Molecular phylogeny of fungus gnats (Diptera: Mycetophilidae) revisited: position of Manotinae,Metanepsiini, and other enigmatic taxa as inferred from multigene analysis

The phylogeny of selected genera from four subfamilies of fungus gnats (Diptera: Mycetophilidae) – Manotinae, Leiinae, Sciophilinae and Gnoristinae (including Metanepsiini) – is reconstructed based on the combined analysis of five mitochondrial (12S, 16S, COI, COII, cytB) and two nuclear (28S, ITS2) gene markers. Results of the different analyses all support Manotinae as a monophyletic group, with Leiinae as the sister group. Allactoneura DeMeijere is nested in the monophyletic and strongly supported clade of Leiinae. The tribe Metanepsiini is revealed as paraphyletic and the genera Metanepsia Edwards and Chalastonepsia Søli do not appear to be closely related. The genera Docosia Winnertz, Ectrepesthoneura Enderlein, Novakia Strobl and Syntemna Winnertz were placed with a group of genera included traditionally in the Gnoristinae. The monophyly of Dziedzickia Johannsen and Phthinia Winnertz is not supported. The genera of Sciophilinae (excluding Paratinia Mik but including Eudicrana Loew) form a monophyletic group in the Bayesian model.     

Ecological differentiation between the Sardinian endemic Maniola nurag and the pan-European M. jurtina

Recently refined evolutionary theories have highlighted that ecological interactions and environmental gradients can play a major role in speciation. This paper reports on a 3‐year field study, in which the ecology of two congeneric butterfly species was used to explore and compare the environmental factors determining their spatial distribution. These data are discussed in the context of possible speciation scenarios between the Sardinian populations of Maniola nurag and M. jurtina. M. nurag is endemic to the island of Sardinia, while M. jurtina is widespread over Europe. In Sardinia, the two species are locally sympatric. Mark–release–recapture experiments were combined with measures of environmental variables in 15 1‐ha plots, established in areas of potential habitat for the butterflies. Constrained linear models were parameterized from mark–recapture data to estimate both individual (survival and capture probabilities) and population (population size and recruitment) parameters. The two species had similar demography, movement patterns, life history, and behaviour. Population sizes developed in a parabolic fashion from beginning to end of the flight season. Differences included local population size, adult phenology, and habitat requirements. Long‐distance movements larger than 1.5 km were observed, suggesting a substantial amount of gene‐flow between populations of the endemic as well as the widespread species. Multivariate analyses revealed four main environmental gradients responsible for the abundance of the butterflies in an area. Both species responded similarly to environmental variables. However, each species’s abundance was correlated with a different environmental gradient determined by vegetation cover and structure. When sympatric, the two species responded to subtle differences in microhabitat structure. This might originally have induced their divergence. This study is an example of how empirical field data on population dynamics, dispersal, and habitat characteristics of two sympatric congeners can further our understanding of how species differentiate despite existing gene‐flow. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 89 , 561–574.     

MAINTENANCE OF IONS,PROTEINS AND WATER IN LENS FIBER CELLS BEFORE AND AFTER TREATMENT WITH NON-IONIC DETERGENTS

If the plasma membrane and its associated transport proteins are solely responsible for maintenance of the asymmetric solute distribution then disruption of the plasma membrane would quickly lead to the symmetric distribution of all unattached inorganic ions between the cell and the extracellular environment. To test this hypothesis fresh pig lenses were incubated in Hanks ’ balanced salt solution in either absence or presence of non-ionic detergents (0.2 % Triton X-100 or 0.2 % Brij 58). Both detergents caused permeabilization of every lens fiber cell as shown by electron microscopy. The flux kinetics of K⁺, Mg^{2 +}, Na⁺, Ca^{2 +}, water and protein out of and into the permeabilized lens fiber cells was measured. Triton X-100 caused a faster flux rate of all solutes than did Brij 58. The Triton X-100 induced flux of solutes and water was associated with a decrease in lens ATP. Incubation of untreated lenses in solutions of different osmotic pressures at 0 °C demonstrated that the major fraction of lens water was osmotically unresponsive. Thus the asymmetric distribution of solutes in lens fiber cells is dependent on an intact plasma membrane and on a co-operative ATP-dependent association between K⁺, Mg^{2 +}, water and cytomatrix proteins.     

Spring migratory routes of eight trans-Saharan passerines through the central and western Mediterranean; results from a network of insular and coastal ringing sites

Detailed information on spring migration routes of songbirds across the Mediterranean is still scanty. Results are presented here from a study on the occurrence of eight Palaearctic-African migrants across the western and central Mediterranean based on ringing data collected during the Progetto Piccole Isole, a co-ordinated project based on standardized mist netting at 21 islands and coastal stations. The species were Melodious Warbler Hippolais polyglotta , Icterine Warbler Hippolais icterina, Bonelli's Warbler Phylloscopus bonelli , Wood Warbler Phylloscopus sibilatrix , Pied Flycatcher Ficedula hypoleuca , Collared Flycatcher Ficedula albicollis , Red-backed Shrike Lanius collurio and Woodchat Shrike Lanius senator . Their capture frequencies were expressed as the frequency of each species relative to the total number of migrants ringed at each station in order to correct for differences in capture effort between stations. For most species, the detailed, quantitative results confirmed the scanty information available so far, whereas in the case of Melodious Warbler and Bonelli's Warbler, evidence of undescribed and unexpected migration routes was found. These results suggest that standardized mist netting provides an important method for studying the migratory routes of small birds, in particular for those species which are not frequently ringed in their breeding and wintering grounds.     

Effect of Ethylene and Culture Environment on Rice Callus Proliferation

Modifications to the gaseous envelope by callus during culturein Petri dishes were shown to reduce growth and promote necrosisof several rice (Oryza sativa L.) cultivars. Incubatingcallusunder a continuous flow of gas mixtures of known compositionsuggested that the inhibition of growth was caused by the accumulationof ethylene, the depletion of oxygen and, to a lesser extent,the accumulation of carbon dioxide. In order to evaluate theimportance of ethylene accumulation aminoethoxyvinylglycine(AVG), 1-aminocyclopropane-l-carboxylic acid (ACC and silvernitrate (AgNO₃), were added to the nutrient medium and ethylenemeasurements performed during callus culture. Ethylene restrictedcallus growth particularly under high (35 °C) as comparedto moderate (25 °C) temperatures and under illuminated ascompared to darkened incubation. Under illuminated incubationat 25 °C AVG (5 mmol m^–3) and AgNO°(50 mmol m^–3)significantly improvedcallus growth (100 and 60% respectively)while ACC (200 mmol m^–3) significantly decreased growth(40%). AVG and AgNO₃ were less effective under dark incubationat 25 °C where ethylene production was lower. Furthermore,callus growth was significantly better in large as comparedto small culture vessels since the ethylene concentration wasdiluted and more oxygen was available for respiration. Bettercontrol of ethylene and increased oxygen availability couldbe a way ofproducing healthy callus for the formation of embryogenictissues of otherwise recalcitrant cultivars of rice (e.g. IndicaIR42) and may be a way of improving manipulation of other cerealspecies. Key words: 1-Aminocyclopropane-1-carboxylic acid, aminoethoxyvinylglycine, callus, ethylene, Oryza sativa, silver nitrate     

Taphonomy of Saint-Vallier (Drôme, France), the reference locality for the biozone MN17 (Upper Pliocene)

Saint-Vallier (Drôme, France) is the biostratigraphical reference locality for the biozone MN17 (Upper Pliocene, between 2.4 and 1.9  Ma). The taphonomic study presented here is based on the results of a recent 6-year campaign of renewed study. Carefully controlled excavations allow the study of the relative position of remains and their orientation inside several fossil clusters. Flowing water is the most likely agent responsible for fossil orientation: bones of the animals were carried by water from distal to proximal areas of deposition, where they were quickly buried by sediments. Taphonomic clues suggest that the causes of mortality were attritional. The study of mortality profiles yields more ambiguous results although not necessarily in contrast with the attritional hypothesis. Such a study shows the importance of the sample size for the interpretation of the age diagrams.     

Circadian regulation of leaf hydraulic conductance in sunflower (Helianthus annuus L. cv Margot)

The circadian regulation of leaf hydraulic conductance (K_leaf) was investigated in Helianthus annuus L. (sunflower). K_leaf was measured with an high pressure flow meter during the light and dark period from plants growing at a photoperiod of 12 h. K_leaf was 4.0 e−4 kg s⁻¹ m⁻² MPa⁻¹ during the light period (LL) and 30–40% less during the dark period (DL). When photoperiod was inverted and leaves were measured for K_leaf at their subjective light or dark periods, K_leaf adjusted to the new conditions requiring 48 h for increasing from dark to light values and 4 d for the opposite transition. Plants put in continuous dark showed K_leaf oscillating from light to dark values in phase with their subjective photoperiod indicating that K_leaf changes were induced by the circadian clock. Several cuts through the minor veins reduced leaf hydraulic resistance (R_leaf) of both LL and DL to the same value (1.0 e + 3 MPa m² s kg⁻¹) that equalled the vascular resistance (R_v). The contribution of the non-vascular leaf resistance (R_nv) to R_leaf was of 71.9% in DL and of 58.4% in LL. The dominant R_nv was shown to be reversibly modulated by mercurials, suggesting that aquaporins play a role in diurnal changes of K_leaf.     

Adipokinetic hormone counteracts oxidative stress elicited in insects by hydrogen peroxide: in vivo and in vitro study

The role of adipokinetic hormone (AKH) in counteracting oxidative stress elicited in the insect body is studied in response to exogenously applied hydrogen peroxide, an important metabolite of oxidative processes. In vivo experiments reveal that the injection of hydrogen peroxide (8 µmol) into the haemocoel of the firebug, Pyrrhocoris apterus L. (Heteroptera: Pyrrhocoridae) increases the level of AKH by 2.8‐fold in the central nervous system (CNS) and by 3.8‐fold in the haemolymph. The injection of hydrogen peroxide also increases the mortality of experimental insects, whereas co‐injection of hydrogen peroxide with Pyrap‐AKH (40 pmol) reduces mortality to almost control levels. Importantly, an increase in haemolymph protein carbonyl levels (i.e. an oxidative stress biomarker) elicited by hydrogen peroxide is decreased by 3.6‐fold to control levels when hydrogen peroxide is co‐injected with Pyrap‐AKH. Similar results are obtained using in vitro experiments. Oxidative stress biomarkers such as malondialdehyde and protein carbonyls are significantly enhanced upon exposure of the isolated CNS to hydrogen peroxide in vitro, whereas co‐treatment of the CNS with hydrogen peroxide and Pyrap‐AKH reduces levels significantly. Moreover, a marked decrease in catalase activity compared with controls is recorded when the CNS is incubated with hydrogen peroxide. Incubation of the CNS with hydrogen peroxide and Pyrap‐AKH together curbs the negative effect on catalase activity. Taken together, the results of the present study provide strong support for the recently published data on the feedback regulation between oxidative stressors and AKH action, and implicate AKH in counteracting oxidative stress. The in vitro experiments should facilitate research on the mode of action of AKH in relation to oxidative stress, and could help clarify the key pathways involved in this process.     

The structure of the seed in some Western Australian species of the genus Hibbertia (Dilleniaceae)

The internal structure of the seed was studied for four Western Australian species of the genus Hibbertia (Fam.: Dilleniaceae). The principal mechanical layer of the seed coat is the endotesta. The large cells of the endotesta are heavily cutinized in potentially viable seeds of all examined species. However, results for H. hypericoides show that the cells of the endotesta are distinctly less cutinized in non-viable seeds. The tegmen consists of only one layer which is spirally thickened. Remains of the nucellus enclose the endosperm. The endosperm is rich in both starch and lipids, and the embryo is underdeveloped and minute.     

Evidence of a complex phylogeographic structure in the common dormouse,Muscardinus avellanarius (Rodentia: Gliridae)

This is the first mitochondrial phylogeography of the common dormouse, Muscardinus avellanarius (Linnaeus, 1758), a hibernating rodent strictly protected in Europe (Habitat Directive, annex IV; Bern Convention, annex III). The 84 individuals of M. avellanarius, sampled throughout the distributional range of the species, have been sequenced at the mitochondrial DNA gene (cytochrome b, 704 base pairs). The results revealed two highly divergent lineages, with an ancient separation around 7.7 Mya and a genetic divergence of 7.7%. Lineage 1 occurs in Western Europe (France, Belgium, and Switzerland) and Italy, and lineage 2 occurs in Central–Northern Europe (Poland, Germany, Latvia, and Lithuania), on the Balkan Peninsula, and in Turkey. Furthermore, these two lineages are subdivided into five sublineages genetically isolated with a strong geographical association. Therefore, lineage 1 branches into two further sublineages (Western European and Italian), whereas lineage 2 contained three sublineages (Central–Northern European, Turkish, and Balkan). We observed low genetic diversity within the sublineages, in contrast to the significant level of genetic differentiation between them. The understanding of genetic population structure is essential for identifying units to be conserved. Therefore, these results may have important implications for M. avellanarius conservation. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 648–664.