THE PHLOEM OF ETAPTERIS LECLERCQII AND BOTRYOPTERIS TRIDENTATA

The phloem of Etapteris leclercqii and Botryopteris tridentata petioles is described from Lower Pennsylvanian coal balls. Petioles of B. tridentata are characterized in transverse section by an omega-shaped xylem trace, a phloem zone which extends from 2-10 cells in width, and 2-parted cortex. Etapteris leclercqii petioles exhibit a 4–9 cell-wide phloem zone surrounding the central clepsydroid xylem mass, and a 3-parted cortex. In both taxa a 1–2 cell layer parenchyma sheath separates the xylem from the extra-xylary tissues. The phloem of both species consists of sieve elements that average about 20 μm in diam by 200 μm in length in Botryopteris, and 100 μm in length in Etapteris, with horizontal-slightly oblique end walls. In transmitted light, the radial walls of the sieve elements form an irregular reticulate pattern enclosing elliptical lighter areas. With the scanning electron microscope, these areas appear as horizontal-slightly oblique furrows on the cell wall, with many small indentations lining the furrows. These indentations, because of their regular occurrence and size (from a few fractions of a micron up to 1.0 μm in diam), are interpreted as sieve pores, and the elliptical areas that enclose them as sieve areas. The phloem of E. leclercqii and B. tridentata is compared with that described for other fossil genera and with that of extant ferns.     

PHLOEM ANATOMY OF THE CARBONIFEROUS COENOPTERID FERNS ANACHOROPTERIS AND ANKYROPTERIS

Phloem histology in the petioles of two genera of Pennsylvanian ferns is detailed from coal balls collected at various localities in North America. Both Ankyropteris and Anachoropteris have primary phloem that completely surrounds the central xylem trace and is separated from it by a parenchymatous sheath. Ankyropteris contains very narrow (about 13.5 μm diam) sieve elements and a few strands of phloem parenchyma. End walls are either horizontal or slightly oblique and sieve areas as well as scattered individual pores have been observed. Anachoropteris phloem contains two different sizes of sieve elements. Small sieve elements that surround the C-shaped trace are similar to those seen in Ankyropteris. Larger elements (approximately 50–120 μm in diam) are present only within the C-shaped trace, and are elongate (up to 2.5 mm) with very oblique end walls. Sieve areas on these large cells are conspicuous, 5–8.5 μm in diam and aggregated into groups. The cell wall within each sieve area appears to be composed of criss-crossed fibrillar material. Phloem anatomy in these two ferns is compared to that previously described in other Carboniferous vascular cryptogams, as well as that known from extant plants.     

PHLOEM STRUCTURE IN THE CARBONIFEROUS FERN PSARONIUS (MARATTIALES)

Phloem anatomy in stems of Psaronius is described from coal ball specimens collected at the Berryville, IL and Lewis Creek, KY localities. Phloem completely surrounds the C-shaped xylem segments, but is more extensively developed on the abaxial side of the trace. The phloem zone consists of a central band of large diameter (approximately 90–120 μm) sieve elements surrounded by a mixed zone of smaller sieve elements and phloem parenchyma. Phloem is separated from the xylem by a parenchymatous xylem sheath. On the abaxial side of the trace, a discontinuous arc of very small diameter cells (7.8 μm) is present between the xylem sheath and the metaxylem. These cells corrrespond in position and size to protophloem cells in living marattialeans. Metaphloem sieve elements exhibit discrete, circular-oval sieve areas on their side and end walls, some of which show evidence of sieve pores. Phloem structure in Psaronius is compared with that known for living members of the Marattiales.     

The distribution of P-Protein in mature sieve elements of celery

Summary The extent of blocking of sieve-plate pores caused by release of cell turgor was investigated by fixing and processing for electron microscopy a long length of celery (Apium graveolens L.) phloem. Differences in distribution of P-protein within the pores were observed between those cells near the two cut ends, and the central cells.To assess the effect of chemical fixation on the distribution of P-protein, strands of celery phloem (fixed or unfixed, and not treated with cryoprotectants) were frozen in Freon 12 and then freeze-substituted. In sieve elements from unfixed tissue there were a greater number of sieve plates displaying partially open pores.Direct freezing of unprotected phloem tissue in Freon 12 resulted in the formation of ice crystals within the lumen of the sieve elements. Freezing of tissue at rates fast enough to avoid the formation of damaging ice crystals resulted in sieve-plate pores having an unoccluded central channel with a peripheral lining of P-protein. In the lumen of the sieve elements the P-protein filaments occurred as discrete bundles ca. 0.5 m in diameter, and as a parietal layer varying in thickness from 0.1 to 0.5 m.     

BATRACHOSPERMUM HETEROCORTICUM SP. NOV. AND POLYSIPHONIA SUBTILISSIMA (RHODOPHYTA) FROM FLORIDA SPRING-FED STREAMS1

Polysiphonia subtilissima Mont. Is reported for the first time from a freshwater environment. The presence of four pericentral cells, subdichotomous branching, apical trichoblasts and rhizoids arising from pericentral cells combined with a lack of cortication and reproductive cells is consistent with marine populations of this species. The range of filament length is 1.4–4.7 cm. Branch diameters are 38–76 μm and pericentral cell lengths are 58–125 μm. Batrachospermum heterocorticum sp. nov. is distinguished primarily by a developmental change in cortical filaments from typical cylindrical cells (5.0–7.9 μm diam in initial stages to enlarged, elliptical cells (12.9–24.1 μm diam) in mature axes. Another unique feature of this species is carpogonia with cylindrical, pedicellate trichogynes on stringht carpogonial branches in mid to outer portions of lateral whorls. Other characteristics of B. heterocorticum include the following: olive-green color, 2–6 cm length, dichotomous to trichotomous fascicles in 4–7 tiers, 385–647 μm whorl diameters, 109–198 μm carpospore diameters and relatively small “chantransia” filaments.     

PHYLOGENETIC RELATIONSHIPS OF CAULERPA (CHLOROPHYTA) BASED ON COMPARATIVE CHLOROPLAST ULTRASTRUCTURE1,2

The ultrastructure of chloroplasts from 28 of the 73 species of Caulerpa Lamouroux (Chlorophyta, Caulerpales) has been studied to aid in interpreting phylogenetic relationships among the 12 recognized sections. Variations of systematic value include pyrenoid occurrence and fine structure, thylakoid architecture and amount of photosynthate storage. Comparisons of field and culture specimens indicate these characters are consistent. Chloroplast thylakoids are grouped into bands, with the distribution of bands differing among species. In the most common arrangement, bands are evenly distributed throughout the chloroplast. A few species show lateral displacement of bands whereas others have a majority of bands arranged at one end of the chloroplast. Starch is stored cither as one or two large grains (> 1 μm diam.) or numerous small grains (< 0.5 μm diam.). Electron-transparent regions are common in other species in which chloroplasts rarely store starch. Simple, embedded pyrenoids are present in several species of section Sedoideae. An opaque region occurs in chloroplasts of C. elongata which may represent an intermediate stage in the evolutionary loss of the pyrenoid. It is suggested that the chloroplast of Caulerpa evolved, from a large, complex, pyrenoid-containing organelle housing both photosynthetic and amylogenic functions, to a small, structurally simpler one, specialized for photosynthesis alone. A phylogeny of the 12 sections of Caulerpa is constructed, based on chloroplast evolution which agrees with an earlier morphology-based hypothesis on the origin and evolution of Caulerpa.     

Mycoplasma-like Organisms in Phloem Elements of Ammobium alatum

Using transmission electron and fluorescent microscopes, Mycoplasma-like organisms (MLOs) were found in phloem cells of stems and leaves of Ammobium alatum. The diameter of these organisms ranged from 0.12–0.73 μm and averaged 0.4 μm. Symptoms induced by MLOs included chlorosis and reddening of leaves and winged stems, plant stunting and flower phyllody. This is the first report of MLOs associated with disease symptoms in A. alatum.     

IN-SITU MORPHOLOGY AND OCCURRENCE OF EUCARYOTIC PHOTOTROPHS OF BACTERIAL SIZE IN THE PICOPLANKTON OF ESTUARINE AND OCEANIC WATERS1

Concentrates of the picoplankton (0.2–2.0 μm) sized fraction from the euphotic zone of estuarine and oceanic waters were examined by transmission electron microscopy. In addition to the numerous phototrophic procaryotes (chroococcoid cyanobacteria) previously reported, small phototrophic eucaryotes were observed in 20 of 25 samples examined. Micromonas pusilla (Butcher) Manton and Parks, a 1 × 1.5 μm flagellate, was abundant in estuarine samples in summer. Similar sized cells of non-flagellated chlorophytes, either Nannochloris Naumann or Chlorella Beijerinck, were observed sporadically in many samples. The most ubiquitous microalga was a scaled, non-flagellated prasinophyte that occurred at 9 of 15 different locations on 15 of 20 sampling dates in water samples from Iceland to the Caribbean Sea, This tiny alga (0.5 to 1.0 μm in diam.) is probably the smallest known photo-trophic eucaryote and has not heretofore been described. Enrichment cultures using conventional techniques on several cruises yielded only the Chlorella-type of green alga, as well as numerous isolates of unicellular chroococcoid cyanobacteria.     

Urceolaria viridis n. sp., a Ciliate (Peritrichida, Mobilina) with Elongate Symbiotic Green Algae

SYNOPSIS. A new peritrichous ciliate containing elongate symbiotic green algae is described from the marine limpet Acmaea digitalis Eschscholtz (1833). Urceolaria viridis n. sp. is characterized as follows: turban-shaped; diam. 40 μ; diam. striated band 30 μ; diam. corona 23 μ; width of corona 2μ; 16–22 denticles; peristome slightly more than one turn; circlet of delicate cirri adoral to ciliary girdle; macronucleus H-shaped with both ends of side pieces widely separated; diam. micronucleus 4.5 μ, endosome 2.5 μ; cytoplasm containing elongate green algae measuring 2–4 by 8–16 μ.     

OCCURRENCE OF TRI-LAMELLAR BODIES IN ISOLATES OF NOSTOC (CYANOPHYCEAE)1

Tri-lamellar bodies were observed in eight of 29 isolates of Nostoc examined. They appeared identical to the previously described bodies in various species of Anabaena. The bodies consist of three discoid lamellae each ca. 0.3 μm diam and 8 nm thick. The outer lamella (closest to the plasma membrane) is separated from the middle lamella by a 12 nm space whereas the middle and inner lamellae are ca. 8 nm apart. Osmiophilic striations 3 nm wide were generally observed running between the lamellae. Osmiophilic β granules were usually associated with the inner lamella. The bodies were most always located close to the plasma membrane along the longitudinal wall near the junction of the cross and longitudinal walls. In three isolates the bodies located near the cross walls were associated with gas vesicles and possessed a slightly different morphology. These tri-lamellar bodies consisted of three discoid lamellae, each ca. 2 nm thick, ca. 25 nm apart with electron dense material between the inner and middle lamellae. Pores 20 nm diam and ca. 60 nm apart were observed in layer 2 of the cell wall adjacent to the tri-lamellar bodies. These wall pores were also observed in isolates lacking tri-lamellar bodies.     

A scanning electron microscope study of the stem structure in the West African Cane Palms (Calamoideae)

The canes of thirteen rattan taxa comprising two Calamus, three Eremospatha, five Laccosperma and three Oncocalamus were studied anatomically by scanning electron microscopy (SEM). Six out of the taxa tagged L. secundiflorum a, b, c; Oncocalamus sp. d, Oncocalmus sp. e and Calamus sp. f were unresolved herbarium voucher specimens many decades old. SEM photomicrographs revealed cell‐tissue specific differences in anticlinal cell wall of the epidermis, fibrous sheaths, sheath parenchyma and phloem strands, leaf traces, and vessel diam‐ eter. Metaxylem vessel diameters varied between 20–212 µm in the outer peripheral zone and 90–310 µm in the central cylinder, except in L. secundiflorium c where the trend is reversed. Regression analysis of the metaxylem diameter values along radial distance shows large interspecific differences from central to outer cylinder. The weak R² values (P ≤ 0.5) in nine of the thirteen species suggests that other unmeasured variables contribute to the observed trend. The structure of the unresolved voucher specimens appear to follow the same general trend in the identified specimens suggestive of phenotypic divergence between po‐ pulations brought about by developmental plasti‐ city. The leaf trace observed in Oncocalamus sp. e is unique and deserved to be examined more closely. A simple arithmetical expression indicat‐ ing structural synergy between protoxylem and metaxylem for possible volume ratio balance is included. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Histopathology of roots of Glycine max (L.) Merrill induced by root-knot nematode (Meloidogyne incognita)

In Glycine max, the second-stage juveniles of Meloidogyne incognita entered the roots through the apical meristem or elongation zone. The juveniles induced giant cells in the zone of vascular strands. Near the head of the nematode and adjacent to the giant cells, the vascular strands exhibited abnormalities in their shapes and structures; both xylem and phloem were found to be affected. The giant cells had dense and granular cytoplasm, and large nuclei with large nucleoli. Some parenchyma cells exhibited hypertrophy, while others exhibited hyperplasia. The distinctive feature of the study is reporting the occurrence of abnormal xylem, abnormal phloem and abnormal parenchyma.     

A rickettsia-like organism associated with diseased white clover

Diseased white clovers (Trifolium repens), with symptoms similar to clover club leaf, were found in England. Electron microscopy of thin sections revealed rickettsia-like organisms, 1–2 μm long by 0–2-0-3 μm diameter, in the phloem cells of these diseased plants. They were not present in healthy clover plants. At 23–30 °C the organisms multiplied rapidly and symptoms became more severe. After treatment with penicillin the organisms degenerated and plants showed remission of symptoms.     

OBSERVATIONS ON THE MORPHOLOGY AND SEXUAL REPRODUCTION OF COOLIA MONOTIS (DINOPHYCEAE)1

The surface morphology of the dinoflagellate Coolia monotis Meunier was compared with the surface morphology of Ostreopsis, The apical pore of C. monotis is similar in architecture to that of Ostreopsis but considerably longer (12 μm) than in O. heptagona (8–9 μm) and O. ovata (6–7 μm). A ventral pore in C. monotis is located on the right ventral margin between apical plate l′ and precingular plate 6″ and is similar in appearance and location to the ventral pore of O. ovata. The longitudinal flagellum (20 μm) in C. monotis is longer than in O. ovata (12 μ). Although Coolia and Ostreopsis appear to be distinctly different and should remain as two separate genera, they appear to be related. Cells of C. monotis divided by binary fission. Doubling time was 3–4 days in the logarithmic phase of growth at 23°C, 12:12 h L:D, 30–90 μE-m^?2·s^?1, and a salinity of 36%. Cultures reached cell densities of 2.5 × 10³ cells·L^?1 after 15 days of growth. The sexual process in C. monotis occurred in Erdschreiber's medium when Danish soil extract was substituted with mangrove sediment extract under the culture conditions described above. Gamete fusion produced large biflagellated planozygotes (70–75 μm diam). Planozygote maturation involved cytoplasmic reorganization, loss of motility, development of a spherical shape (80–90 μm diam), and two to three orange accumulation bodies. The cells at this stage appeared to be thin-walled cysts. Further development included reorganization of cyst contents, emergence of non-motile gametes, and development of chloroplasts, sulcus, and girdle. The nucleus of the newly formed cells occupied 50% or more of the total cell volume. Meiosis occurred in the cyst, but nuclear cyclosis was not observed. Four daughter cells were produced within 36–48 h, and motile gametes developed. The gametes exhibited sexuality for 2 months and completed the sexual life cycle by going through a thin-walled cyst stage.     

THREE NEW BENTHIC SPECIES OF PROROCENTRUM (DINOPHYCEAE) FROM CARRIE BOW CAY,BELIZE: P. SABULOSUM SP. NOV., P. SCULPTILE SP. NOV., AND P. ARENARIUM SP. NOV.1

Three new benthic, sand-dwelling dinqflagellate species, Prorocentrum sabulosum, Prorocentrum scuptile, and Prorocentrum arenarium, from coral rubble are described from scanning electron micrographs. Species were identified based on shape, size, surface micromorphology, ornamentation of thecal plates, and architecture of the periflagellar area and intercalary band. Cells of P. sabulosum are oval with a cell size of 48–50 μm long and 41–48 μm wide. The areolae are round to oval and numerous (332–450 per valve) and range from 1 to 1.6 μm in size. The periflagellar area of P. sabulosum bears a wide V-shaped depression with a flat ridge and lacks ornamentation; it accommodates six pores: one large flagellar pore, an adjacent smaller auxiliary pore, and four pores of unknown function. The flagellar and auxiliary pores are surrounded by a narrow apical collar. The intercalary band of P. sabulosum is smooth. Prorocentrum sculptile cells are broadly oval, 32–37 nm long, and 30–32 μm wide in valve view with a deep-sculptured apical area. The valves are smooth and are marked with shallow depressions (856–975 per valve). Some of these depressions have a small round opening (0.13 μm in diameter). The periflagellar area is V-shaped with a deeply indented depression; it accommodates the two flagella and a thin angled apical plate. The intercalary band is smooth. Prorocentrum arenarium cells are nearly round in valve view 30–32 μm in diameter. Thecal surface is smooth with scattered kidney-shaped valve poroids (65–73 per valve) and marginal poroids (50–57 per valve). Length and width of poroids are 0.62 μm and 0.36 μm, respectively. The periflagellar area is an unornamented, broad triangle into which a large flagellar pore and a smaller auxiliary pore are fitted. Both flagella, longitudinal and transverse, protrude from the flagellar pore. The intercalary band is smooth. The presence of a peduncle-like structure (2–3 μm long) in P. arenarium was observed situated in the flagellar pore.     

SPERMATOGENESIS IN LYCOPODIUM: THE MATURE SPERMATOZOID

The mature spermatozoid of Lycopodium cernuum is a blunt ended, fusiform cell, 8–10 μm long by 4–5 μm wide. A multilayered structure (MLS) and a subtending anterior mitochondrion are located at the anterior of the cell. The MLS is coiled through 1–1.5 gyres in a shallow sinistral helix around the periphery of the cell. The MLS would be triangular in outline if unwound and laid flat, about 1.4 μm wide, 7.5–8 μm long, and 80 nm thick. The MLS comprises four layers, S₁–S₄. The S₁ forms the spline, a supportive sheet of microtubules; the S₂, lamellate in younger stages, is an homogeneous, darkly staining layer in the mature sperm; the S₃ and S₄ retain their lamellate appearance and are delimited by lateral connections. Approximately 200 S₁ microtubules extend posteriorly from the MLS at about 45° to the MLS long axis and form a partial sheath around the nucleus. The two basal bodies are located on opposite sides of the cell external to the MLS. Each is tangential to the curve of the MLS and surrounded by a globular matrix. At their attachment, the axonemes are oriented laterally and are antiparallel to each other. Distally, the flagella, each about 38 μm long, trail behind the cell as it swims. The nucleus is roughly ovoid, about 4 μm diam, and centrally or sometimes laterally located. The greater volume of the nucleus is occupied by condensed, amorphic chromatin. Cavities within the chromatin are often seen to contain spheroidal inclusions that have two differently staining regions. The inclusions are also located at the periphery of the chromatin. The posterior of the cell is occupied by several small mitochondria and an amyloplast, about 2 μm diam containing numerous starch grains.     

Ultrastructure of the heart of the marine mussel,Geukensia demissa

The structure of the heart of Geukensia demissa, a common object of physiological and biochemical investigation, is described by scanning, transmission and freeze-fracture electron microscopy. A single-cell epithelial layer covers the ventricle, but an endothelium is lacking. Myofibers are small (6–7 μm diam.), mononucleate, and tapered. Glycogen is concentrated peripherally. Mitochondria are particularly concentrated under the sarcolemma, near the ends of the nucleus, and in rows between bundles of myofilaments. The myofilaments (6–8nm thin, 30–35 nm thick filament diam.) are loosely arranged into sarcomeres (2–4 μm) by Z bodies. Many of these Z bodies interconnect, and some anchor to the sarcolemma forming attachment plaques. Cells are joined by intercalated discs consisting of fascia adherentes, spot desmosomes, and gap junctions. The gap junctions include intramembrane particles. T tubules are absent. The sarcolemma is coupled to the junctional sarcoplasmic reticulum (JSR) over 357ndash;40% of the cell surface. Tubules extend from the JSR deep into and throughout the cell as an irregularly dispersed network. The SR occupies 1% of the cell volume. A few, small (0.1–1.0 μm) unmyelinated nerves are present, but no neuromuscular junctions were seen. The auricles have fewer and smaller myocytes than the ventricle. The auricles also contain podocytes with pedicels having 20–35 nm slits and containing sieve-like projections. The morphology of the Geukensia heart is similar to that of other bivalves.     

Secondary phloem anatomy of Cycadeoidea (Bennettitales)

Secondary phloem anatomy of several species of Cycadeoidea is described from trunks in the Wieland Collection, Peabody Museum of Natural History. The trunks were collected from the Lakota Formation, Lower Cretaceous, Black Hills of South Dakota. Secondary phloem is extensively developed and consists of alternating, tangential bands of fibers and sieve elements, with rare phloem parenchyma. Uniseriate rays, 2-22 cells high, occur between every one to three files of the axial system. Fibers are long, more than 1200 μm, approximately 26.6-34.2 μm in diameter, and have slit-like apertures on the lateral walls. Sieve elements range from 16-25 μm in diameter and are up to 500 μm long. Elliptical sieve areas appear on both end and radial walls and measure 10 μm across; minute spots, which may represent sieve pores, are present within the sieve areas. Secondary phloem of North American Cycadeoidea is similar in organization (alternating tangential bands) and cell types (sieve cells, fibers, axial parenchyma) to that known in other extant and fossil cycadophytes and some seed ferns. The unusual pattern of cell types and thickness of secondary phloem is discussed in the context of plant habit, phloem efficiency, and potential phylogenetic importance.     

THREE NEW BENTHIC SPECIES OF PROROCENTRUM (DINOPHYCEAE) FROM BELIZE: P. NORRISIANUM SP. NOV., P. TROPICALIS SP. NOV., and P. RETICULATUM SP. NOV.1

Three new benthic, photosynthetic dinoflagellate species, Prorocentrum norrisianum, Prorocentrum tropicalis, and Prorocentrum reticulatum, from floating detritus and coral rubble of Central America are described from scanning electron micrographs. Species were identified based on shape, size, surface micromorphology, thecal plate ornamentation, and architecture of the periflagellar area and intercalary band. Cells of P. norrisianum are ovate with a cell size of 20–25 μm long and 13–16 μm wide. The theca is delicate, its surface smooth, pores species specific with 95 to 105 pores per valve. Pores are round with a diameter of about 0.1 μm. The periflagellar area is V-shaped, located on the right valve in a shallow depression. It has no ornamentation. The flagellar and auxiliary pores are unequal in size. The intercalary band is smooth. Prorocentrum tropicalis cells are ovoid, 50–55 μm long and 40–45 μm wide in valve view with maximum width behind the middle region, narrow at the anterior end. The periflagellar area, situated in the right valve, is a V-shaped wide triangle with a deeply indented depression; the left valve exhibits a flat ridge. The periflagellar area is unornamented, and the flagellar and auxiliary pores are unequal in size. The valve surface is rugose with evenly distributed valve poroids. Each poroid appears to have a small dome in the center. The intercalary band is rimlike around the cell margin, granulated, and horizontally striated. Prorocentrum reticulatum cells are oblong in valve view; cells are 55–60 μm long and 40–45 μm wide. Thecal surface is reticulated; it is composed of a labyrinth of ridges with alternating depressions that vary in size and shape. Each depression has a narrow, oblong-kidney-shaped opening about 0.6 μm long. The periflagellar area is a deep, V-shaped triangle. The right valve of P. reticulatum is excavated, and contains a large flagellar pore and a smaller auxiliary pore surrounded by a narrow apical collar. The left valve margin exhibits a curved flat ridge. The intercalary band is smooth.