Single Integument Research Articles

Morphological and Structural Details of Tomato Seed Coat Formation: A Different Functional Role of the Inner and Outer Epidermises in Unitegmic Ovule

In order to understand how and what structures of the tomato ovule with a single integument form the seed coat of a mature seed, a detailed study of the main development stages of the tomato ovule integument was carried out using the methods of light and electron microscopy. The integument itself it was shown to transform in the course of development into the coat (skin) of a mature seed, but the outer and inner epidermises of the integument and some layers of the integument parenchyma are mainly involved in this process. The outer epidermis cells are highly modified in later stages; their walls are thickened and lignified, creating a unique relatively hard outer coat. The fate of the inner epidermis of integument is completely different. It is separated from the other parenchyma cells of integument and is transformed into an independent new secretory tissue, an endothelium, which fences off the forming embryo and endosperm from the death zone. Due to the secretory activity of the endothelium, the dying inner parenchyma cells of the integument are lysed. Soon after the cuticle covers the endosperm, the lysis of dead integument cells stops and their flattened remnants form dense layers, which then enter the final composition of the coat of mature tomato seed. The endothelium itself returns to the location of the integument inner epidermis.

Floral anatomy, embryology, seed, and fruit development in Cephalanthus (Naucleeae-Rubiaceae), with emphasis on C. glabratus.

Information on the reproductive anatomy in genera of the tribe Naucleeae, particularly Cephalanthus, is scarce and fragmented. Of the six species in the genus, only the mature megagamethophyte of Cephalanthus occidentalis has been described. This study aims to provide information on embryological aspects in flowers of C. glabratus and to analyze the morphology and anatomy of the flowers, fruit, and seed in the six species of the genus. Cephalanthus glabratus have imperfect flowers: pistillate (PF) and staminate (SF). In the PF, the ovules are functional, while in the SF, they atrophy during the formation of the embryo sac. The mature ovule has a single integument, corresponds to the Phyllis type and the embryo sac is a Polygonum type, forming only in the PF. The presence of pollenkitt and secondary presentation of pollen were observed in the SF, as well as in the pollen formation previously described, whereas in the PF, they are absent, due to the collapse of the pollen grains inside the indehiscent anthers. The analysis of the ontogeny of the ovular excrescence in C. glabratus determined its funicular origin, calling it an aril. Its development is a pre-anthesis event, initiated during megasporogenesis. In seeds, the aril is a fleshy, white appendage which almost completely envelops the seeds of Cephalanthus, except for Cephalanthus natalensis where it is noticeably more reduced. Studies of the fruit in Cephalanthus species indicate that the infructescence is a dry schizocarp which separates into uni-seminated mericarps, except in C. natalensis that has fleshy indehiscent fruit.

Floral morphology and morphogenesis in Sanguisorba (Rosaceae): flower diversification despite petal reduction and spatial constraints

Abstract Most Rosaceae flowers are pentamerous and have petals, but subtribe Sanguisorbinae have small tetramerous (or trimerous) flowers without petals, and their floral morphology and morphogenesis remain poorly known. We investigated the floral development of three Sanguisorba spp. using scanning electron microscopy to clarify the relationship between floral development and mature flowers, with emphasis on floral constraints affecting reduction of petals, diversity of androecial patterns and the development of the gynoecium and ovule, and to clarify the pollination mechanisms in the absence of petals. All species have botryoids, with numerous flowers initiated acropetally, and the maturation of flowers follows different directions. All flowers are tetramerous and sepals emerge pairwise. No petal primordia appear. Antesepalous stamens are initiated in a pairwise manner; only S. hakusanensis has a second whorl. The ovary appears inferior by development of a cystiform hypanthium. There is only one ovule with a single integument by reduction of the inner. The space occupied by the larger sepals forces the stamens to develop sequentially. Our data demonstrate that petals are basically absent and that there is a possible shift of pollination mechanisms in Sanguisorba among the mainly wind-pollinated Sanguisorbinae, as both anemophilous and entomophilous characters are found in this genus.

Palissya – absolutely incomprehensible or surprisingly interpretable: a new morphological model, affiliations and phylogenetic insights

Abstract The morphology of the adaxial structures of cones belonging to Palissya Endlicher 1847 emend. nov. are reinterpreted based on exquisitely preserved permineralised material from the Lower Cretaceous of Queensland. Although the material was not found in situ, it likely derives from the Orallo Formation, which is Valanginian in age. The cones have dual vascular bundles in each bract/scale complex, and the different tissue types in the bract and ovule/scale complex support interpretation of the cone as a compound structure. Since the early twentieth century it has been widely accepted that each ovule is surrounded by a cup-shaped structure, but the detailed morphology of the “cup” has hitherto been unclear. These new three-dimensionally preserved specimens with in situ ovules are described as Palissya tillackiorum sp. nov. This study demonstrates that the “cup” is formed from a pair of thin scales that subtend but are not fused to each ovule; each pair of scales comprises a thicker outer and thinner inner scale. The organographic relationships among ovules and scales in Palissya show a high degree of synorganisation. The adaxial surface of the bract/scale complex has 2–6 pairs of erect (orthotropous) ovules. The ovule/scale units are arranged symmetrically in two parallel rows on either side of the midline of the bract/scale. Individual ovule/scale units are comparable to those seen in extant Podocarpaceae and Taxaceae. The ovules are thin-walled and are interpreted to have a single integument and a non-thickened (non-lignified) micropyle. These new insights allow reinterpretation of material previously referred to Palissya. A new species is described from Yorkshire, England, as P. harrisii C.R. Hill ex Pattemore & Rozefelds sp. nov. All species based on well preserved cones are reconsidered herein: P. sphenolepis (Braun 1843) Nathorst 1908 emend. Florin 1958, P. elegans Parris, Drinnan & Cantrill 1995 emend. nov., P. bartrumii Edwards 1934 emend. nov., P. antarctica Cantrill 2000 and P. hunanensis Wang 2012. Palissya ovalis Parris et al. 1995 differs structurally from Palissya and is transferred to Knezourocarpon Pattemore 2000 emend. nov. Representatives of this genus may superficially resemble those of Palissya in compressions and impressions, and their congeneric status has been previously suggested; hence its inclusion in this study. Knezourocarpon has adaxial processes that are positioned in two parallel rows but it lacks ovules and paired lateral scales that formed a cup-shape, and its processes attach directly to a central vascular trace. The improved understanding of Palissya’s morphology allows for definite separation of these genera, although the higher-order affiliation of Knezourocarpon remains unclear.

From Plant Ontology to Gene Ontology and back

Our recent goal for the Plant Ontology (PO) is to have it integrated with the Gene Ontology (GO). The simplest aspect of this is to link morphological and anatomical images of structures for PO terms with genes involved in the development of those structures (GO terms). This, also, would include images of expression analyses by in situ hybridization. By using a process of reciprocal illumination, we will be able to clarify and/or redefine PO terms. In particular, an example of this is the integument development during ovule maturation in seed plants. Gymnosperms have one integument; whereas, Angiosperms (flowering plants) have two integuments, i.e., an inner integument surrounded by an outer integument. The question that arises is which of the two integuments in the Angiosperms is the equivalent of the single integument in the gymnosperms? In Angiosperms, the gene INNER NO OUTER (INO) is involved in the proper development of the outer integument but not in the inner integument and in ino mutants there is no outer integument. INO genes are angiosperm specific as no orthologs have been found in gymnosperms. Thus, it appears that the inner integument of the Angiosperms is equivalent –homologous- to the single integument of the gymnosperms and the PO terms can be revised accordingly.

Embryology of two mycoheterotrophic orchid species, Gastrodia elata and Gastrodia nantoensis: ovule and embryo development.

Background Gastrodia elata, a famous herbal medicine, has been received great attention on its treatments of headache, vertigo and epilepsy. Gastrodia nantoensis is a newly described species from central Taiwan with potential medicinal value. Gastrodia species are fully mycoheterotrophic orchids, and the courses of their seed development are more rapid as compared to the chlorophyllous orchids. A better understanding of their reproductive biology would provide insights into the propagation and conservation of the mycoheterotrophic orchid species.ResultsBased on the histological and histochemical investigations, we observed some notable features in ovule and embryo development. First, only the archesporial cell and/or megasporocyte are present within their ovaries at the time of anthesis. Second, their suspensors consist of a single cell and their mature embryos consist of a gradient of small to large cells. Nile red staining of a globular embryo reveals the presence of cuticular material in the surface wall of embryo proper and the lateral walls of suspensor cell, indicating that the basal wall of suspensor cell is the major route for nutrient supply from maternal tissues to embryo proper. Third, their seed coats are derived from a single integument, and lignin but not cuticular material is present in the outer most layer of seed coat and persists through seed maturation.ConclusionsThe faster seed development of Gastrodia species is due to the speedy courses of ovule and embryo development. In the mature seeds, the presence of a differentiated apical zone in embryo proper suggests the easy-to-germinate character. This study provides basic knowledge for further molecular studies on embryo development and symbiotic germination of Gastrodia species.

Conservation of the role of INNER NO OUTER in development of unitegmic ovules of the Solanaceae despite a divergence in protein function.

BackgroundThe INNER NO OUTER (INO) gene is expressed in the outermost cell layer of the outer integument of bitegmic ovules and is essential for this organ’s growth. The role and cross-species functional conservation of INO orthologs were examined in members of the Solanaceae, which have unitegmic ovules. Unitegmy has evolved several times in disparate angiosperm lineages. INO expression has been observed in the outermost cell layers of all examined unitegmic ovules, but the functional role of INO in unitegmic ovules has not previously been evaluated.ResultsINO orthologs were unambiguously identified in tobacco and tomato by sequence homology. Expression of the tomato INO gene was limited to the outer cell layer of the single integument indicating that this single integument has properties of the outer integument. Expression occurred only after integument initiation, later than observed in ovules of other examined angiosperms. Virus-induced knock-down of expression of the INO ortholog in tobacco inhibited growth of the outer cell layer of the integument leading to a decrease in both integument extension and curvature of the ovule. The altered ovules closely resemble those of the aberrant testa shape (ats) ino mutant combination in Arabidopsis where we see the effect of the ino mutation on a single fused integument produced by the ats mutation. Despite significant sequence identity and similar expression patterns, the tomato INO coding region was not able to complement the Arabidopsis ino mutant.ConclusionsThe similarity of effects of ino mutations on the unitegmic ovules of tobacco and the fused integuments of the Arabidopsis ats mutant show that: 1) INO orthologs play the same role in promoting integument growth in ovules of tobacco and Arabidopsis; and 2) the unitegmic ovules of tobacco (and hence other solanaceous species) are most likely the result of a congenital fusion of two ancestral integuments. Our results further indicate that INO has a conserved role in growth of the outermost cell layer of integuments. The curvature of solanaceous ovules is driven by unequal growth of the outer layers of the single integument that likely correspond to an ancestral outer integument.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0835-z) contains supplementary material, which is available to authorized users.

Westersheimia pramelreuthensis from the Carnian (Upper Triassic) of Lunz, Austria: More Evidence for a Unitegmic Seed Coat in Early Bennettitales

Premise of research. Isolated ovules and dispersed seeds have been obtained from bulk macerations of carbonaceous shales bearing fossil plants from the Carnian (Upper Triassic) flora of Lunz am See, Austria. Through cuticle analysis, these well-preserved specimens were identified as ovules, interseminal scales, and seeds of Westersheimia pramelreuthensis, a peculiar bennettitalean ovuliferous organ. The preservation of several cuticular layers enables for interpretation of the architecture of the ovules and seeds.Methodology. The excellently preserved plant fossils were investigated using LM and epifluorescence microscopy of cuticles. For comparison, the architecture of ovules and seeds of bennettitalean reproductive organs from Scania (Sweden), Jameson Land (Greenland), and Yorkshire (United Kingdom) were reevaluated.Pivotal results. The preserved layers indicate that the nucellus is surrounded by an integument, whose apical end constitutes the micropyle. The single integument constitutes the monolayered...

Transition from two to one integument in Prunus species: expression pattern of INNER NO OUTER (INO), ABERRANT TESTA SHAPE (ATS) and ETTIN (ETT).

While gymnosperm ovules have one integument, in most angiosperms two integuments surround the ovules. Unitegmic ovules have arisen independently several times during the evolution of angiosperms, but the ultimate genetic cause of the presence of a single integument remains elusive. We compared species of the genus Prunus that have different numbers of integuments: bitegmic species, such as Prunus armeniaca (apricot) and Prunus persica (peach), and unitegmic species, such as Prunus incisa, analyzing the expression pattern of genes that are involved in integument development in Arabidopsis thaliana: INNER NO OUTER (INO), ABERRANT TESTA SHAPE (ATS) and ETTIN (ETT). Bitegmic and unitegmic species showed similar INO expression patterns, indicative of the conservation of an outer integument. However, expression of ETT, which occurs in the boundary of the outer and inner integuments, was altered in unitegmic ovules, which showed lack of ETT expression. These results strongly suggest that the presence of a single integument could be attributable to the amalgamation of two integuments and support the role of ETT in the fusion of the outer and inner integuments in unitegmic ovules, a situation that could be widespread in other unitegmic species of angiosperms.

Phylogenetics, ancestral state reconstruction, and a new infrafamilial classification of the pantropical Ochnaceae (Medusagynaceae, Ochnaceae s.str., Quiinaceae) based on five DNA regions

Ochnaceae s.str. (Malpighiales) are a pantropical family of about 500 species and 27 genera of almost exclusively woody plants. Infrafamilial classification and relationships have been controversial partially due to the lack of a robust phylogenetic framework. Including all genera except Indosinia and Perissocarpa and DNA sequence data for five DNA regions (ITS, matK, ndhF, rbcL, trnL-F), we provide for the first time a nearly complete molecular phylogenetic analysis of Ochnaceae s.l. resolving most of the phylogenetic backbone of the family. Based on this, we present a new classification of Ochnaceae s.l., with Medusagynoideae and Quiinoideae included as subfamilies and the former subfamilies Ochnoideae and Sauvagesioideae recognized at the rank of tribe. Our data support a monophyletic Ochneae, but Sauvagesieae in the traditional circumscription is paraphyletic because Testulea emerges as sister to the rest of Ochnoideae, and the next clade shows Luxemburgia+Philacra as sister group to the remaining Ochnoideae. To avoid paraphyly, we classify Luxemburgieae and Testuleeae as new tribes. The African genus Lophira, which has switched between subfamilies (here tribes) in past classifications, emerges as sister to all other Ochneae. Thus, endosperm-free seeds and ovules with partly to completely united integuments (resulting in an apparently single integument) are characters that unite all members of that tribe. The relationships within its largest clade, Ochnineae (former Ochneae), are poorly resolved, but former Ochninae (Brackenridgea, Ochna) are polyphyletic. Within Sauvagesieae, the genus Sauvagesia in its broad circumscription is polyphyletic as Sauvagesia serrata is sister to a clade of Adenarake, Sauvagesia spp., and three other genera. Within Quiinoideae, in contrast to former phylogenetic hypotheses, Lacunaria and Touroulia form a clade that is sister to Quiina. Bayesian ancestral state reconstructions showed that zygomorphic flowers with adaptations to buzz-pollination (poricidal anthers), a syncarpous gynoecium (a near-apocarpous gynoecium evolved independently in Quiinoideae and Ochninae), numerous ovules, septicidal capsules, and winged seeds with endosperm are the ancestral condition in Ochnoideae. Although in some lineages poricidal anthers were lost secondarily, the evolution of poricidal superstructures secured the maintenance of buzz-pollination in some of these genera, indicating a strong selective pressure on keeping that specialized pollination system.

Seed characteristics of Withania somnifera (Solanaceae)

The seed characteristics of Withania somnifera were studied using light and scanning electron microscopy in order to determine the specific features of this species. The seed color is yellow, and the seed shape is reniform measuring between two to three millimeters. The seed of W. somnifera is exarillate and albuminous. The seed coat type is exotestal. The seed coat develops from a single integument. The young seed coat consists of single-layered exotesta, multi-layered mesotesta and single-layered endotesta. However, parenchymatous mesotesta layers are completely compressed at maturity. Therefore, the seed coat was represented by sclerenchymatous exotesta. The primary sculpture on the seed surface is reticulate, and cells are irregular in shape with undulating anticlinal walls. In addition, the seed surface has several characteristic holes between the anticlinal walls.

Seed coat development, anatomy and scanning electron microscopy of Harpagophytum procumbens (Devil's Claw), Pedaliaceae

Seed coat development of Harpagophytum procumbens (Devil's Claw) and the possible role of the mature seed coat in seed dormancy were studied by light microscopy (LM), transmission electron microscopy (TEM) and environmental scanning electron microscopy (ESEM). Very young ovules of H. procumbens have a single thick integument consisting of densely packed thin-walled parenchyma cells that are uniform in shape and size. During later developmental stages the parenchyma cells differentiate into 4 different zones. Zone 1 is the multi-layered inner epidermis of the single integument that eventually develops into a tough impenetrable covering that tightly encloses the embryo. The inner epidermis is delineated on the inside by a few layers of collapsed remnant endosperm cell wall layers and on the outside by remnant cell wall layers of zone 2, also called the middle layer. Together with the inner epidermis these remnant cell wall layers from collapsed cells may contribute towards seed coat impermeability. Zone 2 underneath the inner epidermis consists of large thin-walled parenchyma cells. Zone 3 is the sub-epidermal layers underneath the outer epidermis referred to as a hypodermis and zone 4 is the single outer seed coat epidermal layer. Both zones 3 and 4 develop unusual secondary wall thickenings. The primary cell walls of the outer epidermis and hypodermis disintegrated during the final stages of seed maturation, leaving only a scaffold of these secondary cell wall thickenings. In the mature seed coat the outer fibrillar seed coat consists of the outer epidermis and hypodermis and separates easily to reveal the dense, smooth inner epidermis of the seed coat. Outer epidermal and hypodermal wall thickenings develop over primary pit fields and arise from the deposition of secondary cell wall material in the form of alternative electron dense and electron lucent layers. ESEM studies showed that the outer epidermal and hypodermal seed coat layers are exceptionally hygroscopic. At 100% relative humidity within the ESEM chamber, drops of water readily condense on the seed surface and react in various ways with the seed coat components, resulting in the swelling and expansion of the wall thickenings. The flexible fibrous outer seed coat epidermis and hypodermis may enhance soil seed contact and retention of water, while the inner seed coat epidermis maintains structural and perhaps chemical seed dormancy due to the possible presence of inhibitors.

Is the anthophyte hypothesis alive and well? New evidence from the reproductive structures of Bennettitales

Bennettitales is an extinct group of seed plants with reproductive structures that are similar in some respects to both Gnetales and angiosperms, but systematic relationships among the three clades remain controversial. This study summarizes characters of bennettitalean plants and presents new evidence for the structure of cones and seeds that help clarify relationships of Bennettitales to flowering plants, Gnetales, and other potential angiosperm sister groups. Bennettitales have simple mono- or bisporangiate cones. Seeds are borne terminally on sporophylls. They have a unique structure that includes a nucellus with a solid apex, no pollen chamber, and a single integument, and they are clearly not enclosed by a cupule or other specialized structures. Such features differ substantially from Gnetales, flowering plants, and the seed fern Caytonia, providing no compelling evidence for the origin of the angiospermous carpel. Cladistic tests were performed to assess the strength of the "anthophyte hypothesis" and possible relationships of Bennettitales, Gnetales, and Caytonia to flowering plants. Our results do not support the anthophyte hypothesis for the origin of angiosperms by a transformation of fertile organs that were already aggregated into a cone or flower-like structure. However, the anthophyte topology of the seed plant tree continues to be supported by morphological analyses of living and extinct taxa.

ScORK17, a transmembrane receptor-like kinase predominantly expressed in ovules is involved in seed development

The mRNA expression of the Solanum chacoense Ovule Receptor Kinase 17 (ScORK17), a receptor kinase of the LRR-VI subfamily, is highly specific to the female reproductive tissues. No LRR-VI subfamily members in any plant species have yet been attributed a function. A phylogenetic tree inferred using the kinase domain of LRR-VI subfamily members separated the family into two clades: one containing an average of 8.2 LRR per protein and a second clade containing an average of 2.7. In situ hybridization analyses showed that the ScORK17 signal was mainly detected in the single ovule integument and in the endothelium. Transient expression analysis also revealed that ScORK17 was N-glycosylated in planta. Overexpression of ScORK17 in S. chacoense did not produce plants with an altered phenotype. However, when heterologous transformation was performed with a full-length ScORK17 clone in A. thaliana, the resulting transgenic plants showed reduced seed set, mainly due to aberrant embryo sac development, thus supporting a developmental role for ScORK17 in ovule and seed development.

ABERRANT TESTA SHAPE encodes a KANADI family member, linking polarity determination to separation and growth of Arabidopsis ovule integuments

The Arabidopsis aberrant testa shape (ats) mutant produces a single integument instead of the two integuments seen in wild-type ovules. Cellular anatomy and patterns of marker gene expression indicate that the single integument results from congenital fusion of the two integuments of the wild type. Isolation of the ATS locus showed it to encode a member of the KANADI (KAN) family of putative transcription factors, previously referred to as KAN4. ATS was expressed at the border between the two integuments at the time of their initiation, with expression later confined to the abaxial layer of the inner integument. In an inner no outer (ino) mutant background, where an outer integument does not form, the ats mutation led to amorphous inner integument growth. The kan1kan2 double mutant exhibits a similar amorphous growth of the outer integument without affecting inner integument growth. We hypothesize that ATS and KAN1/KAN2 play similar roles in the specification of polarity in the inner and outer integuments, respectively, that parallel the known roles of KAN proteins in promoting abaxial identity during leaf development. INO and other members of the YABBY gene family have been hypothesized to have similar parallel roles in outer integument and leaf development. Together, these two hypotheses lead us to propose a model for normal integument growth that also explains the described mutant phenotypes.

Mechanisms of Derived Unitegmy among Impatiens Species

Morphological transitions associated with ovule diversification provide unique opportunities for studies of developmental evolution. Here, we investigate the underlying mechanisms of one such transition, reduction in integument number, which has occurred several times among diverse angiosperms. In particular, reduction in integument number occurred early in the history of the asterids, a large clade comprising approximately one-third of all flowering plants. Unlike the vast majority of other eudicots, nearly all asterids have a single integument, with the only exceptions in the Ericales, a sister group to the other asterids. Impatiens, a genus of the Ericales, includes species with one integument, two integuments, or an apparently intermediate bifid integument. A comparison of the development of representative Impatiens species and analysis of the expression patterns of putative orthologs of the Arabidopsis thaliana ovule development gene INNER NO OUTER (INO) has enabled us to propose a mechanism responsible for morphological transitions between integument types in this group. We attribute transitions between each of the three integument morphologies to congenital fusion via a combination of variation in the location of subdermal growth beneath primordia and the merging of primordia. Evidence of multiple transitions in integument morphology among Impatiens species suggests that control of underlying developmental programs is relatively plastic and that changes in a small number of genes may have been responsible for the transitions. Our expression data also indicate that the role of INO in the outgrowth and abaxial-adaxial polarity of the outer integument has been conserved between two divergent angiosperms, the rosid Arabidopsis and the asterid Impatiens.

The ultrastructure of seed coat development inRanunculus sceleratus

Seed-coat development in Ranunculus sceleratus L. has been studied by electron microscopy. Three layers developed from the single integument. The outer epidermis consisted of elongated and flattened cells that were always well attached to each other. The cells were characterized by thin walls, the presence of chloroplasts and small vacuoles. The cells of the middle layer were originally closely packed. Gradually, extensive intercellular spaces were formed. The cells of the inner epidermis elongated initially, until they became cubic and developed a thick wall with numerous wall ingrowths at the side bordering the nucellus. Thus, they give rise to a mechanical layer protecting the inner part of the seed. The elongation of cells, the thickening of cell walls, the formation of wall ridges, and the formation of intercellular spaces each coincided with characteristic configurations of microtubules. Plasmodesmata were originally found between all cells of the integument but their number decreased drastically during development, especially between the three developing seed-coat layers. Well-differentiated chloroamyloplasts, present in all cells of the developing seed-coat, pointed to autotrophy during development. Maturation eventually led to the disappearance of cytoplasm in all cells, the compression of the cells of the outer epidermis and middle layers, and the formation of a mechanical layer from the inner epidermis.

The histogenesis of integuments in some species of menispermanceae

The mode of initiation and development of integuments was investigated in six species of five genera in Menispermanceae, which have bitegmic and unitegmic ovules. The species investigated have similar integumentary structures at maturity in each of the bitegmic and unitegmic ovules. In bitegmic ovules (e.g.Cocculus), both integuments are for the most part two-cell layered. The initiation of inner integument (ii) begins with divisions of dermal cells of the nucellar primordium. The initiation of the outer integument (oi) commences with divisions of subdermal cells. In unitegmic ovules (e.g.Stephania), the integument is initiated by periclinal divisions of dermal cells, and cells of subdermal origin (which may represent the oi in case of bitegmy) form a small swelling on the raphal side and, on the antiraphal side, are included in the base of the single integument. Unitegmy of Menispermanceae (at least in the case of the genera investigated) seems to have been derived through elimination of oi, rather than through “integumentary shifting” (Bouman and Calis, 1977), a process suggested for explanation of unitegmy as in Ranunculaceae.

The ovular integument of Pittosporum undulatum Vent.: Fine structural observations

Abstract The single integument of the anacampylotropous ovule of Pittosporum undulatum Vent. (Pittosporaceae) has been investigated by light and electron microscopy. Three integumentary different regions have been detected: a peripheric layer, a parenchymatous outer region and an inner region. These may be distinguished in the medium antiraphe side on the basis of the cell shape, size and features. The cells of the peripheric layer facing the ovaric hollow appear different from the inner ones adjacent to the nucellar epidermis in their shape, for the thickness of their continuous cuticular layers and for the structure of the cell walls. The outer cells are also covered by an epicuticular layer and show the occurrence of plastids with starch grains, electron dense matrix and Light Stained Membranes. The same cells accumulate lipid bodies and a reserve of protein nature in cytoplasmic areas lacking ribosomes. The inner peripheric cells show thick inner periclinal walls characterized by a loose texture often...

Membranes of the protoplast L-form of Proteus mirabilis

Isolated membranes of the cell wall-less stable protoplast L-form of Proteus mirabilis were characterized by density gradient centrifugation and by assay for their major chemical constituents, proteins, phospholipids and lipopolysacchartide, and for some specific marker enzymes of the cytoplasmic membrane. In most of the analyzed properties the L-form protoplast membrane resembled the bacterial cytoplasmic membrane, with some notable modifications. Considerable amounts of lipopolysaccharide, normally an exclusive constituent of the outer membrane, were found. Furthermore, the L-form membranes contained the functions of the reduced nicotinamide adenine dinucleotide oxidase system, of D-lactate dehydrogenase (EC 1.1.1.28) and of succinate dehydrogenase (EC 1.3.99.1) at specific activities comparable to, or in some cases considerably higher than, those present in cytoplasmic membranes of the bacterial form. Of two peptidoglycan DD-carboxypeptidase/transpeptidases (EC 3.4.17.8 and EC 2.3.2.10). which are normally present in the cytoplasmic membrane of the bacterial form of P. mirabilis, the membrane of the protoplast L-form contained only one. Electron microscopy of thin sectioned L-form protoplasts showed extensive heterogeneity of membraneous structures. In addition to the single membraneous integument, internal membrane-bounded vesicles and multiple stacks of membranes were present, as the result of unbalanced growth and membrane synthesis in the L-form state.