The Small-Subunit rRNA Gene Sequences of Venerids and the Phylogeny of Bivalvia

The complete nucleotide sequence of the 18S subunit of ribosomal DNA (rDNA) was determined for the venerid clams Callista chione (Pitarinae) and Venus verrucosa (Venerinae). Comparison of the new sequences with the published sequences of 1 annelid, 2 gastropods, 2 polyplacophorans, and 19 bivalves showed that when the annelids are used as outgroup the gastropods diverge from the bivalves, which form a cluster including the polyplacophorans. When the gastropods alone were compared with the bivalves, the latter split in two groups corresponding to the two subclasses of Heterodonta and Pteriomorpha. The former include two taxa that diverged early, Galeomma and Tridacna, while the Veneridae and Mactridae form two sister groups. In contrast to previous reports and in line with morphological data, the Ostreidae are included in the Pteriomorphia and form a monophyletic group. Received: 16 May 1998 / Accepted: 11 August 1998     

Characterization of a highly repeated DNA family in tapetinae species (mollusca bivalvia: veneridae)

A repetitive DNA family (phBglII400) was characterized in the clam species Tapes philippinarum (Veneridae Tapetinae). The tandemly repeated sequences are AT-rich and show a mainly pericentromeric localization, as most satellite DNAs do. Sequence analysis of phBglII400 DNA family showed a high level of intraspecific homogeneity. Furthermore a 200 bp subunit motif within the 400 bp monomer was apparent as well as the existence of two main "open reading frames" along the 400 bp sequence.In order to investigate the possible distribution of this DNA family among Veneridae, Southern blot analyses were performed on genomic DNAs of Tapes decussatus, Venerupis aurea and Paphia undulata (Tapetinae), Callista chione (Pitarinae), Chamelea gallina (Chioninae) and Venus verrucosa (Venerinae). The phBglII400 family has been found in two additional Tapetinae, namely V. aurea and P. undulata, but not in T. decussatus or other analyzed species. This pattern of sat-DNA distribution supports the high level of differentiation of T. decussatus observed in the previous gene-allozyme analysis. All of these suggest a better allocation of T. decussatus to a genus different from that of T. philippinarum.     

Differential survival of Venus gallina and Scapharca inaequivalvis during anoxic stress: Covalent modification of phosphofructokinase and glycogen phosphorylase during anoxia

Summary Biochemical mechanisms underlying anaerobiosis were assessed in two Mediterranean bivalve species, Scapharca inaequivalvis and Venus gallina, with widely differing tolerances for oxygen lack. These species displayed LT₅₀ values for anoxic survival at 17–18°C of 17 and 4 d, respectively. Succinate and alanine were the major products of 24 h anaerobic metabolism in both species but only S. inaequivalvis further metabolized succinate to propionate. Both species reduced metabolic rate while anoxic but metabolic arrest was more pronounced in S. inaequivalvis. Calculated ATP turnover rate (MATP) during exposure to N₂-bubbled seawater was only 4.51% of the aerobic rate in S. inaequivalvis but was 12.68% in V. gallina. To counteract a greater load of acid end products, V. gallina foot showed a significantly greater buffering capacity, 23.38±0.20 slykes, compared to 19.6±0.79 slykes in S. inaequivalvis. The two species also differed distinctly in the enzymatic regulation of anaerobiosis. In V. gallina anoxia exposure caused only a small change in PFK kinetic parameters (a decrease in K_a AMP) and had no effect on glycogen phosphorylase. By contrast, S. inaequivalvis foot showed a strong modification of enzyme properties in anoxia. The percentage of glycogen phosphorylase in the a form dropped significantly only in S. inaequivalvis. Other changes included alterations in the properties of PFK leading to a less active enzyme form in anoxia. Compared to the aerobic enzyme form, PFK from anoxic foot showed a reduced affinity for fructose-6-P (K_m increased 2.4-fold), greater inhibition by ATP (I₅₀ decreased 6.8-fold), and an increase in sensitivity to AMP activation (K_a decreased by 50%). These enzyme changes appear to be key to a glycolytic rate depression during anaerobiosis in S. inaequivalvis foot muscle.Abbreviations EDTA ethylenediaminetetraacetic acid - EGTA ethyleneglycol-bis-(2-aminoethyl)-tetraacetic acid - Fructose-2,6-P ₂ fructose-2,6-bisphosphate - Fructose-6-P fructose-6-phosphate - K _a AMP Activation constant (concentration of AMP required to increase the reaction to twice the rate it shows in the absence of AMP) - MATP ATP turnover rate - P _i inorganic phosphate - PCA Perchloric acid - PFK 6-phosphofructo-1-kinase - TCA Trichloroacetic acid     

Bioremediation efficiencies of <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis> cultivated in an enclosed sea area of Hangzhou Bay,China

Due to the discharge of nutrients into the East China Sea, severe eutrophication has appeared in Hangzhou Bay. Therefore, we cultivated Gracilaria verrucosa on a large scale in the Jinshan enclosed sea with an area of 1.72 km² in the northern part of Hangzhou Bay to perform bioremediation. The Fengxian enclosed sea with an area of 2.3 km² and 50 km far from Jinshan was used as the control. The results showed that the Hangzhou Bay was severely eutrophicated before G. verrucosa cultivation. During the period of cultivation between August 2006 and July 2007, the annual growth rate of G. verrucosa was 9.42% day⁻¹, and the sea water quality was improved from worse than grade IV to grades II–III, with the concentration of dissolved inorganic nitrogen (DIN) and PO₄-P significantly lower than that in the Fengxian enclosed sea (p < 0.01). The concentration of NH₄-N, NO₃-N, NO₂-N, and PO₄-P after G. verrucosa cultivation was decreased by 54.12%, 75.54%, 49.81%, and 49.00%, respectively. The density of phytoplankton in the Jinshan enclosed sea with cultivation of G. verrucosa was 6.90 –126.53 × 10⁴ cells m⁻³, which was significantly lower than that in the Fengxian enclosed sea. In addition, species diversity, richness, and evenness was significantly increased after cultivation of G. verrucosa in the Jinshan enclosed sea compared with that in the Fengxian enclosed sea. The density of Skeletonema costatum, Prorocentrum micans, and Prorocentrum donghaiense, which were the usual species of red tides at the coastal sea of China, in the Jinshan enclosed sea with cultivation of G. verrucosa was significantly lower. Based on these results, if the water quality in the Jinshan enclosed sea were to be maintained at grade I (DIN ≤0.20 mg  L⁻¹) or II (DIN ≤0.30 mg  L⁻¹), 21.8 t or 18.0 t fresh weight of G. verrucosa need to be cultivated, respectively. These results indicated that large-scale cultivation of G. verrucosa could play a significant role in the bioremediation of Hangzhou Bay.     

Effects of temperature and salinity on the growth of Gracilaria verrucosa and G. chorda, with the Potential for mariculture in Korea

Effects of temperature and salinity on the growth of the two agarophytes, Gracilaria verrucosa (Hudson) Papenfuss and Gracilaria chorda Holmes were examined in Korea. Both species grew over a wide range of temperatures (10–30 ^∘C) and salinities (5–35‰), and grew well at 17–30 ^∘C and a salinity of 15–30‰. In culture, G. verrucosa grew faster than G. chorda and their maximum growth rates were 4.95% day⁻¹ (30 ^∘C, 25‰) and 4.47% day⁻¹ (at 25 ^∘C, 25‰), respectively. In the field population the maximum growth and fertility of G. chorda were observed in summer. The growth rate of G. verrucosa was slightly higher than that of G. chorda for 2 weeks on the cultivation rope and in culture but it was much lower after being contaminated with epiphytes. The biomass of the epiphytes was 0.82 g dry wt. per host plant in G. verrucosa and 0.001 g in G. chorda. G. chorda exhibited resistance to epiphytism and grew 7 times in length and the dry weight increased 15 times after 55 days. In conclusion, G. chorda appears to be a good agarophyte with a fast growth rate and resistance to epiphytesm, and compared with G. verrucosa, has good potential for commercial cultivation.     

Phylogenetic relationships of Western Mediterranean subterranean Trechini groundbeetles (Coleoptera: Carabidae)

Faille, A., Casale, A. & Ribera, I. (2010). Phylogenetic relationships of Western Mediterranean subterranean Trechini groundbeetles (Coleoptera: Carabidae). —Zoologica Scripta, 40, 282–295. Carabid beetles of tribe Trechini (Coleoptera) are one of the main groups of insects that colonized the subterranean environment. Many species of this group have developed similar morphological modifications related to the subterranean life, resulting in a characteristic Aphaenops‐like phenotype that obscures their phylogenetic relationships (depigmented, blind, elongated body and appendages, narrow head and pronotum). We present here the result of a molecular study using a combination of nuclear (small ribosomal unit, large ribosomal unit) and mitochondrial (cox1, cyb, rrnL, trnL, nad1) genes to investigate the phylogenetic placement of the highly modified subterranean genera of the tribe Trechini from the west Mediterranean area (France, Spain, Morocco and Sardinia). Our results confirm the multiple independent origin of troglomorphism among these genera, and reveal a pattern largely determined by geographical proximity. We discuss the validity of some groups proposed on the base of morphological features, and provide estimates of divergence between subterranean genera and other groups of Trechini, including epigean species of the same area. We compare the estimated age for the origin of the main groups resulting from two different calibrations, using one the standard mitochondrial mutation rate (2.3% divergence per Myr) and the other the separation between Sardinia and mainland 33 Ma. Under the first scenario, the main groups of genera would have a late Miocene origin, with a subsequent colonization of north Africa at the Pliocene–Pleistocene boundary. The assumption that the main groups originated through vicariance due to the separation of the Sardinian plate in the Oligocene results in a Messinian origin of the north African subterranean taxa, and a global mitochondrial rate reduced to 1% divergence per Myr.     

Growth,photosynthesis and agar in wild-type strains of Gracilaria verrucosa and G. conferta (Gracilariales,Rhodophyta), as a strain selection experiment

The local species Gracilaria conferta and the foreign G. verrucosa were grown together under a wide range of photon flux density and temperature conditions. Gracilaria verrucosa showed a higher growth rate, especially under low temperatures, and higher photosynthetic performances as well as higher ribulose-1,5-bisphosphate carboxylase activity as compared with G. conferta. Gracilaria verrucosa also showed a better quality and yield of agar, suggesting that this species could be more suitable than G. conferta for outdoor cultivation in Israel and may improve winter growth in ponds. Growth rate and agar quality (gel strength) were rated as the most suitable characteristics influencing the preference of strains for outdoor cultivation.     

Studies on the ultrastructure and taxonomy of the genusTetraselmis (Prasinophyceae)

Ultrastructural investigations on many isolates ofTetraselmis have revealed that the species have characteristic fine structural features of the pyrenoid and it was proposed (Horiet al., 1982) that the genus be subdivided into four subgenera. In the present study of this series, species of the subgenusPrasinocladia, includingTetraselmis marina, T. verrucosa andT. verrucosa f.rubens, are described in detail.     

Molecular phylogeny and systematics of the Pieridae (Lepidoptera: Papilionoidea): higher classification and biogeography

The systematic relationships of the butterfly family Pieridae are poorly understood. Much of our current understanding is based primarily on detailed morphological observations made 50–70 years ago. However, the family and its putative four subfamilies and two tribes, have rarely been subjected to rigorous phylogenetic analysis. Here we present results based on an analysis of molecular characters used to reconstruct the phylogeny of the Pieridae in order to infer higher‐level classification above the generic level and patterns of historical biogeography. Our sample contained 90 taxa representing 74 genera and six subgenera, or 89% of all genera recognized in the family. Three complementary approaches were employed: (1) a combined analysis of a 30 taxon subset for sequences from four gene regions, including elongation factor‐1 alpha (EF‐1α), wingless, cytochrome oxidase subunit I (COI), and 28S (3675 bp, 1031 parsimony‐informative characters), mainly to establish higher‐level relationships, (2) a single‐gene analysis of the 90 taxon data set for sequences from EF‐1α (1066 bp, 364 parsimony‐informative characters), mainly to establish lower‐level relationships, and (3) an all available data analysis of the entire data set for sequences from the four genes, to recover both deep and shallow nodes. Analyses using maximum parsimony, maximum likelihood and Bayesian inference provided similar results. All supported monophyly for the four subfamilies but not for the two tribes, with the Anthocharidini polyphyletic and the Pierini paraphyletic. The combined and all available data analyses support the following relationships among the subfamilies: ((Pseudopontiinae + Dismorphiinae) + (Coliadinae + Pierinae)), corroborating Ehrlich’s 1958 phenetic hypothesis. On the basis of these analyses, and additional morphological and life history evidence, we propose a reclassification of the subfamily Pierinae into two tribes (Anthocharidini s.s., Pierini s.s.) and two informal groups (Colotis group, Leptosia), with the tribe Pierini s.s. subdivided into three subtribes (Appiadina, Pierina, Aporiina) and three genera (Elodina, Dixeia, Belenois) of uncertain status (incertae sedis). The combined and all available data analyses support the following relationships among the Pierinae: (Colotis group + Anthocharidini s.s. + Leptosia + (Elodina + ((Dixeia + Belenois) + Appiadina + Pierina + Aporiina))). Application of a molecular clock calibrated using fossil evidence and semiparametric rate smoothing suggests that divergence between the Pierina and Aporiina occurred no later than the Palaeocene (> 60 Myr). The minimum estimate for the age of the crown‐group of the Pieridae was 112–82 Myr, with a mean of 95 Myr. A historical biogeographical hypothesis is proposed to explain the present‐day distribution of the clade Pseudopontiinae + Dismorphiinae, which argues for an origin of the two subfamilies in western Gondwana (Africa + South America) during the Late Cretaceous. © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society, 2006, 147 , 239–275.     

GEOLOGICALLY DATED SEA BARRIERS CALIBRATE A PROTEIN CLOCK FOR AEGEAN WATER FROGS

Reliable estimates of phylogenetic relationships and divergence times are a crucial requirement for many evolutionary studies, but are usually difficult because fossils are scarce and their interpretation is often uncertain. Frogs are fresh water animals that generally are unable to cross salt water barriers (their skin is readily permeable to both salt and water). The geologically determined ages of salt water barriers that isolate related frog populations thus provide an independent measure of the minimum date of genetic divergence between pairs of such populations. For the genetically well-studied western Palearctic water frogs (Rana esculenta group), the Aegean region provides an ideal area for determining the relationship between genetic divergence and time of spatial isolation, using a nested set of geologically determined isolation times (12,000 yr, 200,000 yr, 1.8 Myr, 2–3 Myr, and 5.2 Myr). Using 31 electrophoretic loci for 33 pairs of neighboring frog populations, a linear relationship between geologically determined isolation time and Hillis' modified Nei genetic distance was found: D^*_Nei = (0.04 ± 0.01) + (0.10 ± 0.01) isolation time [Myr] corresponding to an average divergence rate (“molecular clock” pace) of 0.10 D^*_Nei/Myr (0.10 D_Nei/Myr). This rate is in the range of previous estimates reported for protein electrophoretic data; the value is conservative because relatively few of the loci used are “fast evolvers” (13%; sAAT, ALB, EST-5, MPI). Removing these fast evolvers from the analysis results in 0.08 D^*_Nei/Myr (0.08 D_Nei/Myr). The confidence limits for estimation of the divergence time given the genetic distance are large, but unusually narrow for this kind of study; they permit us to estimate divergence times during the Pliocene and Miocene. Few previous studies, including sequence analyses, have provided reasonable estimates of divergence time for the Pliocene. A test using the outgroup taxa Rana perezi and Rana saharica (also isolated for 5.2 Myr by the Strait of Gibraltar) fits the calibration well: observed genetic Nei distance D^*_Nei = 0.55, expected D^*_Nei = 0.56. The calculated divergence times, based on this absolute molecular clock, suggest a series of speciation events after the Messinian (5.2 Myr), possibly triggered by the rapid ecological changes accompanying the desiccation and refilling of the Mediterranean Basin.     

Estimating the Productivity of the Black Sea Bivalve Chamelea gallinafrom the Size and Biomass of Its Populations

Characteristics of annual production based on various mass data (whole animal mass; shell mass; and wet, dry, and ash-free dry soft tissue mass), the P/B coefficient, and specific production were estimated for the bivalve mollusk Chamelea gallinasampled from 21 localities in the northwestern and eastern Black Sea. The annual production correlated with population biomass (r= 0.994) and mean individual mass (r= –0.737), while the P/Bcoefficient depended mainly on mean individual mass (r= –0.719). Estimates of the P/Bcoefficient in Ch. gallinafrom the Black Sea were smaller than those of other marine invertebrates found worldwide with the same energy equivalent. Multiple linear regression equations were calculated for empirical relationships of various characteristics of annual production to the population biomass and mean individual biomass of Ch. gallina.     

Health risk perspectives of metal(loid) exposure via consumption of striped venus clam (Chamelea gallina Linnaeus, 1758)

The metal(loid)s are mainly transported to the Black Sea via rivers and affect the coastal area where fisheries are intensive. Chamelea gallina (striped venus clam) is the most abundant mollusk species in the Black Sea and used as a bioindicator for monitoring of metal(loid) pollution. In this respect, the concentrations of As, Cd, Cr, Cu, Pb, and Zn in C. gallina collected from 16 different locations along the Black Sea coast (Turkey, Bulgaria, and Russia) were determined and evaluated for the potential human health risk. The results obtained were found to be below the maximum permissible limits indicated in the food safety guidelines. Similarly, the analyzed metal(loid)s did not pose a potential hazard to humans for consumption of the C. gallina, regarding provisional tolerable weekly intake described by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Moreover, target hazard quotient and hazard index were found to be lower than 1 for not only average consumer but also the consumer who consumed a portion (160 g) once a week. In contrast, consumption of C. gallina could cause health risks for Cd and As in case of increasing portion sizes.     

Characterisation of arbuscular mycorrhizal fungi colonisation in cluster roots of shape Hakea verrucosa F. Muell (Proteaceae), and its effect on growth and nutrient acquisition in ultramafic soil

Ultramafic soils at Bandalup Hill (Western Australia) are characterised by high concentrations of Ni and low levels of P. Amongst the plant species that can sustain such hostile conditions, Hakea verrucosa F. Muell from a non-mycorrhizal family (Proteaceae) would be expected to rely on cluster roots to access P. However, the acidification of ultramafic soils by cluster roots might increase the dissolution of soil Ni, and therefore its availability to plants. Symbiosis with mycorrhizal fungi, on the other hand, might help to reduce the uptake of Ni by H. verrucosa. Therefore, the aim of this study was to investigate the mycorrhizal status of H. verrucosa, and assess any contribution from mycorrhizal fungi to its growth and nutrient status. Seedlings of H. verrucosa were first grown in undisturbed ultramafic soil cores from Bandalup Hill for 8 weeks to assess the presence of mycorrhizal fungi in their roots. In a second experiment, H. verrucosa seedlings were grown in the same ultramafic soil that was either steamed or left untreated. Seedlings were inoculated with an arbuscular mycorrhizal (AM) fungal consortium from Bandalup Hill. Fungal hyphae, vesicles, as well as intracellular arbuscules and hyphal coils were observed in the cluster roots of H. verrucosa in both experiments. In the first experiment, 57% of the root length was colonized by AM fungi. Seedlings had high (between 1.4 and 1.9) shoot to root ratios and their roots had very few root hairs, despite growing in P-deficient soil. Steaming of the ultramafic soil increased the growth of seedlings and their nutrient uptake. Inoculation with AM fungi reduced the seedling growth in steamed ultramafic soil; however, it increased their shoot P and K concentration and also the shoot K content. The shoot Ni concentration of seedlings was not affected by the presence of AM fungi.     

Molecular biogeography: using the Corsica-Sardinia microplate disjunction to calibrate mitochondrial rDNA evolutionary rates in mountain newts (Euproctus)

Mitochondrial DNA (mtDNA) sequence variation was examined in the three species belonging to the newt genus Euproctus: E. asper, E. montanus, and E. platycephalus, and in three other species belonging to the same family: Triturus carnifex, T. vulgaris and Pleurodeles waltl. The Euproctus species inhabit mountain streams in the Pyrenean region, Corsica, and Sardinia, respectively. This vicariant distribution is believed to be a result of the disjunction and rotation of the Sardinia-Corsica microplate from the Pyrenean region and suggested dates for each cladogenetic event are available. A total of 915 bp from 12S and 16S ribosomal rRNA genes were compared for each taxon. These are the first mt-rDNA sequence data for salamanders. Sequences were used to reconstruct phylogenetic trees, investigate evolutionary rates for these genes, calibrate them with absolute time since divergence, and compare rates with published ones. Using P. waltl as the outgroup, all phylogenetic methods used (parsimony, maximum likelihood, and Neighbor Joining) produced trees with identical topologies and similar bootstrap values associated with each node. These sequence data cannot unambiguously resolve the splitting events leading to the main radiation of the genus Triturus and the origin of the genus Euproctus. These events may well have occurred very close in time, consistent with other sorts of data. Although it is unlikely strict linearity holds for all kinds of substitutions, relative rate tests of the molecular clock hypothesis could not reject clock-like behavior of sequence changes along Euproctus lineages. Estimates of absolute rates of base changes are 0.35% per Myr since divergence for all substitutions and 0.14% per Myr for transversions; these estimates are similar to other vertebrate estimates. A comparison with distance measures from allozyme studies agrees quite well with regard to relative divergences of the three Euproctus species.     

The yield,physical and chemical properties of agar gel from Gracilaria species (Gracilariales,Rhodophyta) of the Kenya coast

Six species of Gracilaria, G. corticata J. Agardh, G. crassa Harvey, G. millardetii J. Agardh, G. salicornia (J. Ag.) Dawson, G. verrucosa (Huds.) Papenfuss and Gracilaria sp, collected from different stations along the Kenya coast were studied. The yield of hot water native agar extract ranges from 8.1–30% of dry weight, with G. verrucosa and G. salicornia having the greatest and the least yield, respectively. The gel-strength of 1.5% agar solution was highest in G. verrucosa (220 g cm^–2) and lowest in G. corticata (< 60 g cm – 2) whereas the highest gelling temperature was recorded for Gracilaria sp. (40.4 °C) and the lowest in G. verrucosa (28.9 °C). 3,6 anhydrogalactose content was the highest in G. verrucosa and the lowest in G. corticata while sulphate content was higher in the latter.     

Rates of Nucleotide Substitution in Angiosperm Mitochondrial DNA Sequences and Dates of Divergence Between Brassica and Other Angiosperm Lineages

We obtained 16 nucleotide sequences (∼1400 bp each) of the first intron of the mitochondrial (mt) gene for NADH subunit 4 (nad4) from 10 species of Brassicaceae. Using these new sequences and five published sequences from GenBank, we constructed a phylogenetic tree of the Brassicaceae species under study and showed that the rate of nucleotide substitution in the first intron of nad4 is very low, about 0.16–0.23 × 10⁻⁹ substitution per site per year, which is about half of the silent rate in exons of nad4. The ratios of substitution rates in this intron, ITS, and IGS are approximately 1:23:73, where ITS is the nuclear intergenic spacer between 18S and 25S rRNA genes and IGS is the intergenic spacer of 5S rRNA genes. A segment (335 bp) in the first intron of nad4 in Brassicaceae species that is absent in wheat was considered as a nonfunctional sequence and used to estimate the neutral rate (the rate of mutation) in mtDNA to be 0.5–0.7 × 10⁻⁹ substitution per site per year, which is about three times higher than the substitution rate in the rest of the first intron of nad4. We estimated that the dates of divergence are 170–235 million years (Myr) for the monocot–dicot split, 112–156 Myr for the Brassicaceae–Lettuce split, 14.5–20.4 Myr for the Brassica–Arabidopsis split, and 14.5–20.4 Myr for the Arabidopsis–Arabideae split. Received: 14 July 1998 / Accepted: 1 October 1998     

Biochemical and energy dynamics throughout the reproductive cycle of the striped venus Chamelea gallina (Mollusca,Bivalvia)

The striped venus Chamelea gallina is an important commercial bivalve species in Europe. However, large inter-annual fluctuations in stock abundance and periodic recruitment failure threaten the biological and economic sustainability of this fishery. This study aimed to improve the knowledge of the reproductive cycle and reproductive strategies of this species from the Algarve coast (southern Portugal) in order to contribute to the establishment of management measures and to assess its potential for aquaculture. The reproductive cycle of C. gallina followed a seasonal cycle, significantly influenced by sea surface temperature and food availability. Gametogenesis took place in winter, coinciding with the phytoplankton bloom. Spawning occurred during summer, followed by a short period of sexual inactivity in autumn. Condition index did not reflect the reproductive cycle of C. gallina and generally, followed the same trend of chlorophyll a. Glycogen was positively correlated with gonadal index and chlorophyll a. High total lipid values were recorded throughout gonad ripeness and spawning, but decreased at the end of the spawning and in the rest period. The extended spawning period of C. gallina will allow larvae to be obtained for much of the year by artificial spawning of wild broodstock.     

A molecular phylogeny of antarctic chironomidae and its implications for biogeographical history

The chironomid midges Belgica antarctica, Eretmoptera murphyi (subfamily Orthocladiinae) and Parochlus steinenii (subfamily Podonominae), are the only Diptera species currently found in Antarctica. The relationships between these species and a range of further taxa of Chironomidae were examined by sequencing domains 1 and 3–5 of 28S ribosomal RNA. The resulting molecular relationships between B. antarctica and E. murphyi, within Orthocladiinae, were highly supported by validation analyses, confirming their position within Chironomidae, as generated by classical taxonomy. Within Podonominae, P. steinenii from the Maritime Antarctic was more closely related to material from sub-Antarctic South Georgia than to material from Patagonia. Taking advantage of the availability of a molecular substitution rate calculated for this gene in Diptera, a dating of divergence between our study taxa was tentatively established. The divergence dates obtained were 49 million years (Myr), between B. antarctica and E. murphyi, and 68.5 Myr between these species and the closest Orthocladiinae taxon tested from Patagonia, suggesting that B. antarctica and E. murphyi were representatives of an ancient lineage. As both are endemic to their respective tectonic microplates, their contemporary distribution is, therefore, likely to have been shaped by vicariance rather than dispersal.     

Estimation of molecular clocks for ITS and 28S rDNA in Erysiphales

Based on a comparative phylogenetic analysis of Goloviomyces and their host tribes of the Asteraceae, we speculate that Golovinomyces first acquired parasitism to the Asteraceae after migration of the family into the Northern Hemisphere and before the divergence of the tribe Carduaeae. The divergence time of the Carduaeae is estimated to be 25.2Myr ago based on the molecular clock of rbcL sequences of the Asteraceae. When 25.2Myr is given at the node of the first split of the phylogenetic tree of Golovinomyces, nucleotide substitution rates of the Erysiphales are calculated to be 2.52 × 10^–9 per site per year (0.01D = 3.97Myr) in the ITS region and 6.5 × 10^–10 per site per year (0.01D = 15.4Myr) in the D1 and D2 regions of the 28S rDNA.