Physiological Recordings and RNA Sequencing of the Gustatory Appendages of the Yellow-fever Mosquito Aedes aegypti

Electrophysiological recording of action potentials from sensory neurons of mosquitoes provides investigators a glimpse into the chemical perception of these disease vectors. We have recently identified a bitter sensing neuron in the labellum of female Aedes aegypti that responds to DEET and other repellents, as well as bitter quinine, through direct electrophysiological investigation. These gustatory receptor neuron responses prompted our sequencing of total mRNA from both male and female labella and tarsi samples to elucidate the putative chemoreception genes expressed in these contact chemoreception tissues. Samples of tarsi were divided into pro-, meso- and metathoracic subtypes for both sexes. We then validated our dataset by conducting qRT-PCR on the same tissue samples and used statistical methods to compare results between the two methods. Studies addressing molecular function may now target specific genes to determine those involved in repellent perception by mosquitoes. These receptor pathways may be used to screen novel repellents towards disruption of host-seeking behavior to curb the spread of harmful viruses.     

Exotic plant invasion alters chaparral ecosystem resistance and resilience pre- and post-wildfire

Perturbations such as wildfire and exotic plant invasion have significant impacts on soils, and the extent to which invaded soils are resistant or resilient to these disturbances varies by ecosystem type. Replacement of shrublands by herbaceous exotics pre- and post-wildfire may drastically alter soil chemical and biological properties for an unknown duration. We assessed above and belowground resistance and resilience to exotic plant invasion both before and after a chaparral wildfire. We hypothesized that exotic plant species would change chemical characteristics of chaparral soils by altering litter and microbial inputs, and that controlling exotics and seeding native species would restore chemical characteristics to pre-invaded conditions. We additionally hypothesized that exotic plant species would slow succession above- and belowground, as well as recovery of post-wildfire chaparral structure and function. Plant species composition and soil nutrient pools and cycling rates were evaluated in mature and invaded chaparral pre- and post-wildfire. Exotic plant species were weeded and native species were seeded to assess impacts of exotic competition on native species recovery. Invasion did not impact all soil characteristics before fire, but increased soil C/N ratio, pH, and N cycling rates, and reduced NO₃-N availability. After fire, invasives slowed succession above- and belowground. Removal of exotics and seeding natives facilitated succession and resulted in plant composition similar to uninvaded, post-wildfire chaparral. The chaparral ecosystem was not resistant to impacts of invasion as indicated by altered soil chemistry and C and N cycling rates; however, short-term restoration led to recovery of extractable nitrogen availability indicating resilience of chaparral soils. This suggests that the permanence of exotic plant species, once established, represents a greater ecological challenge than exotic plant impacts on soils.     

The Streamlined Genome of Phytomonas spp. Relative to Human Pathogenic Kinetoplastids Reveals a Parasite Tailored for Plants

Members of the family Trypanosomatidae infect many organisms, including animals, plants and humans. Plant-infecting trypanosomes are grouped under the single genus Phytomonas, failing to reflect the wide biological and pathological diversity of these protists. While some Phytomonas spp. multiply in the latex of plants, or in fruit or seeds without apparent pathogenicity, others colonize the phloem sap and afflict plants of substantial economic value, including the coffee tree, coconut and oil palms. Plant trypanosomes have not been studied extensively at the genome level, a major gap in understanding and controlling pathogenesis. We describe the genome sequences of two plant trypanosomatids, one pathogenic isolate from a Guianan coconut and one non-symptomatic isolate from Euphorbia collected in France. Although these parasites have extremely distinct pathogenic impacts, very few genes are unique to either, with the vast majority of genes shared by both isolates. Significantly, both Phytomonas spp. genomes consist essentially of single copy genes for the bulk of their metabolic enzymes, whereas other trypanosomatids e.g. Leishmania and Trypanosoma possess multiple paralogous genes or families. Indeed, comparison with other trypanosomatid genomes revealed a highly streamlined genome, encoding for a minimized metabolic system while conserving the major pathways, and with retention of a full complement of endomembrane organelles, but with no evidence for functional complexity. Identification of the metabolic genes of Phytomonas provides opportunities for establishing in vitro culturing of these fastidious parasites and new tools for the control of agricultural plant disease.     

Ethological description of a fixed action pattern in a kissing bug (Triatominae): vision,gustation, proboscis extension and drinking of water and guava

Triatomines (Heteroptera, Reduviidae) are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease in America. These true bugs have traditionally been considered to be blood suckers, although some species have been catalogued as being entomophagous. By using their highly specialized mouthparts, these insects have evolved a stereotyped habit which includes lifting up the proboscis, piercing and sucking, when the occasion arises. Most triatomines bite their sleeping and unaware vertebrate or invertebrate hosts, but they can also search for other targets, guided, in part, by visual and chemical stimuli. In this study, we observed that triatomines apparently visually identify a drop of water in the distance, then taste it with their legs, upon which proboscis extension and sucking ensues. This invariant behavior or fixed action pattern, observed in several triatomine species (Rhodnius prolixus, Triatoma infestans and Panstrongylus geniculatus), was also elicited by a dummy drop of water and guava fruit. We discuss evolutionary and ethological aspects of this innate behavior. Digital video images related to this article are available at http://www.momo-p.com/showdetail-e.php?movieid=momo180314rp01a and http://www.momo-p.com/showdetail-e.php?movieid=momo180314rp03a.     

Green leaf volatiles enhance aggregation pheromone of boll weevil, Anthonomus grandis

Enhancement of an insect pheromone response by green leaf volatiles is reported for the first time in the boll weevil, Anthonomus grandis Boh. (Coleoptera: Curculionidae). Single cell recordings from antennal olfactory receptors revealed a class of cells selectively responsive to six carbon alcohols and aldehydes (i.e., green leaf volatiles). Field tests with released weevils demonstrated enhanced trap captures with trans-2-hexen-l-ol, cis-3-hexen-l-ol, or l-hexanol paired with grandlure, the boll weevil aggregation pheromone, when in competition with grandlure alone. Although dose-response curves constructed from electroantennograms were indicative of similar populations of receptor cells for selected six carbon alcohols, one of the compounds tested, cis-2-hexen-l-ol, was inactive in field tests. Trans-2-hexenal was active in single cell recordings, but was also inactive in field tests. In tests in cotton fields with indigenous weevil populations, trans-2-hexen-l-ol not only enhanced pheromone trap captures, but also extended the longevity of attractiveness of pheromone-baited traps. The combined electrophysiological and field data support across-fiber coding of green leaf volatiles by boll weevil olfactory receptors. The results are discussed with regard to the chemistry of the host plant of the boll weevil, cotton (Gossypium hirsutum L.), and potential economic significance for boll weevil eradication/suppression.

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Résumé Pour la première fois, l'augmentation de la puissance de la phéromone d'agrégation sous l'action de l'odeur verte est signalée chez A. grandis. Les enregistrements unitaires de cellules des récepteurs olfactifs des antennes a mis en évidence une catégorie de cellules réagissant sélectivement aux alcools et aux aldéhydes à 6 carbones (c'est-à-dire aux substances volatiles des feuilles vertes). Des essais dans la nature ont montré une augmentation des captures par les pièges de grandlure (phéromone d'agrégation du charançon) renforcés par du trans-2-hexen-l-ol, du cis-3-hexen-l-ol, ou du l-hexanol.Bien que les courbes des réponses en fonction des doses construites à partir des électroantennogrammes indiquent des catégories de cellules réceptrices identiques pour les alcools à 6 carbones examinés, l'une de ces substances, cis-2-hexen-l-ol, s'est révélée inactive dans la nature. Le trans-2-hexenal qui était actif avec des enregistrements unitaires de cellules, était lui aussi inactif dans la nature. Dans des essais dans des champs de coton avec des populations indigènes de charançons, le trans-2-hexen-l-ol a, non seulement augmenté l'efficacité des pièges à phéromone, mais a prolongé leur attractivité. Les résultats d'électrophysiologie combinés aux données de la nature appuient le modelage transversal des substances vertes volatiles des feuilles par les récepteurs olfactifs du charançon. Ces résultats sont interprétés en relation avec la chimie de la plante-hôte de A. grandis, le coton, Gossypium hirsutum L., et leur possibilité d'utilisation économique pour l'éradication du charançon.

     

The oxidation of d- and l-glycerate by rat liver

1. The interconversion of hydroxypyruvate and l-glycerate in the presence of NAD and rat-liver l-lactate dehydrogenase has been demonstrated. Michaelis constants for these substrates together with an equilibrium constant have been determined and compared with those for pyruvate and l-lactate. 2. The presence of d-glycerate dehydrogenase in rat liver has been confirmed and the enzyme has been purified 16–20-fold from the supernatant fraction of a homogenate, when it is free of l-lactate dehydrogenase, with a 23–29% recovery. The enzyme catalyses the interconversion of hydroxypyruvate and d-glycerate in the presence of either NAD or NADP with almost equal efficiency. d-Glycerate dehydrogenase also catalyses the reduction of glyoxylate, but is distinct from l-lactate dehydrogenase in that it fails to act on pyruvate, d-lactate or l-lactate. The enzyme is strongly dependent on free thiol groups, as shown by inhibition with p-chloromercuribenzoate, and in the presence of sodium chloride the reduction of hydroxypyruvate is activated. Michaelis constants for these substrates of d-glycerate dehydrogenase and an equilibrium constant for the NAD-catalysed reaction have been calculated. 3. An explanation for the lowered V_max. with d-glycerate as compared with dl-glycerate for the rabbit-kidney d-α-hydroxy acid dehydrogenase has been proposed.     

Structure of the Proteolipid Protein Extracted from Bovine Central Nervous System Myelin with Nondenaturing Detergents

As a basis for attempts to define the structures of the proteins within myelin, methods have been developed for their extraction and isolation in solutions of non-denaturing detergents. With use of solutions of deoxycholate or Triton X-100, up to 90% of the protein has been extracted from bovine CNS myelin, along with most of the phospholipid. The proteolipid protein has been purified in deoxycholate solutions by chromatography on a blue dye-ligand column, which retained all of the basic protein and 2',3'-cyclic nucleotide-3'-phosphodiesterase, and then on Sephacryl S300, which separated proteolipid protein from phospholipid and high-molecular-weight proteins. The proteolipid protein was isolated from Triton X-100 extracts of myelin by adsorption onto phosphocellulose resin, with subsequent elution by 0.5 M sodium chloride. Gel permeation chromatography was used as the final purification step. Sedimentation equilibrium experiments gave a monomer molecular weight of 134,000 +/- 8000 in deoxycholate and 145,000 +/- 17,000 in Triton X-100 solutions. On the basis of an apparent subunit molecular weight of 23,500 it was deduced that the native protein is probably hexameric. Above 0.2 gL-1 in Triton X-100 solutions and 0.5 gL-1 in deoxycholate solutions the protein aggregated. In deoxycholate solutions the protein adopts the highly helical conformation expected for an intrinsic membrane protein.