The occurrence of a multilayered structure in the sperm of a pteridophyte

Summary A multilayered structure, previously recorded only in bryophytes, is reported in spermatids of three species of Equisetum. It is interpreted as comprising four layers, recalling the Vierergruppe of Marchantia spermatids. Unlike the multilayered structure of bryophyte spermatids, a rather compact organelle, that of Equisetum forms a thin strip extending almost the whole diameter of the cell. As in bryophytes the upper layer of the Equisetum multilayered structure is composed of parallel microtubules, extending far beyond the underlying layers. The microtubular band is considered equivalent to the similar structure seen in a variety of plant spermatozoids, but in Equisetum it is much longer and composed of many more microtubules than reported from any other plant spermatozoid. The morphology of the multilayered structure is related to the large size and short, broad form of Equisetum spermatozoids.     

Carbon,phosphorus and nitrogen budgets of the littoral Equisetum belt in an oligotrophic lake

Stores and flows of carbon, phosphorus and nitrogen in a littoral Equisetum stand were studied in 1978–1980 in the oligotrophic, mesohumic lake Pääjärvi, southern Finland. The major carbon and nutrient stores were sediment and Equisetum. The seasonal cycle of the macrophyte vegetation had a profound influence on the whole littoral ecosystem. In spring, when only dead remains of Equisetum were present above ground, there were few differences in nutrient, chlorophyll a and zooplankton concentrations between the littoral and the open lake; phytoplankton and epiphytes were the major producers.In early June, when new shoots of Equisetum reached the water surface, water exchange between the littoral and the open lake started to diminish, and the characteristic features of a closed macrophyte zone gradually developed: by August the P, Chl a and zooplankton concentrations in the littoral were 5–10 times those in the open lake. From late June until autumn Equisetum was overwhelmingly dominant both in biomass and in production.The measured total primary production and respiration values indicated a high rate of internal cycling of carbon and nutrients. The daily P requirements of plant growth exceeded the total P stored in the water by a factor of 2–4, and also exceeded the release of nutrients in excretion. High N:P ratios in the water (total 10–64, inorganic 18–171) suggested that P was probably always the limiting nutrient.The P content of the annual production of Equisetum in Pääjärvi was 2.3% of the mean annual P load, and 5.3% of the mean total P storage in the water volume of the lake.     

Defining natural factors that stimulate and inhibit cellulose:xyloglucan hetero-transglucosylation

Certain transglucanases can covalently graft cellulose and mixed-linkage β-glucan (MLG) as donor substrates onto xyloglucan as acceptor substrate and thus exhibit cellulose:xyloglucan endotransglucosylase (CXE) and MLG:xyloglucan endotransglucosylase (MXE) activities in vivo and in vitro. However, missing information on factors that stimulate or inhibit these hetero-transglucosylation reactions limits our insight into their biological functions. To explore factors that influence hetero-transglucosylation, we studied Equisetum fluviatile hetero-trans-β-glucanase (EfHTG), which exhibits both CXE and MXE activity, exceeding its xyloglucan:xyloglucan homo-transglucosylation (XET) activity. Enzyme assays employed radiolabelled and fluorescently labelled oligomeric acceptor substrates, and were conducted in vitro and in cell walls (in situ). With whole denatured Equisetum cell walls as donor substrate, exogenous EfHTG (extracted from Equisetum or produced in Pichia) exhibited all three activities (CXE, MXE, XET) in competition with each other. Acting on pure cellulose as donor substrate, the CXE action of Pichia-produced EfHTG was up to approximately 300% increased by addition of methanol-boiled Equisetum extracts; there was no similar effect when the same enzyme acted on soluble donors (MLG or xyloglucan). The methanol-stable factor is proposed to be expansin-like, a suggestion supported by observations of pH dependence. Screening numerous low-molecular-weight compounds for hetero-transglucanase inhibition showed that cellobiose was highly effective, inhibiting the abundant endogenous CXE and MXE (but not XET) action in Equisetum internodes. Furthermore, cellobiose retarded Equisetum stem elongation, potentially owing to its effect on hetero-transglucosylation reactions. This work provides insight and tools to further study the role of cellulose hetero-transglucosylation in planta by identifying factors that govern this reaction.     

Reproductive biology and population dynamics of wild gametophytes ofEquisetum

This is d study of the reproductive biology of wild gametophytes of Equisetum arvense L., E. fluviatile L., and E. palustre L., which form extensive populations during the summer months on previously inundated bare mud along the margins of reservoirs and lakes in Northern England and Wales. The garnetophytes have all the characteristics of pioneer species: they rapidly attain sexual maturity and are adversely affected by competition. Early in the summer mixtures of male and female gametophytes are produced. The former are smaller and their frequency (which is always less) diminishes throughout the season as a result of bryophytic competition. The rarity of bisexual, initially female, gametophytes, in wild populations is related to the absence of metabolite accumulation which mediates this change in rulture. Significantly different sex ratios between populations and species underlines the labile nature of the sex-determining mechanism in Equisetum. The high frequency of females bearing sporophytes indicates that intergarnetophytic fertilization is highly effective in nature. Absence of any correlation between the incidence of fertilized females and the proportions of males, in conjunction with a consideration of the male gamete dispersal distance, suggests that sporophyte formation is restricted by the availability of ripe archegonia. Sexual reproduction in Equisetum is probably limited by the narrow range of conditions under which gametophytes can become established rather than availability of water for fertilization. The majority of females, which bear one sporophyte, are smaller than unfertilized or polyembryonic females. Their small size results from nutrient demands and allelopathic compounds from the sporophytes which also probably prevent the establishment of gametophytes within mature stands of the parent species. Correlations between female diameters and frequencies of males suggest that gametophytes are more likely to produce archegonia under favourable conditions. The natural reproductive biology of Equisetum is in accord with predictions based on an understanding of the mechanism of sex determination in axenic culture. Several striking parallels between sexuality in Equisetum and dioecious angiosperms are revealed. The absence of winter flooding at two of the gametophyte sites led to the establishment of mature rtands of Equisetum, which produced cones after 3–4 years; two hybrids, E. fluviatile×E. arvense and E. fluviatile×E. palustre, were detected.     

Fungal endorhizal associates of Equisetum species from Western and Arctic Canada

We describe endorhizal fungi associated with Equisetum species collected from Ellesmere Island (82°N), Axel Heiberg Island (80°N), and from sites in Yukon Territory and the Prairie Provinces (51–67°N). Fungal colonization was assessed using a multiple quantitation microintersect method for lactofuchsin-stained roots examined with wide-field and confocal epifluorescence microscopy. Equisetum roots host abundant and diverse endorhizal fungal associates. For 85 specimens from 14 sites, total colonization averaged 30 ± 3%, range 0–97%. Colonization rates for wide aseptate hyphae characteristic of arbuscular mycorrhizae (5 ± 1%) was significantly less than for fine endophytes (20 ± 3%) or septate endophytes (17 ± 2%). Equisetum spp. are abundant in tundra and an important understory plant in boreal forests, where they are particularly common on burned or disturbed sites. Endorhizal fungi associated with Equisetum may have broad ecological relevance.     

OCCURRENCE OF INTERCALARY AND UNINTERRUPTED MERISTEMS IN EQUISETUM

Quantitative data on cell length and mitotic index in elongating internodes of seven species of Equisetum are presented as evidence for the occurrence of two patterns of internode development in rhizomes of different species. In rhizomes of three species of subgenus Equisetum (E. arvense, E. diffusum, E. telmateia) uninterrupted meristems are present, characterized by acropetal internode maturation. In rhizomes of four species of subgenus Hippochaetae (E. hyemale, E. variegatum, E. scripoides, and E. laevigatum) intercalary meristems are present. The number and locations of intercalary meristems are described for an aerial shoot of E. diffusum. The absence or diminution of intercalary meristems from rhizomes of a variety of vascular plants with intercalary meristems in aerial shoots is discussed from the standpoint of adaptive significance.     

STRUCTURALLY PRESERVED SPHENOPHYTES FROM THE TRIASSIC OF ANTARCTICA: VEGETATIVE REMAINS OF SPACIINODUM,GEN. NOV.

Sphenophyte remains of Early-Middle Triassic age are described from silicified peat collected in the Transantarctic Mountains of Antarctica. The new sphenophyte, Spaciinodum collinsonii sp. nov., is represented by ribbed, jointed stems with characteristic pith and carinal canals. Stems are relatively small, ranging from 1.8–3.0 mm in diameter, lack secondary tissues, and are characterized by vallecular canals that are restricted to nodal regions. The internodal vascular system consists of 12–18 collateral bundles which alternate between successive internodes. A complete vascular ring is present in the nodal region and is surrounded by a continuous double endodermis. Xylem is endarch and composed of elements ranging from annular to reticulate. The Antarctic sphenophyte is compared with other Gondwana fossil articulates and extant Equisetum. Superficial stomata suggest affinities with modern Equisetum subgenus Equisetum; however, some anatomical differences preclude assignment with living species.     

“Lycostrobus” chinleana,an equisetalean cone from the Upper Triassic of the southwestern United States and its phylogenetic implications

Detailed study of the cone Lycostrobus chinleana Daugherty shows that the fossil was incorrectly attributed to the Lycopodiales by the author and to the quillworts by Retallack and that it actually should be assigned to the Equisetales. The cone, which occurs in the Upper Triassic Chinle Formation at several localities in the southwestern United States, is ∼2.5 cm wide and nearly 6 cm long and consists of a stout axis bearing whorls of peltate sporangiophores. Each sporangiophore is composed of a slender stalk and a hexagonal disk, which typically bears a single, generally long, lanceolate, forward-directed leaf-like umbo tip on the outer surface and several recurrent sporangia on the inner surface. Small round to oval trilete spores occur in the sporangia. Since the leaf-like umbo tip is similar to the sterile leaves found in certain calamite cones and the recurrent sporangia are equisetalean-like, it appears that the cone may represent a intermediate stage between Calamites and modern Equisetum. According to this hypothesis, the nonbracteate Equisetum cone could have developed from a bracteate calamite cone, through reduction and fusion of the bracts and the sporangiophores, rather than by the loss of whorls of bracts of the Calamites cone as suggested earlier by others. As a result of this study the cone is assigned to the new Equisetalean genus Equicalastrobus and redescribed under the name E. chinleana (Daugherty) Grauvogel-Stamm and Ash, n. comb.     

CRYSTALLINE SILICA IN PLANTS

Crystalline silica has been found in Fragaria leaves, Equisetum shoots, diatomite, and tabaschir. Differing proportions of amorphous silica (opal) also occur in each of these. The crystalline X-ray reflections of α-quartz, low tridymite, or α-cristobalite have been found in virtually all specimens. However, unidentified crystalline reflections are noted, particulary in the silica isolated from Fragaria leaves and diatomite.     

Hetero‐trans‐β‐glucanase,an enzyme unique to Equisetum plants,functionalizes cellulose

Cell walls are metabolically active components of plant cells. They contain diverse enzymes, including transglycanases (endotransglycosylases), enzymes that ‘cut and paste’ certain structural polysaccharide molecules and thus potentially remodel the wall during growth and development. Known transglycanase activities modify several cell‐wall polysaccharides (xyloglucan, mannans, mixed‐linkage β‐glucan and xylans); however, no transglycanases were known to act on cellulose, the principal polysaccharide of biomass. We now report the discovery and characterization of hetero‐trans‐β‐glucanase (HTG), a transglycanase that targets cellulose, in horsetails (Equisetum spp., an early‐diverging genus of monilophytes). HTG is also remarkable in predominantly catalysing hetero‐transglycosylation: its preferred donor substrates (cellulose or mixed‐linkage β‐glucan) differ qualitatively from its acceptor substrate (xyloglucan). HTG thus generates stable cellulose–xyloglucan and mixed‐linkage β‐glucan–xyloglucan covalent bonds, and may therefore strengthen ageing Equisetum tissues by inter‐linking different structural polysaccharides of the cell wall. 3D modelling suggests that only three key amino acid substitutions (Trp → Pro, Gly → Ser and Arg → Leu) are responsible for the evolution of HTG's unique specificity from the better‐known xyloglucan‐acting homo‐transglycanases (xyloglucan endotransglucosylase/hydrolases; XTH). Among land plants, HTG appears to be confined to Equisetum, but its target polysaccharides are widespread, potentially offering opportunities for enhancing crop mechanical properties, such as wind resistance. In addition, by linking cellulose to xyloglucan fragments previously tagged with compounds such as dyes or indicators, HTG may be useful biotechnologically for manufacturing stably functionalized celluloses, thereby potentially offering a commercially valuable ‘green’ technology for industrially manipulating biomass.     

Amino acid sequence study of ferredoxin from a club moss,Lycopodium clavatum L

A ferredoxin was purified as the pure state from a club moss (Lycopodium clavatum L.) and sequenced. The ferredoxin was composed of 99 amino acids and had a molecular mass of 10,728, excluding iron and sulfur atoms. The ferredoxin sequence was rather distinct from that fromMarchantia polymorpha, Equisetum andGleichenia japonica. Based on comparison of ferredoxin sequences thus far established, the phylogenetic relationship between lower vascular plants is discussed.     

Bio-degradation and utilization of silica and quartz

Summary A mesophilic bacterium,Proteus mirabilis, which was known to be able to accumulate monomer silicate ions, a thermophilic bacterium,Bacillus caldolyticus, originating from a habitat with high silica concentrations, and a silicautilizing plant,Equisetum arvense, were all found to produce monomer silica from its polymer. The monomer silica, resulting from the mineralysis of either experimentally polymerized silica, or from quartz, is taken up byP. mirabilis cells, and also byEquisetum, which then deposits the silica again as a polymer in its stem and leaves. WithB. caldolyticus, which does not utilize the depolymerized product under the given conditions, we found that the intensity of the mineralysis depends on the growth rate of the organism.     

Biogeography and genome size evolution of the oldest extant vascular plant genus,Equisetum (Equisetaceae)

Background and AimsExtant plant groups with a long fossil history are key elements in understanding vascular plant evolution. Horsetails (Equisetum, Equisetaceae) have a nearly continuous fossil record dating back to the Carboniferous, but their phylogenetic and biogeographic patterns are still poorly understood. We use here the most extensive phylogenetic analysis to date as a framework to evaluate their age, biogeography and genome size evolution.MethodsDNA sequences of four plastid loci were used to estimate divergence times and investigate the biogeographic history of all extant species of Equisetum. Flow cytometry was used to study genome size evolution against the framework of phylogenetic relationships in Equisetum.Key ResultsOn a well-supported phylogenetic tree including all extant Equisetum species, a molecular clock calibrated with multiple fossils places the node at which the outgroup and Equisetum diverged at 343 Mya (Early Carboniferous), with the first major split among extant species occurring 170 Mya (Middle Jurassic). These dates are older than those reported in some other recent molecular clock studies but are largely in agreement with a timeline established by fossil appearance in the geological record. Representatives of evergreen subgenus Hippochaete have much larger genome sizes than those of deciduous subgenus Equisetum, despite their shared conserved chromosome number. Subgenus Paramochaete has an intermediate genome size and maintains the same number of chromosomes.ConclusionsThe first divergences among extant members of the genus coincided with the break-up of Pangaea and the resulting more humid, warmer climate. Subsequent tectonic activity most likely involved vicariance events that led to species divergences combined with some more recent, long-distance dispersal events. We hypothesize that differences in genome size between subgenera may be related to the number of sperm flagellae.     

Alien Grazing: Indirect Effects of Muskrats on Invertebrates

Effects of alien herbivores on plants are quite well known, especially on oceanic islands. Much less is known about the indirect effects of alien herbivores. Via indirect effects indigenous organisms can become more vulnerable to other ecological factors. We studied the effect of dense muskrat population on aquatic invertebrates in patches, which were in different succession phases after grazing. The succession was divided into three phases:(i) open-water area, (ii) mixed floating and submerged vegetation, and (iii) undisturbed Equisetum stands. Intensive muskrat grazing affected especially the size distribution of invertebrates. The proportion of small invertebrates was greatest in clear-cut areas, whereas that of large invertebrates was greatest in the Equisetum-stands. According to activity traps, the clear-cut areas harboured good populations of small perch, which are known as invertebrate predators. The study showed that the indirect effects of alien herbivore can be considerable. In this case, they seemed to be mediated to invertebrates through the change in vegetative habitat structure. This change rendered invertebrates more vulnerable to fish predation.     

THE ROOT APICAL MERISTEM OF EQUISETUM DIFFUSUM: STRUCTURE AND DEVELOPMENT

The root apical meristem of Equisetum diffusum Don has a prominent four-sided pyramidal apical cell with its base (distal face) in contact with the root cap. Derivatives (merophytes) that contribute to the main body of the root are produced from the three proximal faces of the apical cell. The first division of a proximal merophyte is periclinal to the root surface separating a small inner cell from a larger outer cell. The inner cell is the precursor of the vascular cylinder. The larger outer cell is the precursor of the epidermis, cortex, endodermis, and pericycle. Radial sectors, established early in the development of the cortex, alternate with sectors in the vascular cylinder. These developmental steps show quite clearly that early root development in Equisetum is markedly different from that of most ferns.     

Modifications of Cortical Cell Walls in Roots of Seedless Vascular Plants

Forty-three species of seedless vascular plants were assessed for modifications to root cortical cell walls. All species except Lycopodium had an endodermis with distinct Casparian bands. Experiments with the apoplastic tracer berberine hemisulfate showed that walls of all root cortical cells in the two Lycopodium species tested were permeable to this tracer. Although most species examined lacked a hypodermis several Equisetum species had a hypodermis with modified walls. Three Selaginella species had distinct Casparian bands in this cortical cell layer. This layer, therefore, is an exodermis in Selaginella and its presence limited the inward diffusion of the apoplastic tracer berberine hemisulfate.     

Effects of temperature and gibberellic acid on 4-desmethylsterol composition of Avena sativa stem segments

Of the three morphological subunits of Avena sativa stem segments (node, leaf-sheath and internode) examined, internodes constituted the richest source of phospholipids and sterols, yielding almost double the concentration of lipid found in the leaf-sheath. The phospholipid compositions of nodes and internodes were similar, comprising mostly phosphatidylcholine (PC) and phosphatidylethanolamine (PE), with linoleic, linolenic and palmitic acids as the predominant component fatty acids. Leaf-sheath tissue contained mainly PE, with equally high amounts of palmitic, linoleic and linolenic acids. β-Sitosterol and cholesterol were the major 4-desmethylsterols of the internode, while β-sitosterol was predominant in the node and leaf-sheath tissues. The growth temperature of segments prior to isolation produced its major effect on the concentration of stigmasterol, which decreased markedly with temperature. The sitosterol/stigmasterol ratio increased significantly as temperature decreased. Stem segments isolated from plants treated with gibberellic acid (GA₃) for 3 weeks, showed a significant reduction in the amounts of 4-desmethyl sterols on a dry wt basis when compared with control segments. However, when stem segments were incubated with GA₃ for 20 hr, no change in 4-desmethylsterol composition or concentration was observed, even though significant growth in response to GA₃ occurred.     

An Upper Pliocene <Emphasis Type="Italic">Equisetum</Emphasis> (Equisetales) from the Atotonilco El Grande Formation in Central Mexico

The present work documents the occurrence of Equisetum in the Upper Pliocene deposits of Central Mexico, describing and comparing fossil taxa with modern specimens. All taxonomic characters suggest a close affinity to the modern species Equisetum myriochaetum Schldl. et Cham. This work, which records the first Upper Pliocene occurrence of Equisetum in Central Mexico, confirms the wide distribution of the genus in high latitudes and more humid climates.     

Sporogenesis in Eusporangiate Ferns: I. Monoplastidic Meiosis in Angiopteris (Marattiales)

Angiopteris (Marattiales) undergoes the more primitive form of monoplastidic meiosis, while other ferns have evolved the polyplastidic type typical of seed plants. In monoplastidic cell division, the single plastid divides and serves as site of the microtubule organizing center (MTOC) for spindle formation resulting in coordinated division of plastid, nucleus, and cytoplasm. In plants with polyplastidic cell division, the MTOC is diffuse and generally perinuclear. Monoplastidic cell division is seen as a plesiomorphic feature that was inherited from algal ancestors containing a single plastid and modified through evolution. Monoplastidic meiosis occurs in all groups of bryophytes (although in only a few hepatics), Isoetes, Selaginella, certain generic segregates of Lycopodium, and in members of the Marattiales. It is not known to occur in psilophytes, Equisetum, leptosporangiate ferns, or seed plants. Received 30 January 2001/ Accepted in revised form 24 April 2001     

The occurrence of the fossil genus Graminidites in the Maastrichtian Scollard Formation,Alberta, Canada,and its palaeoecological and palaeogeographical significance

Fossil grass pollen is common in Late Tertiary sediments and its rare occurrence in the Early Tertiary led to the belief that its presence is restricted to Tertiary sediments. A literature survey shows that rare occurrences of Graminidites spp. have been reported from Campanian–Maastrichtian strata worldwide. All reported Graminidites spp. have been studied by transmitted light microscopes, which are inadequate to resolve their fine exine sculpture. Graminidites sp. occurs in the Maastrichtian Scollard Formation of Alberta, Canada, and is studied here with light microscopy and scanning electron microscopy. A new species, G. ulkapites, occurs at the initiation of a temperate climate in the area. It occurs in post‐dinosaurian beds locally but in pre‐Cretaceous–Tertiary boundary strata, which implies that grasses could also be present in dinosaur inhabitations. Grass phytoliths, reported from dinosaurian coprolites from the Maastrichtian Deccan Intertrappean sediments of India, indicate that grasses were ingested by dinosaurs even if not included in the dinosaurian diet. A plot of worldwide Senonian occurrences of Graminidites spp. on a late Maastrichtian palaeogeographic map demonstrates that the data are inadequate to reveal the place of origin or migratory pattern. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167 , 235–248.