STUDIES ON FOSSIL AZOLLA: PRIMITIVE TYPES OF MEGASPORES AND MASSULAE FROM THE CRETACEOUS

Fossil Salviniaceae are described from the Claggett Shale and Judith River Formation, late Cretaceous (Campanian stage) of Montana. A new genus, Parazolla, from the Claggett Shale, has megaspores in which the swimming apparatus is composed of a number of elongate floats attached to the megaspore body and invested by coiled hairs. The floats separate at maturity. Massulae (bearing microspores) have simple hair-like glochidia, many of which are knobbed at their tips. Glochidia tend to resemble the perisporial hairs of the megaspore body. This resemblance provides fossil evidence of the homology of these two hair-like structures among living species of Azolla. In Azolla simplex from the Judith River Formation the megaspore has a single cap-like so-called columellate float. Massulae, which have anchor-shaped glochidia, are associated with these megaspores. A. simplex is the oldest species of Azolla and Parazolla the oldest member of the Salviniaceae so far found.     

A CONTRIBUTION TO THE EARLY TERTIARY FOSSIL RECORD OF THE SALVINIACEAE

Megaspores and massulae of Azolla, Azollopsis, and Salvinia are recorded from the Paleocene and Eocene of Montana and the Dakotas. Three new species of Azolla are described and two species of Azolla are redescribed. The columella of the floating apparatus of the megaspore apparatuses of Azolla appears to be a phylogenetic localization of the perispore around the megaspore. A new section of Azolla (section Kremastospora) is created for species whose megaspore apparatuses have many floats and whose massulae have hooked (anchor-shaped) glochidia. Salvinia preauriculata is based only on leaf remains from the Eocene of North America. Megaspores and massulae which probably are conspecific with the leaf remains are described. The megaspores and massulae of the fossil species are much smaller than those of the living species, S. auriculata, with which S. preauriculata has been compared. The similar size of the megaspores and massulae of the fossil species suggests a less specialized condition than in living species, where these two structures differ greatly in size. Azollopsis tomentosa, previously known from the late Cretaceous, has been found in the Eocene.     

EM-studies on Salviniaceae Megaspores from the Middle Miocene Fasterholt Flora,Denmark

Reproductive organs of Salvinia and Azolla were found together with other small plant remains in the Middle Miocene Fasterholt flora, Denmark. They were examined by the use of SEM and TEM; this was done with the three species of Azolla for the first time. Salvinia is represented by one, or possibly two, species. The megaspore of S. cerebrata has a characteristic surface sculpture of irregular, rounded ridges. Some specimens differ from the typical S. cerebrata by having strongly developed verrucae or clavae. They are described as Salvinia sp., but it is possible that they represent a variety of S. cerebrata. Azolla is represented by three species. A. nikitinii and A. ventricosa have nine floats in two tiers; massulae have not been found. A. tuganensis has three floats in one tier, massulae have anchor-shaped glochidia.     

Paleoazolla,a new heterosporous fern from the Upper Cretaceous of Argentina

Megaspore and microspore massulae of Paleoazolla patagonica gen. et sp. nov. are described from the Upper Cretaceous of La Colonia Formation, Chubut Province, Argentina. The new fern possesses megaspore complexes with three to four glochidiate floats attached directly to the megaspore; a columella-like structure appears to be absent. The megaspore wall consists of a two-layered exine that is smooth to irregularly perforate, a two-layered perine with a spongy, densely packed endoperine, and a loosely organized exoperine. Infrafilosum hairs cover the exoperine. Microspore massulae are irregular in size and shape and bear multibarbed glochidia that generally have anchor-shaped tips. A comparison with other azollaceous and salviniaceous genera, particularly Azolla, is provided, together with a discussion of some evolutionary trends within the family.     

Some structures in Azolla megaspores,and an anomalous form

On the assumption that the “float” apparatus of Azolla arose by the “capture” and retention of separate floats developed as massulae round abortive megaspores, a model is proposed, accounting hypothetically for the separate origin of three-floated, nine-floated, fifteen-floated and twenty-four-floated species in different types of cytological behaviour during origin of the functional megaspore. It is considered irrelevant to ask whether nine-floated arose from three-floated or vice versa. Each type arose de novo and possibly more than once, out of the initial cytological situation which is constant for all species.An abnormal form of Azolla imbricata is described, which in terms of the model appears to represent a reversion to a Salvinia-like ancestral form, by loss of linked genes responsible for the Azolla “syndrome”.Anatomical and morphological studies on five modern and two Paleocene species are used to illustrate adaptation of perine as float-retaining mechanisms. A new structure, the manicula, produced from the perispore, is described in A. teschiana and A. velus.     

EMBRYOLOGICAL STUDIES IN THE FAMILY SAXIFRAGACEAE (S.L.). I. DEVELOPMENT OF THE OVULE AND EMBRYO SAC IN SAXIFRAGA FORTUNEI VAR. PARTITA (MAKINO) NAKAI

The development of the ovule, megaspore and megagametophyte in Saxifraga fortunei var. partita (Makino) Nakai was observed. The ovule is anatropous, bitegmic, and crassinucellate. Both integuments originate from the epidermis. The archesporium is considered to be multicellular. The primary sporogenous cell functions as the megaspore mother cell which forms a T-shaped tetrad. The chalazal member of the megaspore tetrad is functional and develops into a Polygonum-type embryo sac. In the pyriform synergids the filiform apparatus is observed, but any hook or indentations could not be recognized. The antipodal cells are detectable until the Helobial endosperm undergoes several nuclear divisions. Secondary multiplication of the nuclei or the cells of the antipodals could not be observed.     

DEVELOPMENT OF THE OVULE AND FEMALE GAMETOPHYTE IN EUSTACHYS PETRAEA AND E. GLAUCA (POACEAE)

The unilocular pistil in Eustachys contains a single ovule with lateral placentation. In E. petraea and E. glauca, the mature ovule is bitegmic, tenuinucellate, and amphitropous with the endostomic micropyle oriented toward the base of the locule. A single hypodermal archesporial cell enlarges to form the megasporocyte. The chalazal dyad member is larger than the micropylar one, and meiosis II is nonsynchronized. Two-thirds of the tetrads are linear and one-third T-shaped. The chalazal megaspore is functional. Initially ovoid, the two-nucleate female gametophyte becomes curved as it enlarges. The four-nucleate stage becomes wider at its extremities and constricted in the center. Synchronous mitotic divisions establish the eight-nucleate stage with four nuclei at each pole separated by a large central vacuole. In E. petraea, the maturation sequence begins with antipodal differentiation, followed by differentiation of the egg apparatus, migration of the polar nuclei to the center, and division of the antipodals to produce twelve cells. The sequence in E. glauca begins with migration of the polar nuclei followed by differentiation of the antipodals, egg apparatus, and antipodal replication to six cells. The polar nuclei fuse to form a secondary nucleus appressed to the egg cell in E. glauca and separated from it by a vacuole in E. petraea. T-tests for length measurements for various stages of development indicate that the functional megaspore and two-nucleate female gametophyte are significantly larger in E. glauca than in E. petraea. There is no significant difference in gametophyte length at the four-nucleate stage, and at the eight-nucleate stage, length in E. petraea surpasses that in E. glauca. This gap widens significantly at the mature stage. Nuclear volumes are significantly greater in E. glauca than in E. petraea in the functional megaspore and two-nucleate stage, but the volumes are similar at the four-nucleate stage. Consideration of the differences in structural complexity between the sporophyte and gametophyte generations leads to the conclusion that the female gametophytes of these species are more distinctive than are the sporophytes.     

Megaspore germination, embryo development and maintenance of the symbiotic association in Azolla filiculoides Lam.

Megaspore germination and embryo development in Azolla filiculoides was examined using SEM and thin–sectioning. Within the released megaspore apparatus, resting cells of the endosymbiont Anabaena azollae Stras. arc located distally to the outside of the mcgasporangial wall and adhering to the inside of the megasporocarp wall. Growth of the female gametophyte displaces the floats pushing this part of the wall (the indusial cap) upwards, so providing access to the archegonia for the multifiagellalc spermalozoids. Embryo development and its inoculation with Anabaena involves a subtly–timed sequence of events resulting in the perpetuation of the symbiosis. Growth of the lunnel–shaped cotyledon leaf ruptures the mcgasporangial wall to provide access and a channelled route between the Anabaena and embryo shoot apex; subsequent leaf development severely restricts such access. During this process, the Anabaena is dislodged by the cotyledon leaf and growth of the first leaf traps the now actively–dividing Anabaena colonv; this becomes established around subapical trichomes from where filaments become incorporated into the cavities of developing leaves. The voung sporophyte rises vertically to the water surface as a result of gas accumulation in intercellular spaces; at no stage do floats endow buoyancy.     

DEVELOPMENT OF THE OVULE AND MEGAGAMETOPHYTE IN SAXIFRAGA HIERACIFOLIA

Wiggins , Ira L. (Stanford U., Stanford, Calif.) Development of the ovule and megagametophyte in Saxifraga hieracifolia. Amer. Jour. Bot. 46(10): 692–697. Illus. 1059.—Buds of Saxifraga hieracifolia collected in the vicinity of Point Barrow, Alaska, fixed, sectioned, and stained by standard methods, revealed that the archesporial cell in the ovule of this species is hypodermal and gives rise to the megaspore mother cell and a small number of parietal cells. Occasionally 2 megaspore mother cells occur within an ovule. Meiosis in the megaspore mother cell produces a linear tetrad of megaspores, the chalazal one of which normally gives rise to a monosporic, Polygonum-type megagametophyte. The polar nuclei fuse near the chalazal end of the megagametophyte and the antipodal cells disintegrate prior to fertilization. A distinct filiform apparatus and a marked lateral “spur” develop on each synergid. Vacuolation in the egg cell and in the synergids follows the usual pattern. Only a single integument surrounds the nucellus.     

THE FEMALE GAMETOPHYTE OF MACHAERANTHERA PATTERSONII (ASTER PATTERSONII) AND OF M. TANACETIFOLIA

The nucellus of Machaeranthera pattersonii (A. Gray) Greene (Aster pattersonii A. Gray) contains only one megaspore mother cell, and the female gametophyte develops from the chalazal megaspore of a row of four, thus conforming to the Polygonum type of development. These observations are contrary to the older work of Palm. Three nuclear divisions produce the typical eight nuclei with the egg apparatus, primary endosperm cell with two polar nuclei, and two antipodal cells, the micropylar one containing two nuclei. Usually no more antipodal cells are formed, although there is further nuclear division, apparently followed by nuclear fusion. The antipodal cells remain about the same size without forming an antipodal haustorium. Cell division accompanies the first division of the primary endosperm nucleus. The early stages of the embryo resemble those of other Compositae. Machaeranthera tanacetifolia (HBK) Nees also shows the Polygonum type of development of the female gametophyte. It is suggested that Palm may have been working on some species of Erigeron that had been wrongly identified, which would account for the difference in observations.     

Two new species ofIsoetes (Isoetaceae) from Jeju Island, South Korea

We name and describe two new species,Isoetes jejuensis and I.hallasanensis, from Jeju Island, South Korea. These taxa are compared with related species in East Asia, including I.coreana andI. sinensis. The rugulate megaspore ornamentation ofI. jejuensis differs from those otherIsoetes species. Furthermore,I. hallasanensis varies fromI. asiatica in its microspore ornamentation and chromosome number, although both species have an echinate megaspore ornamentation. These new species are tetraploid (2n = 44), whereasI. coreana is hexaploid (2n = 66).     

Ultrastructural interpretation of the Late Cretaceous megaspore Glomerisporites pupus and its associated microspores

The ultrastructure of the Late Cretaceous (Santonian–?early Campanian) megaspore Glomerisporites pupus and its associated microspores has been examined in an attempt to resolve a number of problems concerning the interpretation of their morphology. The new observations presented are based on an analysis of entire, fragmentary, and thin-sectioned specimens under scanning and transmission electron microscopes. These add to, and partly correct, previous observations on this taxon. They include the following: (1) The exine of the megaspore consists of thin, homogeneous, outer undulating and inner electron dense layers, with a thicker zone of spongy structure in-between. (2) The perispore (or perine) of the megaspore comprises four layers, in order towards the exterior: loose filamentous, dense filamentous, vacuolate, and columnar. (3) This is completely enclosed by a thick mat of hairs, which appears to be attached to the underlying perisporal layers by means of connections with a few of the “spines” that originate from the dense filamentous zone, and with some elements of the columnar perine. (4) The tripartite neck (acrolamella) of the spore, which is hidden beneath the mat of hairs, is predominantly an extension of the dense filamentous and vacuolate layers, but also involves the columnar layer, especially in the lower part. (5) Some of the numerous small floats that are embedded in the mat have hairs originating from them. (6) Both long tangled and circinate hairs surround the perispore of the microspores. (7) The exine of the microspore was at least partly attached to the perispore when the organ was viable. (8) It comprises four zones that vary in structure and electron density. These facts and comparisons made with other megaspores and their associated microspores confirm evolutionary links between G. pupus and several taxa included within the Salviniaceae (Azolla, Parazolla, Salvinia) and possible ancestors of this group (the parent plants of Ariadnaesporites and Capulisporites).     

Ultrastructural features of megasporogenesis inTorreya nucifera (Taxaceae)

In megasporogenesis ofTorreya nucifera (Taxaceae) more than one product of meiosis can start to germinate, a process previously observed inTaxus. Ultrastructural analysis ofT. nucifera revealed that this behaviour is cytologically determined by the presence of a transfer cell type wall-membrane apparatus and by the presence of uncommonly complex plasmalemmasomes in the megasporocyte. These features may determine a uniform supply of nutrients to the entire megasporocyte cytoplasm, so that after meiosis the germination potential is not restricted to the chalazal megaspore. Even if more than one product of meiosis can originate the female gametophyte in bothTaxus andTorreya, this process involves different ultrastructural aspects in the two species. In conclusion, the extension of the germination potential is a common characteristic to both species but it is most probably the result of some evolutionary convergence.     

宁夏枸杞大孢子发生和雌配子体发育的细胞学研究

采用半薄切片技术和组织化学染色法对宁夏枸杞大孢子发生和雌配子体发育过程中的细胞结构变化及营养物质积累特征进行了观察。结果表明,(1)宁夏枸杞为中轴胎座,多室子房,倒生胚珠,单珠被,薄珠心类型。(2)位于珠心表皮下的孢原细胞可直接发育为大孢子母细胞,减数分裂后形成直线型大孢子四分体,合点端第一个大孢子发育为功能大孢子,胚囊发育类型为蓼型,具有珠被绒毡层。(3)初形成的胚囊外周组织中没有营养物质积累,成熟胚囊时期出现了大量的淀粉粒且呈珠孔端明显多于合点端的极性分布特征。(4)助细胞的珠孔端具有明显的丝状器结构,呈PAS正反应表现出多糖性质,成熟胚囊具有承珠盘结构。     

八角莲大孢子发生和雌配子体形成

首次报道了八角莲(Dysosma versipellis (Hance)M.cheng)大孢子发生和雌配子体形成的过程.结果:双珠被,多为厚珠心胚珠,少数为假厚珠心,胚珠多为横生,少数为弯生;边缘胎座,子房一室,多胚珠,珠孔由两层珠被共同形成,呈"之"字形;多为单孢原,位于珠心表皮下:偶见2～3个孢原细胞位于珠心表皮下;大孢子母细胞有两种发生方式;直线形大孢子四分体,合点端的大孢子发育为功能大孢子,蓼型胚囊;成熟胚囊中,二个极核在受精前合并为次生核;三个反足细胞不发达,较早退化;"品"字形卵器极性明显,其中卵细胞与助细胞极性相反;助细胞发达,其丝状器在不同发育时期形态及大小不同,且具吸器功能.     

Suavitas imbricata gen. et sp. nov., an anatomically preserved seed analogue of putative lycophyte affinities from upper Pennsylvanian marine deposits

A megasporangiate fructification has been discovered in dysoxic, offshore marine shales from Upper Pennsylvanian deposits of northcentral Texas. This specimen, described as Suavitas imbricata gen. et sp. nov., consists of imbricating, helically arranged sporophylls that diverge from a stout axis. The axis is characterized by parenchymatous pith and weakly developed vascular tissue. Sporangia are terminal with a horizontally elongated, apical dehiscence slit. There is one large functional megaspore in each sporangium, and the megaspore wall is constructed of interconnected rodlets. Parsimony analysis indicates that Suavitas may be related to rhizomorphic lycophytes. The unique combination of characters displayed by this species illustrates the central role played by extinct species in characterizing overall patterns of plant relationships, and emphasizes the uniqueness of plant communities from extrabasinal habitats during the late Paleozoic.     

DIVERSITY AND HOST SPECIFICITY OF AZOLLA CYANOBIONTS1

A unique, hereditary symbiosis exists between the water fern Azolla and cyanobacteria that reside within a cavity in the dorsal leaf‐lobe of the plant. This association has been studied extensively, and questions have frequently been raised regarding the number and diversity of cyanobionts (cyanobacterial symbionts) among the different Azolla strains and species. In this work, denaturating gradient gel electrophoresis (DGGE) and a clone library based on the 16S rRNA gene were used to study the genetic diversity and host specificity of the cyanobionts in 35 Azolla strains covering a wide taxonomic and geographic range. DNA was extracted directly from the cyanobacterial packets, isolated after enzymatic digestion of the Azolla leaves. Our results indicated the existence of different cyanobiont strains among Azolla species, and diversity within a single Azolla species, independent of the geographic origin of the host. Furthermore, the cyanobiont exhibited host‐species specificity and showed most divergence between the two sections of genus Azolla, Azolla and Rhizosperma. These findings are in agreement with the recent redefinition of the taxon Azolla cristata within the section Azolla. With regard to the taxonomic status of the cyanobiont, the genus Anabaena of the Nostocaceae family was identified as the closest relative by this work.     

Taxonomic re-evaluation of the Azolla genus in Portugal

ABSTRACT

The Portuguese and the European Flora refer to the presence of two or three Azolla species in Portugal: A. filiculoides Lam., A. caroliniana Willd. and/or Azolla mexicana Presl., the latter included in the last edition of Flora Europaea. In the present work, the taxonomy of Azolla species is reviewed using the two most important characters that can distinguish between these two/three species: papillae in the dorsal leaf lobe and perine architecture of the megaspore apparatus. Other characteristics, such as the hyaline border cells of the dorsal leaf lobes and the number of glochidia septa in microsporangium massulae, are also used. All the Azolla specimens, collected from several locations in Portugal, were identified as Azolla filiculoides Lam. This identification disagrees with previous published reports on Azolla taxonomy in Portugal as well as with herbarium identification.     

Use of artificial substrata by introduced and cryptogenic marine species in Paranaguá Bay,southern Brazil

Abstract

Ports are important locations for the introduction of marine species, while marinas and pontoons often serve as secondary habitats for these species. In a marina near Paranaguá Port, a major international port in southern Brazil, the encrusting community was studied to (i) identify possibly introduced species, and (ii) examine the use of artificial substrata by these species. Samples (20 × 20 cm) were taken from fibreglass floats (boardwalks and boat hulls) and concrete columns. A total of 85 species were found of which 50 were classified into three categories: four introduced, 33 cryptogenic and 13 native. The introduced species were the hydrozoan Garveia franciscana (on floats, boats and submerged concrete), the polychaete Polydora cornuta (more abundant on floats and submerged concrete) and the barnacles Amphibalanus reticulatus (equally abundant on the three substrata) and Striatobalanus amaryllis (only on floats and submerged concrete). Organisms were most abundant on floating boardwalks, while species richness and composition were similar to that on boat hulls (32 and 37, respectively), which are an important vector for intraregional transport. All substrata supported at least three of the four introduced, and many of the cryptogenic species. The proportion of introduced to the total number of species was greater than observed in other ports. This demonstrates that the introduction potential is great in Paranaguá Bay, especially considering that this study was restricted to one site and sampled only hard substrata. None of the introduced species has yet been identified as invasive, but all are generalists with respect to substratum, indicating their invasive potential. The ability to colonise stable concrete walls shows that they could also colonise the natural granite rocky substrata in the bay, and the ability to colonise floating surfaces indicate their capability of spreading in the region on the hulls of recreational boats.     

Embryological studies on Indian Taraxacum Weber. Embryo sac,endosperm and embryo

Abstract

The present paper deals with the embryological studies of ten species of Taraxacum, out of which nine polyploid species are apomictic and one diploid species T. wallichii shows normal sexual behaviour. Occasional formation of normal megaspore tetrads and triads are observed in tetraploid and triploid species.

Autonomous development of endosperm from the secondary nucleus seems to be the rule. The growth of endosperm is relatively slow.