PLASTID INCLUSIONS IN EPIDERMAL CELLS OF BETA

Inclusions were found in epidermal plastids of Beta vulgaris L., by the use of the electron microscope. One type of inclusion resembles the membrane-bound plastid inclusions found by others in various meristematic plant cells; the other does not appear to have been described before. The second type of inclusion is not membrane-bound and appears elongated and fibrillar in longisection. The elongated plastid inclusion sometimes becomes quite large. It is postulated that Beta epidermal cells may store two different proteins under some conditions.     

Structure and Development of Plastids in Epidermal Cells of African Violet (Saintpaulia ionantha Wendl.) in Culture

The structure of plastids in epidermal cells of African violet(Saintpaulia ionantha Wendl.) ‘Marge Winters’ wasexamined using transmission electron microscopy before and afterplacement of leaf tissue in culture. The plastids from matureepidermal cells contained membrane-bound inclusion bodies andprolamellar bodies in various stages of development. Starchgrains and poorly developed granal stacks were observed in asmall number of plastids. After placement of the leaves on suitableculture medium, inclusion bodies decreased in size and the lamellarsystem became more organized. The plastids in the epidermaltissue are believed to be in an arrested state of developmentand are released from this state by placement on a medium inductiveto organogenesis. Saintpaulia ionantha Wendl., African violet, membrane-bound inclusion, tissue culture, plastid development     

DEVELOPMENT OF CHLOROPLAST FINE STRUCTURE IN ASPEN TISSUE CULTURE

Chloroplast ontogeny has been examined in 42-day etiolated triploid aspen callus (Populus tremuloides Michx.) subjected to two different light conditions. White and low-intensity red illumination showed little differences in their stimulatory effects on plastid development, the red light-irradiated plastids developing only slightly more slowly. Asynchronous plastid development was noted in both lighting systems. Etioplasts contained an interconnected tubular net, phytoferritin aggregates, electron-transparent vesicles which seem to invaginate from the inner plastid membrane, membrane-bound homogeneous spheroids and starch grains. Irradiation caused various morphological changes within the proplastids; the tubular complex became transformed into the more ordered prolamellar body-like structure from which radiated membrane-bound sacs filled with electron-dense material. These sacs, characterized as thylakoid precursors, were transformed into a thylakoidal system typical of mature chloroplasts. This ontogenetic scheme represents an additional pathway for the development of photosynthetic lamellae. Other light-induced changes in the developing plastid include disappearance of phytoferritin particles and homogeneous spheroids, decrease in starch content, and appearance of osmiophilic droplets.     

PLASTIDS IN LATICIFERS OF PAPAVER. I. DEVELOPMENT AND CYTOCHEMISTRY OF LATICIFER PLASTIDS IN P. SOMNIFERUM L. (PAPAVERACEAE)

Plastids were observed in all stages of laticifer differentiation in Papaver somniferum L. Plastids in laticifer initials were present as proplastids that later developed electron-dense inclusions, but never possessed the thylakoids or starch grains that characterize chloroplasts in other cells. Electron-dense inclusions in laticifer plastids were membrane-bound and appeared to arise from the accumulation of material within an invagination of the inner plastid membrane. Cytochemical studies of these plastid inclusions indicated that their matrix was not composed of crystalline protein, α-amylose, amylopectin or polysaccharide. The results suggest that the electron-dense, membrane-bound inclusions in laticifer plastids may be composed of lipoprotein.     

The chromoplasts of Or mutants of cauliflower (Brassica oleracea L. var. botrytis)

Summary. The Or mutation in cauliflower (Brassica oleracea L. var. botrytis) leads to abnormal accumulations of -carotene in orange chromoplasts, in tissues in which leucoplasts are characteristic of wild-type plants. Or chromoplasts were investigated by light microscopy of fresh materials and electron microscopy of glutaraldehyde- and potassium permanganate-fixed materials. Carotenoid inclusions in Or chromoplasts resemble those found in carrot root chromoplasts in their optical activity and angular shape. Electron microscopy revealed that the inclusions are made up of parallel, membrane-bound compartments. These stacks of membranes are variously rolled and folded into three-dimensional objects. We classify Or chromoplasts as membranous chromoplasts. The Or mutation also limits plastid replication so that a single chromoplast constitutes the plastidome in most of the affected cells. There are one to two chromoplasts in each cell of a shoot apex. The ability of differentiated chromoplasts to divide in the apical meristems of Or mutant plants resembles the ability of proplastids to maintain plastid continuity from cell to cell in meristems of Arabidopsis thaliana mutants in which plastid replication is drastically limited. The findings are used to discuss the number of levels of regulation involved in plastid replication.     

Occurrence and Structure of Iron Inclusions in Adipocytes of Larval Lampreys

The occurrence and structure of adipocytes in the larvae of two lamprey species, Geotria australis and Petromyzon marinus, were examined by electron microscopy. Adipocytes from both species possessed large electron-dense inclusions which histochemical and energy dispersal X-ray analyses show as containing iron. The greatest concentration of inclusions in adipocytes was found in the nephric fold of G. australis. While some iron is present in the cytoplasmic matrix as ferritin, the majority is seen in large ammocoetes in membrane-bound dense aggregations of haemosiderin. The wide variety of inclusion types seen in smaller larvae may reflect on the method of formation of these inclusions within the cell. Because of the high level of iron loading in the larval lamprey nephric fold, this readily accessible tissue may provide a valuable model for studies of iron metabolism in vertebrates.     

Anucleated cryptophyte vestiges in the gonyaulacalean dinoflagellates Amylax buxus and Amylax triacantha (Dinophyceae)

Cryptophyte vestiges showing selective digestion of nuclei were found in the gonyaulacalean dinoflagellates Amylax buxus (Balech) Dodge and Amylax triacantha (Jörgensen) Sournia. They emitted bright yellow‐orange fluorescence (590‐nm emission) under epifluorescent microscopy and possessed U‐shaped plastids, suggesting the vestiges were active in photosynthesis. Under transmission electron microscopy, the plastid was characterized by a loose arrangement of two to three thylakoid stacks and included a stalked pyrenoid, as in the cryptophyte genus Teleaulax. Indeed, molecular data based on the plastid small‐subunit rRNA gene demonstrated that the vestiges in Amylax originated from Teleaulax amphioxeia. The stolen plastid (kleptoplastids) in Dinophysis is also derived from this cryptophyte species. However, in sharp contrast to Dinophysis, the plastid of the vestige in Amylax was surrounded by a double layer of plastid endoplasmic reticulum, and within the periplastidal area, a nucleomorph was retained. The vestiges also possessed mitochondria with characteristic plate‐like cristae, but lost the cell‐surface structure. The phagocytotic membrane of the dinoflagellates seemed to surround the cryptophytes right after the incorporation, but the membrane itself would probably be digested eventually. Remarkably, only one cryptophyte cell among 14 vestiges in a cell of A. buxus had a nucleus. This is the first recording of possible kleptoplastidy in gonyaulacalean dinoflagellates, and documents the strategy of a dinoflagellate involving the selective elimination of the cryptophyte nucleus.     

Die Siebröhren-Plastiden der Monocotyledonen

Zusammenfassung In vergleichenden feinstrukturellen Beobachtungen an 24 Monocotyledonen aus 21 Familien wird ein für Monocotylen-Siebröhren charakteristischer Plastidentyp näher beschrieben. Neben gelegentlichen Ablagerungen von Siebröhrenstärke enthalten ausdifferenzierte Siebröhren-Plastiden zahlreiche keilförmige, kontrastreiche und proteinhaltige Kristalloide. Sie entstehen in der Matrix der noch amöboiden Formveränderungen unterworfenen Proplastiden; in reifem Zustand werden sie aus gekreuzten Reihen paralleler, gerader und kontrastreicher Filamente (50–60 Å) aufgebaut.Die Siebröhren-Plastiden von Nymphaea alba und Nuphar luteum bilden keine Kristalloide aus, dagegen läßt sich Siebröhrenstärke wie in den übrigen bisher untersuchten Dicotylen nachweisen.

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Sieve-tube plastids of monocotyledonsComparative investigations of the fine structure and distribution of specific plastids

Summary Fine-structural investigations of 24 monocotyledons from 21 families and all but one order succeeded in revealing a plastid with cuneate proteinaceous inclusion bodies as being typical of monocot sieve-tubes. Inclusion bodies originate in large numbers during plastid differentiation; they concentrate in the matrix and aggregate around an invisible centre, that mostly lies at one end of the elongated ameboid proplastid. The inclusion-free part of the young plastid contains countless vesicles and short membranes, presumably invaginations of the inner plastid envelope. Proteinaceous inclusion bodies show a crystal-like structure composed of 50–60 Å subunits in straight and parallel order. Besides these crystal-like inclusion bodies sieve-tube plastids of many monocotyledons also contain starch. — Sieve-tube plastids of Nuphar luteum and Nymphaea alba look like plastids in dicotyledon sieve-tubes, starch being their only inclusion.

     

A predicted plastid rhomboid protease affects phosphatidic acid metabolism in Arabidopsis thaliana

The thylakoid membranes of the chloroplast harbor the photosynthetic machinery that converts light into chemical energy. Chloroplast membranes are unique in their lipid makeup, which is dominated by the galactolipids mono‐ and digalactosyldiacylglycerol (MGDG and DGDG). The most abundant galactolipid, MGDG, is assembled through both plastid and endoplasmic reticulum (ER) pathways in Arabidopsis, resulting in distinguishable molecular lipid species. Phosphatidic acid (PA) is the first glycerolipid formed by the plastid galactolipid biosynthetic pathway. It is converted to substrate diacylglycerol (DAG) for MGDG Synthase (MGD1) which adds to it a galactose from UDP‐Gal. The enzymatic reactions yielding these galactolipids have been well established. However, auxiliary or regulatory factors are largely unknown. We identified a predicted rhomboid‐like protease 10 (RBL10), located in plastids of Arabidopsis thaliana, that affects galactolipid biosynthesis likely through intramembrane proteolysis. Plants with T‐DNA disruptions in RBL10 have greatly decreased 16:3 (acyl carbons:double bonds) and increased 18:3 acyl chain abundance in MGDG of leaves. Additionally, rbl10‐1 mutants show reduced [¹⁴C]–acetate incorporation into MGDG during pulse?chase labeling, indicating a reduced flux through the plastid galactolipid biosynthesis pathway. While plastid MGDG biosynthesis is blocked in rbl10‐1 mutants, they are capable of synthesizing PA, as well as producing normal amounts of MGDG by compensating with ER‐derived lipid precursors. These findings link this predicted protease to the utilization of PA for plastid galactolipid biosynthesis potentially revealing a regulatory mechanism in chloroplasts.     

A single origin of the photosynthetic organelle in different Paulinella lineages

Background  

Gaining the ability to photosynthesize was a key event in eukaryotic evolution because algae and plants form the base of the food chain on our planet. The eukaryotic machines of photosynthesis are plastids (e.g., chloroplast in plants) that evolved from cyanobacteria through primary endosymbiosis. Our knowledge of plastid evolution, however, remains limited because the primary endosymbiosis occurred more than a billion years ago. In this context, the thecate "green amoeba" Paulinella chromatophora is remarkable because it very recently (i.e., minimum age of ≈ 60 million years ago) acquired a photosynthetic organelle (termed a "chromatophore"; i.e., plastid) via an independent primary endosymbiosis involving a Prochlorococcus or Synechococcus -like cyanobacterium. All data regarding P. chromatophora stem from a single isolate from Germany (strain M0880/a). Here we brought into culture a novel photosynthetic Paulinella strain (FK01) and generated molecular sequence data from these cells and from four different cell samples, all isolated from freshwater habitats in Japan. Our study had two aims. The first was to compare and contrast cell ultrastructure of the M0880/a and FK01 strains using scanning electron microscopy. The second was to assess the phylogenetic diversity of photosynthetic Paulinella to test the hypothesis they share a vertically inherited plastid that originated in their common ancestor.     

LATICIFER ULTRASTRUCTURE AND DIFFERENTIATION IN SEEDLINGS OF PAPAVER BRACTEATUM LINDL., POPULATION ARYA II (PAPAVERACEAE)

Laticifers of Papaver bracteatum Lindl., population Arya II, seedlings were examined by electron microscopy. Laticifers were first differentiated in procambium of the radicle associated with phloem about 72 hr after seeds were sown. Proliferation of membrane-bound vesicles of apparent endoplasmic reticulum origin distinguished laticifers from adjacent cells. Vesicles developed electron-dense caps from the internal condensation of small particles. Laticifer initials possessed the usual complement of organelles that became obscured in mature cells by the large, closely packed vesicles. Plastids contained an electron-dense, membrane-bound inclusion, but never developed lamellae or starch grains. Articulation and anastomoses between laticifer elements resulted from gradual removal of wall materials by both cells on opposite sides of the common walls at a perforation site. Differentiation of the laticifer initials and the micromorphology of the protoplast of P. bracteatum is similar to that reported for P. somniferum.     

PLASTID DIVISION IN MALLOMONAS (SYNUROPHYCEAE,HETEROKONTA)1

Laser scanning confocal microscopy and TEM were used to study the morphology of secondary plastids in algae of the genus Mallomonas (Synurophyceae). At interphase, Mallomonas splendens (G. S. West) Playfair, M. rasilis Dürrschm., M. striata Asmund, and M. adamas K. Harris et W. H. Bradley contained a single H‐shaped plastid consisting of two large lobes connected by a narrow isthmus. Labeling of DNA revealed a necklace‐like arrangement of plastid nucleoids at the periphery of the M. splendens plastid and a less‐patterned array in M. rasilis. The TEM of M. splendens and M. rasilis showed an electron‐dense belt surrounding the plastid isthmus in interphase cells; this putative plastid‐dividing ring (PD ring) was adpressed to the inner pair of the four plastid membranes, suggesting that it is homologous to the PD ring of green and red plastids. The PD ring did not contain actin (indicated by lack of staining with phalloidin) and displayed filaments or tubules of 5–10 nm in diameter that may be homologous to the tubules described in red algal PD rings. Confocal microscopy of chl autofluorescence from M. splendens showed that the plastid isthmus was severed as mitosis began, giving rise to two single‐lobed daughter plastids, which, as mitosis and cell division progressed, separated from one another and then each constricted to form the H‐shaped plastids of daughter cells. Similar plastid division cycles were observed in M. rasilis and M. adamas; however, the plastid isthmus of M. striata was retained throughout most of cell division and was eventually severed by the cell cleavage furrow.     

Intranuclear Inclusions in the Ameboid Cells of Protostelium zonatum*

SYNOPSIS. Two morphologically distinct types of intranuclear inclusions are found in ameboid cells of the protostelid mycetozoan Protostelium zonatum. One type of inclusion is a coiled tubular structure which in cross section appears as cisternae and oval to elliptical vesicles 40–60 nm in diameter. These tubular and vesicular structures are formed by a unit membrane that is connected directly with the inner nuclear membrane. The other type of inclusion is a membrane-bound structure that contains amorphous and/or fibrous material. These inclusions usually are present at several locations in a nucleus. No similar structures occur in the cytoplasm.     

The isolation of apparently homoplastidic mutants induced by a nuclear recessive gene in Arabidopsis thaliana

Summary The nuclear recessive gene, chm1, of Arabidopsis thaliana is a imitator that induces a variety of plastid alterations giving rise to mixed cells and variegated leaves. The variegation is maternally transmitted but chm1 is transmitted in a Mendelian fashion (Rédei 1973; Rédei and Plurad 1973). In order to characterize the different types of plastid alterations induced by chm1, isolating homoplastidic lines, each apparently containing one type of mutant plastid in its cells, was essential since such characterization cannot be carried out on mixed cells. We have used two genetic approaches to isolate several apparently homoplastidic mutant lines by the removal of the mutator from the genetic background, and the maternal transmission of the mutant plastids. The rapidity of obtaining homoplastidic lines in the absence of chm1 indicated a non-stochastic sorting-out of plastids in mixed cells. That each of the chm1-free homoplastidic mutant lines was apparently homoplastidic for one type of mutant plastids was confirmed by electron microscopic observations. Here we report, for the first time, the production of different homoplastidic lines in the absence of the nuclear-mutator gene. Such genetically-stable homogeneous material should be a useful tool for studying the molecular mechanism(s) by which chm1 induces a variety of heritable plastid alterations.     

Electron Microscope Observations on the Fine Structure of the Chloroplasts of Algae. II. The Chloroplasts of Caulerpa (Chlorophyceae)

The plastid ultrastructure of 12 species of Caulerpa (Chlorophyceae) was investigated by the electron microscope. A simpler pyrenoid structure was demonstrated in one species. The conspicuous concentric lamellar bodies, which are characteristic of algae with heteroplasty, were present both in the chloroplasts and the amyloplasts of all the species examined. The possible function and taxonomical value of the bodies were discussed.     

A global plastid phylogeny of the fern genus Asplenium (Aspleniaceae)

The infrageneric relationships and taxonomy of the largest fern genus, Asplenium (Aspleniaceae), have remained poorly understood. Previous studies have focused mainly on specific species complexes involving a few or dozens of species only, or have achieved a large taxon sampling but only one plastid marker was used. In the present study, DNA sequences from six plastid markers (atpB, rbcL, rps4, rps4-trnS, trnL and trnL-F) of 1030 accessions (616 of them newly sequenced here) representing c. 420 species of Asplenium (60% of estimated species diversity), 16 species of Hymenasplenium, three Diplaziopsidaceae, and four Rhachidosoraceae were used to produce the largest genus-level phylogeny yet for ferns. Our major results include: (i) Asplenium as broadly circumscribed is monophyletic based on our inclusion of representatives of 32 of 38 named segregate genera; (ii) 11 major clades in Asplenium are identified, and their relationships are mostly well-resolved and strongly supported; (iii) numerous species, unsampled in previous studies, suggest new relationships and numerous cryptic species and species complexes in Asplenium; and (iv) the accrued molecular evidence provides an essential foundation for further investigations of complex patterns of geographical diversification, speciation and reticulate evolution in this family.     

Photo-oxidation of membrane-bound and soluble cytochromec in the green sulfur bacteriumChlorobium tepidum

We studied the photosynthetic electron transfer system of membrane-bound and soluble cytochromec inChlorobium tepidum, a thermophilic green sulfur bacterium, using whole cells and membrane preparations. Sulfide and thiosulfate, physiological electron donors, enhanced flash-induced photo-oxidation ofc-type cytochromes in whole cells. In membranes,c-553 cytochromes with two (or three) heme groups served as immediate electron donors for photo-oxidized bacteriochlorophyll (P840) in the reaction center, and appeared to be closely associated with the reaction center complex. The membrane-bound cytochromec-553 had anE _m-value of 180 mV. When isolated soluble cytochromec-553, which has an apparent molecular weight of 10 kDa and seems to correspond to the cytochromec-555 inChlorobium limicola andChlorobium vibrioforme, was added to a membrane suspension, rapid photo-oxidation of both soluble and membrane-bound cytochromesc-553 was observed. The oxidation of soluble cytochromec-553 was inhibited by high salt concentrations. In whole cells, photo-oxidation was observed in the absence of exogenous electron donors and re-reduction was inhibited by stigmatellin, an inhibitor of the cytochromebc complex. These results suggest that the role of membrane-bound and soluble cytochromec inC. tepidum is similar to the role of cytochromec in the photosynthetic electron transfer system of purple bacteria.     

PERIDINIUM BALTICUM (LEVANDER) LEMMERMANN,AN UNUSUAL DINOFLAGELLATE WITH A MESOCARYOTIC AND AN EUCARYOTIC NUCLEUS123

Light and electron microscopy indicate that Peridinium balticum possesses 2 Feulgen-positive, membrane-bound nuclei which divide synchronously. One nucleus has the typical structure of dinocaryotic dinoflagellates, while the other nucleus has a structure typical of eucaryotic organisms. Connections between each nucleus and the endoplasmic reticulum are common. Membrane-bound vesicles are intimately associated with the nuclear envelope of the eucaryotic nucleus.     

OBSERVATIONS ON PLASTID FINE-STRUCTURE IN THE HOLOPARASITIC ANGIOSPERM EPIFAGUS VIRGINIANA

The ultrastructure of plastids in cortex and phloem parenchyma cells of Epifagus virginiana (L.) Bart. is described. Based upon morphology and content, several distinct plastid types appear to exist. “Tubular” complexes, lipid globules and electron dense inclusions in different arrangements appear to account for the degree of plastid variability. When results obtained with Epifagus are compared with those obtained by others for a closely related genus, a striking parallel is shown to exist.