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Differential expression of the vitellogenin gene in the spruce terminal weevil feeding on resistant versus susceptible host trees.

Reproductive maturation and oviposition of the spruce terminal weevil (Pissodes strobi, Peck) are inhibited by resistant Sitka spruce (Picea sitchensis (Bong) Carr.). Vitellogenin is an egg yolk protein precursor which accumulates during the maturation of eggs. We describe the molecular cloning and characterization of a fragment of the vitellogenin gene from the spruce terminal weevil. The DNA sequence of this fragment has high identity to vitellogenin sequences from other insects and less to nematode and vertebrate vitellogenins. It hybridizes on Northern blots to a single 6.0 kb mRNA that is expressed only in females, and only after they have commenced reproductive development. Vitellogenin gene expression is induced by treatment with juvenile hormone, and is differentially regulated in insects feeding on resistant or susceptible trees. The effects of resistance on vitellogenin gene expression were studied to confirm our earlier findings and to provide a sensitive bioassay for identifying resistance factors.

Amino Acid Sequence↗

Suitability of two root-mining weevils for the biological control of scentless chamomile, Tripleurospermum perforatum, with special regard to potential non-target effects.

The biology and host range of the two root-mining weevils Diplapion confluens Kirby and Coryssomerus capucinus (Beck), two potential agents for the biological control of scentless chamomile Tripleurospermum perforatum (Mérat) Laínz, were studied in the field in southern Germany and eastern Austria, and in a common garden and under laboratory conditions in Delémont, Switzerland from 1993 to 1999. Both weevils were univoltine, and females started to lay eggs in early spring. Diplapion confluens had three and C. capucinus five instars. Larvae of both species were found in the field from mid-April until the end of July; later instars preferentially fed in the vascular cylinder of the shoot base, root crown or root. Although larvae of both species occupy the same temporal and spatial niche within their host plants, they occurred at all investigated field sites together, and showed a similar distribution within sites. No negative or positive interspecific association was detected. Host-specificity tests including no-choice, single-choice, and multiple-choice tests under confined conditions, as well as tests under field conditions with natural and augmented insect densities revealed that both herbivores were specific to plant species in the tribe Anthemideae. However, their development to mature larva or adult on several cultivated plants, as well as on one plant species native to North America, rendered them unsuitable for field release in North America. It was concluded that to investigate non-target effects reliably, host-specificity tests with biological control agents should be carried out under a variety of conditions, particularly with augmented insect densities, as are expected to occur naturally after release.

Animals↗

Elaeolenchus parthenonema n. g., n. sp. (Nematoda: Sphaerularioidea: Anandranematidae n. fam.) parasitic in the palm-pollinating weevil Elaeidobius kamerunicus Faust, with a phylogenetic synopsis of the Sphaerularioidea Lubbock, 1861.

A new nematode, Elaeolenchus parthenonema n. g., n. sp., is described from the palm-pollinating weevil Elaeidobius kamerunicus Faust. The new genus is placed in the Anandranematidae n. fam., which, together with the genus Anandranema Poinar et al., 1993, is characterised by nematodes having only a single autotokous generation in the insect host. This is the first report of a member of this superfamily reproducing only parthenogenetically. The development of E. parthenonema and its effect on the weevil host is discussed, along with a phylogenetic synopsis of the families of the Sphaerularioidea Lubbock 1861. The Beddingiidae n. fam. is proposed for Beddingia Blinova & Korenchenko, 1986, comprising the original Deladenus parasites of Hymenoptera that possess both free-living and parasitic amphimictic generations in their life-cycles. This family is considered to have the most primitive type of development in the superfamily.

Animals↗

Karyotypic analysis of the cotton boll weevil, Anthonomus grandis Boheman.

The diploid chromosome number of the cotton boll weevil, Anthonomus grandis Boheman, is 44. Both C- and N-banding techniques of mitotic cells demonstrated constitutive heterochromatin in the p arm of the eight largest chromosomes, the p arm of the X chromosome, and the centromeric region of autosomal groups A-D. Neither the y nor the group E autosomes appeared to contain constitutive heterochromatin. Supernumerary chromosomes were not found in the boll weevil. Restriction endonuclease banding of primary spermatocytes revealed a rod-shaped Xy tetrad in which the X and y were terminally associated. The p arm of the large, submetacentric X was C-band positive. While two of the autosomal tetrads were typically ring-shaped in primary spermatocytes, the remaining 19 autosomal tetrads were rod-shaped.

Animals↗

Quantitative GC analysis of secondary alcohol pheromones: determination of release rate of red palm weevil, Rhynchophorus ferrugineus, pheromone from lures.

Aliphatic secondary alcohols are components of several aggregation pheromones of important beetle and weevil pests. Some of these pheromones are used frequently for the monitoring and mass trapping of the relevant insects. We encountered severe difficulties in direct GC quantitative analysis of these compounds. Therefore, we developed a simple GC analysis of secondary alcohols convening them to trifluoroacetyl derivatives and using secondary alcohol acetates as internal standards. This method was applied for the quantitative analysis of several secondary alcohols, including the aggregation pheromone components of the almond bark beetle and the red palm weevil. The release rate of the latter pheromone from commercial lures was also determined.

Alcohols↗

Bruchins, plant mitogens from weevils: structural requirements for activity.

Bruchins are 3-hydroxypropanoate esters of long-chain alpha,omega-diols from pea weevils and cowpea weevils that have been shown to initiate callus formation on pea pods at extremely low application rates. Synthetic analogs have been prepared and examined to evaluate structural requirements for inducing this unusual neoplastic response. Chain length (optimum length C22-C24) is important, whereas unsaturation within the chain is relatively unimportant. Difunctionality is required for maximum activity, but the alpha,omega-diols themselves are inactive. Most critical is the ester portion(s) of the molecules; 3-hydroxypropanoate esters are far more active than any analogs examined.

Animals↗

Amplification and sequencing of DNA from a 120-135-million-year-old weevil.

DNA has been successfully isolated from both fossilized plant and animal tissues. The oldest material, dated as 25-40 million years old (Tertiary), was obtained from amber-entombed bees and termites. Tissues from both these insects yielded DNA of good quality, which could be amplified by the polymerase chain reaction (PCR) and subsequently sequenced, including the genes encoding 18S ribosomal RNA and 16S rRNA. We report here the extraction of DNA from a 120-135-million-year-old weevil (Nemonychidae, Coleoptera) found in Lebanese amber, PCR amplification of segments of the 18S rRNA gene and the internal transcribed spacer, and the corresponding nucleotide sequences of their 315- and 226-base-pair fragments, respectively. These sequences were used for preliminary phylogenetic analysis of the nemonychid's sequence with three extant coleopterans: Lecontellus pinicola (Nemonychidae), Hypera brunneipennis (Curculionidae) and the mealworm Tenebrio molitor (Tenebrionidae), and two extant dipterans: the fruitfly Drosophila melanogaster (Drosophilidae) and mosquito Aedes albopictus (Culicidae) for the purpose of ascertaining the origin of the extracted and amplified DNA. The results revealed that the PCR-amplified material is that of the extinct nemonychid weevil. This represents the oldest fossil DNA ever extracted and sequenced, extending by 80 million years the age of any previously reported DNA.

Animals↗

Characterization of a high-affinity binding site for the pea albumin 1b entomotoxin in the weevil Sitophilus.

The toxicity of the pea albumin 1b (PA1b), a 37 amino-acid peptide extracted from pea seeds, for cereal weevils (Sitophilus oryzae, Sitophilus granarius and Sitophilus zeamais) was recently discovered. The mechanism of action of this new entomotoxin is still unknown and potentially involves a target protein in the insect tissues. This work describes the characterization of a high-affinity binding site for PA1b in a microsomal fraction of Sitophilus spp. extracts. Purified PA1b was labeled to a high specific radioactivity (c. 900 Ci.mmol-1) using 125I, and the iodinated ligand was found to be biologically active. Binding of this ligand to the microsomal fraction of S. oryzae extract was found to be saturable and reversible, with an affinity (Kd) of 2.6 nm, and a high maximal binding capacity (Bmax) of 40 pmol.mg-1 of protein. A binding site displaying similar characteristics was detectable in the five susceptible weevils strains tested, as well as in the pea aphid or in the fruit fly. However, no binding activity was detectable in extracts from four S. oryzae strains previously shown to be resistant to the toxin through a recessive monogenic mechanism. Therefore, we suggest that this binding site might be involved in the mechanism of action of PA1b.

Albumins↗

The influence of plant age on tolerance of rice to injury by the rice water weevil, Lissorhoptrus oryzophilus (Coleoptera: Curculionidae).

For most plant species, tolerance to many types of herbivory increases as plants age, but the applicability of this pattern to root herbivory has not been tested. Injury to roots of rice plants by larvae of the rice water weevil, Lissorhoptrus oryzophilus Kuschel, causes severe reductions in yields in the United States. It is generally thought that young rice plants, because their root systems are smaller, are less tolerant than older plants of root feeding by L. oryzophilus. Field experiments were conducted to test this hypothesis. Plots of rice (4.7 to 6.5 m2) were established and subjected to natural infestations of L. oryzophilus larvae. A soil insecticide was applied to plots at different times during the tillering phase of rice in order to manipulate the timing of weevil infestation. The impact of these treatments (timings of insecticide applications) was assessed by comparing relationships between yield loss and larval pressure for each treatment using analysis of covariance. Yield losses ranged from 13% to over 40% in plots not treated with insecticide. Patterns of yield losses from plots treated with insecticide at different times were best explained by the hypothesis that yield loss is determined both by the age of plants infested and by the size of larvae infesting plants. Young plants appear to be less tolerant than older plants, and feeding by large larvae appears to be more deleterious than feeding by smaller larvae. Management practices that delay infestation of rice by L. oryzophilus until plants are older may be an important component of management programmes for this pest.

Animals↗

The influence of habitat and altitude on oxygen uptake in sub-Antarctic weevils.

Whole-organism oxygen uptake rate and its temperature sensitivity (determined using regression analyses and estimates of Q10) were examined in six closely related weevil species (Palirhoeus eatoni, Bothrometopus randi, Bothrometopus elongatus, Bothrometopus parvulus, Ectemnorhinus similis, and Ectemnorhinus marioni) from sub-Antarctic Marion Island over a short time period and using the same methods. Bothrometopus elongatus, B. parvulus, and the two Ectemnorhinus species have populations at both high and low elevations, and pairwise comparisons of these species were made. Regressions of the log of oxygen uptake rate on temperature and Q10 values revealed that the Ectemnorhinus species have a significantly greater thermal sensitivity than do species in the genera Bothrometopus and Palirhoeus. This may be considered an adaptation of the former to their moist lowland habitats and the requirements of angiosperm-feeding in E. similis. It is argued that elevated oxygen uptake rates and reduced slopes of the regression of the log of oxygen uptake rate on temperature in species and populations from high altitudes compared with those from low elevations provide evidence for metabolic cold adaptation. In addition, it seems likely that elevated oxygen uptake rates and their reduced thermal sensitivity within the genera Bothrometopus and Palirhoeus are an adaptation to the cold Neogene environments they evolved in. However, because data on the more basal taxa in the Ectemnorhinus group of weevils are not available, this temperature compensation could not be attributed conclusively to adaptation. Q10's of the lowland populations of all the species were negatively correlated with body water content, and it is suggested that the low temperature sensitivity of metabolism in P. eatoni and the Bothrometopus species may also be due to constraints imposed on them by their dry habitats.

Altitude↗

Mitochondrial DNA in the bark weevils: size, structure and heteroplasmy.

Mitochondrial DNA of higher animals has been described as an example of extreme efficiency in genome structure and function. Where exceptionally large size molecules have been found (greater than 20 kb), most have occurred as rare variants within a species, suggesting that these variants arise infrequently and do not persist for long periods in evolutionary time. In contrast, all individuals of at least three species of bark weevil (Curculionidae: Pissodes) possess a mitochondrial genome of unusually large size (30-36 kb). The molecule owes its large size to a dramatically enlarged A + T-rich region (9-13 kb). Gene content and order outside of this region appear to be identical to that found in Drosophila. A series of 0.8-2.0-kb repeated sequences occur adjacent to the large A + T rich region and have perhaps played a role in the generation of the large size as well as an unprecedented frequency of size variant heteroplasmy. Every weevil sampled in all three species (n = 219) exhibits anywhere from two to five distinct size classes of mtDNA. The persistence of this large amount of size polymorphism through two speciation events combined with the abundant size variation within individuals suggests that these molecules may not be subject to strong selection for small overall size and efficiency of replication. This pattern of variation contrasts strongly with the conservation of gene content and arrangement in the coding region of the molecule.

Animals↗

Sexual reproduction in the white pine weevil (Pissodes strobi [Peck] [Coleoptera: Curculionidae]): implications for population genetic diversity.

Controlled mating experiments in the white pine weevil (Pissodes strobi [Peck]) indicated that female weevils either stored sperm or fertilized eggs from one season to the next, and were able to colonize Sitka spruce (Picea sitchensis [Bong.] Carr.) trees without additional mating events. This was interpreted as being beneficial for the insect, in that population establishment in a new habitat could be initiated by dispersing previously mated females without participation of the male. This makes colonization and population/outbreak development more likely as it reduces the need for mate searching in the second season. Paternity identification, based on microsatellite molecular markers, established that the progeny produced in year 2 by females mated only in year 1, were often fathered by more than one male. Multiple paternity, coupled with a lack of parthenogenesis, which was also demonstrated herein, may help to account for the high degree of genetic diversity evidenced in this species.

Animals↗

Genetic structuring of boll weevil populations in the US based on RAPD markers.

Abstract Randomly amplified polymorphic DNA (RAPD) analysis was performed to infer the magnitude and pattern of genetic differentiation among boll weevil populations from eighteen locations across eight US states and north-east Mexico. Sixty-seven reproducible bands from six random primers were analysed for genetic variation within and between weevil populations. Genetic and geographical distances among all populations were positively correlated, reflecting a pattern of isolation by distance within a larger metapopulation. Gene flow between south-central, western and eastern regions is limited, but migration between locations within regions appears to be relatively frequent up to distances of approximately 300-400 km. However, estimates of effective migration were much lower than those estimated from mtDNA-RFLP data reported previously.

Animals↗

Transfection of cultured cells of the cotton boll weevil, Anthonomus grandis, with a heat-shock-promoter-chloramphenicol-acetyltransferase construct.

Expression of heat shock proteins (hsp) in the BRL-AG-3C cell line from the cotton boll weevil was examined. It was determined that the maximal expression of endogenous hsp occurred at 41 degrees C. Various transfection methods were then compared using this cell line in conjunction with a transiently expressed bacterial gene marker (chloramphenicol acetyltransferase) which was under the control of the Drosophila hsp 70 gene promoter. The cationic lipid preparation Lipofectin was found to be very efficient at transfecting the boll weevil cells. Polylysine and 20-hydroxyecdysone-conjugated polylysine were moderately effective, whereas polybrene and electroporation, under the conditions reported herein, were ineffective at transfecting this cell line.

Animals↗

(-)-olivil and (+)-1-acetoxypinoresinol from the olive tree (Olea europaea Linne; Oleaceae) as feeding stimulants of the olive weevil (Dyscerus perforatus).

Guided by a feeding stimulant activity test on the olive weevil (Dyscerus perforatus), two compounds that showed potent feeding stimulant activity were isolated from the olive tree (Olea europaea). Based on their spectral data and a literature survey, they were identified as (-)-olivil (1) and (+ )-1-acetoxypinoresinol (2). The activities of these minor lignans were significantly higher for the female than for the male weevil.

Animals↗

Effective biological control of Salvinia molesta in the Senegal River by means of the weevil Cyrtobagous salviniae.

The invasion of Salvinia molesta in the Lower Senegal River Delta in Mauritania and Senegal in 1999 posed a serious threat to the socioeconomic conditions of the local people as well as to wetland biodiversity. Eventually, an effective biological control of S. molesta was obtained by means of the weevil Cyrtobagous salviniae, which was introduced in the river in Senegal and Mauritania in May 2000 and in Senegal in April 2001. In October 2001, it became apparent that the weevils were doing a magnificent job. The color of the plants was turning from green to dark-brown or black, and subsequently the plants started to sink to the bottom. Detailed monitoring of the dispersal of C. salviniae in November-December 2001 confirmed the visual observations of the outcome of biological control. In April 2002, it could be concluded that S. molesta was no longer a problem in the Senegal River.

Animals↗

Mortality of life stages of cowpea weevil (Coleoptera: Bruchidae) exposed to low pressure at different temperatures.

Previous studies have shown that low pressure creates a low oxygen controlled atmosphere that can kill stored-product insects. The current study was conducted to determine the mortality of life stages of the cowpea weevil, Callosbruchus maculatus (F.) (Coleoptera: Bruchidae), exposed to different low pressures and temperatures for various exposure periods. The adults were the most susceptible life stage to low pressure; 99% mortality was achieved within 0.8 h at 32.5 mmHg, 30 degrees C. The pupae were the most tolerant life stage to low pressure, requiring exposure periods between 28.98 and 153.20 h at temperatures of 20-35 degrees C to achieve 99% mortality. Mortality increased with exposure time and also with increasing temperature in all life stages. Early stage eggs (3 h old) and late stage eggs (48 h old) experienced higher mortality (values for LT99 of 42.331 and 46.652 h, respectively) compared with intermediate aged eggs (24 h old; LT99 of 74.735 h) under the same conditions of low pressure and temperature. Dried beans, including cowpea, Vigna unguiculata (Walp.), are currently protected with fumigants. Application of low pressure as a pest management tool represents a potential nonchemical alternative to fumigants such as methyl bromide and phosphine for controlling the cowpea weevil and related bruchids.

Animals↗

Emergence trap developed to capture adult large pine weevil Hylobius abietis (Coleoptera: Curculionidae) and its parasite Bracon hylobii (Hymenoptera: Braconidae).

A novel type of emergence trap capable of capturing and separating 'live' insect catches is described. The trap was shown to be 96% efficient at capturing newly emergent adult Hylobius abietis Linnaeus on bare ground and at least 82% efficient over stumps on a weedy Sitka spruce clearfell. The trap was more than 80% efficient at capturing Bracon hylobii Ratzeburg, the most commonly found parasite of H. abietis. It was also shown to be effective at capturing adult H. abietis of unknown age (98%), indicating that it could also be used to trap out H. abietis from known areas to estimate on-site overwintering densities. Fifty-four percent of newly emergent weevils were captured within 12 h of release on bare mineral soils. Forty-two percent of unknown age weevils and 52% of parasites were captured within 1 h of their release within the trap. The rapid rates of capture mean that when traps are checked frequently they can be used to reflect accurately temporal patterns of emergence. Its potential for use in control programmes and ecological studies is discussed.

Animals↗