Using palynomorphological characteristics for the identification of species of Alsinoideae (Caryophyllaceae): a systematic approach

Palynological characters of 18 species belonging to seven genera of the traditional subfamily Alsinoideae including Arenaria, Cerastium, Eremogone, Lepyrodiclis, Minuartia, Sabulina and Stellaria were studied in detail using light and scanning electron microscopy. Pollen grains of subfamily Alsinoideae are subspheroidal or prolate, pantoporate and 20.04 to 51.4 µm in size, prominent and sunken apertures uniformly distributed on the pollen surface. Two types of pollen grain ornamentation were observed, i.e. microechinate-punctate or microechinate-perforate. Echini are present on the surface of the pollen of all investigated species with medium, dense, or sparse echinodensity. These species exhibit variation in polar view, equatorial diameter, number of apertures, exine thickness, diameter of pore, appendages per pore, pore ornamentation, echini arrangement, echinidensity and shape of pollen. Based on qualitative characters, a dichotomous key has been developed for quick and easy identification. The present investigation contributes to the systematic approach using palynological characteristics and correct identification of species for members of subfamily Alsinoideae (Caryophyllaceae).     

Zur Pollenmorphologie der Gattung Waltheria L. (Sterculiaceae)

The pollen grains of 69 specimens representing 44 species or supposed species of the genus Waltheria L. have been studied. Based mainly on differences in exine characters 2 pollen types, a spinulose and a suprareticulate one were pointed out. Both types show ± parallel trends in the development of the aperture system to an increase of aperture number, to a shortening of the colpi and to more distinct bounds and nexine thickenings of the ora. They exemplify important steps of aperture evolution, which in the spinulose type leads from zonocolporate by way of zonopororate to pantopororate and in the suprareticulate type from zonocolporate to pantocolporate grains. The tendencies of sporoderm evolution concern a more distinct morphological and partially also a regional differentiation of the exine elements (bacula, spinulae), which prove to be close correlated with the evolution levels of the apertures. According to the pollen morphological point of view the species of the genus Waltheria are to be arranged in two groups, which may be regarded as two related, but relative independent phyletic entities. These evidences speak in favour of a taxonomic     

Interspecific pollen transfer as a mechanism of competition: Consequences of foreign pollen contamination for seed set in the alpine wildflower,Polemonium viscosum

Summary Earlier studies showed that under natural conditions foreign pollen receipt is correlated with loss of seed set in populations of P. viscosum. Here, we report on experimental pollinations using foreign pollen from two co-flowering species that share pollinators with P. viscosum. Experiments were designed to ascertain whether improper pollen transfer causes reduced seed set under controlled conditions and, if so, to determine the stage at which interference occurs. In flowers pre-treated by pollination with either Mertensia or Castilleja pollen, adherence of subsequently deposited conspecific pollen grains was unaffected. However, conspecific grains had significantly lower germination and ovule fertilization success in flowers receiving foreign pollen than in controls. These results suggest that pollinator infidelity in high alpine plant communities can reduce female reproductive success of P. viscosum by inhibiting the performance of conspecific pollen grains. Negative effects of Mertensia and Castilleja pollen were comparable despite large interspecific differences in pollen grain morphology. Accordingly, susceptibility to foreign pollen interference appears to be an intrinsic property of P. viscosum that is not dependent on the unique structural properties of pollen produced by competitive species.     

STUDIES OF SEED FERN POLLEN: DEVELOPMENT OF THE EXINE IN MONOLETES (MEDULLOSALES)

Monoletes pollen extracted from the seed fern synangium Dolerotheca sclerotica Baxter illustrate four stages in the development of the sporoderm. In the first stage the grains are up to 100 μm long and possess an apparent homogeneous exine in which there is little differentiation between the nexine and sexine. Numerous nexine lamellae and the initiation of sexine expansion mark stage 2 in exine ontogeny. Further expansion of the sexine continues in the third stage until the ratio between the nexine and sexine is approximately 1:5. The final stage in maturation of the sporoderm shows an expanded alveolate sexine with some of the sporopollenin units broken and disorganized. It is at this stage of development that nexine lamellae are most prominent. The formation of sporoderm layers in the fossil grains is compared with pollen grain development in living cycads (Cycadophyta) and a model proposed to account for the apparent early formation of nexine lamellae in Monoletes. The evolution of exine components in early pollen types is discussed.     

In vitro pollination of angiosperm ovules with gymnosperm pollen grains

Summary Pollen grains of the gymnosperm species Ephedra distachya and Pinus wallichiana germinated abundantly on the in vitro cultured placentae of the angiosperm species Nicotiana tabacum, Melandrium album and Allium moly. Some P. wallichiana pollen tubes entered the micropyles of M. album. Embryological observations of cross-pollinated M. album ovules 2 or 3 d after cross-pollination revealed the presence of pollen tube remnants within the embryo sacs. Karyological disturbances in the two- and three-celled proembryos and endosperm nuclei indicated their probable hybrid origin. In some crosses, gynogenetic haploid proembryos were also noticed.     

A revised generic classification of the tribe Sileneae (Caryophyllaceae)

A reclassification of the tribe Sileneae compatible with molecular data is presented. The genus Eudianthe ( E. laeta and E. coeli-rosa ) is restored. Viscaria, Ixoca (= Heliosperma ), and Atocion together form a well supported monophyletic group distinct from Silene and Lychnis , and are recognized at generic level. With Agrostemma and Petrocoptis , the number of genera in the tribe sums up to eight. The new combinations Silene samojedora, Silene ajanensis, Lychnis abyssinica, Atocion asterias, Atocion compacta, Atocion lerchenfeldiana , and Atocion rupestris are made.     

Pollen ultrastructure in <Emphasis Type="Italic">Aristolochia manshuriensis</Emphasis> and <Emphasis Type="Italic">A. contorta</Emphasis> (Aristolochiaceae)

Pollen ultrastructure has been studied in two relict and rare species of the genus Aristolochia, A. contorta Bunge and A. manshuriensis Kom. (Aristolochiaceae). Both species have inaperturate, spheroidal, sometimes distally monocolpate or distally bicolpate pollen grains. The equatorial and polar axes of pollen grain in A. manshuriensis are 48.5 and 44.0 μm, respectively. The percentage of defective pollen grains in A. manshuriensis is 3.4%. The fossulate, perforated exine is up to 2.3 μm in thickness; the sexine and the nexine are almost equal in thickness. In A. contorta, the equatorial axis of pollen grain is 36.6 μm: the defectiveness percentage, 24.5%. The exine is verrucate, up to 0.3 μm in thickness, while the sexine is two to three times thicker than the nexine. The pollen germination experiments have shown that pollen of A. manshuriensis, in contrast to A. contorta, can germinate in 10–20% sucrose at 22°С. These data and the high percentage of pollen defectiveness in A. contorta indicate that the androecium function in this species is reduced. The reduction of the androecium function is evidenced by a small amount of pollen grains in anthers or empty anthers and a high percentage of defective pollen grains.     

POLLEN AND GENE DISPERSAL: THE INFLUENCES OF COMPETITION FOR POLLINATION

Pollinators that forage indiscriminately can transfer pollen from one species to another, reducing the amount that reaches conspecific flowers. I present evidence that the presence of another plant species visited by the same pollinators can also reduce pollen dispersal distances and outcrossing. This has the potential to influence gene flow and reproductive success. Pollen carryover and movement patterns were measured for the shared insect pollinators of Stellaria pubera and Claytonia virginica in North Carolina. Bee flies deposited similar amounts of Stellaria pollen on a series of pistillate Claytonia flowers as on a series of pistillate Stellaria flowers. In arrays of potted plants, flies and solitary bees visited most flowers on a plant before leaving and then flew to a nearby plant chosen independently of species; 95% of moves were to one of 12 nearest neighbors. These measures of pollen carryover and movement patterns were used in a set of computer simulations to predict pollen dispersal distances. The simulations suggested that C. virginica substantially reduces outcrossing and pollen flow in S. pubera. These predictions were tested by tracking dye movement from anthers in populations of potted plants. Addition of C. virginica reduced the mean squared distance moved by dye to receptive S. pubera flowers by 23% and reduced the amount of dye moved by 51%. The estimated pollen component of gene flow was also much lower in a natural population of 5. pubera mixed with C. virginica than in the synthetic single-species populations.     

Evaluation ofHadena perplexa [Lepidoptera: Phalaenidae] as a biocontrol agent of bladder campionSilene vulgaris [Caryophyllaceae] in Canada: Rearing and host specificity

Rearing techniques and results of preliminary host range tests are reported forHadena perplexa (Denis & Schiffermuller) (Lep.: Phalaenidae) a candidate biocontrol agent against the weed bladder campion,Silene vulgaris (Moench) Garcke, in Manitoba, Canada. In the laboratory, it was necessary to pipette a 15% honey solution in water into the flowers as food for the adult moths. When reared singly to avoid cannibalism, 56% of the 1^st instar larvae developed to pupae. Larvae fed on a natural diet for 10 days can then be reared on either one of 2 artificial diets. Choice oviposition tests and no-choice larval feeding tests were conducted with plant species closely related toS. vulgaris in the generaSilene, Dianthus, Gypsophila, Lychnis, Saponaria. Species in 4 of 5 of these genera were accepted for oviposition, and species in all 5 genera supported the development of 1^st instar larvae to the pupal stage.H. perplexa should not be introduced into Canada.      

Beiträge zur Pollenmorphologie und Systematik der Convolvulaceen-Gattungen Maripa und Mouroucoa

Suggested by a previous communication on the occurence of two types of apertures among the pollen grains of Maripa s. l. (Convolvulaceae), 16 species of this genus were investigated palynologically. 3-zonocolpare grains mixed with 4–6-pantocolpate and 6–15-pantocolpate grains were found. The structure of the exine (infratectale bacula with or without supratectale processes) reveals further differences. The combination of these characters in the genus Maripa offers the separation into four types of pollen grains. The genus Mouroucoa is represented by its own type. These facts stress the necessity of the generical separation of the two genera. Consideration of pollen morphology yields some new aspects for subdivision of the genus Maripa.     

POLLEN WALL AND TAPETAL ORBICULAR WALL DEVELOPMENT IN SORGHUM BICOLOR (GRAMINEAE)

Pollen wall development in Sorghum bicolor is morphologically and temporally paralleled by the formation of a prominent orbicular wall on the inner tangential surface of the tapetum. In the late tetrad stage, a thin, nearly uniform primexine forms around each microspore (except at the pore site) beneath the intact callose; concurrently, small spherical bodies (pro-orbicules) appear between the undulate tapetal plasmalemma and the disappearing tapetal primary wall. Within the primexine, differentially staining loci appear, which only develop into young bacula as the callose disappears. Thus, microspore walls are devoid of a visible exine pattern when released from tetrads. Afterwards, sporopollenin accumulates simultaneously on the primexine and bacula, forming the exine, and on the pro-orbicules, forming orbicules. Channels develop in the tectum and nexine, and both layers thicken to complete the microspore exine. Channeled sporopollenin also accumulates on the orbicules. A prominent sporopollenin reticulum interconnects the individual orbicules to produce an orbicular wall; this wall persists even after the tapetal protoplasts degenerate and after anthesis. While the pollen grains become engorged with reserves, a thick intine, containing conspicuous cytoplasmic channels, forms beneath the exine. Fibrous material collects beneath the orbicular wall. The parallel development and morphological similarities between the tapetal and pollen walls are discussed.     

长梗苦草花粉粒的电镜观察

通过扫描电镜和透射电镜首次对沉水植物梗苦草（Vallisneria longipedunculata)的花粉粒进行了观察,其花粉粒的细胞壁非常薄,无萌发孔,但有2－3个具有较质丝的凹穴。花粉粒细胞壁的覆盖层十分不明显。具散生的颗粒;外壁内层较为厚实,但柱状结构分化不明显,整体呈海绵状;花粉内壁较厚。在花粉粒内部,有大量的单粒和复粒的淀粉粒,但未见到半复粒。     

迷果芹(\%Sphallerocarpus gracilis)和红三叶

对迷果芹(Sphallerocarpusgracilis(Bess.)K-Pol.)和红三叶(TrifoliumpratenseL.)进行了染色体计数及核型分析.迷果芹的染色体数目为2n=20,核型公式为K(2n)=2x=20=14m+4sm+2st(SAT);核型类型为2A,为较对称核型,该种植物的染色体数目及核型均为首次报道.红三叶的染色体数目有2n=14、16、28、32等类型,本研究首次报道了2n=14的核型公式为K(2n)=2x=14=2M+12m,核型类型为1B,为较原始的对称核型.     

ULTRASTRUCTURE OF STEWARTIOTHECA (MEDULLOSACEAE) PREPOLLEN

Ultrastructure of prepollen from the medullosan seed fern Stewartiotheca warrenae Eggert and Rothwell has been studied with the scanning electron microscope. Nitrocellulose (“liquid” type) peel preparations of pollen organ sections were placed on millipore filters and the grain fragments were extracted by rinsing the peels with acetone. Prepollen is of the Monoletes-type and is elliptical in polar view, averaging 264 μm long and 183 μm wide. Individual grains possess two deep, crescent-shaped grooves on the distal surface and a monolete suture with a median deflection proximally. The exine is constructed of a thick (1–2 μm), homogeneous nexine layer overlain by a sexine that is characterized by interconnecting spheroidal chambers. Chamber diameter decreases only slightly from the exterior of the sexine, inward, but the frequency and degree of interconnection among the chambers increases noticeably toward the interior of the wall layer. With regard to this latter feature the prepollen of Stewartiotheca appears intermediate between the organizations known for other Monoletes grains. The grains of Stewartiotheca compare most closely with those of Dolerotheca and Sullitheca.     

Isolation of 44 polymorphic microsatellite loci in three host races of the phytopathogenic fungus Microbotryum violaceum

We report the development of 44 microsatellite markers in three host races of the fungus Microbotryum violaceum, a sexually transmitted disease of the Caryophyllaceae. An enrichment protocol was used to isolate microsatellite loci from three strains, collected, respectively, from the plant species Gypsophila repens, Dianthus sylvestris, and Silene vulgaris. Polymorphism and cross‐amplification were explored with 32 isolates of M. violaceum, collected on 12 different plant species in natural populations.     

Pollen morphology of Microstachys (Euphorbiaceae) with emphasis on neotropical species

Abstract

In Euphorbiaceae tribe Hippomaneae is the pantropical genus Microstachys, comprised by 24 species, mostly from Brazil. Palynological studies with several representatives of the genus are scarce. The purpose of this study was to describe the pollen morphology of Microstachys and find possible differentiating characteristics between its species. Ten species of the genus were analysed, represented by 21 herbarium collections. Their pollen grains were acetolysed, measured and photographed under light microscopy (LM) and scanning electron microscopy (SEM). The analysed species possess pollen in monads, of small to medium size, isopolars, amb distincticly trilobed, oblate-spheroidal, prolate-spheroidal or subprolate, tricolporate with narrow and very long colpi, almost fused at the poles sometimes forming an invagination with tapering ends and margo of the colporus wide to very wide and psilate; endoapertures lalongate to very lalongate with tapering ends, presence of a narrow costae distinct in the endoaperture; exine microreticulate homobrochate, walls with simple columella; at the mesocolpus the sexine separates from the nexine forming cavea; and sexine always thicker than the nexine. Microstachys is a stenopalynous genus, despite its species varying in pollen size, shape and width of the cavea, margo of the colporus, and length of the endoapertures, demonstrating taxonomic value, and indicating a contribution towards future systematic analyses.     

Variation of pollen morphology, and its implications in the phylogeny of Clematis (Ranunculaceae)

Clematis s.l. (including Archiclematis and Naravelia) is a genus of approximately 300 species with cosmopolitan distribution. The diversity of its pollen was surveyed in 162 taxa belonging to all infrageneric groups of Clematis s.l. Pollen morphology was investigated by use of scanning electron microscopy to identify useful characters, test taxonomic and systematic hypotheses, and elucidate pollen character evolution on the basis of the molecular phylogeny. Clematis pollen is small to medium (14.8–32.1?μm?×?14.2–28.7?μm), oblate to prolate (P/E?=?0.9–1.4) in shape. The apertures may be tricolpate and pantoporate sometimes with 4-zonocolpate and pantocolpate pollen grains as transitional forms. The tricolpate pollen grains are predominant and occur in all the sections of the genus, whereas pantoporate pollen grains can be found in sect. Tubulosae, sect. Viorna, sect. Viticella, and Naravelia only. Phylogenetic mapping of aperture types reveals that the pantoporate pollen type may be the apomorphy in the genus and evolved several times. The surface ornamentation in all taxa studied is similar and characterized by microechinae evenly distributed on the microperforate tectum. The size and density of spinules on the tectum vary greatly but successive in the whole genus. According to the character syndromes of the ornamentation, separating sect. Brachiata from sect. Meclatis is supported. Though pollen morphology may contribute to investigation of problematic taxa, the taxonomic value of pollen morphology is limited at the species level.     

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.     

Hedgerows as an environment for forest plants: a comparative case study of five species

Many areas in Europe are dominated by agricultural land use, and as a consequence, many typical forest plant species suffer from habitat loss and fragmentation. Hedgerows, one of the common elements of rural landscapes, have been considered as potential refuges for these species. The main objective of this study was to examine whether forests and hedgerows differ in environmental conditions, and whether important life-history attributes of the populations differ between the two habitat types. We selected five species commonly found in the region in both forests and hedgerows (Adoxa moschatellina, Anemone nemorosa, Circaea lutetiana, Polygonatum multiflorum and Stellaria holostea), and sampled data on 10 populations of each species in each habitat type, including measurements of light and various soil factors. Hedgerows had higher relative light availability and tended to have higher soil nutrient contents and lower soil water values than forests. The comparison of plant performance values between habitat types did not show consistent patterns across species. Anemone and Polygonatum performed equally well in hedgerows and forests, whereas Stellaria appeared to have a higher fitness in hedgerows. In contrast, Circaea showed a higher reproduction under forest conditions. For Adoxa, the results were somewhat contradictory: whereas the reproductive output of this species was higher in forests, population density was higher in hedgerows. The abiotic factors most closely related to the performance values were relative light and soil water availability. The majority of plant performance values did not differ between hedgerows and forests. We therefore conclude that the tested forest species are capable of growing also in hedgerows and will survive equally well in forest and its “surrogate” habitat.     

THE ULTRASTRUCTURE OF SAHNIA POLLEN (PENTOXYLALES)

The micromorphology and fine structure of in situ pentoxylalean pollen are described from the holotype of Sahnia laxiphora Drinnan and Chambers 1985 collected from the Lower Cretaceous (Valanginian-Aptian) of Victoria, southeastern Australia. Pollen grains are ovoid, monosulcate, and relatively small, averaging 26 μm in length. Exine ornamentation is psilate. The sporoderm is two-parted with the sexine staining lightly throughout and approximately six times the thickness of the more darkly staining nexine. The exine over the sulcus is typically strongly invaginated, and may or may not include an extremely thin sexine layer. The outer part of the sexine is homogeneous, while the inner part is composed of relatively large granules separated by irregular lacunae of various sizes; lacunae are most pronounced at the sexinenexine interface. Faint lamellae characterize the nexine in both apertural and nonapertural regions. Granular orbicules are often associated with the exine surfaces and also occur appressed to pollen sac walls along with lamellated tapetal membranes. Sporoderm ultrastructure is compared to that of nonsaccate pollen of other groups, and particularly to pollen of Bennettitales, Gnetales, angiosperms, and similar plants, to which the Pentoxylales have been thought to be closely related. Although Sahnia laxiphora pollen is not identical to that of any of these taxa, the strongest similarity is with pollen of Bennettitales.