Genetic differentiation ofDrosophila melanogaster populations as assessed by two-dimensional electrophoresis

Seven populations ofDrosophila melanogaster, representing a worldwide distribution, were compared using two-dimensional protein gel electrophoresis. A total of 611 protein spots was scored, which probably represent a sample of over 500 loci that were surveyed. Of the protein spots scored, 521 spots were found to be invariant, but another 90 spots were found to be variable among the populations. Of these variable protein spots, 12 were found to be present in only one population. All the populations, except one, had at least one protein spot restricted to itself. However, the Japanese population had by far the most, with five protein spots restricted to this one population, which has been observed in previous studies of private alleles in oriental populations. The mean genetic similarity (F) found among the seven populations was 0.965, with a range of between 0.956 and 0.977. This is similar to previous reports of lower variation found in population genetic surveys using two-dimensional electrophoresis. It was found that the historical relationships among these populations was somewhat congruent with the geographic distribution of the populations, but as in previous studies, it was not exactly coincident.     

HOST EFFECTS ON FERTILITY AND REPRODUCTIVE SUCCESS OF DENDROCTONUS PONDEROSAE HOPKINS (COLEOPTERA: SCOLYTIDAE)

Mountain pine beetles from lodgepole and limber pine in western Canada were crossbred. We compared data about reproductive success and fecundity of parents as well as development, mortality, and fertility of their progeny to determine whether there was reproductive isolation among beetle populations in these hosts. Three factors, directly or indirectly related to the host, influenced reproductive performance of parents (reproductive success, egg gallery length, fecundity, and number of eggs laid per centimeter of gallery) as well as the mortality, dry weight, and fat content of the progeny: (1) the host species in which progeny were reared, (2) the host species in which the female parent was reared, and (3) whether both parents originated from the same or different host species. Limber pine appears to be a better host for Dendroctonus ponderosae reproduction and survival than lodgepole pine. Nonetheless, beetles reared from lodgepole and limber pine can reproduce in either host and will mate with each other. Progeny of all crosses were fertile. Thus, there is no apparent barrier to prevent beetles from the two host species from interbreeding in the field.     

Met-enkephalin effects on associations formed in utero

Rat fetuses exposed to an odor stimulus and an aversive stimulus in utero showed an aversion to the odor when tested 16 days postnatally. Fetuses that also received 80 μg/kg Met-enkephalin showed a greater aversion to the odor stimulus than those subjects that did not receive the peptide. The difference between these groups was marginally significant. Control subjects did not show an aversion. But, subjects exposed to the odor and Met-enkephalin without the aversive stimulus, when tested, showed a significant preference for the odor over other control groups. These data show that associative learning in rat fetuses at 20 days of gestation may be enhanced by administration of Met-enkephalin.     

Engineering enzymes

Could we design and construct enzymes to catalyse any desired reaction? Compared with organic chemical catalysts, enzymes are highly specific and work in dilute aqueous solutions at ambient temperatures. Substrates are brought together from solution to precise orientations in the active site of an enzyme and the amino acid side-chains of the enzyme may assist catalysis by attacking or destabilizing substrate bonds. In principle, a novel enzyme could be constructed de novo or from pre-existing enzymes. Altering enzymes by recombinant DNA techniques offers most chance of success.     

Review of Canadian species of the genera Gnathusa Fenyes,Mniusa Mulsant & Rey and Ocyusa Kraatz (Coleoptera,Staphylinidae, Aleocharinae)

Four species of Gnathusa Fenyes (G. alfacaribou Klimaszewski & Langor, G. caribou Lohse, G. eva Fenyes, and G. tenuicornis Fenyes) occur in the Nearctic and in Canada. Three species of Ocyusa Kraatz (O. asperula Casey, O. californica Bernhauer, O. canadensis Lohse), and three species of Mniusa Mulsant and Ray (M. minutissima (Klimaszewski & Langor), M. yukonensis (Klimaszewski & Godin), and M. odelli Klimaszewski & Webster, sp. n.), are known from the Nearctic and all but O. californica occur in Canada. The recently described Gnathusa minutissima Klimaszewski and Langor and Ocyusa yukonensis Klimaszewski and Godin, are transferred here to the genus Mniusa Mulsant & Rey. New provincial and state records are reported for: G. eva (Alberta), G. tenuicornis (Alberta, Oregon, and New Brunswick), O. canadensis (New Brunswick and Newfoundland), M. minutissima (New Brunswick), and M. yukonensis (Nova Scotia, New Brunswick, Quebec, and British Columbia). The female of M. yukonensis was discovered and is illustrated for the first time. The genus Mniusa is reported for the first time from Canada and represents the first confirmed generic record for North America. Keys for identification of all Canadian species, images of body and genital structures, maps showing distribution mainly in Canada, and new bionomics data are provided.     

Intraspecific trait variability shapes leaf trait response to altered fire regimes

Background and AimsUnderstanding impacts of altered disturbance regimes on community structure and function is a key goal for community ecology. Functional traits link species composition to ecosystem functioning. Changes in the distribution of functional traits at community scales in response to disturbance can be driven not only by shifts in species composition, but also by shifts in intraspecific trait values. Understanding the relative importance of these two processes has important implications for predicting community responses to altered disturbance regimes.MethodsWe experimentally manipulated fire return intervals in replicated blocks of a fire-adapted, longleaf pine (Pinus palustris) ecosystem in North Carolina, USA and measured specific leaf area (SLA), leaf dry matter content (LDMC) and compositional responses along a lowland to upland gradient over a 4 year period. Plots were burned between zero and four times. Using a trait-based approach, we simulate hypothetical scenarios which allow species presence, abundance or trait values to vary over time and compare these with observed traits to understand the relative contributions of each of these three processes to observed trait patterns at the study site. We addressed the following questions. (1) How do changes in the fire regime affect community composition, structure and community-level trait responses? (2) Are these effects consistent across a gradient of fire intensity? (3) What are the relative contributions of species turnover, changes in abundance and changes in intraspecific trait values to observed changes in community-weighted mean (CWM) traits in response to altered fire regime?Key ResultsWe found strong evidence that altered fire return interval impacted understorey plant communities. The number of fires a plot experienced significantly affected the magnitude of its compositional change and shifted the ecotone boundary separating shrub-dominated lowland areas from grass-dominated upland areas, with suppression sites (0 burns) experiencing an upland shift and annual burn sites a lowland shift. We found significant effects of burn regimes on the CWM of SLA, and that observed shifts in both SLA and LDMC were driven primarily by intraspecific changes in trait values.ConclusionsIn a fire-adapted ecosystem, increased fire frequency altered community composition and structure of the ecosystem through changes in the position of the shrub line. We also found that plant traits responded directionally to increased fire frequency, with SLA decreasing in response to fire frequency across the environmental gradient. For both SLA and LDMC, nearly all of the observed changes in CWM traits were driven by intraspecific variation.     

Development of a multi-excitation fluorescence (MEF) imaging method to improve the information content of benthic coral reef surveys

Benthic surveys are a key component of monitoring and conservation efforts for coral reefs worldwide. While traditional image-based surveys rely on manual annotation of photographs to characterise benthic composition, automatic image annotation based on computer vision is becoming increasingly common. However, accurate classification of some benthic groups from reflectance images presents a challenge to local ecologists and computers alike. Most coral reef organisms produce one or a combination of fluorescent pigments, such as Green Fluorescent Protein (GFP)-like proteins found in corals, chlorophyll-a found in all photosynthetic organisms, and phycobiliproteins found in red macroalgae, crustose coralline algae (CCA) and cyanobacteria. Building on the potential of these pigments as a target for automatic image annotation, we developed a novel imaging method based on off-the-shelf components to improve classification of coral and other biotic substrates using a multi-excitation fluorescence (MEF) imaging system. We used RGB cameras to image the fluorescence emission of coral and algal pigments stimulated by narrow-waveband blue and green light, and then combined the information into three-channel pseudocolour images. Using a set of a priori rules defined by the relative pixel intensity produced in different channels, the method achieved successful classification of organisms into three categories based on the dominant fluorescent pigment expressed, facilitating discrimination of traditionally problematic groups. This work provides a conceptual foundation for future technological developments that will improve the cost, accuracy and speed of coral reef surveys.

     

Conserved and divergent aspects of Robo receptor signaling and regulation between Drosophila Robo1 and C. elegans SAX-3

The evolutionarily conserved Roundabout (Robo) family of axon guidance receptors control midline crossing of axons in response to the midline repellant ligand Slit in bilaterian animals including insects, nematodes, and vertebrates. Despite this strong evolutionary conservation, it is unclear whether the signaling mechanism(s) downstream of Robo receptors are similarly conserved. To directly compare midline repulsive signaling in Robo family members from different species, here we use a transgenic approach to express the Robo family receptor SAX-3 from the nematode Caenorhabditis elegans in neurons of the fruit fly, Drosophila melanogaster. We examine SAX-3’s ability to repel Drosophila axons from the Slit-expressing midline in gain of function assays, and test SAX-3’s ability to substitute for Drosophila Robo1 during fly embryonic development in genetic rescue experiments. We show that C. elegans SAX-3 is properly translated and localized to neuronal axons when expressed in the Drosophila embryonic CNS, and that SAX-3 can signal midline repulsion in Drosophila embryonic neurons, although not as efficiently as Drosophila Robo1. Using a series of Robo1/SAX-3 chimeras, we show that the SAX-3 cytoplasmic domain can signal midline repulsion to the same extent as Robo1 when combined with the Robo1 ectodomain. We show that SAX-3 is not subject to endosomal sorting by the negative regulator Commissureless (Comm) in Drosophila neurons in vivo, and that peri-membrane and ectodomain sequences are both required for Comm sorting of Drosophila Robo1.