Pollen morphology inParis and its related genera

Pollen grains ofParis, Medeola, andScoliopus have been examined with light, scanning electron, and transmission electron microscopies.P. quadrifolia L.,P. verticillata Bieb.,P. delavayi Franch. andP. polyphylla Smith have monosulcate pollen characterized by psilate, foveolate or reticulate exine sculpture. In contrast to the former species,P. japonica (Fr. et Sav.) Franch. andP. tetraphylla A. Gray have monosulcate pollen with gemmate (rarely rugulate) exine.Medeola has monosulcate pollen with reticulate exine that is distinct from that ofParis. Scoliopus has monosulcate pollen characterized by a peculiar reticulate exine pattern. The palynological evidence suggests thatParis andTrillium are closely related to each other, andMedeola andScoliopus should be separated fromParis andTrillium.     

A new pollen type,C-banded and fluorochrome counterstained chromosomes,and evolution inGuatteria and related genera (Annonaceae)

Guatteria, Guatteriopsis, Guatteriella andHeteropetalum share the same conspicuous pollen type which is new for theSpermatophyta. It is zonoaperturate with a folded aperture region and an extremely reduced exine. First chromosome counts and karyotype analyses forGuatteriopsis (4 species investigated) andGuatteriella (1 species) are identical with those ofGuatteria (19 species seen): 2n = 28. The genome is characterized by diploidization and partly telocentric chromosomes. Sequentially Giemsa C- and fluorochrome banded chromosomes and interphase nuclei are described. The cuticular folding pattern is distinct forHeteropetalum only. Growth forms and ecology are reported for many species. The evolutionary pattern of theGuatteria group is discussed and compared with other genera and families.     

The systematic relationships ofAegialitis (Plumbaginaceae) as revealed by pollen morphology

Light and first SEM observations on the pollen ofAegialitis indicate that the two species of the genus are clearly distinguishable from all other genera of the tribeStaticeae to whichAegialitis presently is relegated. Intraspecific exine or aperture dimorphism which appears frequently in this tribe, is not recorded in this genus.A. annulata andA. rotundifolia have distinct monomorphic pollen and show a great resemblance in exine features with the tribePlumbagineae, particularly species ofPlumbago. Considering these and other features, separate subtribal status forAegialitis within thePlumbagineae is proposed.     

Dimorphic exine sculpturing in three distylous species ofLinum (Linaceae)

DistylousLinum grandiflorum, L. mucronatum, andL. pubescens have dimorphic pollen grains. In short-styled flowers the exine has monomorphic processes each with a ring of papillae. In long-styled flowers the exine has two types of processes: small processes terminating in a spinule and larger processes with a central spinule surrounded by a ring of spinuels, papillae or buttresses. The distylousLinum species of four different sections, in which morph-specific differences in the wall structure of stigmatic papillae have been reported, also exhibit exine dimorphism. The patterns of these dimorphisms are similar in all four sections in which distyly is present.     

Correlation between pollen morphology and pollination mechanisms in the Hydrocharitaceae

The pollen morphology of 11 genera and 11 species of the Hydrocharitaceae and one species of the Najadaceae was studied using scanning and transmission electron microscopies, and the exine structures and sculptures are discussed in relation to pollination mechanisms and the molecular phylogeny. The pollen grains of the Hydrocharitaceae are spherical, inaperturate, and form monads or tetrads, while those of the Najadaceae are elliptical, inaperturate, and form monads. The entomophilous genera Egeria, Blyxa, Ottelia, Stratiotes, and Hydrocharis share pollen grains that have projections like spines or bacula. The anemophilous genus Limnobium has reticulate pollen grains. The hypohydrophilous genera Thalassia and Najas are characterized by pollen grains with reduced exine structures. The pollen-epihydrophilous genera Elodea and Hydrilla have tightly arranged small spinous pollen grains, and the male flower-epihydrophilous genera Enhalus and Vallisneria have reduced reticulate or gemmate exines. Character state reconstruction of the exine structures and sculptures using a molecular phylogenetic tree suggests that variation in the exine is generally correlated with the pollination mechanism; the selective pressures acting on the pollination mechanisms have reduced the exine structure in hypohydrophilous plants and resulted in various exine sculptures that are adapted to the different pollination mechanisms in entomophilous, anemophilous, and pollen-epihydrophilous plants.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Sippen derOleaceae,Scrophulariaceae, Plantaginaceae undAsteraceae

Different members of the Angiosperm familiesOleaceae, Scrophulariaceae, Plantaginaceae andAsteraceae with variable modes of pollination were studied in relation to ultrastructure of pollenkitt and exine. In entomophilous representatives the mostly electron-dense and homogeneous pollenkitt is located on the outside layer of the exine, making the pollen sticky. In related anemophilous taxa the pollenkitt mostly is electrontransparent and inhomogeneous; it becomes inactive by remaining in the loculus or sinking to the bottom of the exine caves; this makes the pollen powdery. According to such criteria,Fraxinus ornus andRhinanthus serotinus are entomophilous, whileFraxinus excelsior is anemophilous. The pollen ofLathraea squamaria eventually becomes powdery and suited for wind dispersal. The broad transition field between entomo- and anemophily is demonstrated byPlantago andArtemisia. Plantago lanceolata is anemophilous, whilePl. serpentina andPl. media are amphiphilous.Artemisia nitida andA. mutellina are entomophilous,A. absinthium, A. pedemontana andA. gabriellae are amphiphilous, but tend towards entomo- (A. absinthium) or anemophily (A. gabriellae), whileA. chamaemelifolia and especiallyA. vulgaris are anemophilous.

     

Chloroplast DNA restriction site variation in theVernonieae (Asteraceae), an initial appraisal of the relationship of New and Old World taxa and the monophyly ofVernonia

Chloroplast DNA restriction site variation was examined for 35 taxa in theVernonieae and four outgroup tribes, using 17 restriction enzymes mapped for ca. 900 restriction sites per species; 139 mutations were found to be phylogenetically informative. Phylogenetic trees were constructed using Wagner and weighted parsimony, and evaluated by bootstrap and decay analyses. Relationships of Old and New World taxa indicate complex geographical relationships; there was no clear geographic separation by hemisphere. The relationships between Old and New World Vernonias found here support prior morphological analyses. The sister group to all New and most Old World taxa was composed of a small group of Old World species including yellow-flowered, trinervate-leaved species previously postulated to be basal in the tribe. The majority of both New and Old World taxa are derived from a lineage beginning with the monotypic genusStokesia, an endemic of the southeastern United States. The genusVernonia was also found to be paraphyletic within both the New and Old World. Available data do not support either the separation ofVernonia or the tribeVernonieae into geographically distinct lineages. The pattern of relationships within theVernonieae for taxa from North America, Asia, Africa, Central and South America is most similar to that of several other groups of both plants and animals with a boreotropical origin, rather than an origin in Gondwanaland. Such a pattern of distribution suggests more ancient vicariant events than are routinely postulated for theAsteraceae.     

Antibodies to pollen exines

A polyclonal antiserum and monoclonal antibodies have been prepared to purified pollen exines of Calocedrus decurrens Florin. The location of the antigen is in the exine, as shown by light-and electron-microscopic immunocytochemistry. The greatest reduction in antibody binding follows treatment of the exine with chemicals known to alter sporopollenin. These results provide evidence that sporopollenin is antigenic. Exines of ten species of gymnosperms and angiosperms also bound the polyclonal antiserum, indicating similarity of sporopollenin structure.     

Pollen evolution and systematics inAnnonaceae with special reference to the disulcate Australian endemic genera

All genera ofAnnonaceae endemic in Australia (Ancana, Fitzalania, Haplostichanthus) show almost exactly the same type of disulcate (disulculate) pollen with intact exine extending over the sulci. Tetrad stages inHaplostichanthus andAncana reveal a latudinal subequatiorial orientation of the two sulci at the proximal hemisphere. Sometimes they fuse into a ±zonosulcate aperture.Fissistigma pollen grains are ±globose and have a flattened pole with a central elevation and a concentric groove, covered by a somewhat reduced exine. This palynological characters give further support for separating the generaAncana andFissistigma. Germination was observed inHaplostichanthus where the pollen tube emerges at one of the two sulci and inFissistigma where the flattened part breaks up during germination. The aperture types described here are obviously transitional stages between aperturate and inaperturate pollen grains and are discussed in regard to pollen evolution.     

Zur Ultrastruktur und Funktion pollenverbindender Fäden beiEricaceae und anderen Angiospermenfamilien

Viscin threads and other pollen connecting threads of some angiosperm families were investigated, especially those ofEricaceae. According to the definition adopted, viscin threads are ± long exinous processes which consist of exinous material and connect pollen grains or tetrads. Such viscin threads are found within theOnagraceae, Caesalpiniaceae, Ericaceae, andMimosaceae only. While they differ in structure and composition, they always consist of sporopollenin and exhibit a very strong stickiness, even after all viscid substances have been removed by acetolysis. In contrast, the pollen connecting scleroprotein threads ofOrchidaceae and the cellular threads ofStrelitzia reginae Aiton. (Musaceae) are not connected with the exine surface, are destroyed by acetolysis, and thus do not correspond to viscin threads.

     

Therps16 intron and the phylogeny of theRubioideae (Rubiaceae)

The phylogeny of the subfamilyRubioideae (Rubiaceae) was estimated from sequence variation in therps16 intron (cpDNA) in 143 ingroup and 5 outgroup taxa. The analysis largely confirms a recent one based onrbcL sequences, but branch support is often much stronger. Three of the traditional subfamilies are supported,Rubioideae, Cinchonoideae s. str., andIxoroideae s. l. while there is no support forAntirheoideae. TheRubioideae are the sister group of all otherRubiaceae and comprise the tribesAnthospermeae, Coccocypseleae, Cruckshanksieae, Coussareeae, Gaertnereae, Hedyotideae, Knoxieae, Morindeae, Ophiorrhizeae, Paederieae, Pauridiantheae, Perameae, Psychotrieae, Rubieae, Spermacoceae, Theligoneae, andUrophylleae. TheHamelieae andHillieae belong to theCinchonoideae. Rachicallis andSiemensia should be transferred from theHedyotideae to theCinchonoideae. ThePauridiantheae, Urophylleae, Ophiorrhizeae, andRaritebe form the basalmost subclade of theRubioideae. The second basalmost clade consists of the generaLasianthus andPerama. The third basalmost clade consists of the tribesCoussareeae, Coccocypseleae andCruckshanksieae, and the generaDeclieuxia andHindsia. The tribesKnoxieae, Anthospermeae, Argostemmateae, Paederieae, Theligoneae, Rubieae, Hedyotideae, andSpermacoceae are members of one clade. TheKnoxieae are monophyletic ifOtiophora, Otomeria, andPentas are included. The tribeAnthospermeae is supported as monophyletic, but its subtribes are not. ThePaederieae, together withTheligonum, form a paraphyletic grade basal to theRubieae. TheHedyotideae, includingSchismatoclada, form a grade at the base of theSpermacoceae. TheGaertnereae are monophyletic and distinct from thePsychotrieae. TheMorindeae are monophyletic and includeDamnacanthus andMitchella. Schradera is the sister group of theMorindeae. ThePsychotrieae are monophyletic when theGaertnereae, Lasianthus, andDeclieuxia are excluded. The recognition of a subtribeHydnophytineae leaves the rest of thePsychotrieae paraphyletic.Psychotria is paraphyletic with respect to all other genera of the tribe. Approximately 50 genera are here classified for the first time based on molecular data.     

The tubular exine ofLauraceae andHernandiaceae,a novel type of exine structure in seed plants

The pollen grains ofLauraceae andHernandiaceae are characteristic in having a tubular exine of ± microfibrillar structure. This is coated with a layer of medium electron-dense globules and very peculiar spines, made up of a substance differing from that of the exine. Such a pollen wall structure is otherwise unknown in seed plants and thus adds to the list of odd features erratically present in various taxa of the Ranalean complex.     

Ultrastruktur und systematische Bedeutung des Pollens beiBocageopsis,Ephedranthus, Malmea undUnonopsis (Annonaceae)

The four genera investigated show solitary sulcate pollen grains. The structure and sculpture of the sporoderm is very similar inBocageopsis andUnonopsis, and supports the close relationship of both genera; their flower morphology also is very similar. In contrast, the sporoderm ofEphedranthus, and especially that ofMalmea, is different in some characters and suggests more remote relationships; this is also confirmed by differences in flower morphology. Within this group of genera a distinct exine progression can be recognized from non-columellate (Malmea) to granular (Unonopsis guatterioides), further to somewhat irregularly (Bocageopsis, someUnonopsis species), and finally to very regularly columellate (Ephedranthus). The sculpture of the tectum varies from a reticulum with large lumina (Malmea) to one with small performations (Ephedranthus). Within theAnnonaceae the genusMalmea is among the most primitive in respect to pollen structure. The sulcus of the four genera is very large and runs over 1/3 of the pollen grain. It is characterized by a reduction of the exine and a bulgy thickening of the intine.

     

Phylogenetic analysis ofUlmaceae

A phylogenetic analysis of theUlmaceae, Cannabaceae, Barbeyaceae, andBroussonetia of theMoraceae produced nine equally parsimonious trees with 127 steps. TheUlmoideae (Ulmaceae, sensuGrudzinskaya) are a monophyletic group and distinct from theCeltidoideae. The genusAmpelocera occupies an isolated taxonomic position among the celtidoids. The similarity ofAmpelocera to the fossil celtidoid flowerEoceltis of North America suggests thatAmpelocera posesses an archaic suite of characters, and occupies a primitive position among the celtidoids, theCannabaceae and theMoraceae. The relationships among the other celtidoid taxa,Cannabaceae, andBroussonetia are problematic. TheCannabaceae andBroussonetia of theMoraceae are nested within the celtidoids suggesting that this is a paraphyletic group. The close, but unresolved, relationship of the celtidoids to theMoraceae andCannabaceae observed in this analysis, and the appearance of the celtidoids in the fossil record prior to the ulmoids suggests that the evolutionary relationship of the ulmoids and celtidoids may be more distant than current taxonomic treatments reflect.     

Ultrastruktur und Verteilung des Pollenkitts in der insekten- und windblütigen GattungAcer (Aceraceae)

Several insect- and wind-pollinated species of the genusAcer have been investigated and compared in regard to pollen stickiness. The considerable amount of very inhomogeneous pollenkitt inA. negundo remains on the loculus wall or is deposited inside the exine cavities; thus the pollen is powdery. The small amount of non-homogeneous, granular pollenkitt inA. campestre mostly disappears into the exine cavities; only small droplets appear on the tectum surface; the pollen stickiness therefore is only moderate. On the other hand,A. pseudoplatanus andA. opalus produce an average amount of granular pollenkitt, which is deposited partially inside the exine and partially as a slender film on the tectum surface; in both the pollen is sticky.A. platanoides contains a great deal of granular and homogeneous pollenkitt; it does not only fill up the exine cavities but also extends as a thick, ± homogeneous, non-granular layer of pollenkitt over the tectum surface; therefore, the pollen is very sticky.—The characteristics of pollen agglutination together with other aspects of floral biology illustrate the wide spectrum between unequivocal entomophily and anemophily within the genusAcer: WhileA. negundo is anemophilous andA. platanoides is entomophilous, the remaining species investigated have a pollination syndrome with entomophilous and anemophilous features and are thus amphiphilous. The evolution withinAcer is tending not only towards dioecy, but also towards anemophily.

Herrn Prof. Dr.L. Geitler zum 80. Geburtstag gewidmet.     

Dimorphic exine sculpturing in two distylous species ofDyerophytum (Plumbaginaceae)

Light and SEM observations on the pollen ofDyerophytum africanum andD. indicum have revealed marked differences in exine features. These distylous species also have dimorphic pollen. In the short-styled individuals of both species, the sexine and nexine are of equal thickness, and the clava-like sexinous processes are short without marked projections. In the long-styled individuals, the sexine is thicker than the nexine, the clavae are higher than broad with an apical spinule. Pollen size and apertures are identical in both morphs. — Palynological evidence is presented for relationships betweenDyerophytum andCeratostigma, Plumbago andAegialitis. Moreover, the genusDyerophytum exhibits pollen morphological similarities with some species ofLinum (Linaceae).     

Comparative ultrastructure of pyrenoids inTrebouxia (Microthamniales,Chlorophyta)

Pyrenoid ultrastructure has been investigated from cultures of all 26 species ofTrebouxia with the aim of establishing pyrenoids as a taxonomic character. Different arrangements and forms of thylakoid lamellae within the pyrenoid matrix allow eight pyrenoid types to be distinguished. Each type is characteristic of a group of species. Thegigantea- andimpressa-type are similar, differing only in the form of the tubules: short, branched tubules mark thegigantea-type; ± long and straight invaginations theimpressa-type. Thearboricola-type is characterized by meandering pyrenoid membranes developing from lamellae parallel with each other in young autospores. Pyrenoids of thegelatinosa-type are traversed by thin parallel-arranged tubules. Few thylakoids with a curved profile are typical of theirregularis-type. Thecorticola-type is different from all others in having a distinct starch sheath closely connected with the pyrenoid matrix and no pyrenoglobuli being associated with the pyrenoid membranes. No true pyrenoids have been found inT. magna andT. erici. Within the chloroplast, they have indistinct areas with pyrenoglobuli, but without differentiated thylakoids. Pyrenoid morphology is stable in culture on different media as well as in phycobionts within lichen thalli. Comparing the pyrenoid of a lichenizedTrebouxia with that from cultured species, the identification of the phycobiont within the lichen thallus is possible, without the need of culturing the algae. This has been shown in species ofParmelia andHypogymnia. New aspects for the taxonomy and systematics ofTrebouxia are discussed.Dedicated to Prof. DrLothar Geitler on the occasion of the 90th anniversary of his birthday.     

Polarity, aperture condition and germination in pollen grains ofEphedra (Gnetales)

Pollen grain polarity, aperture condition and pollen tube formation were examined inEphedra americana, E. foliata, E. rupestris, E. distachya, andE. fragilis using LM, SEM and TEM. In the characteristic oblate pollen, as seen in situ in the tetrad configuration, the polar axis is the minor one and the equatorial plane runs between the two narrow ends of the microspore. The intine is thick in fresh fixed mature pollen but we have seen no indication of regions having an exceptionally thick intine that could be considered associated with an aperture or apertures. About three minutes after transferring fresh pollen to the germinating medium the ridged exine splits and twists away from the intine and its enclosed protoplast. The shed exine spreads out and curls into a scroll-like configuration that is as distinctive as that of the pollen shape had been but now having the ridges and valleys perpendicular to the long axis. The pollen tube develops, in our experience with more than a hundred germinating pollen grains, near one of the narrow tips of the pollen grain's equatorial plane. The location of the pollen tube initiation probably is related to the position of the tube cell nucleus. The pollen tube starts to grow about one hour after the exine was shed. The pollen tube emerges close to the narrow end (equator) of the gametophyte. This end emerged first as the exine is shed and is opposite to the prothallial cells. The stout pollen tube is c. 10µm in diameter grown in vitro on agar. In our germination medium the stout tube continued to elongate for about 24 hours reaching a length of c. 100 µm. With respect to exine morphology the aperture condition could be considered as inaperturate. The pollen tube, however, is formed in a germination area near one end of the exineless gametophyte.     

Paleobotanical evidence on the early radiation of nonmagnoliid dicotyledons

Paleobotanical studies indicate that several isolated and systematically depauperate groups of extant woody dicotyledons originated in the Mid Cretaceous. TheChloranthaceae had probably differentiated into insect-pollinated (Chloranthus andSarcandra) and wind-pollinated (Ascarina andHedyosmum) forms by the end of the Albian, and leaves referable to theTrochodendrales are known from the Albian and Cenomanian. In the latest Cretaceous and Early Tertiary, extinct representatives of theTrochodendrales includedNordenskioldia and theJoffrea-Nyssidium complex. ThePlatanaceae also differentiated before the end of the Albian and initially had insect-pollinated, unisexual flowers with five carpels or stamens. Some of these features persisted in the platanoid lineage until the Early Tertiary, and during the Paleocene and Eocene thePlatanaceae included forms with elliptical, palmate and pinnate foliage. The history of thePlatanaceae suggests that several features of the reproductive morphology of extant taxa may have arisen in association with a trend toward wind pollination. In the Mid Cretaceous, platanoid foliage partially intergrades with pinnateSapindopsis and pedateDebeya-Dewalquea leaves suggesting a close relationship betweenPlatanaceae andRosidae andFagaceae respectively. TheChloranthaceae, Trochodendrales, andPlatanaceae all occupy a somewhat intermediate position between theMagnoliidae andHamamelidae and are of considerable interest with respect to their role in the initial radiation of nonmagnoliid (higher) dicotyledons.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen:Ranunculaceae,Hamamelidaceae, Platanaceae undFagaceae

Transfer of fresh pollen either by wind or by insects is strongly related to the degree of its stickiness. The sticky substances involved should be understood as pollenkitt. Ultrastructural investigations indicate that the sticky pollen of entomophilous angiosperms (in this publicationHamamelis vernalis andH. virginiana, Corylopsis platypetala) results from the deposition of usually electron-dense, homogeneous pollenkitt mainly on the outside layer of the exine. In contrast, the pollen of anemophilous angiosperms (in this publicationParrotia persica, Platanus orientalis, Thalictrum flavum andQuercus robur), but also of some entomophilous angiosperms with special pollination ecology (e.g.Aquilegia vulgaris) is rather dry and powdery. In these cases the pollenkitt often is electron-transparent and inhomogeneous and is mainly removed into the cavities of the exine. Ultrastructure and deposition of pollen and pollenkitt in ambophilous angiosperms (in this publicationThalictrum minus, Castanea sativa) are ± intermediate.