Native bradyrhizobial symbionts of Lupinus mariae-josephae, a unique endemism thriving in alkaline soils in Eastern Spain

Lupinus mariae-josephae (Lmj) es una especie de lupino endémica de una pequeña y específica área de Comunidad Valenciana (Este de España), donde prospera en suelos alcalinoscalcáreos, un hábitat singular para los altramuces, que crecen preferentemente en suelos ácidos o neutros. Esto hace de Lmj una especie de lupino única. Cuando se inició este trabajo, la extensión conocida de este endemismo abarcaba unos 700 kilómetros cuadrados, confinados en la provincia de Valencia. En esta área, Lmj prospera en pequeñas poblaciones aisladas que contienen un número reducido de plantas por lo que se la consideró una especie en peligro de extinción. Todos los esfuerzos, utilizando estrategias clásicas dirigidas a ampliar el área de crecimiento de Lmj y garantizar su conservación, han tenido un éxito limitado. El trabajo que se presenta está dirigido a mejorar el conocimiento de la ecología de Lmj, en particular la interacción simbiótica que establece con bacterias del suelo denominadas rizobios y se centra en la caracterización fenotípica, filogenética y genómica de esos rizobios. También se investiga la posible contribución de la simbiosis en mejorar la conservación de Lmj. Para este fin, se han estudiado diferentes aspectos que se describen a continuación. El primero objetivo se centró en aislar y estudiar de la diversidad genética de las bacterias endosimbióticas de Lmj. . Se realizó un análisis filogenético de genes esenciales que mostró que las cepas de Lmj pertenecen al género Bradyrhizobium y que presentan una gran diversidad con características fenotípicas y simbióticas diferentes de cepas de Bradyrhizobium que nodulan otras especies de lupinos nativos de España (cepas ISLU). Las cepas estudiadas se dividieron en dos grupos (Clado I y Clado II). El Clado I, incluye a las cepas Lmj, definiendo un nuevo linaje, filogenéticamente relacionado con otras especies de Bradyrhizobium, como B. jicamae y B. elkanii. El Clado II contiene cepas ISLU relacionadas con cepas de B. canariense y B. japonicum que establecen simbiosis con lupinos de suelos ácidos. Otro análisis filogenético basado en genes simbióticos, distribuyó las cepas de Lmj en sólo dos grupos diferentes. La singularidad y gran diversidad de estas cepas en una pequeña área geográfica, hacen de este, un atractivo sistema para el estudio de la evolución y adaptación de las bacterias simbióticas a su respectiva planta huésped. Adicionalmente, se estudio la presencia de bacterias capaces de nodular Lmj en suelos básicos de Chiapas, México. Sorprendentemente, estos suelos contienen bacterias capaces establecer interacciones simbióticas eficientes con Lmj en ensayos de invernadero. A continuación se investigó la taxonomía de los endosimbiontes de Lmj analizando la secuencia de cuatro genes esenciales (16S rRNA, recA, glnII y atpD) y el promedio de identidad de nucleótidos de genomas completos de algunas cepas representativas de la diversidad (ANIm). Se identificaron nuevas especies de Bradyrhizobium dentro del Clado I y se definió una de ellas: 'Bradyrhizobium valentinum' sp. nov (cepa tipo LmjM3T = CECT 8364T, LMG 2761T). También se abordó cómo conservar Lmj en su hábitat natural mediante inoculación con alguna de las cepas aisladas. Se demostró la ausencia de bacterias capaces de nodular Lmj en suelos rojos alcalinos o ‘‘terra rossa’’ de la Península Ibérica y Baleares. Dos cepas, altamente eficientes en cuanto a la fijación de nitrógeno, LmjC y LmjM3T, fueron seleccionadas para ser empleadas como inoculantes. Dos experimentos de campo llevados a cabo en años consecutivos en áreas con características edafoclimáticas similares a las que presentan las poblaciones de Lmj, lograron la reproducción exitosa de la planta. Se concluyó que un ciclo reproductivo exitoso de Lmj es absolutamente dependiente de la inoculación con sus simbiontes naturales y que la simbiosis debe ser considerada un factor esencial en estrategias de conservación de leguminosas en peligro. La obtención de varias secuencias genómicas de cepas aisladas de Lmj y de otras cepas de Bradyrhizobium reveló una alta similitud entre los genomas de las cepas del Clado I, y permitió la identificación de cinco posibles nuevas especies. Además, se estudiaron tres agrupaciones de genes relacionados con la simbiosis (nod, nif y fix) definiendo un nuevo linaje para las cepas de Lmj, diferente del symbiovar “genistearum” de B. canariense y B. japonicum. La baja diversidad encontrada en el análisis filogenético de los genes simbióticos contrasta con la gran diversidad asociada a genes esenciales. La presencia de plásmidos en cepas del género Bradyrhizobium ha sido descrita en muy pocas ocasiones, sin embargo el análisis de la secuencia genómica de la cepa ISLU101, aislada de Lupinus angustifolius, reveló la presencia de un origen de replicación extracromosómico homólogo al operón repABC, presente en el plásmido de Bradyrhizobium sp BTAi1. Gracias a esta secuencia se identificaron genes homólogos en 19 de 72 cepas ISLU. Filogenéticamente, las secuencias de repABC se agruparon en un grupo monofilético con las de pBTAi1 y separadas de los rizobios de crecimiento rápido. Finalmente, se identificaron sistemas de secreción de proteínas de tipo III (T3SS) en nueve genomas de cepas de Lmj. Los T3SS pueden inyectar proteínas efectoras al interior de células vegetales. Su presencia en rizobios se ha relacionado con la gama de hospedador que pueden nodular y puede tener un efecto beneficioso, neutro o perjudicial en la simbiosis. Los T3SS de las cepas de Lmj codifican para una proteína efectora similar a NopE, un efector dependiente de T3SS descrito en B. diazoefficiens USDA 110T. La proteína NopE de la cepa LmjC se ha caracterizado bioquímicamente. ABSTRACT Lupinus mariae-josephae (Lmj) is a lupine species endemic of a unique small area in Valencia region (Eastern Spain) where the lupine plants thrive in alkaline-limed soils, which preferentially grow in acid or neutral soils. This is the type of soils native lupines of Spain. When this work was initiated, the extension of the endemic area of Lmj was of about 700 squared kilometers confined to the Valencia province. In this area, Lmj thrives in small, isolated patches containing a reduced number of plants, and points to an endemism that can easily became endangered or extinct. Consequently, the Valencia Community authorities gave a ‘‘microreserve” status for conservation of the species. All efforts, using classical strategies directed to extend the area of Lmj growth and ensure its conservation have been so far unsuccessful. The work presented here is directed to improve our knowledge of Lmj ecology and it is centered in the characterization of the rhizobial symbiosis by phenotypic, phylogenetic and genomic analysis as well as in investigate the potential contribution of the symbiosis to improve its conservation. To this end, five different topics have been studied, and results are briefly described here. Extensive details can be followed en the attached, published articles. The first topic deals with the indigenous rhizobial symbionts of the Lmj endemism, and its genetic diversity was investigated. The Lmj root symbionts belong to the Bradyrhizobium genus, and phylogenetic analysis based on core genes identified a large diversity of Bradyrhizobium strains with phenotypic and symbiotic characteristics different from rhizobia nodulating other Lupinus spp. native of Spain. The strains were split in two clades. Clade II contained strains close to classical B. canariense and B. japonicum lineages that establish symbioses with lupines in acid soils of the Mediterranean area. Clade I included Lmj strains that define a new lineage, close to other Bradyrhizobium species as B. jicamae and B. elkanii. The phylogenetic analysis based on symbiotic genes identified only two distinct clusters. The singularity and large diversity of these strains in such a small geographical area makes this an attractive system for studying the evolution and adaptation of the rhizobial symbiont to the plant host. Additionally, the presence of bacteria able to nodulate Lmj in basic soils from Chiapas, Mexico was investigated. Surprisingly, these soils contain bacteria able to effectively nodulate and fix nitrogen with Lmj plants in greenhouse assays. In the second topic, the taxonomic status of the endosymbiotic bacteria of Lmj from Valencia endemism and Chiapas was investigated. Results from phylogenetic analysis of core genes and Average Nucleotide Identity (ANIm) using draft genomic sequences identified new Bradyrhizobium species within strains of Clade I of Lmj endosymbiotic bacteria. Only one of these potentially new species has been defined, meanwhile the others are under process of characterization. The name ‘Bradyrhizobium valentinum’ sp. nov. was proposed for the defined species (type strain LmjM3T= CECT 8364T, LMG 2761T). The third topic was directed to conservation of endangered Lmj in its natural habitat. The relevant conclusion of this experimentation is that the symbiosis should be considered as a relevant factor in the conservation strategies for endangered legumes. First, we showed absence of bacteria able to nodulate Lmj in all the inspected ‘‘terra rossa’’ or alkaline red soils of the Iberian Peninsula and Balearic Islands. Then, two efficient nitrogen fixing strains with Lmj plants, LmjC and LmjM3T, were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural Lmj populations, and successful reproduction of the plant was achieved. The relevant conclusion from these assays was that the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia The forth topic deep into the analysis of the genomic of Lmj representative strains. To this end, draft genomic sequences of selected Lmj strains and type strains of Bradyrhizobium spp. were assembled. The comparison analysis of the draft genomic sequences of Lmj strains and related Bradyrhizobium species grouped in Clade I, revealed a high genomic homology among them, and allowed the definition of five potentially new species of Lmj nodulating bacteria. Also, based on the available draft genomic sequences, only three clusters of nod, fix and nif genes from Lmj strains were identified and showed to define a new symbiotic lineage, distant from that of B. canariense and B. japonicum bv. genistearum. The low diversity exhibited by the phylogenetic analysis of symbiotic genes contrast with the large diversity of strains as regards the housekeeping genes analyzed. Besides, the genomic analysis of a Lupinus angustifolius strain ISLU101, revealed the presence of an extrachromosomal replication origin homologous to repABC cluster from plasmid present in Bradyrhizobium spp BTAi1. This repABC cluster gene sequence allowed the identification of extrachromosomic replication origin in 19 out of 72 Bradyrhizobium strains from Lupinus spp., a highly significant result since the absence of plasmids in the Bradyrhizobium genus was traditionally assumed. The repABC gene sequences of these strains grouped them in a unique monophyletic group, related to B. sp. BTAi1 plasmid, but differentiated from the repABC gene cluster of plasmids in fast growing rhizobium strains. The last topic was focused on characterization of type III secreted effectors present in Lmj endosymbiotic bacteria. Type III secretion systems (T3SS) are specialized protein export machineries which can deliver effector proteins into plant cells. The presence of T3SS in rhizobia has frequently been related to the symbiotic nodulation host-range and may have a beneficial or detrimental effect on the symbiosis with legumes. In this context, the presence of T3SS in genomes of nine Lmj strains was investigated, and it was shown the presence of clusters encoding NopE type III-secreted protein similar to the NopE1 and NopE2 of B. diazoefficiens USDA 110T. The putative NopE protein of LmjC strain is at present being characterized regarding its structure and function.

Late Quaternary extinction of a tree species in eastern North America

Widespread species- and genus-level extinctions of mammals in North America and Europe occurred during the last deglaciation [16,000–9,000 yr B.P. (by 14C)], a period of rapid and often abrupt climatic and vegetational change. These extinctions are variously ascribed to environmental change and overkill by human hunters. By contrast, plant extinctions since the Middle Pleistocene are undocumented, suggesting that plant species have been able to respond to environmental changes of the past several glacial/interglacial cycles by migration. We provide evidence from morphological studies of fossil cones and anatomical studies of fossil needles that a now-extinct species of spruce (Picea critchfieldii sp. nov.) was widespread in eastern North America during the Last Glacial Maximum. P. critchfieldii was dominant in vegetation of the Lower Mississippi Valley, and extended at least as far east as western Georgia. P. critchfieldii disappeared during the last deglaciation, and its extinction is not directly attributable to human activities. Similarly widespread plant species may be at risk of extinction in the face of future climate change.

Corais carboníferos da Amazônia

O presente trabalho compreende o estudo de corais carboníferos da Amazônia. Baseou-se fundamentalmente, em material de Bom Jardim e Monte Cristo, pertencente à Universidade de São Paulo. Contou o autor, ainda, com material de Miritituba, pertencente às coleções do Museu Paraense Emílio Goeldi. Está dividido em duas partes. A primeira inclui: introdução; método de trabalho; terminologia; distribuição geográfica dos corais e fauna associada; considerações sobre os corais registrados para a Formação Itaituba; paleoecologia; significado das faunas de corais, idade e correlação. A segunda versa sobre a taxinomia. A relação dos corais descritos é a seguinte: Tetracoralla: Stereostylus mendesi Pinto, sp. nov.; Stereostylus leinzi Pinto, sp. nov.; Lomphamplexus sp.; Amplexizapherentis petrii Pinto, sp. nov.; Dibunophylloides duncanae Pinto, sp. nov.; Dibunophylloides geiseli Pinto, sp. nov. Tabulata - Multithecopora milanoi Pinto, sp. nov.. Dos registros anteriores, as espécies assinaladas por Katzer, 1903, não foram encontradas. Pelo exame externo do material por ele coletado, e depositado no Museu Paraense Emílio Goeldi, parecem não pertencer aos gêneros assinalados e algumas certamente pertencerão às espécies novas aqui descritas. Quanto à espécie assinalada por Duarte (1938) certamente não pertence ao gênero mencionado. O único coral dos grupos estudados, assinalado para o Grupo Tarma, não corresponde a nenhuma das espécies aqui estudadas. O presente estudo confirma idade Westfaliano C ou D (Desmoinesiano) para os afloramentos de Bom Jardim e talvez pouco mais recente para os de Monte Cristo.

Chronostratigraphic summary and raw counts of selected dinoflagellate cysts and architarch taxa of ODP Hole 151-907A

A detailed dinoflagellate cyst investigation of the almost continuous Middle Miocene through Pliocene of Ocean Drilling Program Hole 907A in the Iceland Sea has been conducted at 100-kyr resolution. The investigated section is well constrained by magnetostratigraphy, providing for the first time an independent temporal control on a succession of northern high-latitude dinoflagellate cyst bioevents. Based on the highest/lowest occurrences (HO/LO) and highest common occurrence (HCO) of 20 dinoflagellate cyst taxa and one acritarch species, 26 bioevents have been defined and compared with those recorded at selected DSDP, ODP, and IODP sites from the North Atlantic and contiguous seas, and in outcrops and boreholes from the onshore and offshore eastern U.S.A., and the North Sea and Mediterranean basins. Comparisons reveal near-synchronous HOs of the dinoflagellate cysts Batiacasphaera micropapillata (3.8-3.4 Ma, mid-Pliocene) and Reticulatosphaera actinocoronata (4.8-4.2 Ma, Lower Pliocene) across the Nordic Seas and North Atlantic, highlighting their value on a supraregional scale. This probably applies also to Hystrichosphaeropsis obscura (upper Tortonian), when excluding ODP Hole 907A where its sporadic upper stratigraphic range presumably relates to cooling in the early Tortonian. Over a broader time span within the upper Tortonian, the HO of Operculodinium piaseckii likely also permits correlation across the Nordic Seas and North Atlantic, and the HO of Labyrinthodinium truncatum appears useful in the Labrador and Nordic Seas. Biostratigraphic markers useful for regional rather than supraregional correlation are the HOs of Batiacasphaera hirsuta (c. 8.4 Ma, upper Tortonian) and Unipontidinium aquaeductus (c. 13.6-13.9 Ma, upper Langhian), the HCO of the acritarch Decahedrella martinheadii (c. 6.7-6.3 Ma, Messinian), and possibly the LO of Cerebrocysta irregulare sp. nov. (c. 13.8 Ma, uppermost Langhian) across the Nordic Seas. Since Habibacysta tectata, B. micropapillata, R. actinocoronata and D. martinheadii have been observed in the Arctic Ocean, they are potentially useful for high latitude correlations in the polar domain. The LOs of Habibacysta tectata and Unipontidinium aquaeductus suggest a mid- to late Langhian age (15.1-13.7 Ma) for deposits at the base of Hole 907A, thus providing new constraints on the age of basalts at the base of ODP Hole 907A. The stratigraphically important dinoflagellate cysts Cerebrocysta irregulare sp. nov., and Impagidinium elongatum sp. nov. are formally described.

Early Cretaceous large, few-chambered Globigerinelloides from the Tethys

New findings of well-preserved Early Cretaceous planktonic foraminiferal assemblages from the Cismon core (NE Italy), Calabianca (NW Sicily), Lesches en Diois (SE France) and DSDP Site 545 (off Morocco) sections allow a better understanding of the morphological features of several taxa. This paper deals with the revision of the small, planispiral individuals that several authors include in the genus Blowiella Krechmar and Gorbachik. Comparison of morphological characteristics between Blowiella and the genus Globigerinelloides Cushman and ten Dam has resulted in retention of the latter as senior synonym of Blowiella. In fact, the morphological differences (i.e. the number of chambers in the outer whorl, the width of the umbilical area, and size and spacing of pores) used to distinguish Blowiella from Globigerinelloides cannot, in our opinion, be used in discriminating genera, but can only be applied at species level. The small, few-chambered species of the genus Globigerinelloides retained here are Globigerinelloides blowi(Bolli), Globigerinelloides duboisi (Chevalier), Globigerinelloides maridalensis (Bolli), and Globigerinelloides paragottisi sp. nov. (=Globigerinelloides gottisi auctorum). Stratigraphically, in the sections studied Globigerinelloides blowi and Globigerinelloides paragottisi sp. nov. are first recorded from the mid-Upper Barremian in the Cismon core and Calabianca section, while rare individuals belonging to Globigerinelloides maridalensis and Globigerinelloide duboisi occur intermittently from the Barremian/Aptian boundary and from the Lower Aptian, respectively. All of these taxa become more frequent and abundant just above the Selli Level (OAE1a, Lower Aptian), within the Leupoldina cabri Zone (Upper Aptian). Based on the DSDP Site 545 succession, all four globigerinelloidid taxa range up to the Ticinella bejaouaensis Zone (uppermost Aptian), with Globigerinelloides maridalensis disappearing at the base of the zone, followed in close succession by the disappearance of G. blowi, G. paragottisi and finally G. duboisi.

Absolute abundances of benthic foraminifers and stable isotopes from the Alvin dives 3709-3712

The presence of gas hydrates on the Blake Ridge diapir, northeastern Atlantic Ocean, offers an opportunity to study the impact of methane seepage on the ecology and geochemistry of benthic foraminifera in the late Holocene. Three push cores, covering a time span of ~ 1000 yrs, were retrieved from three distinct microhabitats at the top of the diapir at a water depth of ~ 2150 m: (i) sediments away from seepage (control core), (ii) sediments overlain by clusters of methanotrophic and thiotrophic bivalves, and (iii) chemoautotrophic microbial mats. The foraminiferal assemblages at the two seep sites are marked by a reduction in benthic foraminiferal species diversity, coupled with a near-absence of agglutinated species. However, an opportunistic population rise in CH4- or H2S-tolerant calcareous species (e.g., Globocassidulina subglobosa and Cassidulina laevigata) that utilize the abundant trophic resources at the seeps has led to an increase in the overall assemblage density there. The delta18O and delta13C values of three species of benthic foraminifera - Gyroidinoides laevigatus, Globocassidulina subglobosa, and Uvigerina peregrina - and the planktonic species Globorotalia menardii were acquired from all three cores. The benthic species from methane seeps yield delta13C values of 0.1 to - 4.2 (per mil VPDB), that are distinctly more 13C-depleted relative to the delta13C of 0.4 to - 1.0 (per mil VPDB) at the control (off seep) site. The species from a mussel-bed site exhibit more negative delta13C values than those from microbial mats, possibly reflecting different food sources and higher rate of anaerobic oxidation of methane. The positive delta13C values in the paired planktonic species suggest that authigenic carbonate precipitation did not overprint the observed 13C depletions. Hence the probable cause of negative delta13C of benthic foraminifera is primary calcification from Dissolved Inorganic Carbon (DIC) containing mixed carbon fractions from (a) highly 13C-depleted, microbially-oxidized methane and (b) a seawater source.

Mechanisms of Thermal Adaptation Revealed From the Genomes of the Antarctic Archaea Methanogenium frigidum and Methanococcoides burtonii

We generated draft genome sequences for two cold-adapted Archaea, Methanogenium frigidum and Methanococcoides burtonii, to identify genotypic characteristics that distinguish them from Archaea with a higher optimal growth temperature (OGT). Comparative genomics revealed trends in amino acid and tRNA composition, and structural features of proteins. Proteins from the cold-adapted Archaea are characterized by a higher content of noncharged polar amino acids, particularly Gin and Thr and a lower content of hydrophobic amino acids, particularly Leu. Sequence data from nine methanogen genomes (OGT 15degrees-98degreesC) were used to generate IIII modeled protein structures. Analysis of the models from the cold-adapted Archaea showed a strong tendency in the solvent-accessible area for more Gin, Thr, and hydrophobic residues and fewer charged residues. A cold shock domain (CSD) protein (CspA homolog) was identified in M. frigidum, two hypothetical proteins with CSD-folds in M. burtonii, and a unique winged helix DNA-binding domain protein in M. burtonii. This suggests that these types of nucleic acid binding proteins have a critical role in cold-adapted Archaea. Structural analysis of tRNA sequences from the Archaea indicated that GC content is the major factor influencing tRNA stability in hyperthermophiles, but not in the psychrophiles, mesophiles or moderate thermophiles. Below an OGT of 60degreesC, the GC content in tRNA was largely unchanged, indicating that any requirement for flexibility of tRNA in psychrophiles is mediated by other means. This is the first time that comparisons have been performed with genome data from Archaea spanning the growth temperature extremes. from psychrophiles to hyperthermophiles

Phylogenetic and physiological characterization of a heterotrophic, chemolithoautotrophic Thiothrix strain isolated from activated sludge

The sheathed filamentous bacterium known as strain CT3, isolated by micromanipulation from an activated sludge treatment plant in Italy, is a member of the genus Thiothrix in the gamma-Proteobacteria according to 16S rDNA sequence analysis. The closest phylogenetic neighbours of strain CT3 are strains I and Q(T), which were also isolated from activated sludge and belong to the species Thiothrix fructosivorans. These strains have respectively 99.2 and 99.4 % similarity to CT3 by 16S rDNA sequence comparison. CT3 shows 63-67 % DNA-DNA hybridization with strain I, which is the only currently viable strain of T. fructosivorans. CT3 is the second strain in the genus Thiothrix that has been shown to be capable of growing autotrophically with reduced sulfur compounds as the sole energy source; autotrophy was also confirmed in strain I. The first reported chemolithoautotrophic isolate of this genus was a strain of 'Thiothrix ramosa' that was isolated from a hydrogen sulfide spring and is morphologically distinguishable from all other described strains of Thiothrix, including CT3. CT3 is an aerobic organism that is non-fermentative, not capable of denitrification and able to grow heterotrophically. Autotrophy in the genus Thiothrix should be investigated more fully to better define the taxonomy of this genus.

A new crocodilian from the Lower Cretaceous Crato Formation of north-eastern Brazil

A new mesosuchian crocoddian from the Nova Olinda Member of the Crato Formation (Lower Cretaceous, Aptian) of north-eastern Brazil is described. Susisuchus anatoceps gen. et sp. nov. is the first crocodillan to be reported from this formation. It is represented by an incomplete, partially articulated skeleton: the skull and mandible, partial postcranial axial skeleton, forelimbs and portions of the osteodermal skeleton. Preservation of soft tissues includes the skin surrounding both forelimbs and the digits of the right hand. The state of preservation of the specimen suggests that it was incorporated into the basin as a desiccated carcass. Susisuchus anatoceps is one of the oldest crocodilians with a eusuchian-type dorsal shield, comprising a tetraserial paravertebral shield and, either side of this, two sagittal rows of accessory osteoderms. It also possesses amphicoelous thoracic, lumbar and caudal vertebrae. This combination of postcranial features have never before been seen in a crocodilian and warrant the erection of a new family within Mesosuchia: Susisuchidae. Taxonomically, S. anatoceps is similar to a number of Lower Cretaceous mesosuchians previously considered to have given rise to eusuchians, most notably the Glen Rose crocodilian and a new, but as yet undescribed crocodillan from the Lower Cretaceous Winton Formation of western Queensland, Australia. Preliminary preparation of the Winton crocodilian indicates that it may belong to Susisuchidae, supporting the hypotheses of interchange between the vertebrate faunas of South America and Australia during the Lower Cretaceous.

Diplacanthid acanthodians from the Aztec Siltstone (late Middle Devonian) of southern Victoria Land, Antartica

One articulated and several partial, semi-articulated specimens of acanthodians were collected in 1970 from the freshwater deposits of the Aztec Siltstone (Middle Devonian; Givetian), Portal Mountain, southern Victoria Land, Antarctica, during a Victoria University of Wellington Antarctic Expedition. The Portal Mountain fish fauna, preserved in a finely laminated, non-calcareous siltstone, includes acanthodians, palaeoniscoids, and bothriolepid placoderms. The articulated acanthodian specimens are the most complete fossil fish remains documented so far from the Aztec assemblage, which is the most diverse fossil vertebrate fauna known from Antarctica. They are described as a new taxon, Milesacanthus antarctica gen. et sp. nov., which is assigned to the family Diplacanthidae. Its fin spines show some similarities to spine fragments named Byssacanthoides debenhami from glacial moraine at Granite Harbour, Antarctica, and much larger spines named Antarctonchus glacialis from outcrops of the Aztec Siltstone in the Boomerang Range, southern Victoria Land. Both of these are reviewed, and retained as form taxa for isolated spines. Various isolated remains of fin spines and scales are described from Portal Mountain and Mount Crean (Lashly Range), and referred to Milesacanthus antarctica gen. et sp. nov. The histology of spines and scales is documented for the first time, and compared with acanthodian material from the Devonian of Australia and Europe. Distinctive fin spines from Mount Crean are provisionally assigned to Culmacanthus antarctica Young, 1989b. Several features on the most complete of the new fish specimens - in particular, the apparent lack of an enlarged cheek plate - suggest a revision of the diagnosis for the Diplacanthidae.

A new Late Devonian acanthodian fish from the Hunter Formation near Grenfell, New South Wales

The ischnacanthid acanthodian Grenfellacanthus zerinae gen. et sp. nov. is described on the basis of two large jaw bones from the Late Devonian (late Famennian) Hunter Formation, near Grenfell, N.S.W. The new species is the youngest known ischnacanthid, and the largest ischnacanthid from Gondwana. As for many ischnacanthids, the structure of the jaws and teeth indicate that Grenfellacanthus was probably an ambush predator.

A new species of Glaphyromorphus (Reptilia : Scincidae) from Mt Elliot, north-eastern Queensland

Glaphyromorphus clandestinus, sp. nov., is described from granite-slab habitat on Mt Elliot, north-eastern Queensland. This species can be distinguished from its congeners by a combination of the following characters: large size (SVL 72 mm), adpressed limbs of adult separated by noticeably more than the length of the forelimb, 26 mid-body scale rows, and flanks patterned with dark flecks forming a series of longitudinal lines. The distribution, habitat preferences and habits of this species are poorly known. Currently G. clandestinus is known from a single locality where individuals have been found in an exposed area of exfoliating granite, set in a mosaic of rainforest and eucalyptus woodland. The discovery of this species brings to three the number of vertebrate species known to be endemic to Mt Elliot and highlights the evolutionary significance of this southerly outlier to the mountainous rainforest of the Wet Tropics.

Australian microhylid frogs (Cophixalus and Austrochaperina): phylogeny, taxonomy, calls, distributions and breeding biology

Despite a considerable surge in herpetological research in Australia over the last couple of decades the Australian microhylid frogs (Cophixalus and Austrochaperina) remain relatively poorly known. Herein I present the results of extensive fieldwork and molecular, morphological and call analysis with the aim of resolving taxonomy, call variation and distributions, and increasing our understanding of breeding biology. Analysis of 943 base pairs of mitochondrial 16S rRNA and 12S rRNA provides a well supported phylogeny that is largely consistent with current taxonomy. Levels of divergence between species are substantial and significant phylogeographic structuring is evident in C. ornatus, C. neglectus and C. aenigma, sp. nov. The description of C. concinnus was based on a mixed collection of two species from Thornton Peak and a new species is described to resolve this. C. aenigma, sp. nov., is described from high-elevation (>750 m) rainforest across the Carbine, Thornton, Finnigan and Bakers Blue Mountain uplands, north-east Queensland. C. concinnus is redescribed as a highly distinct species restricted to rainforest and boulder fields at the summit of Thornton Peak (>1100 m). Despite protection in Daintree National Park in the Wet Tropics World Heritage Area, predictions of the impact of global warming suggest C. concinnus to be of very high conservation concern ( Critically Endangered, IUCN criteria). The mating call of two species ( C. mcdonaldi and C. exiguus) is described for the first time and high levels of call variation within C. ornatus, C. neglectus, C. hosmeri, C. aenigma and Austrochaperina fryi are presented. Such variation is often attributable to genetically divergent lineages, altitudinal variation and courtship; however, in some instances ( particularly within C. hosmeri) the source or function of highly distinct calls at a site remains obscure. Molecular, morphological and call analyses allow the clarification of species distributions, especially in the northern mountains of the Wet Tropics. Notes are presented on the breeding biology of C. aenigma, C. bombiens, C. concinnus, C. exiguus, C. infacetus, C. mcdonaldi, C. monticola, C. neglectus, C. ornatus and C. saxatilis, which are largely consistent with previous accounts: small terrestrial clutches usually attended by a male. Courtship behaviour in C. ornatus is described and the first records of multiple clutching in Australian microhylids are presented (for C. ornatus and C. infacetus).

An evolutionary radiation of beeflies in semi-arid Australia: systematics of the Exoprosopini (Diptera : Bombyliidae)

Almost half of the 4822 described beeflies in the world belong to the subfamily Anthracinae, with most of the diversity found in three cosmopolitan tribes: Villini, Anthracini, and Exoprosopini. The Australian Exoprosopini previously contained three genera, Ligyra Newman, Pseudopenthes Roberts and Exoprosopa Macquart. Pseudopenthes is an Australian endemic, with two species including Ps. hesperis, sp. nov. from Western Australia. Two new species of the exoprosopine Atrichochira Hesse, Atr. commoni, sp. nov. and Atr. paramonovi, sp. nov., are also described from Australia, extending the generic distribution from Africa. Cladistic analysis clarified the phylogenetic relationships between the recognised groups of the Exoprosopini and determined generic limits on a world scale. Inclusion of 18 Australian exoprosopines placed the Australian species in the context of the world fauna. The Exoprosopini contains six large groups. The basal group I contains species previously included in Exoprosopa to which the name Defilippia Lioy is applied. Group II contains Heteralonia Rondani, Atrichochira, Micomitra Bowden, Pseudopenthes, and Diatropomma Bowden. Colossoptera Hull is newly synonymised with Heteralonia. Group III is a paraphyletic assemblage of Pterobates Bezzi and Exoprosopa including the Australian Ex. sylvana ( Fabricius). Ligyra is paraphyletic, forming two well-separated clades. The African clade is described as Euligyra Lambkin, gen. nov., which, together with Litorhina Bezzi and Hyperalonia Rondani, form group IV. The Australian group V is true Ligyra. The remaining monophyletic lineage of exoprosopines, group VI, the Balaana-group of genera, shows evidence of an evolutionary radiation of beeflies in semi-arid Australia. Phylogenetic analysis of all 42 species of the Balaana-group of genera formed a basis for delimiting genera. Seven new genera are described by Lambkin & Yeates: Balaana, Kapua, Larrpana, Munjua, Muwarna, Palirika and Wurda. Four non-Australian species belong to Balaana. Thirty two new Australian species are described: Bal. abscondita, Bal. bicuspis, Bal. centrosa, Bal. gigantea, Bal. kingcascadensis, K. corusca, K. irwini, K. westralica, Lar. collessi, Lar. zwicki, Mun. erugata, Mun. lepidokingi, Mun. paralutea, Mun. trigona, Muw. vitreilinearis, Pa. anaxios, Pa. basilikos, Pa. blackdownensis, Pa. bouchardi, Pa. cyanea, Pa. danielsi, Pa. decora, Pa. viridula, Pa. whyalla, W. emu, W. impatientis, W. montebelloensis, W. norrisi, W. patrellia, W. skevingtoni, W. windorah, and W. wyperfeldensis. The following new combinations are proposed: from Colossoptera: Heteralonia latipennis (Brunetti); from Exoprosopa: Bal. grandis (Pallas), Bal. efflatounbeyi (Paramonov), Bal. latelimbata ( Bigot), Bal. obliquebifasciata ( Macquart), Bal. tamerlan (Portschinsky), Bal. onusta ( Walker), Def. busiris (Jaennicke), Def. efflatouni ( Bezzi), Def. eritreae (Greathead), Def. gentilis ( Bezzi), Def. luteicosta ( Bezzi), Def. minos (Meigen), Def. nigrifimbriata ( Hesse), Def. rubescens ( Bezzi), K. adelaidica ( Macquart), Lar. dimidiatipennis ( Bowden), Muw. stellifera ( Walker), and Pa. marginicollis ( Gray); from Ligyra: Eu. enderleini ( Paramonov), Eu. mars ( Bezzi), Eu. monacha (Klug), Eu. paris ( Bezzi), Eu. sisyphus ( Fabricius), and Eu. venus (Karsch).