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.     

The origin of the ovule

Of several theories for the origin of the ovule advanced in this century and based largely on fossil evidence, the telomic concept is widely considered the most plausible. Its principal tenet is the evolution of the integument through fusion of sterile branches or telomes around a terminal megasporangium. The only point of agreement in these theories is that the entire nucellus is a megasporangium that retains a single megaspore and the endosporic female gametophyte. Their differences center on the origin of the integument. A new concept offered here on evidence from ovules of both Paleozoic and extant seed plants significantly alters the telomic theory. It proposes that the nucellus is a sporangiophore of stem origin that bears a terminal megasporangium; that at least some of the fused integumentary telomes were fertile; and that among all features cited to characterize ovules, the unique nature of the retained megaspore alone defines the seed habit. Changes in the seed plant megaspore that extended the period of nutrient absorption over the whole course of female gametophyte development, along with complex physiological changes in the nucellus, were probably achieved along a single phylogenetic line beginning in a Late Devonian population of progymnosperms. For such a combination of events to have occurred more than once is highly unlikely, and, therefore, a monophyletic origin for seed plants is proposed. Several primitive features in ovule structure, some not evidenced since the Lower Carboniferous Period, occur in a mutant form of Arabidopsis thaliana isolated from genetically transformed plants. Their recurrence provides additional support for the proposed concept of ovule origin and also suggests that the genetic mechanisms for expression of primitive features in advanced taxa could be initiated in each case by mutation of a single homeotic gene.     

Permineralized Cardiocarpalean Ovules in Wetland Vegetation from Early Permian Volcaniclastic Sediments of China

In this paper we report the first occurrence of permineralized plant fossils in volcaniclastic lithologies from China and in doing so describe a new species of cardiocarpalean ovule permineralized within tuffaceous sediments from a recently discovered locality near Pingquan, Hebei Province, North China. The tuff is part of the Early Permian Taiyuan Formation which contains a diverse fossil plant assemblage that includes cordaitean spermatophytes, lepidodendralean lycophytes, equisetophytes, and filicalean ferns, all of which were typical of Permian floras of northern China at this time. Specimens of Cardiocarpusdabiziae sp. nov. have the characteristic platyspermic shape and vascularization of cardiocarpalean ovules, and display prominent protuberances on the exterior of the integument. The integument of the ovule is composed of three layers; a thick sarcotesta, a thin and comparatively dense sclerotesta, and a single layer of large endotesta cells. The pollen chamber produces a slender nucellar beak. In several specimens the megaspore membrane contains well-developed tissue of the megagametophyte. The ovules have prominent external integumentary protuberances which suggest that well-developed systems of plant/animal interactions were operative in Early Permian wetland biotas of the Cathaysian realm.     

ONTOGENY OF THE PALEOZOIC OVULE,CALLOSPERMARION PUSILLUM

The ontogeny of the upper Pennsylvanian age gymnospermous ovule, Callospermarion pusillum, is described from petrifaction specimens collected at the Berryville locality in Illinois. Ovules exhibit a wide range of dimensional and structural features that indicate an extensive developmental sequence. Specimens range from ovules with indistinct zonation of the thin-walled integument to those with thick-walled cells of the sclerotesta. The apex of the fleshy nucellus in some specimens is preserved as a cellular mound, while in others a well-formed cellular pollen chamber is present; still other ovules are characterized by a papery-thin nucellus and pollen chamber wall. The megagametophyte of most specimens is represented by a hollow megaspore membrane that may be restricted to the base of the nucellus, or occupy the entire seed cavity. In a few specimens cellular gametophytes are preserved, and in one ovule archegonia with supposed eggs are also present. Variability in each of the features is compared with ontogenetic changes in comparable structures of living gymnospermous ovules and is correlated with ovule size. A developmental sequence for the fossil ovules is proposed.     

Re-evaluation of Halle’s fertile pteridosperms from the Permian floras of Shanxi Province, China

In 1927, T. G. Halle published an extensive synthesis of the Permian fossil plant assemblages from China’s Shanxi Province that included five genera and species of seed plant that either had ovules in organic attachment or closely and repeatedly associated with foliage. These fertile pteridosperms were the subject of a later publication (1929) that provided additional information on each species, but since that time only Nystroemia pectiniformis Halle has been subjected to detailed investigation based on the original materials. Reinvestigation of the remaining four species has yielded additional examples and new information for each species, and has confirmed the pteridospermalean affinity for three of the four species. Alethopteris norinii Halle shows organic attachment of a pollen organ similar to Wittleseya (Newberry) Halle and is reinterpreted as a microsporangiate medullosan pteridosperm. Emplectopteris triangularis Halle and Sphenopteris tenuis (Schenk) Halle both bear numerous ovules attached directly to the leaf laminae, with Emplectopteris conforming with the gross-organisation of callistophytalean pteridosperms and S. tenuis being interpreted as a derived lagenostomalean pteridosperm. However, discovery of the counterpart to the key specimen of Pecopteris wongii Halle that Halle interpreted as bearing ovules demonstrates this species to be a sterile marattialean frond that agrees with its assignment to Pecopteris. In addition, two further specimens are demonstrated to be fertile organs of previously unknown pteridosperms; Norinosperma shanxiensis gen. et sp. nov. bears cardiocarpalean ovules abaxially on the proximal portions of the frond with ovules positioned directly on the veins of a leaf lamina; Norinotheca shanxiensis gen. et sp. nov. bears large pollen organs abaxially and singly on the vein of the leaf lamina. Collectively Halle’s pteridosperms remain important today as they are some of the few specimens that actually demonstrate organic connection of fertile organs to foliage, helping to characterise the morphology and organisation of this seed plant group. These species also demonstrate the persistence of seed plant families and genera in the Permian of China that became regionally extinct at the end of the Carboniferous in Euramerica, and show that pteridosperms remained an important component of Permian as well as Carboniferous floras globally.     

Additional observations on Rhynchosperma quinnii (Medullosaceae): a permineralized ovule from the Chesterian (Upper Mississippian) Fayetteville Formation of Arkansas

New ovules from the Fayetteville Formation (Upper Mississippian) of Arkansas expand our knowledge of the morphology and anatomy of Rhynchosperma and suggest it was produced by a medullosan seed fern. Rhynchosperma has been described as radially symmetrical with a two-layered integument and vascularization in the integument only. The apical portion of the integument is ribbed; the nucellus is fused to the integument and apically differentiated into a dome-shaped pollen chamber. The vascular system is incompletely known and apparently restricted to the base of the integument. The new specimens are like Rhynchosperma in external shape, size, number of ribs, and numerous histological features. However, new data reveal that the nucellus is vascularized by a sheath of tracheids, the integument is vascularized by discrete bundles, the pollen chamber has a nucellar beak, and the nucellus is attached to the integument for a variable distance from the base. In addition, the integument is tripartite with an elaborate apical region; ribs formed by the integument are more pronounced at the apex; and internally open, hollow lobes form a stellate micropylar canal. The presence of a tripartite integument, the nature of the vascular system, the nucellus-integument attachment, the pollen chamber structure, symmetry, and the association with medullosan vegetative remains suggest medullosan affinity for these ovules and strengthens the evidence for the origin of the family before the end of the Lower Carboniferous.     

The embryo sac of Cynara cardunculus: ultrastructure of the development and localisation of the aspartic proteinase cardosin B

Cynara cardunculus is a native plant with flowers that are used traditionally in the manufacture of ewe’s cheese in the Iberian Peninsula. Milk clotting ability of the plant is attributed to the high concentrations of aspartic proteinases (APs), named cardosins, found in the flowers. Although these enzymes are well characterised on a molecular and biochemical basis, the biological role of the majority of plant APs is yet unassigned. We suspected APs play an important role in ovule function, and we characterised the maturation of the ovules of C. cardunculus and its Polygonum-type embryo sacs. The internal layer of the integument differentiates into an endothelium as described for other Asteraceae, with differentiation of two nucellar layers, a podium and a hypostase coinciding with the onset of pollen receptivity. In flowering plants, programmed cell death (PCD) events are essential for the success of nucellar maturation and consequent differentiation of a fully functional embryo sac. In C. cardunculus, nucellar PCD is integral to the maturation of the embryo sac, which in turn is closely correlated with the accumulation of the AP cardosin B specifically in the hypostase. The onset of cardosin B expression temporally coincides with the degeneration of nucellar cells. In fully mature embryo sacs, cardosin B is localised in both the hypostase and epistase, two regions that differentiate through PCD. Thus, cardosin B localisations closely correlate with events of PCD in the nucellus of C. cardunculus suggesting involvement in ovule and embryo sac development and further suggest the biological significance of APs like cardosin B, in this particular process. This work contributes new data to the plant AP research field and indicates an involvement of cardosin B in the PCD-dependent degeneration of the nucellus.     

OVULE ABORTION IN ‘NONPAREIL’ ALMOND (PRUNUS DULCIS [MILL.] D. A. WEBB)

The earliest indication of ovule abortion in almond (Prunus dulcis [Mill.] D. A. Webb ‘Nonpareil‘) is the deposition of callose (as indicated by aniline blue fluorescence) 2 days after pollination which is 2 days before clear histological symptoms of ovule degeneration are evident and 6 days before fertilization of the viable ovule. Callose deposition begins in the chalazal region of the nucellus where the funicular trace enters the ovule and ramifies into the integuments. As ovule abortion progresses, callose deposition in the inner integument extends as a ring around the nucellus. Movement of the fluorescent dye disodium fluorescein (uranin) indicated that translocation from the vascular trace into abortive ovules becomes blocked at the chalazal position. The dye freely penetrates and diffuses into viable ovules but fails to penetrate abortive ovules. Lack of, or delayed and irregular, megagametophyte development was another characteristic of abortive ovules. Biochemical and histochemical analyses of abortive and viable ovules indicated that carbohydrate depletion parallels ovule abortion. These observations lead to the conclusion that ovule abortion is accompanied by blockage in metabolite supply although whether this blockage is the primary cause or a consequence of ovule abortion is uncertain.     

Embryology of tribeDrypeteae, an enigmatic taxon ofEuphorbiaceae

The tribeDrypeteae, whose traditional assignment inPhyllanthoideae ofEuphorbiaceae is now doubtful, is studied embryologically on the basis of a literature survey and examination of six additional species in two of the four constituent genera.Drypeteae are characterized by having several embryological features that are unknown in otherPhyllanthoideae, such as a two- or three-celled ovule archesporium; a thin, two cell-layered parietal layer in the nucellus; no nucellar beak or cap; an early disintegrating nucellar tissue; thick, multiplicative, inner and outer integuments; an endothelium; a few discrete vascular bundles in the outer integument; and a fibrous exotegmen (or its derived state). EmbryologicallyDrypeteae do not fit within thePhyllanthoideae and, as available nucleotide sequence data from therbcL gene suggest, are rather placed nearErythroxylaceae, Rhizophoraceae, Chrysobalanaceae, andLinaceae. Drypeteae share with those families a combination of the fibrous exotegmen, the endothelium, and the thick, multiplicative inner integument.     

Expression of ovule and integument‐associated genes in reduced ovules of Santalales

SUMMARY Santalales comprise mainly parasitic plants including mistletoes and sandalwoods. Bitegmic ovules similar to those found in most other angiosperms are seen in many members of the order, but other members exhibit evolutionary reductions to the unitegmic and ategmic conditions. In some mistletoes, extreme reduction has resulted in the absence of emergent ovules such that embryo sacs appear to remain embedded in placental tissues. Three santalalean representatives (Comandra, Santalum, and Phoradendron), displaying unitegmic, and ategmic ovules, were studied. Observed ovule morphologies were consistent with published reports, including Phoradendron serotinum, which we interpret as having reduced ategmic ovules, consistent with earlier reports on this species. For further understanding of the nature of the ovule reductions we isolated orthologs of the Arabidopsis genes AINTEGUMENTA (ANT) and BELL1 (BEL1), which are associated with ovule development in this species. We observed ovular expression of ANT and BEL1 in patterns largely resembling those seen in the integumented ovules of Arabidopsis. These genes were found to be expressed in the integument of unitegmic ovules and in the surface layers of ategmic ovules, and in some cases, expression of BEL1 was also observed in the surrounding carpel tissue. We hypothesize that ategmic ovules derive from a fusion of the integuments with the nucellus or that the nucellus has taken on some of the characteristics confined to integuments in ancestral species.     

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.     

Gynoecium diversity and systematics of the Laurales

Carpel and ovule structure was comparatively studied in representatives of all eight families of the Laurales: Amborellaceae, Calycanthaceae, Chloranthaceae, Gomortegaceae, Hernandiaceae, Lauraceae, Monimiaceae, and Trimeniaceae. In all representatives the carpels are closed at anthesis. As in Magnoliales/winteroids, closure takes place in three different modes: (1) by postgenital fusion of the stylar (and ovarial) ventral slit (Calycanthaceae, Gomortegaceae, Lauraceae, Hernandiaceae); (2) by occlusion of the inner space by secretion (Amborellaceae, Chloranthaceae, Trimeniaceae, Mollinedioideae of Monimiaceae), all having extremely ascidiate carpels; (3) by a combination of (1) and (2), whereby the ventral slit in the style is postgenitally fused but a central canal remains open, which is filled by secretion (Monimiaceae except Mollinedioideae). The carpels have a single ovule in ventral median placentation; only Calycanthaceae have two lateral ovules, although the upper ovule degenerates. In contrast to Magnoliales/winteroids, several representatives have orthotropous or almost orthotropous ovules (Amborellaceae, Chloranthaceae, Gomortegaceae). Mature ovules vary in length between 425 μm (some Monimiaceae) and 1500 urn (some Calycanthaceae, Trimeniaceae). Although all ovules are crassinucellar, nucellus breadth varies between 60 μm (Chimonanthus, Calycanthaceae) and 500 μm (Hemandia, Hernandiaceae). In almost all representatives the single ovule (two in Calycanthaceae) tightly fills out the ovarial cavity. The micropyle is mostly formed by the inner integument. In a few cases there is no micropyle and the nucellar apex makes direct contact with the inner ovary surface or the funicle (Lauraceae p.p., Calycanthaceae p.p., Hernandiaceae p.p., Monimiaceae p.p.). The ovule is pachychalazal (or perichalazal) in Lauraceae, some Hernandiaceae, and Gomortegaceae. Both integuments are variously lobed or unlobed. The outer integument is semiannular or annular, and this may vary within a family (Calycanthaceae, Hernandiaceae, Monimiaceae); it is also exceedingly diverse in thickness (2–23 cell layers). Gynoecial traits support the association of Chloranthaceae, Trimeniaceae, and Amborellaceae, and also separately Gomortegaceae, Hernandiaceae, and Lauraceae. In addition, affinities of the first group with Schisandraceae, Illiciaceae and Austrobaileyaceae may also be supported.     

Development of ovule,fruit and seed of Xyris (Xyridaceae,Poales) and taxonomic considerations

The development of the ovule, fruit and seed of Xyris spp. was studied to assess the embryological characteristics of potential taxonomic usefulness. All of the studied species have (1) orthotropous, bitegmic and tenuinucellate ovules, with a micropyle formed by both the endostoma and exostoma; (2) a cuticle in the ovules and seeds between the nucellus/endosperm and the inner integument and between the inner and outer integuments; (3) helobial, starchy endosperm; (4) a reduced, campanulate and undifferentiated embryo; (5) a seed coat formed by a tanniferous endotegmen, endotesta with thick‐walled cells and exotesta with thin‐walled cells; and (6) a micropylar operculum formed from inner and outer integuments. The pericarp is composed of a mesocarp with cells containing starch grains and an endocarp and exocarp formed by cells with U‐shaped thickened walls. The studied species differ in the embryo sac development, which can be of the Polygonum or Allium type, and in the pericarp, which can have larger cells in either endocarp or exocarp. The Allium‐type embryo sac development was observed only in Xyris spp. within Xyridaceae. Xyris also differs from the other genera of Xyridaceae by the presence of orthotropous ovules and a seed coat formed by endotegmen, endotesta and exotesta, in agreement with the division of the family into Xyridoideae and Abolbodoideae. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 619–628.     

Self-incompatibility in Acacia retinodes: Site of pollen-tube arrest is the nucellus

In self-incompatible Acacia retinodes Schldl. var. uncifolia J.M. Black there is no inhibition of self pollen tubes before entry into the ovule, but the frequency of fertilized embryo sacs observed after self pollination is only 0.09–0.24 of that observed after outcrossing. Fluorescence- and light-microscope studies of sectioned, squashed or cleared whole ovules indicate that most self pollen tubes are arrested within the first or second layer of cells of the nucellus. The probability that nucellar arrest represents a primitive feature of self-incompatibility is discussed.     

Ovule and gametophyte development in Nicotiana glauca Graham (Solanaceae)

Abstract

The development of ovule and megagametophyte is examined in Nicotiana glauca, using light microscopy. The ovules proved unitegmic, tenuinucellate and endothelial as in all the Solanaceae so far studied. The ovule primordia are of the three-zonate type. The integument, which is of dermal origin, is at first two-layered but later produces additional intermediate cells whose origin is not constant. The nucellus, whose initial curvature bears no relation to the origin of the integument, has, like other Solanaceae, a one or two-celled archesporium from which a single meiocyte develops. The gametophyte is confirmed to be bisporic in origin and its development follows the Allium type. Furthermore, the hypostase, which is rare in the family, is observed below the antipodal cells.     

北美香柏雌球果的发育

用扫描电镜(SEM)观察了北美香柏 Thuja occidentalis 雌球果的发育过程。在北京,北美香柏的雌球果是在八月初由营养芽转变而来,雌球果一般有4~6对苞片,中间2~3对可育,每一苞片腋部着生两枚胚珠,在可育苞片腋部最先观察到一扁平的隆起,并在其上分化出两个胚珠原基,接着分化出珠被和珠心,最后形成扁平而两侧对称的胚珠。在北美香柏雌球果发育过程中,约一半的雌球果在2~3对可育苞片中位于下面的1~2对的腋部产生3个胚珠原基,中间一个较小,并在以后的发育中逐渐退化。由此推测北美香柏的雌球果可能是由祖先类群中每一苞片具多于2个胚珠的雌球果演化而来。在光镜下对雌球果维管系统的观察发现,传粉前幼小雌球果的苞片内仅有一束维管束,传粉后随着苞片基部的居间生长,有4—8束维管束在苞片内形成,但是新发育的维管束木质部和韧皮部相对位置与正常叶性器官一致,这与在以往报道的柏科植物成熟雌球果的苞片中均有反向维管束的发育不同。北美香柏雌球果早期发育和维管束分析结果支持傅德志和杨亲二提出的解释裸子植物生殖器官形态演化的“苞鳞-种鳞复合体”理论。关键词北美香柏;雌球果的发育;胚珠分化;SEM     

Postpollination-prezygotic ovular secretions into the micropylar canal inPseudotsuga menziesii (Pinaceae)

Ovular morphology was examined ultrastructurally inPseudotsuga menziesii to determine the effects of the ovule on pollen development. Vesicles containing lipid-like substances traverse cell walls of the inner epidemis of the integument and release their contents at the integument surface to form the integumentary membrane. A major aqueous secretion from the integument into the micropylar canal is proposed to occur by the movement of the integumentary membrane and its invaginations towards the center of the micropylar canal. The cellular degeneration of the nucellar apex results from the breakdown of vacuoles. After this degeneration, electron-dense substances move from the prothallial cells of the female gametophyte towards the nucellus, and many morphological changes in the nucellus, prothallial cells, and micropylar canal take place simultaneously. We interpret these changes to result from another major secretion from the prothallial cells. Egg cytoplasm appears to disorganize for a short time. Simultaneously, substantial amounts of electron dense-substances in the prothallial cells and lipid-like substances in surface cell walls of the female gametophyte move towards the nucellus as components of the third major secretion.