Resolving the systematic and phylogenetic position of isolated ovules: a case study on a new genus from the Permian of China

Isolated ovules occur in many fossil plant assemblages, where they provide important insights into seed‐plant diversity and evolution. However, in many cases, the ovules cannot be attributed to individual groups of seed plants, restricting systematic and evolutionary assessments that can be made from otherwise well‐characterized fossil taxa. In the present paper, we describe a new kind of ovule discovered in tuffaceous sediments from the Permian‐aged Xuanwei Formation in Guizhou Province, China. This ovule has 180° rotational symmetry and an integument comprising a variably thick sarcotesta, a uniformly thick sclerotesta and a uniformly thin endotesta. The nucellus is attached to the integument at least basally and contains a collapsed seed megaspore; a nucellar apex is absent. Both the integument and nucellus are vascularized by paired bundles in the major plane of the ovule; the integumentary bundles are considerably larger than the nucellar bundles and the nucellar bundles emerge from a conical vascular pad. Generation of a three‐dimensional reconstruction based on serial peels revealed the gross morphology and organization of the ovule and highlighted the presence of features consistent with cardiocarpalean‐type ovules (ovule shape, histological features of the integument) and also features more typical of lagenostomalean‐ and trigonocarpalean‐type ovules (large integumentary bundles, presence of nucellar bundles). To assess the affinity and evolutionary significance of the ovule, it has been included in a cladistic matrix of cardiocarpalean‐, lagenostomalean‐ and trigonocarpalean‐type ovules. Results place the ovule within the cardiocarpalean group of ovules known to have been produced by several plant groups, including cordaitean coniferophytes, pteridosperms and Palaeozoic conifers. The cladistic topology supports generic level distinction of the present species, requiring the establishment of Muricosperma guizhouensis Seyfullah & J.Hilton gen. & sp. nov . Lagenostomalean ovules produced by hydrasperman pteridosperms form a basal paraphyletic grade, whereas trigonocarpalean ovules produced by medullosan pteridosperms form a monophyletic group in which Stephanospermum is paraphyletic with respect to Rhynchosperma and Pachytesta. The results also place the Mississippian ovule Mitrospermum bulbosum apart from all of the Pennsylvanian species of Mitrospermum that form a strongly supported clade. Consequently, M. bulbosum is transferred to the new genus Whitaddera Seyfullah & J.Hilton as W. bulbosa (Long) Seyfullah & J.Hilton. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 164 , 84–108.     

Callospermarion ovules from the Early Permian of northern China: palaeofloristic and palaeogeographic significance of callistophytalean seed-ferns in the Cathaysian flora

A distinctive kind of anatomically preserved cardiocarpalean ovule is described from the Early Permian Taiyuan Formation of northern China. Ovules are small, have 180° rotational symmetry, and possess variably thick integuments with prominent secretory cavities that may be empty, filled with resinous materials and in several instances appear to contain animal coprolites. Comparisons show that (where known) these features conform to those of Callospermarion undulatum (Neely) Rothwell, to which they are assigned, previously only known from the Pennsylvanian of Euramerica. These fossils represent the first indisputable occurrence of the genus Callospermarion in the Early Permian Cathaysian floras, and show the presence of callistophytalean seed-ferns in this palaeofloristic realm for the first time. These data combined with results from previous investigations now support the Early Permian northern Cathaysian flora including the Taiyuan Formation having evolved from the Late Carboniferous and earliest Permian Euramerican flora, with which it shares far too many generic level similarities for these co-occurrences to be coincidental. Our hypothesis is therefore that the Early Permian flora of the northern Cathaysian realm represents the continued evolution of wetland Euramerican-type coal-swamp floras, and as such is likely to present a model for evolutionarily driven floral change as opposed to the climatically driven floral changes observed in the Euramerican flora after the demise of coal-swamp environments. The distribution of coprolites in and immediately around glandular cavities in this species suggests specialised syndromes of herbivory existed in Early Permian Cathaysian ecosystems, with herbivores preferentially selecting these areas of the ovule integument.     

An Early Permian plant assemblage from the Taiyuan Formation of northern China with compression/impression and permineralized preservation

A small but diverse fossil flora is described from the Early Permian Taiyuan Formation occurring at the Yangshuling mine in Pingquan district of Hebei Province, northern China. Fossils occur as compression/impressions within mudrocks and fine-grained sandstones and also as carbonate permineralizations within volcaniclastic tuffs. All are fragmentary and contain lycopsids, sphenopsids, ferns and seed plants, and include several new species. In the compression assemblage sphenopsid and pteridosperm foliage accounts for the majority of the fossils recognised with only a few other kinds of plant organs present. In contrast, the permineralized assemblage is dominated by cordaitaleans with a composition similar to that occurring in coal-ball assemblages elsewhere in the Taiyuan Formation. From the taxonomic synthesis presented it is apparent that the Yangshuling permineralized assemblage contains many of the plant taxa diagnostic of the northern realm of the Early Permian Cathaysian flora, and preserves a representative sample of the wetland coal-swamp vegetation of this time. The permineralized assemblage at Yangshuling represents the first example of anatomically preserved plants from volcaniclastic lithologies from the Palaeozoic of China, raising the possibility of similarly preserved plant-fossil assemblages elsewhere in the Cathaysian realm.     

Ovular development and morphology in some magnoliaceae species

Floral phenology and ovular development ofLiriodendron tulipifera are described. The ovule primordia are initiated in December, followed by prominent development in March, and the ovules are mature in May. The inner integument is formed as an annular rim on the incurving ovule primordia, but the outer integument develops as a semi-annular rim interrupted on the concave side of the funicle. Later, an outgrowth, which is interpreted here as an obturator, arises on the concave side of the funicle. The funicular outgrowth arises far from the inner integument, while the outer integument is close to the inner. The outer integument and the funicular outgrowth together form an envelope complex. Later the outer integument produces two distal lobes, which disappear at maturity. Mature ovules of the threeMagnolia species examined have similar lobes. It is suggested that the hood-shaped outer integument is primitive in angiosperms.     

Developmental morphology of the ovules of Amborella trichopoda (Amborellaceae) and Chloranthus serratus (Chloranthaceae)

The developmental morphology of the outer integument in the pendent orthotropous ovules of Amborella trichopoda (Amborellaceae) and Chloranthus serratus (Chloranthaceae) was studied. In both species the outer integument is semiannular at an early stage and becomes cup-shaped but dorsiventrally somewhat asymmetric at later stages. The outer integument, which is initiated first on the concave and lateral sides of the ovule, differs from that of the anatropous ovules of other basal families with the outer integument semiannular at an early stage or throughout development. The bilateral symmetry of the outer integument is shared by these orthotropous and anatropous ovules. The developmental pattern of the outer integument and ovule incurving characterize the ovule of the Amborellaceae and Chloranthaceae, which is not equivalent to typical orthotropous ovules of eudicots. A phylogenetic analysis of ovule characters in basal angiosperms suggests that anatropous ovules with cup-shaped outer integuments and orthotropous ovules were derived independently in several clades and that the ovules of Amborella and Chloranthus might also be derivative.     

Anatomically preserved cardiocarpalean ovules in the late Palaeozoic Angara floral province of Russia

A new species of Cardiocarpus from the Sim River Basin in the southern Urals of Russia is the first anatomically preserved cardiocarpalean ovule from the Permian of the Angara floral province. Specimens are preserved in a soft organic limestone with large numbers of juvenile bactritoid protoconchs. Ovules are roughly ellipsoidal in the major plane, up to 6.0–7.5 mm in length and 5.3 mm in width, with a diminutive wing and a rounded chalaza. The outer surface of the ovules is smooth. In longitudinal sections, two vascular tissue channels traverse the sclerotesta at the chalaza, and appear to extend to near the apex within each wing. There is a simple, dome-shaped pollen chamber at the apex, which lies below a narrow micropyle. The sclerotesta consists of one or two layers of longitudinally orientated cells at the interior, and sclereids that are radially elongated towards the periphery. The sarcotesta is incompletely preserved, but displays an outer layer of cells that are somewhat elongated towards the long axis of the ovule. Numerical cladistic analysis reveals little correlation between the structure of cardiocarpalean morphotaxa and the phylogenetic relationships of the plants that produced them, where affinities are known. Cardiocarpus angarensis sp. nov. enriches our understanding of Permian spermatophytes in temperate climates of the Northern Hemisphere, and reveals that ovules similar to those of Euramerican and Cathaysian provinces characterized an Angaran spermatophyte.  Journal compilation © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 297–305. No claim to original US government works.     

A structural investigation of the ovule in sugar beet, Beta vulgaris: Integuments and micropyle

The micropyle and the integuments of sugar beet (Beta vulgaris) ovules have been investigated by light and electron microscopy during differentiation and maturation of the ovule. The micropyle itself is formed by the inner integument which is surrounded by the outer integument at its base. The micropyle containts a fibrillar PAS+ substance and is often covered by a thin sheet or hymen. Both integuments are cuticle-covered thin sheets, each 2-few cell layers in thickness. In the outer integument an increase in starch accumulation occurs during ovule maturation and probably functions as nutrient storage for embryo development. The inner epidermis of the inner integument differentiates as the most conspicuous cell layer of the beet ovule. During growth and maturation of the ovule a system of small perinuclear vacuoles containing dense material increases steadily in these cells. At maturity this system fills up more than half of each cell and very dense material has accumulated in each vacuole. This vacuole content is highly refractive and contains tannins and/or polyphenols.     

Permineralized seed fern cupules from the Triassic of Antarctica: implications for cupule and carpel evolution

In this paper we describe the first anatomically preserved Mesozoic seed fern cupule–Petriellaea. The multiovulate cupules were produced singly at the end of a short dichotomizing axis. Cupules are bilateral with a dorsal groove and transverse narrow ventral opening. The vascular system of the cupule consists of a series of traces that extend up the dorsal surface of the cupule and down the ventral face. Ovules are orthotropus, sessile, and borne on the adaxial surface of the leaflike cupule either singly or in multiple rows. They are up to 1.5 mm long, triangular in transverse section, and characterized by a multilayered integument. Nucellus and integument are fused throughout their length, but no pollen chamber is present. In the chalaza is a small vascular disc of transfusion tracheids that represents the extent of the ovule vascular system. Ovules are interpreted as being fossilized at a prepollination stage, although a few possess some evidence of a cellularized megagametophyte. These permineralized cupules indicate that in at least one Mesozoic seed fern group, ovule enclosure resulted from the transverse folding (tip to petiole) of a megasporophyll bearing adaxial ovules. Cupule morphology and ovule enclosure in other Late Paleozoic and Mesozoic seed ferns is discussed.     

Cytological Observation on Megasporogenesis and Embryo Sac Development of Regenerated Ovule in Hyacinth

The morphogenesis of regenerated ovule and cytological changes of its megasporogenesis and embryo sac development were studied. Results showed as follows: 1. the differentiation of the regenerated ovule had followed a normal process in the order of inner integument , outer integument and then funiculus. But the form of the regenerated ovules in vitro was quite different from that of ovule in vivo. Most of the regenerated ovules were orthotropous and hemianatropous , only a few were anatropous which are the same with that in vivo. 2. the megasporogenesis and the embryo sac development also had normal cytological process ,and the Polygonum type-embryo sac consisted of one egg, two synergids , one central cell and three antipodals could be seen in mature regenerated ovule. These ex-perimental results make clear that the regenerated ovule differentiated directly from explant could accomplish the complex processes of megasporogenesis and embryo sac development. By this fact ,authors infer that once the differentiation of ovule primordium, the complex biochemical programs for the megasorogenesis and embryo sac development can be controlled by the ovule itself and need no more information from flower bud and /or plant.     

The Arabidopsis SUPERMAN Gene Mediates Asymmetric Growth of the Outer Integument of Ovules

Arabidopsis superman (sup, also referred to as floral mutant10) mutants have previously been shown to have flowers with supernumerary stamens and reduced carpels as a result of ectopic expression of the floral homeotic gene APETALA3 (AP3). Here, we report that sup mutations also cause specific alterations in ovule development. Growth of the outer integument of wild-type ovules occurs almost exclusively on the abaxial side of the ovule, resulting in a bilaterally symmetrical hoodlike structure. In contrast, the outer integument of sup mutant ovules grows equally on all sides of the ovule, resulting in a nearly radially symmetrical tubular shape. Thus, one role of SUP is to suppress growth of the outer integument on the adaxial side of the ovule. Genetic analyses showed that the effects of sup mutations on ovule development are independent of the presence or absence of AP3 activity. Thus, SUP acts through different mechanisms in its early role in ensuring proper determination of carpel identity and in its later role in asymmetric suppression of outer integument growth.     

Temporal and Spatial Expression of MADS Box Genes, FBP7 and FBP11, During Initiation and Early Development of Ovules in Wild Type and Mutant Petunia hybrida

Abstract: The temporal and spatial distribution of the Petunia Floral Binding Proteins 7 and 11 (FBP7/11) were determined immunocytochemically during ovule initiation and development. In wild type plants, FBP7/11 were first detected in the placenta before ovule primordia were formed. At ovule primordium stage, FBP7/11 levels increased in the placenta and appeared in ovule primordia at the sites where integument primordia developed. At the megagametogenesis stage, FBP7/11 were present at high levels in the placenta, funicle and integument, but not in the nucellus or gametophyte. Transgenics with cosuppression of FBP7/11 formed normal ovule primordia on the placenta from which both normal ovules and carpel-like structures developed. The amount of FBP7/11 was low in the ovules and undetectable in the carpel-like structures. Plants with ectopic expression of FBP7/11 developed normal ovules on the placenta and, in addition, ovule- and carpel-like structures on sepals. Placental and sepal ovules showed the same labeling pattern as observed in wild type ovules. FBP7/11 levels were, however, low or undetectable in the carpel-like structures. The results indicate that FBP7/11 only have indirect roles in ovule primordium initiation. However, at least small quantities are needed for proper ovule differentiation. Thus, the amount of FBP7/11 is related to the type of development after primordium formation, i.e., towards the formation of real ovules or carpel-like structures.     

Ovule morphogenesis in Ranunculaceae and its systematic significance

BACKGROUND AND AIMS: Ranunculaceae has a prominent phylogenetic position in Ranunculales which appears at the base of eudicots. The aims of the present paper are to reveal the features of ovule morphogenesis in different taxa and gain a better understanding of the systematics of Ranunculaceae. METHODS: Flowers of 17 species from three subfamilies, nine tribes and 16 genera of Ranunculaceae, at successive developmental stages, were collected in the wild and studied with a scanning electron microscope. KEY RESULTS: The integuments in the unitegmic ovules in Helleborus, Ranunculus and Oxygraphis, as well as the inner integuments in the bitegmic genera, initiate annularly and eventually become cup-shaped. However, the integuments in the unitegmic ovules in Anemone and Clematis, as well as the outer integuments in the bitegmic genera, arise semi-annularly and eventually become hood-shaped. Different kinds of appendages appear on the ovules during development. In Coptis of subfamily Coptidoideae, a wrap-shaped appendage arises outside the ovule and envelopes the ovule entirely. In the genera of subfamily Thalictroideae and tribe Anemoneae of subfamily Ranunculoideae, appendages appear on the placenta, the funicle or both. In tribe Helleboreae of subfamily Ranunculoideae, an alary appendage is initiated where the integument and the funicle join and becomes hood-shaped. CONCLUSIONS: Ovule morphogenesis characteristics are significant in classification at the levels of subfamilies and tribes. The initiation patterns of the integuments and the development of appendages show diversity in Ranunculaceae. The present observations suggest that the bitegmic, hood-shaped outer integument and endostomic micropyle are primitive while the unitegmic, cupular-shaped outer integument and bistomic micropyle are derivative.     

Interactions among Genes Regulating Ovule Development in Arabidopsis Thaliana

The INNER NO OUTER (INO) and AINTEGUMENTA (ANT) genes are essential for ovule integument development in Arabidopsis thaliana. Ovules of ino mutants initiate two integument primordia, but the outer integument primordium forms on the opposite side of the ovule from the normal location and undergoes no further development. The inner integument appears to develop normally, resulting in erect, unitegmic ovules that resemble those of gymnosperms. ino plants are partially fertile and produce seeds with altered surface topography, demonstrating a lineage dependence in development of the testa. ant mutations affect initiation of both integuments. The strongest of five new ant alleles we have isolated produces ovules that lack integuments and fail to complete megasporogenesis. ant mutations also affect flower development, resulting in narrow petals and the absence of one or both lateral stamens. Characterization of double mutants between ant, ino and other mutations affecting ovule development has enabled the construction of a model for genetic control of ovule development. This model proposes parallel independent regulatory pathways for a number of aspects of this process, a dependence on the presence of an inner integument for development of the embryo sac, and the existence of additional genes regulating ovule development.     

SOME HISTOCHEMICAL,ULTRASTRUCTURAL, AND NUTRITIONAL ASPECTS OF THE OVULE OF QUERCUS GAMBELII

Histochemical analyses of the ovule of Quercus gambelii show that the major food reserves (starch grains and lipids) are located almost exclusively within the outer integument. Vascular traces are present only within this integument which contains numerous, well-developed plasmodesmata. The inner integument is virtually devoid of any food reserves and has very few plasmodesmata. The ovule has a persistent chalazal extension of residual nucellar cells (called the postament) which projects into the embryo sac. Due to the above information and the fact that the synergids rarely contain starch and no plasmodesmata are present in the walls of any of the cells of the egg apparatus (Mogensen, 1972), it is concluded that the synergids play little or no role in embryo sac nutrition. Rather, it is proposed that the pathway of available food materials in the young ovule is from the outer integument to the chalaza and through the postament into the embryo sac.     

Homeotic Transformation of Ovules into Carpel-like Structures in Arabidopsis

Ovules are specialized reproductive organs that develop within the carpels of higher plants. In Arabidopsis, mutations in two genes, BELL1 (BEL1) and APETALA2 (AP2), disrupt ovule development. In Bel1 ovules, the inner integument fails to form, the outer integument develops abnormally, and the embryo sac arrests at a late stage of megagametogenesis. During later stages of ovule development, cells of the outer integument of a Bel1 ovule sometimes develop into a carpel-like structure with stigmatic papillae and second-order ovules. The frequency of carpel-like structures was highest when plants were grown under conditions that normally induced flowering and was correlated with ectopic expression in the ovule of AGAMOUS (AG), an organ-identity gene required for carpel formation. Together, these results suggested that BEL1 negatively regulates AG late in ovule development. Likewise, mutants homozygous for the strong AP2 allele ap2-6 sometimes displayed structures with carpel-like features in place of ovules. However, such abnormal Ap2 ovules are much less ovulelike in morphology and form earlier than the Bel1 carpel-like structures. Because one role of the AP2 gene is to negatively regulate AG expression early in flower development, it is possible that AP2 works in a similar manner in the ovule. A novel ovule phenotype observed in Bel1/Ap2-6 double mutants suggested that BEL1 and AP2 genes function independently during ovule development.     

STRUCTURALLY PRESERVED FOSSIL PLANTS FROM ANTARCTICA. IV. TRIASSIC OVULES

Small, anatomically preserved ovules are described from specimens collected at Fremouw Peak in the central Transantarctic Mountains. The ovules occur within a silicified peat in the upper part of the Fremouw Formation, which is considered to be Early to Middle Triassic. Ovules are radially symmetrical and ovoid, with an integument that consists of a narrow endotesta and a complex sclerotesta. The bilayered nucellus has a characteristic scalloped appearance and is attached to the integument only at the base. Of 43 ovules examined, 50 to 60% contain cellular megagametophyte tissue. One specimen contains a possible archegonium with embryo. Although the ovules have not been found attached, their possible affinities are discussed in relation to the known flora from this locality and other, comparable-aged floras from Gondwana.     

Translocation of Uranin within the Living Ovules of Vanilla

In the ovules of Vanilla (Vanilla planifolia Andr.) before fertilization, outer integument surrounded the lower part of ovule. Uranin got into ovule through funiculus, forming, the first center of fluorescence at the chalaza zone of ovule. Then uranin was transported to micropyle end along inner integument, forming the second center of fluorescence at micropyle end of inner integument. Soon, fluorescence appeared in the egg apparatua. After fertilization, the outer integument ovule extended upward, forming micropyle ogerber with inner integument. After getting into ovule through funiculus, uranin spreads to- ward several directions: l. transported to outer integument at the entrance of micropyle; 2. transported downward to chalaza zone along outer integument at the side of funiculus; 3. extended from chalaza zone to the inside and to the outer integument at the side far from funiculus The ovules of Vanilla had no vascular bundles. On transporting in inner integument, however, the cells in inner layer next to the embryo sac appeared to be the major passage. In mature embryo sac, there was cuticle between inner integument and embryo sac at the half of micropyle end. But between embryo sac at the half of chalaza end and nucellus, cuticle was absent. Nutrient could get into embryo sac from chalaza end undoubtedly. As egg apparatus showed the fluorescence after formation of fluorescence center of inner integument at micropylar end, the possibility that nutrient got into embryo sac from micropyle could not be excluded.     

MITROSPERMUM VINCULUM SP. NOV., A CARDIOCARPALEAN OVULE FROM THE UPPER PENNSYLVANIAN OF OHIO

Numerous anatomically preserved ovules assignable to the genus Mitrospermum have been discovered in Upper Pennsylvanian sediments of Eastern Ohio. Although basically similar to Mitrospermum compressum, the newly discovered specimens exhibit several consistent differences. Ovules are strongly platyspermic, up to 4.2 mm long, 4.0 mm wide, and 0.6 mm thick. In the minor plane, ovules are broadest at the base and taper toward the micropyle. The integument exhibits three topographic regions: endotesta, sclerotesta, and sarcotesta. The sarcotesta is extremely broad in the major plane, where it forms two membranous wings. A single terete vascular bundle enters the base of the ovule, traverses the integument, and divides to form two integumentary bundles and a conspicuous nucellar platform. Integumentary bundles extend toward the tip of the ovule at the margin of the sarcotesta and sclerotesta. A pollen chamber with a prominent nucellar beak is delimited at the tip of the nucellus. Consistent differences in vascularization, size, nature of the seed base, features of the pollen chamber, and the Late Pennsylvanian age demonstrate that the specimens represent a distinct species. The discovery of these ovules extends the stratigraphic range of Mitrospermum to the Upper Pennsylvanian of Ohio.