SPORE MORPHOLOGY IN THE CYATHEACEAE. II. THE GENERA LOPHOSORIA,METAXYA, SPHAEROPTERIS,ALSOPHILA, AND NEPHELEA

Scanning electron microscopy and transmitted light microscopy are used in a palynological study of Lophosoria, Metaxya, Sphaeropteris, Alsophila, and Nephelea of the tree fern family Cyatheaceae. The monotypic American genera Lophosoria and Metaxya each have a unique spore morphology which reinforces the taxonomic distinctness of these genera as indicated by their other characters. All investigated paleotropical species of Sphaeropteris develop a single type of perine characterized by coarse, pointed projections. In the neotropics, the Sphaeropteris horrida group shares this perine type, whereas all other neotropical Sphaeropteris species appear to have a different kind of perine with fine hair-like processes. The exine in paleotropical Sphaeropteris appears uniformly unsculptured, whereas in the neotropics several exine morphologies are found. In Alsophila all investigated neotropical species and the vast majority of the paleotropical species are characterized by a basically ridged perine morphology and an unsculptured exine. In several paleotropical Alsophila species, however, a perine with hair-like processes similar to those in neotropical Sphaeropteris is found, and the exine in several species is variously pitted. In at least one paleotropical Alsophila species, the porate exine morphology is indistinguishable from that in the neotropical genus Cnemidaria. The spores of the American genus Nephelea are similar to those of the majority of Alsophila species in ridged perine morphology and unsculptured exine. Several new instances of atypical spore numbers per sporangium are reported in Sphaeropteris and Alsophila. These and the palynological data are discussed in a taxonomic framework. The spore morphology in these genera is consistent with Tryon's recent generic revision of the family.     

SPORE MORPHOLOGY IN THE CYATHEACEAE. I. THE PERINE AND SPORANGIAL CAPACITY: GENERAL CONSIDERATIONS

The literature on cyatheaceous spore morphology relative to the presence of a perine layer is reviewed, and evidence based on a sodium-hydroxide assay is presented indicating that the outer scultpine layer in certain cyatheaceous spores is perine. Perine so defined characterizes Metaxya, paleotropical and certain neotropical species of Sphaeropteris, nearly all species of Alsophila, all species of Nephelea, and certain species of Trichipteris and Cyathea. It is lacking in Lophosoria, many species of Trichipteris and Cyathea, and all species of Cnemidaria. Two major patterns of spore number per sporangium in the family are reported. Lophosoria, Sphaeropteris, Trichipteris, Cyathea, Cnemidaria, and probably Metaxya are characterized by 64-spored sporangia, whereas most species of Alsophila and all species of Nephelea are characterized by 16-spored sporangia. The congruence of this generic distribution of sporangial-capacity types with Tryon's phyletic arrangement of cyatheaceous genera supports the naturalness of his system. The intrasporangial germination of spores retained in dehisced and dispersed sporangia supports the suggestion that decreased spore number per sporangium in Alsophila and Nephelea may relate to the role of the sporangia as dispersal units. The decreased number of spores per sporangium is associated with a trend toward increase in the number of sporangia per sores, with the highest known count approaching 1000 sporangia per sorus. The Alsophila-Nephelea evolutionary line has probably not been ancestral in the phylogeny of the more advanced groups of ferns.     

SPORE MORPHOLOGY OF ANEMIA,MOHRIA, AND CERATOPTERIS (FILICALES)

Spores of the ferns Anemia and Mohria (Schizaeaceae) and Ceratopteris (Pteridaceae) are surveyed with light microscopy and scanning and transmission electron microscopy. In each genus the spores are trilete with radially symmetrical exine sculpture comprising three sets of parallel or near-parallel muri. Anemia has six spore types. One is reticulate (A. wrightii-type), and the other five types have either cicatricose or canaliculate sculpture that reflect a basic form, i.e., three mural sets that have mutual anastomoses in each radial region. The cicatricose A. mexicana- and the canaliculate A. dregeana-types represent the simplest expressions of this pattern. Specializations include cicatricose, auriculate (A. raddiana-type) and canaliculate, ornate (A. oblongifolia-, A. phyllitidis-types). Exine structure is homogeneous or differentially microporate; the enveloping two-layered perine has granulate structure and a granulate to spiculate and/or pitted surface. Mohria spores have a stratified, granulate to rugulose perine and cicatricose exine sculpturing consistent with the A. mexicana-type pattern, but the muri are hollow. The canaliculate spores of Ceratopteris differ from those of Anemia and Mohria in that the three mural sets are discrete and separated from each other by a stria in each radial region; exine structure is homogeneous and the thin perine is granulate. Within Anemia the A. raddiana-type is exclusive to subgen. Coptophyllum and the A. oblongifolia- and A. phyllitidis-types to subgen. Anemia. Three spore types are shared by two subgenera; i.e., A. wrightii- and A. mexicana-types in Coptophyllum and Anemirhiza, and the A. dregeana-type in Anemirhiza and Anemia. Spore polymorphism is indicated in several species of subgen. Anemia, and smooth immature spores are recorded from all three subgenera.     

SPORE MORPHOLOGY IN THE DICKSONIACEAE. I. THE GENERA CYSTODIUM,THYRSOPTERIS, AND CULCITA

The most mature spores available in herbarium specimens of the dicksoniaceous genera Cystodium, Thyrsopteris, and Culcita were studied by scanning electron microscopy and light microscopy, and representative specimens were analyzed to determine the number of spores produced per sporangium. Thyrsopteris and Culcita feature 64-spored sporangia, but Cystodium is consistently 32-spored. Spores were analyzed both in their native state as found on the specimens and in a perine-free state achieved by treatment with sodium hydroxide or acetolysis mixture. The sodium hydroxide assay demonstrated the presence of a perine in Cystodium, Thyrsopteris, and Culcita subgenus Culcita, but no evidence of a perine so defined was found in Culcita subgenus Calochlaena. Spores of Cystodium feature a nearly psilate exine overlain by a striate inner perine and a granular outer perine and are in several respects similar to those of Metaxya in the cyatheoid-dicksonioid complex and to those of Saccoloma in the dennstaedtioids. The most mature Thyrsopteris spores available may not have been fully mature. They feature a sparsely distributed, granular perinous layer over a microverrucate sculptine. The latter topography is taken as largely perinous since treatment with sodium hydroxide left a nearly psilate exine. The spore morphologies of Cystodium and Thyrsopteris reinforce the taxonomic distinctness of these monotypic genera indicated by their other characters. The two subgenera of Culcita are very dissimilar in their spore morphologies. The exine in subgenus Culcita ranges from psilate to slightly microverrucate proximally and distally, with varying margo development. Spores of subgenus Calochlaena are strongly differentiated from those of subgenus Culcita by their exine of broad spinules which vary in their degree of lateral fusion to each other and in the granular appearance of their distal surfaces. Spore morphology in Culcita strongly supports the argument of those who would raise its subgenera to generic rank.     

Ultrastructure of the spore in four Japanese species of Ptychomitrium Fürnr. (Musci)

The ultrastructure of the mature spore in four Japanese species of the acrocarpous moss genus Ptychomitrium is presented. In all species the spores have a similar pattern: there is no recognisable aperture nor sporoderm polarity, exine and perine are poorly developed, cytoplasm only occasionally shows polarity, and plastids have a well developed inner membrane system. The presence of frequent intine protrusions is a remarkable feature of this genus. A multilaminar structure of the exine, already observed in Grimmia, occurs also in these species although here it is less pronounced. The significance of these features is discussed within these species, as well as in comparison to other taxa, especially the genus Grimmia.     

SPORE WALL FORMATION AND CHLOROPLAST DEVELOPMENT DURING SPOROGENESIS IN THE MOSS FISSIDENS LIMBATUS

The sequence of wall formation in spores of Fissidens limbatus Sullivant is as follows: The exine is formed around the protoplasts after the sporocyte has undergone meiosis. The fully enlarged spores then become coated by the perine; this is followed by intine formation. The source of the intine and exine appears to be from within the spore, but the perine is of an apparent exogenous origin. Ornamentation of the spore is due solely to deposition of the perine. Each spore originally has a single plastid. Plastids increase in number by fission, resulting in mature spores with numerous plastids with well differentiated lamellae.     

Palynology of selected species of Fissidens (Hedw.)

Spores from 19 species of the subgenus Aloma Kindb. of the moss genus Fissidens were analyzed by light and scanning electron microscopy. Aloma is the largest subgenus of Fissidentaceae and is characterized by the presence of a peristome of scariosus type. The spores of the subgenus Aloma are present in monads, size small to very small, heteropolar, plano-convex, with a proximal aperture region, and the sporoderm is formed by a perine, exine, and intine. The intine is not stratified, the exine is psilate, and the perine granulated. The ornamentation elements may occur singly or grouped on the surface of the spore. The aperture region shows irregular contours, ranging from circular to elongated, due to the weakness of the sporoderm proximal pole. The observed variations among species are related to different patterns of distribution of the sporoderm granules and nanogranules. Quantitative analysis combined with qualitative results did not allow all species of the subgenus Aloma to be distinguished. The results of this study demonstrate that the spore is a useful tool for taxonomic studies, and suggest that its characters be included in phylogenetic analyses, to assist in the reconstruction of the evolutionary history of mosses.     

Pollen morphology of the subtribe Cuspariinae (Rutaceae)

The neotropical subtribe Cuspariinae (Rutaceae) comprises as many as 26 genera and over 125 species. Pollen grains from 111 collections representing 71 species and 24 genera were examined by LM, SEM, and TEM. The pollen morphology of this subtribe is very diverse. Grains are mostly 3–6-aperturate and colporate, rarely porate (Spiranthera) or pantocolporate (Almeidea). Exine sculpturing is most commonly reticulate, sometimes perforate, foveolate-perforate, foveolate, foveolate-reticulate, reticulate, striate-reticulate, echinate, clavate, or baculate. The exine structure is columellate and tectate-perforate, columellate and semitectate, or intectate and is stratified into ektexine and endexine. The exine ofLeptothyrsa is distinctive in that the ektexine of the mesocolpium is longitudinally deeply ridged. The pollen ofHortia, characterized by a psilate exine with rare perforations, a very thick foot-layer, and reduced columellae, is unlike that of any member of the Cuspariinae and offers no support for the transfer of this genus from the Toddalioideae. The pollen data correlate with macromorphological characters and are taxonomically useful.     

Pollen morphology of the genus Gagea (Liliaceae) in Iran

Pollen grains of 26 species of the genus Gagea distributed in Iran were examined by light and scanning electron microscopy. Detailed pollen morphological characteristics are given for these species. Among the studied species, the newly described G. iranica together with G. olgae and G. graminifolia possess the smallest pollen grains, and the widely distributed G. lutea the largest ones. Our studies show that the sculpturing of exine provides valuable characters for separating the species, sometimes even for closely related ones, and delimitation of natural groups within the genus. The exine of the genus Gagea is in most cases perforated upon tectum or rarely tectate-columellate. The muri are solid or structured, compound, simpli-, dupli- or pluricolumellate. It seems that the structure of muri is very important in recognizing natural groups within the genus. Tectal perforations vary from <0.2 to 2.0 μm in diameter among the studied species. Regarding sculpturing of the exine in proximal face, four basic types of pollen grains can be distinguished: reticulate, microreticulate, foveolate and perforate. Within the reticulate type there is sufficient variation in exine structure at distal face to describe three subtypes: reticulate, microreticulate and perforate. A diagnostic key is given for all studied taxa based on palynomorphological characters. For a limited number of populations of selected Iranian species of Gagea, further aspects of pollen biology were studied. It seems that populations with ploidy levels other than diploidy, show a low percentage of pollen fertility. Moreover, the rate of pollen fertility is correlated with the manner of the reproduction in certain species.     

Pollenmorphologie Von Borago Officinalis L.

The pollen morphology of the 16 recognized species of the genus Asphodelus L. (Asphodelaceae) has been investigated by light and scanning electron microscopy. Four pollen types can be distinguished on the basis of variation in size of the polar, equatorial longitudinal and transverse axes, as well as in exine structure and sculpturing, which correlate with the subdivision of the genus into sections. Correlations between pollen size and some biological characters are made. Phylogenetic implications are suggested, and trends of pollen evolution in the genus are indicated.     

FINE STRUCTURE OF MISTLETOE POLLEN. III. LARGE-FLOWERED NEOTROPICAL LORANTHACEAE AND THEIR AUSTRALIAN RELATIVES

Pollen of all large-flowered neotropical loranthaceous genera and related Australian taxa (Nuytsia, Atkinsonia) were examined in the light, scanning, and transmission electron microscopes. Trilobate, deeply concave, peroblate or oblate pollen shapes are basic within the complex; triangular, slight convex, suboblate shapes are more derived features. The non-fixiform pollen of Atkinsonia is unique within the family. Most of the large-flowered genera possess exclusively syncolpate aperture types. However, among Psittacanthus species tricolpate, diplosynrugate and diplorugate apertures are also present, representing derived types for the complex. The inaperturate pollen of Atkinsonia appears to have developed independently within the family. Pollen sculpturing is typically non-uniform, i.e., there are pronounced sculpturing differences in polar and equatorial regions. Uniformly sculptured pollen is restricted to Atkinsonia, Ligaria and some species of Psittacanthus. Ultrastructurally, most exine modifications have occurred in the equatoral ektexine. Here the basic organization ranges from essentially columellaless to columellate, the latter the more derived condition. Endexine is typically thick and stratified in polar areas, thin and lamellate in peripheral and apertural regions. Similar pollen morphologies of Nuytsia and Gaiadendron support the idea of a transoceanic evolutionary connection between the Old and New Worlds. Pollen characters show Gaiadendron to be the most primitive and Psittacanthus the most advanced among the large-flowered neotropical genera.     

Pollen morphology inLinum sect.Macrantholinum

Pollen of the two distylous species which make upLinum sect.Macrantholinum differs from that of other distylous species in the genus in being multiporate and in having much more modest differences in exine sculpturing between grains from long- and short-styled plants. Pollen morphology does not help in relating the two species to others in the genus but does support their retention in a separate section.     

PpASCL,the Physcomitrella patens Anther-Specific Chalcone Synthase-Like Enzyme Implicated in Sporopollenin Biosynthesis,Is Needed for Integrity of the Moss Spore Wall and Spore Viability

Sporopollenin is the main constituent of the exine layer of spore and pollen walls. The anther-specific chalcone synthase-like (ASCL) enzyme of Physcomitrella patens, PpASCL, has previously been implicated in the biosynthesis of sporopollenin, the main constituent of exine and perine, the two outermost layers of the moss spore cell wall. We made targeted knockouts of the corresponding gene, PpASCL, and phenotypically characterized ascl sporophytes and spores at different developmental stages. Ascl plants developed normally until late in sporophytic development, when the spores produced were structurally aberrant and inviable. The development of the ascl spore cell wall appeared to be arrested early in microspore development, resulting in small, collapsed spores with altered surface morphology. The typical stratification of the spore cell wall was absent with only an abnormal perine recognisable above an amorphous layer possibly representing remnants of compromised intine and/or exine. Equivalent resistance of the spore walls of ascl mutants and the control strain to acetolysis suggests the presence of chemically inert, defective sporopollenin in the mutants. Anatomical abnormalities of late-stage ascl sporophytes include a persistent large columella and an air space incompletely filled with spores. Our results indicate that the evolutionarily conserved PpASCL gene is needed for proper construction of the spore wall and for normal maturation and viability of moss spores.     

Spore morphology of Serpocaulon A.R.Sm. and related taxa from Brazil (Polypodiaceae)

The epiphytic fern genus Serpocaulon (Polypodiaceae) comprises 42 species nested within the neotropical clade of the family together with genera such as Campyloneurum, Grammitis, Microgramma, Niphidium, Pleopeltis, Pecluma, Polypodium and Terpsichore. Although Serpocaulon is a well-supported genus, its intra-specific relationships are still not completely resolved. The morphological dataset for Serpocaulon and related genera is therefore expanded here with information on spore morphology to provide additional information for phylogenetic consideration. Spores have been collected from herbarium vouchers and acetolysed for light and scanning electron microscopy. All members of the neotropical clade have monolete spores, except for Grammitis and Terpsichore, which have trilete spores. Both verrucate and papillate ornamentation types are recognised. The palynological analysis shows close similarity between spores of Serpocaulon, Campyloneurum, Pleopeltis and Polypodium and supports close phylogenetic relationships between these taxa. Species of the grammitid clade, although nested within the Polypodiaceae, have spore morphology that supports a separate position.     

SPORE WALL ULTRASTRUCTURE IN FOUR SPECIES OF THE LIVERWORT RICCIA

Mature spores of Riccia californica, R. campbelliana, R. sorocarpa, and R. trichocarpa (Marchantiales, Ricciaceae) were examined with both scanning and transmission electron microscopes. The thick-walled spores have species-specific surface sculpturing and similar internal structure. Major surface features include ridges and depressions in areolate or irregular configurations. Finer surface features such as tubercles or papillae may ornament the ridges or depressions. Internally, the sporoderm consists of a thin intine and a three-layered, lamellate exine. The innermost exine lamellae are thin and closely spaced, giving this layer a fibrillar appearance. Lamellae of the middle region are thick in section and have a relatively thick, electron-dense coating. The outermost lamellae are of intermediate thickness and also have a relatively thick electron-dense coating. Lamellae of all regions frequently split, branch and anastomose. The outermost lamella forms the surface features of the spore. Interior lamellae follow the contour of the outermost lamella. Pores usually occur in these four species at the junctures of the triradiate ridge arms and equatorial rim. Pores do not penetrate the entire exine, and they represent localized areas where the outermost group of lamellae have not been formed.     

Pollen Morphology and Taxonomic Position of the Genus Pentaphragma Wall. (Pentaphragmataceae)

The pollen morphology of some Pentaphragma Wall, species has been studied by light- and electron microscopy. To improve the rehydration of the dried material used, compound fixatives have been applied. Intine-held vesicles and fibrillar network in gaps and spaces of the exine stain positively for protein. The pollen morphology of the genus differs strongly from that of Campanulaceae, and shows distinctive characters of its own. Hence from a palynological point of view it appears correct to place the genus in a separate family.     

Pollen morphology of Astragalus section Hymenostegis (Fabaceae) and evaluation of its systematic implications

Astragalus is with nearly 3000 described species the largest genus of flowering plants. So far analyses of pollen characters have only been conducted for a few species of the groups within the genus. Here we analyse pollen grains of 22 species representative for Astragalus section Hymenostegis using scanning electron microscopy. We found the basic shape of the pollen grains to be oblate-spheroidal and apertures to be tricolpate as for other eudicots. The sculpturing pattern of the exine is micro-reticulate. Pollen grains show low morphological variation among different species of this section, but differences occur between sections of the genus. We conclude that the vast morphological differentiation that occurred during the rapid radiation of section Hymenostegis was not accompanied by comparable differentiation in pollen morphology.     

Pollen diversity in the genus Ptilostemon (Asteraceae,Cardueae) from Italy and its taxonomic and palynoecological implications

The results of a study on pollen morphology of the species of the genus Ptilostemon growing in Italy: Ptilostemon niveus, P. greuteri, P. gnaphaloides, P. casabonae, P. strictus, P. stellatus by light and scanning electron microscopy were presented and discussed including in relation to their taxonomic position. The exine shows two different ornamentation patterns: echinae and scabrae. This last ornamentation pattern together with other features was previously described in the genus Ptilostemon. Of 6 species analysed, P. stellatus is the only one that shows a scabrate ornamentation of exine. Two pollen types were recognized through the exine ornamentation, length of ectocolpus and polar outline: P. stellatus type and P. niveus type. A multivariate analysis (principal component analysis and linear discriminant analysis) was carried out with the aim of examining potential morphological characters which could be used to identify taxa. The data suggest that several characteristics can be used to delimit the species in particular the exine ornamentation. The results of our studies support the actually subgeneric classification, and several features of the pollen grains analysed seem to have a palynoecological role and an important taxonomic significance. A dichotomous key based on palynological data is also given.