Characterization of 11 microsatellite loci derived from genomic sequences of polychaete Capitella capitata complex

Eleven microsatellite loci derived from the genomic sequence data of Capitella capitata were characterized using 30 samples. The observed number of alleles per locus ranged from four to 36. The levels of observed and expected heterozygosities for polymorphic loci were from 0.10 to 0.87 and from 0.37 to 0.98, averaging 0.52 and 0.77, respectively. Analyses of Hardy–Weinberg equilibrium and genotypic linkage disequilibrium suggest the possible presence of both null alleles and Wahlund effect. One of the 11 loci was difficult to amplify for genotyping. Therefore, the rest 10 loci are good molecular markers for population genetic analysis.     

Patterns of protein synthesis and accumulation during oogenesis in the polychaete,Pseudopotamilla occelata Moore

Summary

Proteins synthesized and accumulated during oogenesis or Pseudopotamilla occelata were analyzed by two-dimensional Polyacrylamide gel electrophoresis and fluorography. Significant changes in the patterns of synthesis and accumulation occur during oogenesis, paticularly between previtellogenic and vitellogenic stages, although many of the proteins are represented throughout the process. The changes may be generally dependent upon the variation in the gene expression affecting autosynthesis of proteins. However, appearance of proteins, which occur only at the vitellogenic stages but apparently not synthesized within the oocytes, indicates that heterosynthetic proteins are stage-specifically transported into oocytes.     

Twelve microsatellites for two deep sea polychaete tubeworm species,Lamellibrachia luymesi and Seepiophila jonesi,from the Gulf of Mexico

The vestimentiferan tubeworms Lamellibrachia luymesi and Seepiophila jonesi are found at hydrocarbon seeps in the Gulf of Mexico. Primers for polymorphic microsatellite loci were developed from genomic libraries of L. luymesi (five loci) and from S. jonesi tissue (eight loci) and were used to screen individuals collected from nine northern Gulf of Mexico hydrocarbon seep sites. Loci had from four to more than 50 alleles with high expected levels of heterozygosity. Cross‐species amplification, tested on seven vestimentiferan species including both hydrothermal vent and cold seep species, was generally strong in similar species but weak in more genetically distant species.     

Characterization of six polymorphic microsatellites for the polychaete tubeworm Hydroides elegans and cross‐species amplification in the congener Hydroides hexagonus

We isolated and characterized six polymorphic microsatellite loci for the polychaete tubeworm, Hydroides elegans. Two additional loci were not reliably scorable and estimates of heterozygosity were obtained for the other six. In addition, cross‐species amplification was successful for two loci using the congener H. hexagonus. Given that few microsatellite loci are available for polychaetes, these markers will be useful in assessing dispersal and gene flow in H. elegans and probably also other polychaetes.     

Behavioural responses of the yellow emitting annelid Tomopteris helgolandica to photic stimuli

In contrast to most mesopelagic bioluminescent organisms specialised in the emission and reception of blue light, the planktonic annelid Tomopteris helgolandica produces yellow light. This unusual feature has long been suggested to serve for intraspecific communication. Yet, this virtually admitted hypothesis has never been tested. In this behavioural study of spectral colour sensitivity, we first present an illustrated repertoire of the postures and action patterns described by captive specimens. Then video tracking and motion analysis are used to quantify the behavioural responses of singled out worms to photic stimuli imitating intraspecific (yellow) or interspecific (blue) bioluminescent signals. We show the ability of T. helgolandica to react and to contrast its responses to bioluminescent‐like blue and yellow light signals. In particular, the attractive effect of yellow light and the variation of angular velocity observed according to the pattern of yellow stimuli (flashes versus glows) support the intraspecific communication hypothesis. However, given the behavioural patterns of T. helgolandica, including mechanically induced light emission, the possibility that bioluminescence may be part of escape/defence responses to predation, should remain an open question.     

Vismiaquinone,a Δ1-isopentenyl substituted anthraquinone from Vismia reichardtiana

1,8-Dihydroxy-7-(Δ¹-isopentenyl)-6-methoxy-3-methylanthraquinone (vismiaquinone) was isolated from the leaves of Vismia reichardtiana. its structure was established through chemical and spectral means. A hypothesis for the biogenesis of the uncommon Δ¹-isopentenyl side chain is presented.     

Comparison of the thermostability of recombinant luciferases from Brazilian bioluminescent beetles: Relationship with kinetics and bioluminescence colours

Firefly luciferases have been used extensively as bioanalytical reagents and their cDNAs as reporter genes for biosensors and bioimaging, but they are in general unstable at temperatures above 30°C. In the past few years, efforts have been made to stabilize some firefly luciferases for better application as analytical reagents. Novel luciferases from different beetle families, displaying distinct bioluminescence colours and kinetics, may offer desirable alternatives to extend the range of applications. In the past years, our group has cloned the largest variety of luciferases from the three main families of bioluminescent beetles (Elateridae: P. termitilluminans, F. bruchi, P. angustus; Phengodidae: P. hirtus, P. vivianii; and Lampyridae: A. vivianii, C. distinctus and Macrolampis sp2) occurring in Brazilian biomes. We compared the thermostability of these recombinant luciferases and investigated their relationships with bioluminescence spectra and kinetics. The most thermostable luciferases were those of Pyrearinus termitilluminans larval click beetle (534 nm), Amydetes vivianii firefly (539 nm) and Phrixotrix vivianii railroad worm (546 nm), which are the most blue‐shifted examples in each family, confirming the trend that the most blue‐shifted emitting luciferases are also the most thermostable. Comparatively, commercial P. pyralis firefly luciferase was less thermostable than P. termitilluminans click beetle and A. vivianii firefly luciferases. The higher thermostability in these luciferases could be related to higher degree of hydrophobic packing and disulfide bond content (for firefly luciferases).     

Description and relationships of Chaetopterus pugaporcinus, an unusual pelagic polychaete (Annelida, Chaetopteridae)

An extraordinary new species, Chaetopterus pugaporcinus, is described from eight specimens collected from deep mesopelagic waters off Monterey Bay, California, by remotely operated vehicles. All specimens exhibit a consistent combination of both adult and larval characteristics, leaving in question the maturity of the specimens. All specimens lack ciliated larval bands and the stout, modified chaetae (cutting spines) typically found in segment A4 of chaetopterids. If the specimens described here are larvae, they are remarkable for their size, which ranged from 10 to 21 mm total length, nearly twice the length of the largest polychaete larvae previously reported and 5 to 10 times larger than known chaetopterid larvae. Then too, their lack of segment addition prior to settlement would be atypical. If adult, they are particularly unusual in their habitat choice and body form. Morphology of the uncini and comparison to larval morphology indicated a close relationship to either Chaetopterus or Mesochaetopterus. However, the lack of cutting spines and typical adult morphology made it impossible to determine to what genus this species should be allied. Thus, we carried out the first molecular phylogenetic analysis of the Chaetopteridae in order to appropriately place and name the new species. Three partial genes were sequenced for 21 annelid species. The sequencing also provides the first molecular evidence that Chaetopterus variopedatus sensu Hartman (1959) is not a single cosmopolitan species. The question of C. pugaporcinus being a delayed larva or a genuine holopelagic chaetopterid is discussed.     

Comparison of the spectral emission of lux recombinant and bioluminescent marine bacteria

The purpose of the present paper was to study the influence of bacteria harbouring the luciferase‐encoding Vibrio harveyi luxAB genes upon the spectral emission during growth in batch‐culture conditions. In vivo bioluminescence spectra were compared from several bioluminescent strains, either naturally luminescent (Vibrio fischeri and Vibrio harveyi) or in recombinant strains (two Gram‐negative Escherichia coli::luxAB strains and a Gram‐positive Bacillus subtilis::luxAB strain). Spectral emission was recorded from 400 nm to 750 nm using a highly sensitive spectrometer initially devoted to Raman scattering. Two peaks were clearly identified, one at 491–500 nm (± 5 nm) and a second peak at 585–595 (± 5 nm) with the Raman CCD. The former peak was the only one detected with traditional spectrometers with a photomultiplier detector commonly used for spectral emission measurement, due to their lack of sensitivity and low resolution in the 550–650 nm window. When spectra were compared between all the studied bacteria, no difference was observed between natural or recombinant cells, between Gram‐positive and Gram‐negative strains, and growth conditions and growth medium were not found to modify the spectrum of light emission. Copyright © 2003 John Wiley & Sons, Ltd.     

Effects of feeding on zinc and cadmium accumulation by the polychaete Neanthes arenaceodentata

Abstract

The effects of feeding on the kinetics of accumulation and depuration of ⁶⁵Zn and ¹⁰⁹Cd and on the subcellular distribution of these metals has been studied in the polychaete Neanthes arenaceodentata (Moore). Feeding increases the rate of accumulation of Zn and Cd and decreases the length of time for the metals to reach a steady state concentration in the animals. These effects can be attributed to the adsorption of metals from the medium by the algal food. Feeding does not have a marked effect on the depuration of either of the metals. Both metals are lost in a biphasic manner. The initial phase probably represents the elimination of unassimilated metals from the gut lumen.

Feeding does not dramatically alter the subcellular distribution of the metals. Zinc is found primarily in the 200 g pellet while Cd is primarily found in the cytosol associated with two Cd- binding ligand pools with apparent molecular weights of 9.7 and 5.0 kDa. The former co-elutes with purified metallothionein. The latter contains the majority of the Cd in the cytosol and is prominent in fed organisms. The use of these ligands as subcellular indicators of metal induced stress is discussed.     

Early Middle Ordovician scolecodonts from north‐western Argentina and the emergence of labidognath polychaete jaw apparatuses

Scolecodonts provide fossil evidence of the evolution and diversification of jaw‐bearing polychaetes from the latest Cambrian onwards. However, their record before the Darriwilian (Middle Ordovician) is scarce worldwide, which limits our understanding of key evolutionary events. One such event is the emergence of taxa possessing the asymmetrical labidognath‐type jaw apparatus architecture, which became common in the Middle Ordovician and is often dominant throughout the Palaeozoic. Here, we document a small collection of Dapingian scolecodonts from the Capillas section, Sierras Subandinas, north‐western Argentina. The isolated elements recovered allowed us to reconstruct the distinctive jaw apparatus, and to introduce a new taxon, Andiprion paxtonae gen. et sp. nov. The maxillary apparatus of Andiprion is intermediate between the symmetrognath type of the Early Ordovician Kadriorgaspis and the labidognath type that is present in polychaetaspids and related taxa. The apparatus architecture of Andiprion corresponds best to the labidognath type, but the morphology of the individual jaws suggests that it may be the most primitive representative of this lineage currently known. We propose that Andiprion‐like forms were ancestral to polychaetaspids, polychaeturids and ramphoprionids. The Capillas collection provides supporting evidence for the evolutionary homology of the ‘basal plate’ and the left first maxilla. Thus the labidognath‐type asymmetry, with an unpaired left maxilla III, developed as a result of gradual reduction in size of the first right jaw (‘basal plate’) in front of the carriers, instead of loss or fusion of anterior maxillae.     

Calcium-containing granules in myoepithelial cells of the polychaete syllis spongiphila: Possible ionic modulators

Myoepithelial cells of the polychaete Syllis spongiphila have a central core containing abundant membrane-bound granules. Examination of isolated and in situ granules using radioactive labelling, X-ray energy dispersive spectrometry, and scanning and transmission electron microscopy indicated that the granules contain high levels of Ca and P, and take up free Ca ions with a time course of 15-30 min. The granules appear to contribute to the dynamic calcium metabolism of the myoepithelial cells.     

Microtiter plate tests for segregation of bioluminescent bacteria

It has been recently shown that bioluminescence imaging can be usefully applied to provide new insights into bacterial self‐organization. In this work we employ bioluminescence imaging to record images of nutrient rich liquid cultures of the lux‐gene reporter Escherichia coli in microtiter plate wells. The images show that patterns of inhomogenous bioluminescence form along the three‐phase contact lines. The paper analyzes the dependencies of the average number of luminous aggregates (clouds) on various environmental factors. In particular, our results show that optimal (neutral) pH and high aeration rates determine the highest mean number of clouds, and that spatiotemporal patterns do not form in the pH buffered suspensions. In addition, a sigmoidal (switch‐like) dependence of the number of aggregates on the rate of aeration was observed. The obtained bioluminescence imaging data was interpreted by employing the Keller–Segel–Fisher (KSF) model of chemotaxis and logistic growth, adapted to systems of metabolically flexible (two‐state) bacteria. The modified KSF model successfully simulated the observed switch‐like responses. The results of the microtiter plate tests and their simulations indicate that the segregation of bacteria with different activities proceeds in the three‐phase contact line region. Copyright © 2015 John Wiley & Sons, Ltd.     

A luxCDABE-based bioluminescent bioreporter for the detection of phenol

A bioluminescent reporter strain, Acinetobacter sp. DF4-8, was constructed for the detection of phenol by inserting a mopR-like promoter upstream of the Vibrio fischeri bioluminescent luxCDABE gene cassette in a modified mini-Tn5 construct. When introduced into the chromosome of Acinetobacter sp. DF4, the bioreporter produced a sensitive bioluminescent response to phenol at concentrations ranging from 2.5 to 100 ppm. This response was linear (R ²=0.986) in the range from 20 to 90 ppm. A significant bioluminescent response was also recorded when strain DF4-8 was incubated with slurries from aged, phenol-contaminated soil. Received 13 February 2002/ Accepted in revised form 11 July 2002     

Isolation and Characterization of Highly Polymorphic Microsatellites from the Polychaete Pectinaria koreni

We report on the isolation of dinucleotide microsatellites from the polychaete Pectinaria koreni using (GT)₁₀ and (CT)₁₀ olignonucleotide probes. Compound and particularly imperfect microsatellites are predominant in this species. In most cases the associated element is (AT)_n leading to AT microsatellites being the most frequent class after GT repeats. Obtaining single-copy polymerase chain reaction products in the expected size range proved to be difficult, and only 24% of the primer pairs tested produced polymorphic single-locus markers. We obtained five highly variable loci with 16 to 41 alleles and an observed heterozygosity ranging from 0.29 to 0.81. Amplifications have also been obtained from the earliest postlarval stage. These highly informative markers will allow studies of the population structure of P. koreni at fine spatial and temporal scales. Received March 19, 1999; accepted September 10, 1999.     

The role of the heart-body and of the extravasal tissue in disposal of foreign cells in two polychaete annelids

Sheep red cells injected into the coelom of the polychaetes Neoamphitrite figulus and Arenicola marina are phagocytosed by coelomocytes and conveyed to the heart-body and extravasal ('chloragogen') tissue respectively. In the former case the phagocytes creep into the vessels, penetrate the heart-body tissue and bequeath the sheep red cells to the heart body cells. In Arenicola the phagocytes accumulate on the tips of the blind ending vessels and the red cells are similarly transferred to the extravasal tissues. These observations confirm that the heart-body and extravasal cells function not only in production of haemoglobin but in sequestering foreign material.     

A bioluminescent assay for the determination of phosphoenolpyruvate carboxykinase activity in nanogram-sized tissue samples

A highly specific and sensitive assay for the determination of phosphoenolpyruvate carboxykinase (PEPCK) in nanogram-sized tissue samples is described. This test system is based on the stoichiometric transformation of phosphoenolpyruvate into ATP. In a subsequent step ATP is quantified by bioluminescent techniques. The applicability of this assay system is shown by measurements in liver samples with normal and high PEPCK activity levels.