Proterozoic phytoplankton and timing of Chlorophyte algae origins

Abstract: Morphological and reproductive features and cell wall ultrastructure and biochemistry of Proterozoic acritarchs are used to determine their affinity to modern algae. The first appearance datum of these microbiota is traced to infer a minimum age of the divergence of the algal classes to which they may belong. The chronological appearance of microfossils that represent phycoma‐like and zygotic cysts and vegetative cells and/or aplanospores, respectively, interpreted as prasinophyceaen and chlorophyceaen microalgae is related to the Viridiplantae phylogeny. An inferred minimum age of the Chlorophyte origin is before c. 1800 Ma, the Prasinophyceae at c. 1650 Ma and the Chlorophyceae at c. 1450 Ma. These divergence times differ from molecular clock estimates, and the palaeontological evidence suggests that they are older.     

Wall ultrastructure of an Ediacaran acritarch from the Officer Basin, Australia

Well-preserved organic-walled microfossils referred to as acritarchs occur abundantly in Ediacaran deposits in the Officer Basin in Australia. The assemblages are taxonomically diverse, change over short stratigraphical intervals and are largely facies independent across marine basins. Affinities of this informal group of fossils to modern biota are poorly recognized or unknown, with the exception of only a few taxa. Morphological studies by use of transmitted light microscopy, geochemical analyses and other lines of evidence, suggest that some Precambrian acritarchs are related to algae (including prasinophytes, chlorophytes, and perhaps also dinoflagellates). Limitations in magnification and resolution using transmitted light microscopy may be relevant when assessing relationships to modern taxa. Scanning electron microscopy reveals details of morphology, microstructure and wall surface microelements, whereas transmission electron microscopy provides high-resolution images of the cell wall ultrastructure. In the light of previous ultrastructural studies it can be concluded that the division of acritarchs into leiospheres (unornamented) and acanthomorphs (ornamented) is entirely artificial and has no phylogenetic meaning. Examination of Gyalosphaeridium pulchrum using transmission electron microscopy reveals a vesicle wall with four distinct layers. This multilayered wall ultrastructure is broadly shared by a range of morphologically diverse acritarchs as well as some extant microalgae. The chemically resistant biopolymers forming the comparatively thick cell, together with the overall morphology support the interpretation of the microfossil as being in the resting stage in the life cycle. The set of features, morphological and ultrastructural, suggests closer relationship to green algae than dinoflagellates.     

Authigenic anatase within 1 billion‐year‐old cells

The siliciclastic ~1 Ga‐old strata of the Torridon Group, Scotland, contain some of the most exquisitely preserved three‐dimensional organic‐walled microfossils (OWMs) of the Precambrian. A very diverse microfossil assemblage is hosted in a dominantly phosphatic and clay mineral matrix, within the Diabaig and the Cailleach Head (CH) Formations. In this study, we report on several microfossil taxa within the CH Formation (Leiosphaeridia minutissima, Leiosphaeridia crassa, Synsphaeridium spp. and Myxococcoides spp.) that include populations of cells containing an optically transparent and highly refringent mineral, here identified using electron microscopy as anatase (TiO₂). Most anatase crystals occur entirely within individual cells, surrounded by unbroken carbonaceous walls. Rarely, an anatase crystal may protrude outside a cell, interpreted to correspond to zones where the cell wall had broken down prior to anatase precipitation. Where an anatase crystal entombs an organic intracellular inclusion (ICI), the ICI is large and well preserved. These combined observations indicate that the intracellular anatase is an authigenic sedimentary phase, making this the first report of in situ precipitated anatase intimately associated with microfossils. The ability of anatase to preserve relatively large volumes of intracellular and cell wall organic material in these cells suggests that the crystallisation of anatase entombed cellular contents particularly quickly, soon after the death of the cell. This is consistent with the strong affinity of Ti for organic material, the low solubility of TiO₂, and reports of Ti occurring in living organisms. With the data currently available, we propose a mineralisation pathway for anatase involving Ti complexation with organic ligands within specific cells, leading to localised post‐mortem anatase nucleation inside these cells as the complexes broke down. Further overgrowth of the anatase crystals was likely fuelled by very early diagenetic mobilisation of Ti that had been bound to more labile organic material nearby in the sediments.     

The nature and origin of nucleus‐like intracellular inclusions in Paleoproterozoic eukaryote microfossils

The well‐known debate on the nature and origin of intracellular inclusions (ICIs) in silicified microfossils from the early Neoproterozoic Bitter Springs Formation has recently been revived by reports of possible fossilized nuclei in phosphatized animal embryo‐like fossils from the Ediacaran Doushantuo Formation of South China. The revisitation of this discussion prompted a critical and comprehensive investigation of ICIs in some of the oldest indisputable eukaryote microfossils—the ornamented acritarchs Dictyosphaera delicata and Shuiyousphaeridium macroreticulatum from the Paleoproterozoic Ruyang Group of North China—using a suite of characterization approaches: scanning electron microscopy (SEM), transmission electron microscopy (TEM), and focused ion beam scanning electron microscopy (FIB‐SEM). Although the Ruyang acritarchs must have had nuclei when alive, our data suggest that their ICIs represent neither fossilized nuclei nor taphonomically condensed cytoplasm. We instead propose that these ICIs likely represent biologically contracted and consolidated eukaryotic protoplasts (the combination of the nucleus, surrounding cytoplasm, and plasma membrane). As opposed to degradational contraction of prokaryotic cells within a mucoidal sheath—a model proposed to explain the Bitter Springs ICIs—our model implies that protoplast condensation in the Ruyang acritarchs was an in vivo biologically programmed response to adverse conditions in preparation for encystment. While the discovery of bona fide nuclei in Paleoproterozoic acritarchs would be a substantial landmark in our understanding of eukaryote evolution, the various processes (such as degradational and biological condensation of protoplasts) capable of producing nuclei‐mimicking structures require that interpretation of ICIs as fossilized nuclei be based on comprehensive investigations.     

Epitope detection chromatography: a method to dissect the structural heterogeneity and inter‐connections of plant cell‐wall matrix glycans

Plant cell walls are complex, multi‐macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. Even though the chemistry of cell‐wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. Here we describe in detail a method called epitope detection chromatography analysis of cell‐wall matrix glycan sub‐populations and inter‐connections. The method combines chromatographic separations with use of glycan‐directed monoclonal antibodies as detection tools. The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell‐wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub‐populations and their modulation in A. thaliana organs.     

CELL‐WALL POLYMER MAPPING IN THE COENOCYTIC MACROALGA CODIUM VERMILARA (BRYOPSIDALES,CHLOROPHYTA)1

Cell walls in the coenocytic green seaweed Codium vermilara (Olivi) Chiaje (Bryopsidales, Chlorophyta) are composed of ～32% (w/w) β‐(1→4)‐d‐mannans, ～12% sulfated polysaccharides (SPs), and small amounts of hydroxyproline‐rich glycoprotein‐like (HRGP‐L) compounds of the arabinogalactan proteins (AGPs) and arabinosides (extensins). Similar quantities of mannans and SPs were reported previously in the related seaweed C. fragile (Suringar) Hariot. Overall, both seaweed cell walls comprise ～40%–44% of their dry weights. Within the SP group, a variety of polysaccharide structures from pyruvylated arabinogalactan sulfate and pyruvylated galactan sulfate to pyranosic arabinan sulfate are present in Codium cell walls. In this paper, the in situ distribution of the main cell‐wall polymers in the green seaweed C. vermilara was studied, comparing their arrangements with those observed in cell walls from C. fragile. The utricle cell wall in C. vermilara showed by TEM a sandwich structure of two fibrillar‐like layers of similar width delimiting a middle amorphous‐like zone. By immuno‐ and chemical imaging, the in situ distribution of β‐(1→4)‐d‐mannans and HRGP‐like epitopes was shown to consist of two distinct cell‐wall layers, whereas SPs are distributed in the middle area of the wall. The overall cell‐wall polymer arrangement of the SPs, HRGP‐like epitopes, and mannans in the utricles of C. vermilara is different from the ubiquitous green algae C. fragile, in spite of both being phylogenetically very close. In addition, a preliminary cell‐wall model of the utricle moiety is proposed for both seaweeds, C. fragile and C. vermilara.     

First record of organic-walled microfossils from the Tonian Shiwangzhuang Formation of the Tumen Group in western Shandong,North China

Organic-walled microfossils offer important information on the biospheric evolution in pre-Cryogenian and provide biostratigraphic implications for many Proterozoic fossiliferous sequences that are poorly age constrained for the lack of reliable radiometric date. Recently, macroscopic carbonaceous compression fossils have been reported for the first time from the Tonian Shiwangzhuang Formation of the Tumen Group in western Shandong, North China. However, organic-walled microfossils have never been discovered from this formation up till now. To improve our knowledge about Proterozoic biodiversity in North China, we conducted a micropaleontological survey on the argillaceous limestone samples of the Shiwangzhuang Formation, which also contain macroscopic carbonaceous compression fossils, from the Baishicun section in Anqiu, western Shandong, North China. Our investigation shows that the Shiwangzhuang microfossil assemblage is dominated by smooth-walled sphaeromorphic acritarchs and cyanobacterium-like filaments and relatively low abundance of other acritarchs, including 16 taxa, such as Polysphaeroides filliformis, Ostiana microcystis, Simia annulare, ?Jacutianema sp., Arctacellularia tetragonala, Pellicularia tenera, Polythrichoides lineatus, and Navifusa actinomorpha. The Shiwangzhuang organic-walled microfossil assemblage, although consisting of long-ranging and not age diagnostic taxa, is consistent with a Tonian age suggested by macroscopic carbonaceous compression fossils, including the Chuaria-Tawuia and Sinosabellidites-Protoarenicola-Pararenicola assemblages, revealed from the same fossiliferous horizon of the Shiwangzhuang Formation and by organic-walled microfossil assemblage, including the late Mesoproterozoic to Tonian index fossil Trachyhystrichosphaera aimika, from the underlying Tongjiazhuang Formation. However, it is also worth noting that a Cryogenian or Ediacaran age cannot be completely excluded based just on the Shiwangzhuang microfossils because of their limited biostratigraphic utility.     

Fluorescence intensity studies of Triassic acritarchs from the Yanchang Formation in Ordos basin, northwestern China

Fluorescence properties of Early Cambrian acritarchs were investigated using Leica das Mikroskop (DM) microscopy with a mercury lamp. Well-preserved autoflurescence properties show a correlation between acritarchs morphology and the intensity of emitted fluorescence. In accordance with the fluorescence intensity of organic cell walls, two groups of microfossils were distinguished. Results of observation in this study, which are consistent with those of the previous foreign studies, are in good agreement with regular difference in autofluorescence intensity among palynomorphs reported by McPhilemy (1998). Spores and algae, including Botryococcus, have very bright fluorescence while acritarchs often show less intense fluorescence. Dark brown microfossils have been reworked, and have little or no fluorescence. __________ Translated from Acta Micropalaeontologica Sinica, 2006, 23(3): 309–312 [译自: 微体古生物学报]     

An ultrastructural comparison of the cell envelopes of selected strains of Nocardia asteroides and Nocardia brasiliensis

Growth curves were determined for three strains each ofNocardia asteroides andNocardia brasiliensis. Two strains ofN. brasiliensis and one strain ofN. asteroides had longer lag periods of growth than the remaining three strains. All strains had generation times of approximately 5.5 hours.The ultrastructure of the cell envelope of eachNocardia strain in early stationary phase growth was also examined. All the strains had typical trilaminar cell walls and cell membranes. The thickness of the cell wall layers, especially the inner peptidoglycan layer, varied from strain to strain. The inner layer of two strains ofN. brasiliensis and one strain ofN. asteroides was 12 nm or more in thickness, while that of the remaining three strains was 7 nm thick. These observed differences in growth patterns and/or thickness of the cell wall layers could be correlated to the varying degress of virulence as well as the divergent pathologies exhibited by these organisms.     

Ultrastructural and chemical changes in the cell wall of Haematococcus pluvialis (Volvocales,Chlorophyta) during aplanospore formation

Changes in the ultrastructure and chemistry of the cell wall of the unicellular volvocalean green alga Haematococcus pluvialis were investigated during the transformation of flagellates into aplanospores. The motile biflagellated state exhibited a distinct gelatinous extracellular matrix. Its ultrastructure resembled the typical volvocalean multilayered architecture with a median tripartite crystalline layer. The transformation into the non-motile cell state was characterized by formation of a new layer, a primary wall, within the extracellular matrix. During this process, the initial extracellular matrix remained intact except for the outer layers of the tripartite crystalline layer, which decomposed. Further morphogenesis of the aplanospore resulted in the formation of a voluminous multilayered cell wall. A trilaminar sheath was formed inside the primary wall and the innermost and thickest part was an amorphous secondary wall, consisting mostly of a mannan. Results obtained by staining with the fluorescent dye primuline as well as by acetolysis suggest the occurrence of sporopollenin-like material (algaenan) within the trilaminar sheath of the aplanospore cell wall. The primary wall and the outer remnants of the extracellular matrix disintegrated as the aplanospores aged, and were completely absent in the resting cell state.     

Rice cellulose synthase‐like D4 is essential for normal cell‐wall biosynthesis and plant growth

Cellulose synthase‐like (CSL) proteins of glycosyltransferase family 2 (GT2) are believed to be involved in the biosynthesis of cell‐wall polymers. The CSL D sub‐family (CSLD) is common to all plants, but the functions of CSLDs remain to be elucidated. We report here an in‐depth characterization of a narrow leaf and dwarf1 (nd1) rice mutant that shows significant reduction in plant growth due to retarded cell division. Map‐based cloning revealed that ND1 encodes OsCSLD4, one of five members of the CSLD sub‐family in rice. OsCSLD4 is mainly expressed in tissues undergoing rapid growth. Expression of OsCSLD4 fluorescently tagged at the C‐ or N‐terminus in rice protoplast cells or Nicotiana benthamiana leaves showed that the protein is located in the endoplasmic reticulum or Golgi vesicles. Golgi localization was verified using phenotype‐rescued transgenic plants expressing OsCSLD4–GUS under the control of its own promoter. Two phenotype‐altered tissues, culms and root tips, were used to investigate the specific wall defects. Immunological studies and monosaccharide compositional and glycosyl linkage analyses explored several wall compositional effects caused by disruption of OsCSLD4, including alterations in the structure of arabinoxylan and the content of cellulose and homogalacturonan, which are distinct in the monocot grass species Oryza sativa (rice). The inconsistent alterations in the two tissues and the observable structural defects in primary walls indicate that OsCSLD4 plays important roles in cell‐wall formation and plant growth.     

The ichnogenus Palmiraichnus Roselli for fossil bee cells

The ichnogenus Palmiraichnus was erected by Roselli in 1987 to include his formerly described ich‐nospecies castellanosi, and a new one, minor, herein considered a junior synonym of the former. Shape, lining and closure clearly show that these traces are fossil bee cells. Celliforma, the other ichnogenus for non‐clustered fossil bee cells, includes two ichnospecies, bedfordi (=septata) (n. syn.) and pinturensis, which share with Palmiraichnus castellanosi the presence of antechambers and hardened cell walls. This combination of features is characteristic of certain groups of short‐tongued bees and justifies the inclusion of these ichnospecies in Palmiraichnus. Microscopic structures of cell walls and closures are adopted here for ichnotaxonomic purposes and for comparing fossil with extant cells revealing that they are useful ichnotaxobases with respect to the problematic fossil bee cell ichnotaxonomy. The micro‐structure of the cell walls and closures of the three ichnospecies of Palmiraichnus are compared. The presence of antechambers and particularly of the thickened cell rears suggest a relationship between Palmiraichnus castellanosi and Uruguay, an ichno‐taxon also attributable to bees. However, in Uruguay ichnospecies. cells are grouped in well‐defined clusters, whereas in Palmiraichnus castellanosi, cells occur isolated in paleosols, and no intermediate morphologies are known between the two ichnogenera. P. castellanosi, the focus of our study, is also compared with oxaeid cells revealing similarities in size and wall microstructure and differences in microstructure of closures and cell rears.     

Cell wall and cell growth characteristics of giant‐celled algae

Because of their large sizes and simple shapes, giant‐celled algae have been used to study how the structural and mechanical properties of cell walls influence cell growth. Here we review known relationships between cell wall and cell growth properties that are characteristic of three representative taxa of giant‐celled algae, namely, Valonia ventricosa, internodal cells of characean algae, and Vaucheria frigida. Tip‐growing cells of the genus Vaucheria differ from cells undergoing diffuse growth in V. ventricosa and characean algae in terms of their basic architectures (non‐lamellate vs. multilamellate) and their dependence upon pH and Ca²⁺ for cell wall extensibility. To further understand the mechanisms controlling cell growth by cell walls, comparative analyses of cell wall structures and/or associated growth modes will be useful. The giant‐celled algae potentially serve as good models for such investigations because of their wide variety of developmental processes and cell shapes exhibited.     

Thermally-induced structural and chemical alteration of organic-walled microfossils: an experimental approach to understanding fossil preservation in metasediments

The identification and confirmation of bona fide Archean–Paleoproterozoic microfossils can prove to be a challenging task, further compounded by diagenetic and metamorphic histories. While structures of likely biological origin are not uncommon in Precambrian rocks, the search for early fossil life has been disproportionately focused on lesser thermally altered rocks, typically greenschist or lower‐grade metamorphism. Recently, however, an increasing number of inferred micro‐ and macrofossils have been reported from higher‐grade metasediments, prompting us to experimentally test and quantify the preservability of organic‐walled microfossils over varying durations of controlled heating and under two differing redox conditions. Because of their relatively low‐intensity natural thermal alteration, acritarchs from the Mesoproterozoic Ruyang Group were chosen as subjects for experimental heating at approximately 500°C, with durations ranging from 1 to 250 days and in both oxic (normal present day conditions) and anoxic conditions. Upon extraction, the opacity, reflectivity, color, microchemistry, and microstructures of the heated acritarchs were characterized using optic microscopy, scanning electron microscopy, Raman spectroscopy, and X‐ray photoelectron spectroscopy. The results differ for acritarchs prepared under oxic vs. anoxic conditions, with the anoxic replicates surviving experimental heating longer and retaining biological morphologies better, despite an increasing degree of carbonization with continuous heating. Conversely, the oxic replicates show aggressive degradation. In conjunction with fossils from high‐grade metasediments, our data illustrate the preservational potential of organic‐walled microfossils subjected to metamorphism in reducing conditions, offer insights into the search for microfossils in metasediments, and help to elucidate the influence of time on the carbonization/graphitization processes during thermal alteration.     

A microalgal‐based preparation with synergistic cellulolytic and detoxifying action towards chemical‐treated lignocellulose

High‐temperature bioconversion of lignocellulose into fermentable sugars has drawn attention for efficient production of renewable chemicals and biofuels, because competing microbial activities are inhibited at elevated temperatures and thermostable cell wall degrading enzymes are superior to mesophilic enzymes. Here, we report on the development of a platform to produce four different thermostable cell wall degrading enzymes in the chloroplast of Chlamydomonas reinhardtii. The enzyme blend was composed of the cellobiohydrolase CBM3GH5 from C. saccharolyticus, the β‐glucosidase celB from P. furiosus, the endoglucanase B and the endoxylanase XynA from T. neapolitana. In addition, transplastomic microalgae were engineered for the expression of phosphite dehydrogenase D from Pseudomonas stutzeri, allowing for growth in non‐axenic media by selective phosphite nutrition. The cellulolytic blend composed of the glycoside hydrolase (GH) domain GH12/GH5/GH1 allowed the conversion of alkaline‐treated lignocellulose into glucose with efficiencies ranging from 14% to 17% upon 48h of reaction and an enzyme loading of 0.05% (w/w). Hydrolysates from treated cellulosic materials with extracts of transgenic microalgae boosted both the biogas production by methanogenic bacteria and the mixotrophic growth of the oleaginous microalga Chlorella vulgaris. Notably, microalgal treatment suppressed the detrimental effect of inhibitory by‐products released from the alkaline treatment of biomass, thus allowing for efficient assimilation of lignocellulose‐derived sugars by C. vulgaris under mixotrophic growth.     

Morphological and ultrastructural studies of some acritarchs from the Lower Cambrian Lükati Formation, Estonia

Six acritarch species from the Lükati Formation were studied using a combination of techniques, including transmitted light, scanning electron (SEM) and transmission electron (TEM) microscopy. New details of wall ultrastructure, surface microsculpture and internal morphology of the vesicle and processes significantly add to the previously known morphological features and increase the understanding of the form-genera Archaeodiscina, Globosphaeridium, Comasphaeridium, Skiagia, Tasmanites and Leiosphaeridia. Examination of microfossils using TEM revealed a substantial variation in wall ultrastructure among acritarchs. The diversity includes four structural types of vesicle wall in addition to their single- and multi-layered structure and the variable thickness of the wall. These are: electron-tenuous and fibrous; electron-dense and homogeneous; electron-dense and homogeneous but perforated by radial canals; and composite laminated structure. Morphologically recognised groupings of acritarchs (acanthomorphic, disphaeromorphic, sphaeromorphic) and tasmanitid taxa appear to be characterised by particular features of the wall structure, although the wall structure in itself may not be directly indicative of systematic relationships. Structurally diverse vesicle walls are observed in Tasmanites and Leiosphaeridia, taxa that both have been interpreted, based on other lines of evidence, to be of prasinophycean (green algal) affinities. The distinct wall ultrastructure of the Leiosphaeridia studied is similar to that of extant green algal genera, which provides evidence that some Cambrian leiosphaerids were chlorophycean algae, probably related to the Order Chlorococcales. Previous research and interpretations of the wall ultrastructure are also briefly discussed.     

Diverse small spinose acritarchs from the Ediacaran Doushantuo Formation, South China

The Ediacaran Doushantuo Formation in South China is underlain by the Cryogenian Nantuo Formation (glacial rocks) and overlain by the late Ediacaran Dengying Formation. It is characterized by well-preserved, large (normally >100 μm in size) spinose acritarchs (LSAs), which have been shown to be probably the only useful biostratigraphic tool for the global correlation of the early- and middle-Ediacaran. Acritarchs are organic microfossils normally known as single-celled eukaryotic organisms (protists). Although recent research suggests that some large spinose acritarchs may represent diapause egg cysts of metazoans, the biological affinities of the Ediacaran spinose acritarchs, especially for those displaying remarkable size ranges, are still debatable.Recently, smaller specimens of the Ediacaran spinose acritarchs have been found in cherts and phosphorites of the Doushantuo Formation in South China. Many described Ediacaran spinose acritarch taxa display large size variation (from tens to hundreds of microns in vesicle diameter), but some taxa only have smaller (<70 μm) specimens. The morphological comparison with Paleozoic counterparts indicates that some Ediacaran spinose acritarchs may have phylogenetic affinity to eukaryotic algae. More evidence, including wall ultra-structure, geochemical analysis and comparison with modern analogs, is needed to understand the biological affinity of the Ediacaran spinose acritarchs. The remarkable radiation of planktonic protists, characterized by abundant, diverse spinose acritarchs, occurred as early as in the late Neoproterozoic, i.e., 40–60 million years earlier than previously thought.     

Early Cambrian small carbonaceous fossils (SCFs) from an impact crater in western Finland

We describe an assemblage of small carbonaceous fossils (SCFs) and acritarchs from cored siltstones of the Lappajärvi impact structure, west‐central Finland. Previous studies had detected a depauperate acritarch biota ascribed to a deep Proterozoic origin—this age, however, was based on recovery of long‐ranging poorly age‐diagnostic sphaeromorphs. To resolve the age and provenance of these crater sediments, we applied low‐manipulation processing techniques optimized for retrieval of larger organic‐walled microfossils. Our study revealed a previously undetected assemblage containing numerous metazoan SCFs consisting of flattened ‘protoconodonts’ (grasping spines assignable to total group Chaetognatha) and a distinctive fossilised chaeta, possibly representing the oldest known annelid remains. Phylogenetically problematic fossils include various acritarchs (large Leiosphaeridia sp., Tasmanites tenellus, smaller sphaeromorphs, Synsphaeridium, Archaeodiscina and Granomarginata) and filamentous forms (Palaeolyngbya‐ and Rugosoopsis‐like filaments, Siphonophycus), likely representing prokaryotic or protistan grades of organisation. As well as adding new diversity to an emerging SCFs record, these data substantially refine the age of these sediments by more than half a billion years, to an early Cambrian Terreneuvian age. More specifically, the assemblage is equivalent to that of the Lontova Formation from the Baltic States and northwest Russia, but is previously unreported from Finland. Identification of Lontova‐type SCFs/organic‐walled microfossils at Lappajärvi further constrains the poorly resolved extent of maximum flooding during the early Cambrian in Baltica. Renewed attention should be directed to strata that have thus far produced only biostratigraphically long‐ranging or ambiguous palynological assemblages—‘SCF‐style’ processing can reveal hitherto undetected, age‐informative microfossils that are otherwise selectively removed in conventional palynological studies.     

Method for hull‐less barley transformation and manipulation of grain mixed‐linkage beta‐glucan

Hull‐less barley is increasingly offering scope for breeding grains with improved characteristics for human nutrition; however, recalcitrance of hull‐less cultivars to transformation has limited the use of these varieties. To overcome this limitation, we sought to develop an effective transformation system for hull‐less barley using the cultivar Torrens. Torrens yielded a transformation efficiency of 1.8%, using a modified Agrobacterium transformation method. This method was used to over‐express genes encoding synthases for the important dietary fiber component, (1,3;1,4)‐β‐glucan (mixed‐linkage glucan), primarily present in starchy endosperm cell walls. Over‐expression of the HvCslF6 gene, driven by an endosperm‐specific promoter, produced lines where mixed‐linkage glucan content increased on average by 45%, peaking at 70% in some lines, with smaller increases in transgenic HvCslH1 grain. Transgenic HvCslF6 lines displayed alterations where grain had a darker color, were more easily crushed than wild type and were smaller. This was associated with an enlarged cavity in the central endosperm and changes in cell morphology, including aleurone and sub‐aleurone cells. This work provides proof‐of‐concept evidence that mixed‐linkage glucan content in hull‐less barley grain can be increased by over‐expression of the HvCslF6 gene, but also indicates that hull‐less cultivars may be more sensitive to attempts to modify cell wall composition.     

Palaeoenvironmental implications of the ichnology and geochemistry of the Westbury Formation (Rhaetian), Westbury‐on‐Severn,south‐west England

Abstract: The Westbury Formation (Rhaetian) beds of Westbury Garden Cliff, Westbury‐on‐Severn, west of Gloucester, Britain, show an unusual combination of features. Both deep water and emergent characteristics are present within the sediments and the trace fossils. The ichnoassemblage consists of abundant Selenichnites, Planolites beverlyensis and Lockeia with rarer Oniscoidichnus, Chondrites, Rhizocorallium irregulare, Taenidium serpentium, an unusual form of Walcottia and Merostomichnites‐like traces. These trace fossils display an interesting relationship with the sediments: low‐energy Cruziana ichnofacies is found within high‐energy sandstones. The sandstones are interbedded with laminated mudstones, apparently deposited in deep water, but some aspects of the ichnoassemblage, preservation and sedimentation indicate shallower water. One new trace fossil, Radichnus allingtona igen. et isp. nov., closely resembles the traces of modern fiddler crabs and imply emergence, by analogy. This ichnofauna is similar to early stage disaster colonisation in recent experiments in Long Island Sound (south of Connecticut, USA) and with storm‐influenced deposits within the Cardium Formation (Seebe, Alberta, Canada). This indicates a lagoonal environment with influxes of sand and oxygen. Total organic carbon levels were found to fluctuate greatly between stratigraphic layers but remained relatively high. This implies low oxygen conditions. The abundance of sulphur (in pyrite) also supports an interpretation of anoxic conditions, and low sedimentation rates within the shale layers. A restricted shallow basin or lagoonal environment is proposed for the palaeoenvironment, with fluctuating oxygen influencing diversity.