Cytologic and Genetic Characteristics of Endobiotic Bacteria and Kleptoplasts of Virgulinella fragilis (Foraminifera)

The benthic foraminifer Virgulinella fragilis Grindell and Collen 1976 has multiple putative symbioses with both bacterial and kleptoplast endobionts, possibly aiding its survival in environments from dysoxia (5–45 μmol‐O₂/L) to microxia (0–5 μmol‐O₂/L) and in the dark. To clarify the origin and function of V. fragilis endobionts, we used genetic analyses and transmission electron microscope observations. Virgulinella fragilis retained δ‐proteobacteria concentrated at its cell periphery just beneath the cell membranes. Unlike another foraminifer Stainforthia spp., which retains many bacterial species, V. fragilis has a less variable bacterial community. This suggests that V. fragilis maintains a specific intracellular bacterial flora. Unlike the endobiotic bacteria, V. fragilis klepto‐plasts originated from various diatom species and are found in the interior cytoplasm. We found evidence of both retention and digestion of kleptoplasts, and of fragmentation of the kleptoplastid outer membrane that likely facilitates transport of kleptoplastid products to the host. Accumulations of mitochondria were observed encircling endobiotic bacteria. It is likely that the bacteria use host organic material for carbon oxidation. The mitochondria may use oxygen available around the δ‐proteobacteria and synthesize adenosine triphosphate, perhaps for sulfide oxidation.     

PHYLOGENETIC POSITION OF SYMBIODINIUM (DINOPHYCEAE) ISOLATES FROM TRIDACNIDS (BIVALVIA), CARDIIDS (BIVALVIA), A SPONGE (PORIFERA), A SOFT CORAL (ANTHOZOA), AND A FREE-LIVING STRAIN

The genus Symbiodinium is the commonly observed symbiotic dinoflagellate (zooxanthellae) that forms mutual associations with various marine invertebrates. Numerous studies have revealed that the genus is comprised of a group of diverse taxa, and information on the phylogenetic relationships among the genus’ members is increasing. In this study, small subunit (SSU) ribosomal RNA (ssrRNA) gene sequences were determined for 15 more Symbiodinium strains from 12 relatively unstudied host taxa (Indo-Pacific tridacnids, cardiids, sponge, and soft coral), 1 hitherto unreported free-living Symbiodinium strain, and 4 other Symbiodinium strains from four other host taxa (Indo-Pacific zoanthid, foraminifer, jellyfish, and mid-Pacific hard coral). Their respective phylogenetic positions were inferred, and strains that are either closely related to or distinct from previously reported Symbiodinium taxa were revealed. The cultured Symbiodinium strains isolated from individuals of six species of tridacnids and three species of cardiids all had identical ssrRNA gene sequences, are closely related to S. microadriaticum Freudenthal, and are indistinguishable from the RFLP Type A strain previously reported. However, the ssrRNA gene sequences of clam symbionts that were obtained via gene cloning were different from those of the cultured isolates and represent strains that are close to the RFLP Type C strains. The Symbiodinium-like dinoflagellate from the Indo-Pacific sponge Haliclona koremella De Laubenfels is distinct from any of the Symbiodinium taxa studied and may be similar to the symbiont previously isolated from the stony coral Montipora patula Quelch. The isolates from the soft coral Sarcophyton glaucum Quoy et Gaimard and from the zoanthid Zoanthus sp. are both very closely related to S. pilosum Trench et Blank. The free-living Symbiodinium isolate is very closely related to the symbiont isolated from the Indo-Pacific foraminifer Amphisorus hemprichii Ehrenberg, which in turn is distinct from the Red Sea strain isolated from a similar host. Theisolate from Cassiopeia sp. is different from S. microadriaticum F., the type species harbored by Cassiopeia xamachana Bigelow, and is instead very closely related to S. pulchrorum Trench isolated from a sea anemone. The symbiont from the stony coral M. verrucosa Lamarck is a sister taxon to the symbionts isolated from the foraminifera Marginopora kudakajimensis Gudmundsson and Sorites orbiculus Forskål. These data suggest that polymorphic symbioses extend from cnidarians to some bivalve, foraminifer, and jellyfish host species.     

Phylogenetic incongruence between dinoflagellate endosymbionts (Symbiodinium) and their host foraminifera (Sorites): small-subunit ribosomal RNA gene sequence evidence

Phototrophic dinoflagellate zooxanthellae commonly occur as endosymbionts in many planktic and certain benthic foraminifera (soritids). Many taxonomic issues and specific identities of foraminiferal dinoflagellates are not yet resolved. To assess taxonomic affinities among other dinoflagellates, we have determined the complete nucleotide sequence of the small-subunit rRNA coding region from Symbiodinium sp., an endosymbiotic dinoflagellate of the larger foraminifer Sorites orbiculus. The poly merase chain reaction was adopted for the in vitro amplification of ribosomal DNA, utilizing primers complementary to conserved regions. PCR-amplified DNA was directly sequenced and the sequence was aligned to all complete 18S-rDNA dinoflagellate sequences currently available through GenBank. Apicomplexan, ciliate, chromistacean, and rhodophycean sequences were added to infer across-kingdom phylogenetic relationships. Phylogenetic analysis of aligned nucleotide sequences produced a single most parsimonious tree (generated by the branch and bound method of PAUP). The inferred phylogeny indicates that the dinoflagellate extracted from the foraminifer Sorites orbiculus is a sister taxon to the symbiont present in the larger foraminifera Marginopora kudakajimaensis, but only distantly related to the dinoflagellate isolated from the soritid Amphisorus hemprichii. The sequence heterogeneity demonstrates a high degree of genetic diversity among Symbiodinium-like zooxanthellae and re-emphasizes that they are a variety of distinct entities.The inferred molecular phylogenetic relationships among symbiotic dinoflagellates are not congruent with the foraminiferal phylogeny based on cladistic methodology. The lack of correlation between the evolutionary history of dinoflagellate symbionts and their foraminiferal hosts argues against co-evolution. This lack of co-evolution implies that flexible recombinations among hosts and symbionts are evolutionarily favorable over permanently associated lineages, at least in these benthic foraminifera.     

The distribution of <Emphasis Type="Italic">Symbiodinium</Emphasis> diversity within individual host foraminifera

While one-to-one specificity between reef-dwelling hosts and symbiotic dinoflagellates of the genus Symbiodinium may occur, detailed examination of some hosts reveals that they contain multiple symbiont types. Individuals of the foraminifer Amphisorus hemprichii living in Papua New Guinea contained mixed communities of Symbiodinium dominated by symbiont types in clades C and F. Moreover, the types showed a distinct pattern in their distribution across the radius of the foraminifer, with clade F Symbiodinium more prevalent in the center of the host cell. The mixed community of symbionts and their pattern of distribution within the foraminifer is likely the result of processes happening both inside the foraminifer and in its external environment. Persistent mixed symbiont communities in foraminifera may be stabilized through benefits conferred by maintaining multiple symbiont lineages for symbiont shuffling. Alternatively they may be stabilized through a heterogeneous internal host environment, partitioning of symbiont functional roles or limitation of symbiont reproduction by the host. Six factors generally determine the presence of any particular symbiont type within a foraminifer: mode of transmission, availability from the environment, recognition by the host, regulation by the host, competition between lineages, and fitness of the holobiont. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The oldest record of a boring foraminifer: Early Permian of New Mexico,USA

The bioeroding foraminifer Troglotella incrustans Wernli and Fookes (Bolletino della Societa Paleontologica Italiana 31, 1992, 95), is widely reported from Bajocian?, and Oxfordian to Lower Cenomanian (with a Late Jurassic acme) shallow‐water limestones of the Tethyan realm. A single specimen of a boring foraminifer, assigned to T. incrustans, has now been observed from the Lower Permian (Sakmarian) Community Pit Formation of the Doña Ana Mountains, New Mexico, USA. Surviving the end‐Permian mass extinction, T. incrustans might be a Lazarus taxon that persisted in refuges. This finding represents the oldest record of a foraminifer exhibiting an euendolithic way of life. Boring foraminifera have not been previously recorded from strata older than the Jurassic. Boring traces of potentially foraminiferan origin, however, have been already reported from the Lower Carboniferous (?Ordovician).     

Upper Albian planktic foraminifera and radiolarian biostratigraphy (Nebeur – northern Tunisia)

Preliminary lithostratigraphic and biostratigraphic data recovered from Upper Albian pelagic successions of the Jebel Srassif area (Tunisian trough) reveal the presence of a moderately well preserved radiolarian fauna. This fauna is fossilized in blackshale beds of Rotalipora subticinensis, R. ticinensis and the lower part of the R. appenninica foraminifer zones. Thirty-five radiolarian morphotypes were identified. A correlation established with foraminifer planktic biochronology allows us to distinguish three radiolarian assemblages. The close association between organic-rich deposits and the radiolarian bloom can be interpreted as productivity proxy triggered by semi-enclosed basin configuration and an intensified upwelling currents provided by a possibly halokinetic-induced hydrothermalism.     

The Process of Chamber Formation in the Foraminifer Rosalina floridana (Cushman)

SYNOPSIS. The foraminifer Rosalina floridana builds a chambered, calcareous test which is periodically enlarged by the addition of new chambers. R. floridana begins to form a chamber by constructing an algal growth cyst which covers the dorsal side of the animal and is cemented to the substrate. The pseudopods that build the cyst coalesce to form a cytoplasmic template or anlage on which the chamber walls will be secreted. Electron micrographs reveal the anlage cytoplasm to be a “froth” of nearly empty vesicles which contain mitochondria, fibrillar material, and electron dense granules. The organic lining of the new chamber is secreted on the anlage by pseudopods extending thru it. After the organic lining is completed, cytoplasm from within the test flows into the forming chamber and forces the anlage cytoplasm out thru the new aperture. This “frothy” anlage cytoplasm forms a sheath over the dorsal surface of the test; while it is in place, a layer of calcite is deposited on the walls of the new chamber and over the rest of the test. When calcification is completed, the sheath breaks up and is incorporated into newly formed pseudopods as the foraminifer gradually moves out of the growth cyst and begins normal feeding. The production of vesiculated cytoplasm in normal pseudopods and in the anlage is viewed as a method of greatly increasing cytoplasmic volume with a resultant very small loss in cytoplasmic mass. In the anlage during the production of the organic lining the vesiculated cytoplasm apparently acts only as a support for the membranes being secreted by the pseudopods and presumably does not take part in the secretory process. This same cytoplasm forms the sheath that is present during calcification. The view is advanced that, altho the sheath could be active during calcification with the mitochondria within the vesicles actively transporting calcium to sites of crystal growth, its more probable function is to form a partition between the parts to be calcified and the environment. The foraminifer could then secrete CaCO₃ from pooled reserves in the cytoplasm into the area between the sheath and the chamber membranes. The basal membrane is considered the nucleating agent during calcification and is responsible for the ordering of the crystals so their C axes are perpendicular to the surface of the chamber.     

Siphodinarella costata n. gen., n. sp., a new benthic foraminifer from the Coniacian of the Adriatic Carbonate Platform (Slovenia, Croatia)

A new benthic foraminifer is described as Siphodinarella costata n. gen., n. sp. from Coniacian shallow-water platform-interior carbonates of Slovenia and Croatia. The new foraminifer is found in skeletal wackestone in association with small benthic foraminifera, thaumatoporellaceans, and calcimicrobes (Decastronema, Girvanella-type tubes). The existence of an internal siphon in Siphodinarella n. gen. is interpreted as an entosolenian tube and discussed in terms of its generic and suprageneric importance.     

New Nemonychidae,Brentidae and Curculionidae (Coleoptera: Curculionoidea) from the Turonian of Kzyl-Dzhar (Kazakhstan)

Five new genera, Turononemonyxn. gen. (type species: Turononemonyxsamsonovin. sp.) (Nemonychidae: ? Cretonemonychinae: ? Cretonemonychini), Falsotanaosn. gen. (type species: Falsotanaos convexusn. sp.), Pretanaosn. gen. (type species: Pretanaosocularisn. sp.), Longotanaosn. gen. (type species: Longotanaosrasnitsynin. sp.) from Brentidae: Apioninae: Tanaini) and Turonerirhinusn. gen. (type species: Turonerirhinuskaratavensisn. sp.) from Curculionidae (Erirhininae: Erirhinini), and seven new species, Falsotanaosconvexusn. sp., Paratanaos samsonovin. sp., Pretanaos ocularisn. sp., Longotanaos rasnitsynin. sp., Turonerirhinus karatavensisn. sp., Turonerirhinuspunctatusn. sp. and Turonerirhinus poinarin. sp., are described from Kzyl-Dzhar locality (Kazakhstan, Upper Cretaceous, Turonian).http://www.zoobank.org/urn:lsid:zoobank.org:pub:13E0316E-C229-471A-90AA-2D71253B12F9     

The Cyatheaceae (Polypodiopsida) of Peru

A summary is presented of all Peruvian species of scaly tree ferns (Cyatheaceae). A total of 83 species is native to Peru (Sphaeropteris 1 sp., Alsophila sect. Alsophila 9 spp., Alsophila sect. Gymnosphaera 1 sp., Cyathea 72 spp.), with 13 endemic species in Cyathea and one in Alsophila. The following new species are described and illustrated: Alsophila gastonyi, Cyathea chontilla, C. nephele, C. polliculi, and C. praetermissa. The genera Cnemidaria and Trichipteris are no longer recognized and merged in Cyathea. Necessary combinations are made for Cyathea alatissima, C. nervosa, and C. uleana. Cyathea serpens is presented in detail and its relationship is discussed. An artificial key to the Peruvian species of Cyatheaceae is provided.     

Recurrent origin of peripheral,coastal (sub)species in Mediterranean Senecio (Asteraceae)

Background: It is argued that coastal endemic taxa may evolve in parallel at the periphery of the distributional range of a widespread species.

Aims: We tested this hypothesis for the origins of three peripheral, coastal isolates of Senecio, S. glaucus ssp. glaucus (Israel), S. g. ssp. coronopifolius p.p. (Sicily), and S. hesperidium (Morocco), from widespread S. glaucus ssp. coronopifolius. We also determined the relative roles of selection vs. genetic drift in shaping phenotypic divergence in ssp. glaucus and S. hesperidium, using Lande’s test of neutral morphological change.

Methods: We surveyed morphological and/or allozyme variation in the three peripheral isolates and mainly inland populations of S. g. ssp. coronopifolius.

Results: Genetic data supported independent origins of the coastal taxa from nearby populations of ssp. coronopifolius. These descendant and ancestral populations showed pronounced morphological but weak genetic differentiation. Phenotypic similarities between ssp. glaucus (Israel) and S. hesperidium (Morocco) in plant height and floral traits may have resulted from parallel divergent selection from ssp. coronopifolius, though drift remains an alternative cause in S. hesperidium.

Conclusions: Our results indicate parallel ecotype formation and (sub)speciation in Senecio in which primarily selective vs. neutral determinants promoted the recurrent origin of coastal types in, respectively, Israel and Morocco.     

Two new species of Tipula (Lunatipula) Edwards (Diptera: Tipulidae) from Turkey

Two new species in the acuminata group of subgenus Tipula (Lunatipula), Tipula (Lunatipula) oosterbroeki sp. n. and T (L) jaroslavi sp. n., were described and illustrated from Southwest part of Asian Turkey.     

Philopotaminae of Madagascar (Trichoptera: Philopotamidae)

The Malagasy fauna includes two genera of Philopotaminae: Wormaldia McLachlan 1865 and an endemic new genus, Ranarijaodes n. gen. The Malagasy and African (sensu stricto) lineages of Wormaldia are distinct. Apomorphic characters (the shape of the phallic apparatus for the African species and the splitting of the median lobe of the tenth tergite for the Malagasy species) confirm their separation. New captures are reported for two of the four previously known species of Wormaldia. Seven new species are described: Wormaldia ambahibeae n. sp., W. faharoa n. sp., W. fahatelo n. sp., W. randriamasimananai n. sp., W. fahefatra n. sp., W. legrandi n. sp. and W. fahadimy n. sp. Ranarijaodes n. gen. is close to Dolophilodes Ulmer 1909. It is described for the three new species Ranarijaodes fiana n. sp., R. angapi n. sp. and R. atsimo n. sp. The 14 Malagasy species of Philopotaminae are endemics of the island. Their geographical distributions and ecological preferences are described and discussed.     

Effective Cytochrome P450 (CYP) Inhibitors Isolated from Tarragon (Artemisia dracunculus)

Two effective cytochrome P450 (CYP) inhibitors were isolated from tarragon, Artemisia dracunculus. Their structures were spectroscopically identified as 2E,4E-undeca-2,4-diene-8,10-diynoic acid isobutylamide (1) and 2E,4E-undeca-2,4-diene-8,10-diynoic acid piperidide (2). Both compounds had dose-dependent inhibitory effects on CYP3A4 activity with IC₅₀ values of 10.0 ± 1.3 µM for compound 1 and 3.3 ± 0.2 µM for compound 2, and exhibited mechanism-based inhibition. This is the first reported isolation of effective CYP inhibitors from tarragon (Artemisia dracunculus) purchased from a Japanese market.     

Preparation of enantiomerically pure (R)-(1-hydroxyethyl)dimethyl(phenyl)silane using resting cells of Saccharomyces cerevisiae (DHW S-3) as biocatalyst

The prochiral sila-ketone acetyldimethyl-(phenyl)silane (1) was reduced enantioselectively into (R)-(1-hydroxyethyl)dimethyl(phenyl)silane [(R)-2] using resting cells of the commercially available yeast Saccharomyces cerevisiae (DHW S-3) as the biocatalyst. The bioconversion was performed on a 2.0-g scale in a 5-1 bioreactor. Starting with a substrate (1) concentration of 0.4 g·1^–1, the highest production rate measured for this bioconversion was about 45–55 mol (R)-2·1^–1·min^–1. After an incubation time of 1 h, all substrate in the medium had been converted, either biocatalytically reduced to (R)-2 or (probably chemically) converted into dimethyl(phenyl)silanol (Me₂PhSiOH). After extraction of the cell-free medium with ethyl acetate/dichloromethane and subsequent purification of the extract by Kugelrohr distillation and chromatography on silica gel (medium-pressure liquid chromatography), 800 mg (yield 40%) of the bioconversion product (R)-2 was isolated. As shown by HPLC studies (cellulose triacetate as the chiral stationary phase) and ¹H-nuclear magnetic resonance experiments (after derivatization of the bioconversion product with a chiral auxiliary agent), compound (R)-2 was almost enantiomerically pure (> 99% enantiomeric excess).This article is dedicated to Prof. Dr. Fritz Wagner on the occasion of his 65th birthday     

Total Synthesis of ( + )-Methyl Phaseates

( ± )-Methyl phaseates were synthesized from ( ± )-4-(6′-acetoxymethyl-2 ′,6′-dimethyl-1′-cyclohexen-1′-y1)-but-3-en-2-one (20), which was prepared from a useful terpenoid building block, ( ± )-2-hydroxymethyl-2,6-dimethyl-1-cyclohexanone (11a and 11b). Photooxidation of the cyclohexadiene intermediate (22), followed by alkaline hydrolysis and methylation, gave four stereoisomers of ( ± )-methyl phaseates: (2Z,4E)-cis form (2), (2E,4E)-cis form (24), (2Z,4E)-trans form (25) and (2E,4E)-trans form (26).     

Fertility tracers and monsoon forcing at an equatorial site of the Somali Basin (Northwest Indian Ocean)

Temporal changes in biological and isotopic tracers have successfully been used to monitor monsoon variability in the Arabian Sea. Convincing evidence that fertility indicators, such as the relative abundance of the planktic foraminifer Globigerina bulloides, and SiO₂ and Ba fluxes, can monitor monsoon changes, has also been published. Time series of the ¹³C content of the thermocline foraminifer Neogloboquadrina dutertrei, and of an Upwelling Radiolarian Index (URI), have been used to reconstruct upwelling changes in the Somali Basin at 5 °N for the last 160 kyr. In order to establish a reference site for comparison with other upwelling and/or fertility records from the same area, a time series of 8 climatic and fertility proxies (¹³C content of N. dutertrei and Globorotalia menardii, percentages of G. bulloides and of a thermocline foraminiferal group, minimal δ¹⁸ values of Globigerinoides sacculifer, N. dutertrei and G. menardii) are investigated in the spectral domain for the last 360 kyr, at a site located at the equator and outside the Somali upwelling centers (Core MD 85668).Chronostratigraphies for these records are developed by correlation of oxygen isotope record of G. sacculifer to the Martinson standard stacked benthic oxygen isotope record. The temporal resolution of the data, is between 2 and 5 kyr.Cross spectral comparisons with ETP (Eccentricity + Tilt + Precession-composite signal) were used to estimate coherences in conjunction with phase relationships and to quantify relationships between fertility and climatic indicators. The timing of fertility indicator cycles at the equator shows little (obliquity band) or no (other orbital bands) coincidence with monsoon proxies of the Arabian Sea. At the equator, fertility responses are dissociated, and present important leads or lags with ETP signal at the three orbital bands (with the exception of the thermocline foraminifera, in phase with maximum eccentricity). Due to significant leads with ETP in the timing of ¹³C minima and URI cycles, no relationships with the Northern Hemisphere radiation can be deduced. At the obliquity and precessional frequency bands, thermocline foraminifera cycles display significant lag with ETP and ice volume minima, which suggests that radiation is not the sole forcing mechanism. Global climatic and ice volume changes may be an important forcing mechanism for these fertility proxies. The G. bulloides record is more complicated with cycles in phase with Arabian Sea monsoon proxies at the obliquity band, and ¹⁸O minima at the precession frequency.It appears that, at the equator, the southwest monsoon winds are not the main force driving radiolarian and foraminiferal productivity.     

Novelties in Mascagnia (Malpighiaceae)

Seven new species ofMascagnia are described:M. tomentosa from southern Mexico and Central America;M. arenicola from the Guianas;M. riparia, M. tucuruensis, andM. velutina from Brazil;M. boliviana from Bolivia; andM. australis from Argentina and Paraguay. These novelties were previously included in the “M. sepium-complex,” an omniumgatherum comprising superficially similar yellow-flowered taxa.     

2-Alkyl(aryl)-quinazolin-4(3H)-thiones, 2-R-(quinazolin-4(3H)-ylthio)carboxylic acids and amides: synthesis,molecular docking,antimicrobial and anticancer properties

In this study, a series of novel 2-alkyl(aryl)-quinazolin-4(3H)-thiones, 2-R-(quinazolin-4(3H)-ylthio)carboxylic acids and amides were synthesized and evaluated for antimicrobial and anticancer activities. Their structure was confirmed by elemental analysis and spectral data (FT-IR, LC-MS, ¹H-NMR). Antimicrobial activity was tested in vitro against Staphylococcus aureus, Enterococcus faecalis, Enterobacter aerogenes, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumonia, Candida albicans and NCI in vitro preliminary anticancer activity against nine different cancer types. The most active antibacterial and antifungal compounds were: 2.1, 2.2 and 2.4. The introduction of the carboxylic acid or amide residue into the fourth position of quinazolin-4(3H)-thione resulted in the absence of antimicrobial activity. Substance 3.8 inhibited renal cancer UO-31 line and 2.18 – leukemia CCRF-CEM. The results of in silico molecular docking for DHFR and CK2 kinase had no correlation with in vitro properties, proposing the presence of other biological activity pathways.