructed. Autapomorphies of the superfamily Curculionoidea include more or less pronounced snout, clubbed antennae, and partially sclerotized or completely membranous male tergite 9. Weevil families can be divided into three groups. The first group includes the most primitive family Nemonychidae. The second group includes nine families (Anthribidae, Belidae, Oxycorynidae, Eccoptarthridae, Allocorynidae, Rhynchitidae, Attelabidae, Ithyceridae, and Brentidae). The third ("higher") group includes six families (Brachyceridae. Cryptolaryngidae, Dryophthoridae, Curculionidae, Scolytidae, and Platypodidae).

Animals↗

PubMed2006

Effects of carbon dioxide on the searching behaviour of the root-feeding clover weevil Sitona lepidus (Coleoptera: Curculionidae).

The respiratory emission of CO2 from roots is frequently proposed as an attractant that allows soil-dwelling insects to locate host plant roots, but this role has recently become less certain. CO2 is emitted from many sources other than roots, so does not necessarily indicate the presence of host plants, and because of the high density of roots in the upper soil layers, spatial gradients may not always be perceptible by soil-dwelling insects. The role of CO2 in host location was investigated using the clover root weevil Sitona lepidus Gyllenhall and its host plant white clover (Trifolium repens L.) as a model system. Rhizochamber experiments showed that CO2 concentrations were approximately 1000 ppm around the roots of white clover, but significantly decreased with increasing distance from roots. In behavioural experiments, no evidence was found for any attraction by S. lepidus larvae to point emissions of CO2, regardless of emission rates. Fewer than 15% of larvae were attracted to point emissions of CO2, compared with a control response of 17%. However, fractal analysis of movement paths in constant CO2 concentrations demonstrated that searching by S. lepidus larvae significantly intensified when they experienced CO2 concentrations similar to those found around the roots of white clover (i.e. 1000 ppm). It is suggested that respiratory emissions of CO2 may act as a 'search trigger' for S. lepidus, whereby it induces larvae to search a smaller area more intensively, in order to detect location cues that are more specific to their host plant.

Animals↗

PubMed1996

Response of digestive cysteine proteinases from the Colorado potato beetle (Leptinotarsa decemlineata) and the black vine weevil (Otiorynchus sulcatus) to a recombinant form of human stefin A.

The effects of the cystatins, human stefin A (HSA) and oryzacystatin I (OCI) on digestive cysteine proteinases of the Colorado potato beetle (CPB), Leptinotarsa decemlineata, and the black vine weevil (BVW), Otiorynchus sulcatus, were assessed using complementary inhibition assays, cystatin-affinity chromatography, and recombinant forms of the two inhibitors. For both insects, either HSA and OCI used in excess (10 or 20 microM) caused partial and stable inhibition of total proteolytic (azocaseinase) activity, but unlike for OCI the HSA-mediated inhibitions were significantly increased when the inhibitor was used in large excess (100 microM). As demonstrated by complementary inhibition assays, this two-step inhibition of the insect proteases by HSA was due to the differential inactivation of two distinct cysteine proteinase populations in either insect extracts, the rapidly (strongly) inhibited population corresponding to the OCI-sensitive fraction. After removing the cystatin-sensitive proteinases from CPB and BVW midgut extracts using OCI- (or HSA-) affinity chromatography, the effects of the insect "non-target" proteases on the structural integrity of the two cystatins were assessed. While OCI remained essentially stable, HSA was subjected to hydrolysis without the accumulation of detectable stable intermediates, suggesting the presence of multiple exposed cleavage sites sensitive to the action of the insect proteases on this cystatin. This apparent susceptibility of HSA to proteolytic cleavage may partially explain its low efficiency to inactivate the insect OCI-insensitive cysteine proteinases when not used in large excess. It could also have major implications when planning the use of cystatin-expressing transgenic plants for the control of coleopteran pests.

Animals↗

PubMed1997

Characterization and partial cloning of ecdysteroid receptor from a cotton boll weevil embryonic cell line.

A cell line derived from the embryos of the cotton boll weevil, Anthonomus grandis (BRL-AG-2), was used to study morphological and biochemical responses to 20-hydroxyecdysone (20E). The cells respond to 10(-6) M 20E by inhibition of cell growth and enhanced production of some secreted proteins. Crude nuclear extracts containing the ecdysteroid receptor complex proteins consisting of the ecdysteroid receptor (EcR) and ultraspiracle (USP) bound ponasterone A with a Kd of 6.1 nM. Bound radiolabeled ponasterone A was displaced by both 20E and the lepidopteran-specific non-steroidal ecdysteroid agonist, RH-5992, with 41- and about 1,900-fold higher Kd values, respectively. We identified the ecdysteroid receptor components in this cell line, using monoclonal antibodies against the Drosophila ecdysteroid receptor (DmEcR) and ultraspiracle (DmUSP) proteins. A predominant band of about 70 kDa was-detected with anti-EcR, and multiple bands ranging from 50-55 kDa were detected with anti-USP in the A. grandis extracts. Using degenerate primer RT-PCR, we isolated a 450 bp cDNA fragment of the putative AgEcR. Using this fragment as a probe, we identified a large mRNA of ca. 10 kb by Northern blot analysis. These results demonstrate the usefulness of this cell line for the study of ecdysone response and the isolation of the receptor components in A. grandis.

Animals↗

PubMed2003

Proteolytic gut activities in the rice water weevil, Lissorhoptrus brevirostris Suffrian (Coleoptera: Curculionidae).

Digestive endoprotease activities of the rice water weevil, Lissorhoptrus brevirostris Suffrian (Coleoptera: Curculionidae), were characterized based on the ability of gut extracts to hydrolyze specific synthetic substrates, optimal pH, and hydrolysis sensitivity to protease inhibitors. Larvae of this species were found to use a complex proteolytic system that includes cathepsin D-, cathepsin B-, trypsin-, and chymotrypsin-like activities. Trypsin-like activity was evenly distributed among the anterior, middle, and posterior portions of the gut, whereas cathepsin B- and cathepsin D-like activities were mainly located in the anterior and middle sections, and the chymotrypsin-like activity was highest in the middle and posterior sections. Gelatin-containing native-PAGE gels indicated the presence of several aspartyl, cysteine, and serine protease forms and confirmed the spatial organization of the proteolytic digestive process.

Animals↗

PubMed1992

Storage proteins of the larval root weevil Diaprepes abbreviatus (Coleoptera: Curculionidae): riboflavin binding and subunit isolation.

Proteins present at high concentrations in hemolymph of the larval weevil Diaprepes abbreviatus were previously shown to bind a synthetic coumarin, 7-amino-3-phenyl coumarin (coumarin-10). One of the two native proteins previously identified (protein I) is now shown to separate into two distinct bands (proteins Ia and Ib) using native gradient pore-limiting electrophoresis. The high concentration of proteins Ia, Ib, and II in larval hemolymph, their disappearance from hemolymph upon pupation, and an apparent hexameric structure shown by chemical crosslinking identify them as hexameric storage proteins (hexamerins). At least one chromatographic form of Ib isolated by anion exchange HPLC is now shown to bind riboflavin (Rb). Binding was also demonstrated by quenching of Rb fluorescence by a partially isolated mixture of the storage proteins. Lipophorin did not quench Rb fluorescence. Rb was heat-extracted from whole hemolymph and identified by its fluorescence spectra and by reverse phase HPLC with fluorescence detection. The two subunits shared by the three holoproteins have been isolated by sequential density gradient ultracentrifugation, gel permeation HPLC, and reverse phase HPLC. All three holoproteins shared the alpha subunit (M(r) 75,000), while the beta subunit (M(r) 71,000) was lacking from one of the three. Repeated passage through an anion exchange column yielded two of the three proteins (Ib and II) in homogeneous form. Chemical crosslinking with dimethylsuberimidate indicated a hexameric structure for the holoproteins. All subunits and holoproteins stained as high mannose glycoproteins when probed with biotinylated concanavalin A on PVDF membranes. The alpha subunit was high in Met, His, and Thr, and the beta subunit was high in Lys. Both were high in Pro and had approximately 16% Phe+Tyr. Sequences of the 20 N-terminal amino acid residues of each subunit showed 45-60% homology between subunits. These coleopteran proteins also showed some sequential homology but no immunological cross-reactivity with storage proteins from the lepidopterans Galleria mellonella and Heliothis virescens.

Amino Acid Sequence↗

PubMed2003

Resource assessment and clutch size in the bean weevil, Acanthoscelides obtectus.

Bean weevil (Acanthoscelides obtectus (Say)) females were found to use seeds (discrete resource patches) differentially when different sizes were offered in multiple-choice tests. Females, either as a group or as individuals, laid significantly (two to six times) more eggs on large seeds than on those of five times smaller mass. In contrast, seed shape (flattened or spherical) did not contribute to clutch-size adjustment. Thus, A obtectus females seem to measure only relative seed size when a comparison is possible. Nevertheless, females overload seeds with eggs and this can result in larval competition, so that, whereas resource size assessment and a robust egg-load adjustment indicate a trade-off between resource use and female fitness, it does not seem to provide much benefit for the progeny in stored dry beans. Several features, eg the use of oviposition markers and its consequences, may counterbalance the possible negative effects. It is assumed that, due to life cycle differences, females in the bean field may realise different fitness gains in comparison with those living in stores.

Animals↗

PubMed2004

Persistence and residual activity of an organophosphate, pirimiphos-methyl, and three IGRs, hexaflumuron, teflubenzuron and pyriproxyfen, against the cowpea weevil, Callosobruchus maculatus (Coleoptera: Bruchidae).

Three insect growth regulators (IGR), the chitin synthesis inhibitors (CSI) teflubenzuron and hexaflumuron and the juvenile hormone mimic (JHM) pyriproxyfen, as well as the organophosphate (OP) pirimiphos-methyl, were evaluated for their activity against the cowpea weevil, Callosobruchus maculatus (F), in cowpea seeds stored for up to 8 months post-treatment. The initial activity data showed that, based on LC50 level, teflubenzuron had strong ovicidal activity (LC50 = 0.056 mg kg(-1)) followed by pirimiphos-methyl (1.82 mg kg(-1)) and pyriproxyfen (91.9 mg kg(-1)). The residual activity data showed that none of the IGRs tested had strong activity when applied at 200 mg kg(-1) in reducing the oviposition rates of C maculatus at various storage intervals up to 8 months post-treatment. However, teflubenzuron reduced adult emergence (F1 progeny), achieving control ranging from 96.2% at 1 month to 94.3% at 8 months. Hexaflumuron showed a similar trend in its residual activity, ranging between 93.8% control at 1 month to 88.2% control at 8 months post-treatment. However, pyriproxyfen was more active than the CSIs tested and caused complete suppression (100% control) of adult emergence at all storage intervals. Unlike the IGRs tested, pirimiphos-methyl applied at 25 mg kg(-1) was more effective in reducing oviposition rates of C maculatus up to 8 months post-treatment. A strong reduction of adult emergence was also observed at various bimonthly intervals (98.6% control at 1 month to 91.6% control at 8 months post-treatment). The persistence of hexaflumuron and pirimiphos-methyl in cowpea seeds was also studied over a period of 8 months. The loss of hexaflumuron residue in treated cowpeas (200 mg kg(-1)) was very slow during the first month post-treatment (4.43%). At the end of 8 months, the residue level had declined significantly to 46.4% of the initial applied rate. The loss of pirimiphos-methyl residue in treated cowpeas (25 mg kg(-1)) was relatively high during the first month post-treatment (36.7%) and increased to 81.6% after 8 months.

Animals↗

PubMed1997

A molecular aspect of symbiotic interactions between the weevil Sitophilus oryzae and its endosymbiotic bacteria: over-expression of a chaperonin.

Specific proteins of symbiosis were analyzed by the comparison of two-dimensional electrophoresis protein patterns of symbiotic and aposymbiotic strains of the weevil Sitophilus oryzae. One protein was shown to be exclusively expressed in the aposymbiotic strain and three proteins, including a chaperonin, were characterized in the symbiotic strain pattern. The groE-like operon, encoding the two chaperonins groES and GroEL-like proteins of the endocytobiotes, was sequenced. It was found to be very similar to the groE operon of Escherichia coli (82% identity). In vitro and ex vivo experiments of protein labelling demonstrated that almost 40% of the endocytobiote protein synthesis ex vivo is focused on the GroEL-like protein. Finally, we showed by northern blotting that heat shock at 38 degrees C results in groEL mRNA accumulation inside the endocytobiotes. This work supports the hypothesis that chaperonins could have an essential physiological function in the maintenance of the symbiotic association.

Amino Acid Sequence↗

PubMed1977

Esterases of laboratory-reared and field-collected cotton boll weevils, Anthonomus grandis Boh.: polymorphism of adult esterases and formal genetics of esterase II.

The esterases of the cotton boll weevil were separated by polyacrylamide gel electrophoresis into four major regions. These were named Est I-IV in order of migration from anode to origin. Polymorphism was observed in all regions. The Est II region was shown to consist of no more than two bands (fast and slow). The inheritance of the fast and slow bands of Est II was demonstrated to be controlled by codominant autosomal alleles. Analysis of the gene frequency of the Est II region showed that one field population was consistent with the Hardy-Weinberg law (P = 0.995), while a second field population was not at equilibrium (P less than 0.001).

Animals↗

PubMed1986

Distribution and probable physiological role of esterases in reproductive, digestive, and fat-body tissues of the adult cotton boll weevil, Anthonomus grandis Boh.

Polyacrylamide gel electrophoresis was used to examine gut, Malpighian tube, fat-body, testes, and ovarioles tissues of the adult cotton boll weevil, Anthonomus grandis Boh. Esterases for which the inheritance has been reported previously by Terranova using whole-body homogenates were detected in dissected tissues and the probable physiological function of each allozyme is suggested. EST-1 occurs most frequently in ovarioles and female fat bodies. EST-2 is most often found in fat bodies and may be important in lipid turnover. No sex difference was observed. EST-3S is found in fat bodies and reproductive tissue, while EST-3F is always located in gut tissues, indicating that EST-3 is not controlled by a single autosomal locus with two codominant alleles as previously reported. EST-4, the most abundant esterase, can be detected in gut tissue at any age and is probably involved in digestion. EST-5 contains four allozymes which appear most frequently in testes and may be important during reproduction.

Adipose Tissue↗

PubMed1998

Molecular characterization of the principal symbiotic bacteria of the weevil Sitophilus oryzae: a peculiar G + C content of an endocytobiotic DNA.

The principal intracellular symbiotic bacteria of the cereal weevil Sitophilus oryzae were characterized using the sequence of the 16S rDNA gene (rrs gene) and G + C content analysis. Polymerase chain reaction amplification with universal eubacterial primers of the rrs gene showed a single expected sequence of 1,501 bp. Comparison of this sequence with the available database sequences placed the intracellular bacteria of S. oryzae as members of the Enterobacteriaceae family, closely related to the free-living bacteria, Erwinia herbicola and Escherichia coli, and the endocytobiotic bacteria of the tsetse fly and aphids. Moreover, by high-performance liquid chromatography, we measured the genomic G + C content of the S. oryzae principal endocytobiotes (SOPE) as 54%, while the known genomic G + C content of most intracellular bacteria is about 39.5%. Furthermore, based on the third codon position G + C content and the rrs gene G + C content, we demonstrated that most intracellular bacteria except SOPE are A + T biased irrespective of their phylogenetic position. Finally, using the hsp60 gene sequence, the codon usage of SOPE was compared with that of two phylogenetically closely related bacteria: E. coli, a free-living bacterium, and Buchnera aphidicola, the intracellular symbiotic bacteria of aphids. Taken together, these results show a peculiar and distinctly different DNA composition of SOPE with respect to the other obligate intracellular bacteria, and, combined with biological and biochemical data, they elucidate the evolution of symbiosis in S. oryzae.

Animals↗

PubMed2003

Identification of plant odours activating receptor neurones in the weevil Pissodes notatus F. (Coleoptera, Curculionidae).

Plants release complex mixtures of volatiles important in the interaction with insects and other organisms. In the search for compounds that contribute to the perception of odour quality in the weevil Pissodes notatus, single olfactory receptor neurones on the antennae were screened for sensitivity to naturally produced plant volatiles by the use of gas chromatography linked to single cell recordings. We here present 60 olfactory neurones responding to 25 of the numerous compounds released by host and non-host plants. All the neurones show high selectivity and are classified into 12 distinct types. The two most abundant types respond to alpha-pinene, beta-pinene, and 3-carene ( n=17), and to isopinocamphone and pinocamphone ( n=17), respectively. Other neurone types respond to limonene ( n=9), beta-phellandrene ( n=3), and fenchone ( n=4). Responses to beta-caryophyllene ( n=1) and to ethanol ( n=4) are also shown. Except for two pairs, the neurone types do not show overlap of the molecular receptive range. The active compounds are present in the host, Pinus pinaster, as well as in non-hosts, supporting the idea that plant odour quality is mediated by the ratio of the compounds rather than specific odorants.

Animals↗

PubMed2005

Attractive properties of an isoflavonoid found in white clover root nodules on the clover root weevil.

The clover root weevil, Sitona lepidus, frequently feeds on N2 fixing rhizobial root nodules of white clover (Trifolium repens), which may contain isoflavonoids with defensive and plant regulatory properties. This study investigated the isoflavonoids present in N2 fixing (active) root nodules, root nodules that were not fixing N2 (inactive), and roots without nodules, and tested the behavioral responses of neonatal S. lepidus larvae to aglycones of the identified compounds. Formononetin concentrations were higher in the active nodules compared with inactive nodules and roots alone. Moreover, there was a statistically significant attraction to formononetin by S. lepidus in arena experiments, whereas the other isoflavonoids were unattractive. It is suggested that S. lepidus may have become tolerant to the toxic effects of formononetin with repeated exposure, and that it may play a role in root nodule location. Such coevolutionary relationships are widely reported for aboveground insects and plants, but the present study suggests they may also occur belowground.

Animals↗