Microwave incubation improves lipolytic enzyme preservation for ultrastr uctural cytochemistry

Standard methods for the ultrastructural detection of lipase and sphingomyelinase activities in the skin result in considerable loss of structural preservation, often interfering with accurate delineation of enzyme localization in association with specific organelles. Moreover, poor preservation occurs, even after extensive aldehyde prefixation, owing to the prolonged incubation times needed to detect residual enzyme activity, which often require non- physiological conditions. A modified incubation protocol is described here, which uses microwave irradiation in conjunction with drastically shortened incubation times, resulting in both superior ultrastructural preservation and excellent localization in mammalian epidermis. This method should be useful generally not only for the study of lipase localization in skin, but also in conjuction with the cytochemical detection of a variety of enzymes in various types of tissue. This revised version was published online in November 2006 with corrections to the Cover Date.     

Evidence of alarm pheromones in the venom of Polistes dominulus workers (Hymenoptera: Vespidae)

Abstract.  The active and coordinating capacity of defending the nest is a key feature of social insects. The present study investigates the presence of alarm pheromones in the venom of workers of the social wasp, Polistes dominulus . Laboratory experiments were performed with caged colonies of P. dominulus using a wind tunnel apparatus to test the behavioural response of workers to venom released by other workers and to venom extracts. Contrary to that previously reported for European paper wasps, the present results show that the venom is the source of alarm pheromones. Field experiments combining a visual (black target) and a chemical stimulus (venom extract) were performed to test the effect of the venom on the reaction of colonies. Wasps leave the nest, land on the visual target and attack the target significantly more once exposed to venom extract plus target than to solvent plus target. This work shows that the venom of P. dominulus workers elicits an alarm response, reduces the threshold for attack and acts as an attractant on targets. These results using P. dominulus indicate that, in both American and European species, colony defence is based on the same features, suggesting that chemical alarm is a widespread trait in the genus Polistes .     

Melanins in Physiological Conditions Protect Against Lipoperoxidation. A Study on Albino and Pigmented Xenopus

The susceptibility to lipoperoxidation in liver of albino and pigmented Xenopus laevis Daudin, has been studied. Albino Xenopus liver was richer in polyunsaturated fatty acids (PUFAs) than the pigmented one; moreover, it was also richer in mitochondrial superoxide dismutase (MnSOD) and in reduced glutathione (GSH). The thiobarbituric acid-reactive substances (TBARS) values were more abundant in the albino tissue compared to the pigmented tissue both during spontaneous and Fe⁺⁺ induced lipoperoxidation. Therefore, when isolated and purified melanin, in physiological quantities, was added to albino tissue, the TBARS values drastically decreased. Thus, melanin, in our experimental conditions, protects the albino tissue even more than SOD and GSH do. Melanin, in our opinion, acts as an antioxidant, because it is able to scavenge O₂^?.     

Light induces jasmonate‐isoleucine conjugation via OsJAR1‐dependent and ‐independent pathways in rice

The bioactive form of jasmonate is the conjugate of the amino acid isoleucine (Ile) with jasmonic acid (JA), which is biosynthesized in a reaction catalysed by the GH3 enzyme JASMONATE RESISTANT 1 (JAR1). We examined the biochemical properties of OsJAR1 and its involvement in photomorphogenesis of rice (Oryza sativa). OsJAR1 has a similar substrate specificities as its orthologue in Arabidopsis. However, osjar1 loss‐of‐function mutants did not show as severe coleoptile phenotypes as the JA‐deficient mutants coleoptile photomorphogenesis 2 (cpm2) and hebiba, which develop long coleoptiles in all light qualities we examined. Analysis of hormonal contents in the young seedling stage revealed that osjar1 mutants are still able to synthesize JA‐Ile conjugate in response to blue light, suggesting that a redundantly active enzyme can conjugate JA and Ile in rice seedlings. A good candidate for this enzyme is OsJAR2, which was found to be able to catalyse the conjugation of JA with Ile as well as with some additional amino acids. In contrast, if plants in the vegetative stage were mechanically wounded, the content of JA‐Ile was severely reduced in osjar1, demonstrating that OsJAR1 is the most important JA‐Ile conjugating enzyme in the wounding response during the vegetative stage.     

Fruit size preference in the New Zealand pigeon (Hemiphaga novaeseelandiae)

Abstract We investigated whether the New Zealand pigeon Hemiphaga novaeseelandiae (Columbidae) exhibits size‐based preferences for fruits. We tested the hypothesis that in small‐fruited species, pigeons would prefer larger fruits, but in larger‐fruited species, this preference would reverse as the pigeons become increasingly limited by their gape size. We collected undispersed fruits and bird‐dispersed seeds of 10 plant species, some over several sites or years (13 datasets in total). We estimated the fruit size of dispersed seeds by fitting regressions of fruit diameter to seed diameter in intact fruits. We were able to predict fruit diameter from seed diameter in 12 of the 13 populations, although the relationship was stronger in single‐seeded species than in multi‐seeded species. Seven of the 12 populations tested showed a significant difference in seed diameter among undispersed and dispersed seeds. However, our results showed no consistent pattern in fruit size preference by the New Zealand pigeon and did not support our hypothesis. The large‐bodied New Zealand pigeon is generally not gape limited and fruit size preferences appear to be independent of mean fruit size.     

Inter- and intraspecific differences in the life history and growth of Caridina spp. (Decapoda: Atyidae) in Hong Kong streams

1. The life‐history pattern of three populations of Caridina cantonensis (Atyidae) and one population of Caridina serrata were studied in four Hong Kong streams: Pak Ngau Shek (PNS), Kap Man Hang (KMH), Pak Tam Chung (PTC), and Lung Fu Shan (LFS). Caridina cantonensis occurred at PNS, KMH and PTC while C. serrata was present at LFS only. Monthly quantitative sampling was carried out at each study site over 2 years to investigate life‐history patterns and cohort‐specific growth rates. 2. Breeding by both species was mainly restricted to the wet season, and there was an increase in the incidence of ovigerous females as water temperatures rose at the start of the summer monsoon. The number of cohorts and the frequency of recruitment of C. cantonensis (one to three times each year) were highly site‐specific, but consistent between years. 3. Shrimp growth was size‐dependent, and decreased with increasing body size. Caridina cantonensis at PTC had the highest cohort‐specific growth rates (0.05–0.14 mg ash‐free dry weight (AFDW) mg^?1 day^?1), followed by C. serrata at LFS (6% lower), and C. cantonensis at KMH and PNS (16–39% lower). Shrimp life spans exceeded 1 year (17–22 months), depending on site and species. 4. Sexual maturity occurred at an earlier age and at a smaller size in populations with higher growth rates. Shrimps at PTC and LFS matured at approximately 4 months old; this was 3 months earlier than at KMH and 5 months earlier than at PNS. Females of C. cantonensis at PTC and C. serrata at LFS may have bred twice during each breeding season, while females in the other two populations bred once only. The number of cohorts produced by each population in each year did not vary between the years of the study. Most sexually mature individuals only survived long enough (for 10–12 months after maturation) to breed during one season. 5. The mean brood size of C. cantonensis varied among streams and was 25–28% larger at PNS and KMH than at PTC. Egg size did not differ among populations. Larger brood sizes at PNS and KMH may have been a consequence of increased availability of algal food in these unshaded stream sites. Caridina serrata had similar egg and brood sizes to C. cantonensis, but females were smaller and reproductive investment was higher. Thus, the degree of intraspecific variation in atyid breeding in Hong Kong was at least equal to, if not greater than, the extent of interspecific variation. Interspecific differences in life history may reflect the frequency of droughts and spate intensity experienced by C. serrata at LFS.     

Sibling species in the Chrysis ignita complex: molecular,morphological and trophic differentiation of Baltic species,with a description of two new cryptic species (Hymenoptera: Chrysididae)

Cryptic species complexes cause major challenges for taxonomists and alter understanding of species diversity. In Northern Europe, the Chrysis ignita species group is one such complex with numerous sympatric sibling species. The objective of this paper is to assess the taxonomy of 15 species from this group using three different approaches: molecular, morphological and trophic differentiation. The analysed set of molecular markers included a 7400‐bp‐long sequence of the mitochondrial genome covering complete sequences of CO1, CO2, ATP8, ATP6, CO3, ND3, 16S and 12S rRNA, nine tRNAs and a partial sequence of CytB, as well as a 3880‐bp‐long sequence of the nuclear DNA covering a part of 18S rRNA, the ITS1, 5.8S rRNA, ITS2 and a part of 28S rRNA. Discrete diagnostic characters of each species sequence were retrieved using the Characteristic Attribute Organisation System algorithm and a molecular identification key was compiled. The study revealed a higher evolutionary rate of the genes ATP8, ATP6, CO3, ND3 and CytB compared to that of CO1, CO2 and 16S; the studied nuclear markers demonstrated a lower evolutionary rate than the mitochondrial markers. A consensus tree compiled based on the combined mtDNA and nuclear markers with a strongly supported topology resolved the position of the C. schencki – C. parietis sp.n. clade as sister to the C. ignita – C. impressa clade and supported the monophyly of the C. angustula – C. longula clade. We compiled a morphometric species identification key applying linear discriminant equations. The trophic differentiation was assessed using data on host preferences of ten Chrysis species reared from trap‐nests; the analysis demonstrated that most of them are specialists exploiting a single or a few taxonomically related host species. In most cases, all three approaches supported the distinct status of the included species. Moreover, two previously undescribed species were consistently supported by the molecular methods. Therefore, we describe these as new, namely C. horridula sp.n. and C. parietis sp.n. Only C. mediata and C. solida were not clearly distinguished using the molecular phylogeny reconstruction methods. However, based on distinctive niche divergence, the presence of molecular characters and morphometric differences, we consider them as phylogenetically young but distinct species. In view of the weak morphological and molecular differentiation, the widely overlapping distribution areas and often similar habitat preferences and the trophic specialization, the C. ignita complex presents a possible model for studies of sympatric cryptic speciation. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:1EBAF0E1‐5FB7‐4CF4‐A595‐C11982448360 .     

Forecasting ecological and evolutionary strategies to global change: an example from habitat selection by lemmings

Ecologists and evolutionary biologists must develop theories that can predict the consequences of global warming and other impacts on Earth's biota. Theories of adaptive habitat selection are particularly promising because they link distribution and density with fitness. The evolutionarily stable strategy that emerges from adaptive habitat choice is given by the system's habitat isodar, the graph of densities in pairs of habitats such that the expectation of fitness is the same in each. We illustrate how isodars can be converted into adaptive landscapes of habitat selection that display the density‐ and frequency‐dependent fitness of competing strategies of habitat use. The adaptive landscape varies with the abundance of habitats and can thus be used to predict future adaptive distributions of individuals under competing scenarios of habitat change. Application of the theory to three species of Arctic rodents living on Herschel Island in the Beaufort Sea predicts changes in selection gradients as xeric upland increases in frequency with global warming. Selection gradients will become more shallow for brown lemming (Lemmus trimucronatus) and tundra vole (Microtus oeconomus) strategies that preferentially exploit mesic habitat. Climate change will cause selection gradients for the alternative strategy of using mostly xeric habitat to become much steeper. Meanwhile, the adaptive landscape for collared lemmings (Dicrostonyx groenlandicus), which specialize on xeric Dryas‐covered upland, will become increasingly convex. Changes in the adaptive landscapes thus predict expanding niches for Lemmus and Microtus, and a narrower niche for Dicrostonyx. The ability to draw adaptive landscapes from current patterns of distribution represents one of the few methods available to forecast the consequences of climate change on the future distribution and evolution of affected species.     

Coevolution of daily activity timing in a host–parasite system

Coevolutionary theories applied in the study of host–parasite systems indicate that lineages exhibit progressive trends in response to reciprocal selective pressures. Avian brood parasites have generated intense interest as models for coevolutionary processes. Similar to avian cuckoos, Polistes wasp social parasites usurp a nest and exploit the parental care of a congeneric species to rear their own brood. In the present study, we show a coevolutionary arms race in the daily activity pattern in a Polistes host–parasite pair. We measured the daily activity rate, in constant laboratory conditions, of both host and parasite females during the period in which nest usurpations occur. The parasites showed a hyperkinesis in the middle of the day. As the field observations suggested, this mid-day activity is used to perform host nest usurpation attempts. Timing the usurpations allows the parasite to maximize its usurpation attempts during daytime when the host defence is lower. A field comparison of host presence on the nest in two populations with different parasitism rates showed that populations under strong parasitic pressure exhibit timing counteradaptations to optimize nest defence. This study provides the first example of a mutual coadaptation in timing activity in a parasite–host system.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 399–405.