Endosymbiont phylogenesis in the dryophthoridae weevils: evidence for bacterial replacement.
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Intracellular symbiosis is widespread in the insect world where it plays an important role in evolution and adaptation. The weevil family Dryophthoridae (Curculionoidea) is of particular interest in intracellular symbiosis evolution with regard to the great economical and ecological features of these invasive insects, and the potential for comparative studies across a wide range of host plants and environments. Here, we have analyzed the intracellular symbiotic bacteria of 19 Dryophthoridae species collected worldwide, representing a wide range of plant species and tissues. All except one (Sitophilus linearis) harbor symbiotic bacteria within specialized cells (the bacteriocytes) assembled as an organ, the bacteriome. Phylogenetic analysis of the 16S rDNA gene sequence of the Dryophthoridae endosymbionts revealed three endosymbiotic clades belonging to gamma3-Proteobacteria and characterized by different GC contents and evolutionary rate. The genus name Candidatus Nardonella was proposed for the ancestral clade infesting Dryophthoridae 100 MYA and represented by five of nine bacterial genera studied. For this clade showing low GC content (40.5% GC) and high evolutionary rate (0.128 substitutions/site per 100 Myr), a single infection and subsequent cospeciation of the host and the endosymbionts was observed. In the two other insect lineage endosymbionts, with relatively high GC content (53.4% and 53.8% GC), competition with ancestral pathogenic bacteria might have occurred, leading to endosymbiont replacement in present-day last insects. (+info)
Molecular cloning of a cysteine proteinase cDNA from the cotton boll weevil Anthonomus grandis (Coleoptera: Curculionidae).
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The cotton boll weevil (Anthonomus grandis) causes severe cotton crop losses in North and South America. This report describes the presence of cysteine proteinase activity in the cotton boll weevil. Cysteine proteinase inhibitors from different sources were assayed against total A. grandis proteinases but, unexpectedly, no inhibitor tested was particularly effective. In order to screen for active inhibitors against the boll weevil, a cysteine proteinase cDNA (Agcys1) was isolated from A. grandis larvae using degenerate primers and rapid amplification of cDNA ends (RACE) techniques. Sequence analysis showed significant homologies with other insect cysteine proteinases. Northern blot analysis indicated that the mRNA encoding the proteinase was transcribed mainly in the gut of larvae. No mRNA was detected in neonatal larvae, pupae, or in the gut of the adult insect, suggesting that Agcys1 is an important cysteine proteinase for larvae digestion. The isolated gene will facilitate the search for highly active inhibitors towards boll weevil larvae that may provide a new opportunity to control this important insect pest. (+info)
Five types of olfactory receptor neurons in the strawberry blossom weevil Anthonomus rubi: selective responses to inducible host-plant volatiles.
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Plants release hundreds of volatiles that are important in the interaction with herbivorous animals, but which odorants are detected by which species? In this study, single receptor neurons on the antenna of the oligophagous strawberry blossom weevil Anthonomus rubi were screened for sensitivity to naturally produced plant compounds by the use of gas chromatography linked to electrophysiological recordings from single cells. The narrow tuning of the neurons was demonstrated by responses solely to a few structurally related sesquiterpenes, aromatics or monoterpene hydrocarbons out of hundreds of plant constituents tested. We present five olfactory receptor neuron types, identified according to one primary odorant i.e. the compound to which the neurons are most sensitive. These odorants, (-)-germacrene D, (-)-beta-caryophyllene, methyl salicylate, E-beta-ocimene and (3E)-4,8-dimethyl-1,3,7-nonatriene, present in the intact strawberry plant, are induced in higher amounts by weevil feeding. This suggests that these compounds can provide information about the presence of conspecifics. We used protocols especially designed to allow comparison with previously investigated species. Striking similarities, but also differences, are demonstrated between receptor neuron specificity in the strawberry weevil and moths. (+info)
A microfluorometric method for quantifying RNA and DNA in terrestrial insects.
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Evidence is accumulating for a mechanistic linkage between body phosphorus content and growth and reproduction of individual organisms, due in part to variation in allocation of resources to ribosomal RNA. Testing this connection requires reliable methods of quantifying the nucleic acid content of individual organisms. Although methods for quantifying nucleic acids are available for a wide array of organisms, adaptation of such methods for study of insects has been neglected. Sensitive stains and high throughput fluorometric measurements are now available that substantially improve past methodologies. Here we present methods for the extraction and quantification of insect RNA and DNA based on the use of N-lauroylsarcosine and sonication for extraction, the nucleases RNase and DNase, and the use of microplate fluorescent assays to quantify nucleic acids as percent of body weight in insects. We illustrate the method using Drosophila and curculionid weevils. (+info)
Iridovirus in the root weevil Diaprepes abbreviatus.
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Invertebrate iridescent virus 6 (IIV6) was evaluated for mode of transmission and ability to cause infection in the root weevil, Diaprepes abbreviatus (L.). This is the first evidence of IIV6 infection in D. abbreviatus, which caused both patent and sub-lethal covert infections in both larvae and adults. Adults and larvae were successfully infected with IIV6 by puncture, injection and per os. Transmission of IIV6 was demonstrated between infected and healthy individuals regardless of gender. Virus was detected in egg masses produced by virus-infected females suggesting IIV6 is transmitted transovarially. Virus particles were observed in the cytoplasm of weevil cells, and were shown to infect fat bodies, muscle, and nerve tissues, as visualized using transmission electron microscopy. Patent infections resulted in death of individuals within 3 to 4 days post infection. Individuals with covert infections tested positive for virus infection on day 7 by polymerase chain reaction analysis. Sequencing of PCR amplicons confirmed virus infection. Discovery of new pathogens against root weevils may provide new management tools for development of control strategies based on induced epizootics. This is the first report of a virus infecting D. abbreviatus. (+info)
Polygalacturonase from Sitophilus oryzae: possible horizontal transfer of a pectinase gene from fungi to weevils.
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Endo-polygalacturonase, one of the group of enzymes known collectively as pectinases, is widely distributed in bacteria, plants and fungi. The enzyme has also been found in several weevil species and a few other insects, such as aphids, but not in Drosophila melanogaster, Anopheles gambiae, or Caenorhabditis elegans or, as far as is known, in any more primitive animal species. What, then, is the genetic origin of the polygalacturonases in weevils? Since some weevil species harbor symbiotic microorganisms, it has been suggested, reasonably, that the symbionts' genomes of both aphids and weevils, rather than the insects' genomes, could encode polygalacturonase. We report here the cloning of a cDNA that encodes endo-polygalacturonase in the rice weevil, Sitophilus oryzae (L.), and investigations based on the cloned cDNA. Our results, which include analysis of genes in antibiotic-treated rice weevils, indicate that the enzyme is, in fact, encoded by the insect genome. Given the apparent absence of the gene in much of the rest of the animal kingdom, it is therefore likely that the rice weevil polygalacturonase gene was incorporated into the weevil's genome by horizontal transfer, possibly from a fungus. (+info)
Iridovirus infection of cell cultures from the Diaprepes root weevil, Diaprepes abbreviatus.
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We here report the development and viral infection of a Diaprepes root weevil cell culture. Embryonic tissues of the root weevil were used to establish cell cultures for use in screening viral pathogens as potential biological control agents. Tissues were seeded into a prepared solution of insect medium and kept at a temperature of 24 degrees C. The cell culture had primarily fibroblast-like morphology with some epithelial monolayers. Root weevil cells were successfully infected in vitro with a known insect virus, Invertebrate Iridescent Virus 6. Potential uses of insect cell cultures and insect viruses are discussed. (+info)
Protein-enriched pea flour extract protects stored milled rice against the rice weevil, Sitophilus oryzae.
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Studies were conducted to evaluate the effect of a protein-enriched pea (Pisum sativum var. Bonneville) flour extract against the rice weevil, Sitophilus oryzae in its repellency, toxicity, effect on fecundity, stability and sensory properties. Milled rice admixed with pea flour extract at 1% concentration significantly repelled S. oryzae. Mortality of S. oryzae was found to increase and fecundity was markedly suppressed, in rice treated with 1% pea flour extract. The toxicity and reproductive effects of the pea protein-enriched rice were found to be stable for a period of 5 months. The sensory characteristics of stored rice when eaten were not affected by the treatment with pea flour extract. This study indicates that the protein-enriched flour extract obtained from the Bonneville pea may be feasible to protect stored milled rice from insect attack. (+info)