The pollen morphology of Vicia (Leguminosae)

The pollen morphology of 32 species of the genus Vicia was studied using scanning and transmission electron microscope (SEM and TEM). The interstitia of the pollen of the examined species were found to be either regular columellate, irregular columellate, or granular. The granular pattern was recognized to be apomorphy against the columellate patterns in the tribe Vicieae by comparison with the pollen in the tribes Vicieae, Cicereae, Galegeae, Hedysareae, and Trifolieae. The group of Vicia species having the apomorphy was congruent with that having a known apomorphy of the pistil character. This group with such synapomorphies may be monophyletic, though it is treated as polyphyletic in the present infrageneric system of Vicia.     

Pollen morphology of the Dimorphandra group (Leguminosae,Caesalpinioideae)

The Dimorphandra group, as traditionally circumscribed, is a rather diverse assemblage of genera in Leguminosae subfamily Caesalpinioideae that share certain morphological characteristics with the basally branching lineages of subfamily Mimosoideae. It currently comprises 51 species in seven genera: Burkea (1 species), Dimorphandra (26 species), Erythrophleum (10 species), Mora (6 species), Pachyelasma (1 species), Stachyothyrsus (2 species) and Sympetalandra (5 species). This study investigates the pollen morphology of 25 samples from 19 species of all seven genera. Pollen of the Dimorphandra group is small, isopolar, trizonocolporate and released in monads. Apertures are almost equal to polar length, with correspondingly small apocolpial areas. The shape of the aperture apices varies from acute to wide and rounded. Surface ornamentation is psilate, perforate, microreticulate, or perforate‐rugulate. The wall structure is usually columellate with a well developed foot layer. The pollen is small and unspecialised, agreeing with a previously noted pattern of more fixed and homogenous pollen structure in the more derived clades in subfamily Caesalpinioideae, compared with the great diversity of pollen types found in the basally branching lineages.     

The pollen of Dysoxylum (Meliaceae): the demise of Pseudocarapa

An account of the pollen morphology found in the genera Dysoxylum and Pseudocarapa (Meliaceae) is presented. Pollen grains are illustrated by means of electron and some light micrographs. The majority of taxa release grains as tetra-colporate monads. Tetrads have previously only been known in Pseudocarapa. They are also recorded here in three species of Dysoxylum. The taxonomic implications of this are discussed and Pseudocarapa is reduced to a synonym of Dysoxylum.     

Towards unlocking the deep nodes of Leguminosae: Floral development and morphology of the enigmatic Duparquetia orchidacea (Leguminosae, Caesalpinioideae)

Duparquetia orchidacea (Caesalpinioideae-Cassieae-Duparquetiinae) is a monotypic liana from tropical West Africa. Its highly unusual, zygomorphic flowers, the unique pollen morphology, and the lack of vestured pits in the wood correspond with previous phylogenetic studies that resolved the position of the species to an isolated position among the early-branching Leguminosae. Here we present a detailed analysis of floral morphology and development to clarify open questions of its floral organization. We provide new data that can be useful in clarifying phylogenetic relationships among early branching Leguminosae and improve our understanding of floral evolution in this large and important plant family. For comparison, we also present developmental data for other Fabales. Our analysis reveals some unusual and in parts unique developmental patterns, such as strict acropetal organ formation, loss and suppression of floral organs, and early petal enlargement. We interpret alternating left-right symmetries in floral development as clues to a spiral organ formation in ancestral taxa. Early asymmetry of the young carpel helps to interpret enantiostyly of other Leguminosae as an example of imprinted shape. Finally, we show that cochlear-descending petal aestivation in Duparquetia and in Papilionoideae is based on different ontogenetic patterns and therefore is most probably nonhomologous.     

The metareticulate pollen morphology of Alternanthera Forssk. (Gomphrenoideae,Amaranthaceae) and its taxonomic implications

Alternanthera (Amaranthaceae) is a diverse genus largely restricted to the American Tropics that belongs to the alternantheroid clade containing C₄ and C₃–C₄ intermediate species. This research focuses on the study of pollen characters by studying 13 species, representatives of the two major clades and subclades of Alternanthera. General palynological comparisons were conducted with light microscopy (LM), scanning electron microscopy (SEM) and with confocal laser scanning microscopy (CLSM) for exine ultrastructure. Twenty-five characters were measured and described for Alternanthera and among these, 14 pollen characters were used to discriminate pollen groups using cluster analysis and canonical analysis of principal coordinates (CAP). Pollen form and ornamentation, pores number, spines length, number of ektexinous bodies and nanospines on the ektexinous bodies on pore membranes, arrangement of nanopores and spines on structural elements, and metareticula form were taxonomically important and therefore used to construct the first palynological key to the alternantheroid clade species. Our study indicates that the seemingly subtle morphological variation of pollen is useful for recognising three main pollen types within Alternanthera. The much needed palynological terminology for describing the mesoporium in the metareticulate pollen of Amaranthaceae is provided.     

Inflated trichornes in flowers of Bauhinia (Leguminosae: Caesalpinioideae)

TUCKER, S. C., RUGENSTEIN, S. R. & DERSTINE, K. S., 1984. Inflated trichomes in flowers of Bauhinia (Leguminosae: Caesalpinioideae). An unusual kind of bifid multicellular inflated trichome is reported to occur on floral organs of II species of Bauhinia (Leguminosae-Caesalpinioideae), They occur on sepals, petals, stamens, carpels, receptacle, leaves, and stems, but not on all organs in each species. Their development in B. malabarica is described.     

Pollen morphology and structure of Tropical and Subtropical American genera of the Piptadenia -group (Leguminosae: Mimosoideae)

A survey of the pollen morphology of the American taxa of the Piptadenia -group (Mimoseae) was done including all together 384 species of the following genera: Piptadenia , Parapiptadenia , Pseudopiptadenia , Piptadeniopsis , Microlobius (= Goldmania ), Stryphnodendron , Adenopodia , Anadenanthera and Mimosa . Because of a high morphological and structural variation the definition and characterization of pollen types turned out to be very difficult. Nevertheless the presence and durability of some pollen characters allow a provisional grouping: (1) always tetrads or polyads, (2) small size 6 (20) 40 mum, (3) unstable shape but currently ovoidal and, if biconvex, ellipsoidal in the outline, (4) number of pollen grains of the polyads very variable and unstable; the most frequent numbers are 8-12-16, (5) pollen grains irregularly united in polyads, with homomorphous exine and, (6) pores always located in the angles of the pollen grains in distal-subdistal position, rarely subdistal, (7) without or with pseudocolpi or subpseudocolpi (never colpi) in the distal face, (8) endexine forming an annulus around the endoaperture with ends of free lamellae with a central white line, (9) the tectal stratum above the pores forming a small vestibular space (except in Anadenanthera ). These palynological characters or most of them confirm the affinity among genera in the Piptadenia -group and its distinction from other taxa of the subfamily Misosoideae with tetrads or polyads.     

Evolution of pollen,stigmas and ovule numbers at the caesalpinioid–mimosoid interface (Fabaceae)

In this study we examine the pollen, stigmas and ovaries from 62 collections of herbarium material representing 16 genera, using light and scanning electron microscopy. The caesalpinioid Dimorphandra group (Burkea, Dimorphandra, Erythrophleum, Mora, Pachyelasma, Stachyothyrsus and Sympetalandra) pollen grains are small, tricolporate monads, with perforate or psilate ornamentation. Dinizia, Pentaclethra and Aubrevillea have morphological characters that have suggested either a mimosoid or caesalpinioid placement. Dinizia pollen is in permanent tetrads with clavate ornamentation. Pentaclethra pollen grains are monads, two species have tricolporate pollen and the third is porate. Aubrevillea has tricolporate, finely reticulate monads. All ten genera have variable, non‐predictable stigma type and ovule number. The mimosoid Adenanthera group (Adenanthera, Tetrapleura, Amblygonocarpus, Pseudoprosopis, Calpocalyx and Xylia) pollen grains are in 8‐ to 16‐grain polyads. In all Adenanthera group species, the stigmatic cavity is only large enough to accommodate one polyad. In addition, the number of ovules present matches the number of pollen units in one polyad. Polyads have porate, operculate apertures that differ in layout, aperture morphology and development when compared with caesalpinioid and other eudicot pollen. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162 , 594–615.     

Floral development in Tribe Detarieae (Leguminosae: Caesalpinioideae): Amherstia, Brownea, and Tamarindus

Floral development was compared among three taxa in caesalpinioid tribe Detarieae sensu lato: Amherstia nobilis and Tamarindus indica have racemose, helically arranged inflorescences, while Brownea latifolia has cauliflorous capitate flower clusters that arise as racemes. All have acropetal flower order; initiation and development are sequential in all except Brownea, which is synchronous. All have paired persistent showy bracteoles. Floral symmetry is dorsiventral (zygomorphic) in all except Brownea, with radial symmetry at anthesis. Sepals initiate helically on a circular floral apex, starting with a median abaxial sepal, in all. Petals are initiated helically in Brownea, and unidirectionally in Amherstia and Tamarindus. Stamens are initiated unidirectionally in each stamen whorl in all except Amherstia, in which the outer whorl is bidirectional. The carpel initiates concurrently with the petals in Brownea, and with the outer stamens in the other taxa. The two upper (adaxial) sepal primordia become fused during development in all, so that the calyx appears tetramerous. Some reduced petals occur in Amherstia and Tamarindus, and some reduced stamens occur in all. All produce a hypanthium by zonal growth, and all except Tamarindus have the gynoecium attached adaxially to the hypanthial rim.     

Pollen morphology of Adenandra (Rutaceae: Diosminae) and its taxonomic implications

The pollen morphology of Adenandra Willd. (Rutaceae: Diosminae) was investigated to determine its taxonomic significance. Pollen of 27 of the 30 infrageneric taxa (representing 16 of the 18 species) was investigated by LM, SEM and TEM. Adenandra differs from all other Diosminae in having 4-colporate rather than 3-colporate pollen grains. This supports the alleged monophyletic status of the genus. Exine morphology and structure, however, is extremely diverse considering the size of the genus, with eight distinct pollen types and four subtypes being discerned. In some members the grains have uniform macroreticulate, striate or striato-reticulate sculpturing, whereas in others the poles have striate, striato-reticulate or reticulate-perforate sculpturing with various types of reticulate or rugulate sculpturing at the mesocolpia. Taxonomic groupings revealed by the pollen characters suggest elationships between taxa that were not previously apparent. It is suggested that evolutionary diversification among species of Adenandra is often more strikingly reflected by pollen morphology than by macromorphology, hence the eurypalynous state of the genus.     

In vivo assessment of mechanical properties during axolotl development and regeneration using confocal Brillouin microscopy

In processes such as development and regeneration, where large cellular and tissue rearrangements occur, cell fate and behaviour are strongly influenced by tissue mechanics. While most well-established tools probing mechanical properties require an invasive sample preparation, confocal Brillouin microscopy captures mechanical parameters optically with high resolution in a contact-free and label-free fashion. In this work, we took advantage of this tool and the transparency of the highly regenerative axolotl to probe its mechanical properties in vivo for the first time. We mapped the Brillouin frequency shift with high resolution in developing limbs and regenerating digits, the most studied structures in the axolotl. We detected a gradual increase in the cartilage Brillouin frequency shift, suggesting decreasing tissue compressibility during both development and regeneration. Moreover, we were able to correlate such an increase with the regeneration stage, which was undetected with fluorescence microscopy imaging. The present work evidences the potential of Brillouin microscopy to unravel the mechanical changes occurring in vivo in axolotls, setting the basis to apply this technique in the growing field of epimorphic regeneration.     

The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology

Wilts, E.F., Wulfken, D., Ahlrichs, W.H. and Martínez Arbizu, P. 2012. The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology.—Acta Zoologica (Stockholm) 93 : 14–27. The monogonont rotifer Squatinella rostrum was investigated with light, scanning electron and confocal laser scanning microscopy to reveal new morphological data on its inner and outer anatomy. In total, the visualized somatic musculature displays five paired longitudinal muscles (musculi longitudinales I–V) and nine circular muscles (musculi circulares I–IX). Compared to other species, S. rostrum is characterized by the absence of several longitudinal and circular muscles (e.g. musculus longitudinalis capitis, corona sphincter and pars coronalis). A reconstruction of the mastax musculature revealed a total number of seven paired and two unpaired mastax muscles. Possibly homologous somatic and mastax muscles in other, thus far investigated rotifers are discussed. Moreover, we provide a phylogenetic evaluation of the revealed morphological characters and suggest possible autapomorphic characters supporting Squatinella and Lepadellidae. Finally, we refer to some striking similarities in the morphology, ecology and way of movement of Squatinella and Bryceella that may indicate a closer relationship of both taxa.     

Enantiostyly in Chamaecrista ramosa (Fabaceae‐Caesalpinioideae): floral morphology,pollen transfer dynamics and breeding system

Enantiostyly is a form of reciprocal herkogamy, in which floral morphs present reciprocal differences in the position of sexual elements, and occurs in monomorphic and dimorphic forms. This polymorphism maximises cross‐pollination and reduces self‐pollination, being very common within the subtribe Cassiinae (Fabaceae). Nevertheless, few studies have investigated the functionality of enantiostyly, particularly in this plant group. The present study aimed to investigate enantiostyly and its functionality in Chamaecrista ramosa, a monomorphic enantiostylous shrub, in an area of coastal vegetation in northeast Brazil. Pollen deposition and capture on the body of floral visitors, the relationship of these data with floral biology and breeding system, and morph ratio were evaluated. Pollen deposition and capture occurred in specific sites of the floral visitor body, showing the functionality of enantiostyly. The floral architecture, associated with the floral visitor behaviour, resulted in indirect pollen deposition on the floral visitor body. This occurred through a loop made by the pollen upon the inner petal surface, similar that generally reported for other Cassiinae. Chamaecrista ramosa is self‐compatible, although no fruit set was observed through spontaneous self‐pollination. The occurrence and number of floral morphs was similar within clumps. Enantiostyly seems to be advantageous for this species, as it results in efficient pollen capture and deposition, reduces the chances of autogamy and maximises intermorph pollen flow.     

The musculature of three species of gastrotrichs surveyed with confocal laser scanning microscopy (CLSM)

The muscular system of gastrotrichs consists of circular, longitudinal and helicoidal bands that when analysed with confocal laser scanning microscopy, provide new insights into their functional organization and phylogenetic importance. We therefore undertook a comparative study of the muscle organization in three species of Gastrotricha from the orders Macrodasyida (Paradasys sp., Lepidodasyidae; Turbanella sp., Turbanellidae) and Chaetonotida (Polymerurus nodicaudus, Chaetonotidae). The general muscle organization of the marine interstitial macrodasyidans, Paradasys and Turbanella, not only confirms earlier observation on other species but also adds new details concerning the organization and number of helicoidal, longitudinal and other muscle bands (e.g. semicircular band). The freshwater, epibenthic–epiphytic chaetonotid, Polymerurus nodicaudus, has a similar muscular organization to other species of Chaetonotidae, especially species of Chaetonotus, Halichaetonotus and Lepidodermella. Perhaps unique to Polymerurus is the combined presence of an unbranched Rückenhautmuskel (also in Halichaetonotus and Lepidodermella) and a specialized dorsoventral caudal muscle, which flank the splanchnic component of the longitudinal muscles (only in Chaetonotus and Lepidodermella). This combination, together with the presence of splanchnic dorsoventral muscles, known only in Xenotrichulidae, implies a unique phylogenetic position for Polymerurus, and indicates a potential basal position of this taxon among the Chaetonotidae studied so far (i.e. Aspidiophorus, Chaetonotus, Halichaetonotus and Lepidodermella).     

The relationships of the genus Cicer L. (Leguminosae): the evidence from pollen morphology

The genus Cicer has traditionally been placed in the Vicieae, although it has a number of morphological characters which suggest that it is out of place in that tribe and that its affinities lie elsewhere. A survey of pollen morphology in Cicer , the other genera of the Vicieae, the Trifolieae and the Ononideae has been carried out to help determine the relationships of Cicer. The tribe Vicieae (without Cicer) is very homogeneous and is characterized by its long, rectangular pollen grains with small, heavily thickened endoapertures. The Trifolieae is more variable in its pollen morphology but, in general, has rather similar grains to the Vicieae although the structure of the endoaperture is different. The Ononideae and Cicer have nearly spheroidal pollen with very large, unthickened endoapertures. It therefore appears that as far as pollen characters are concerned, Cicer does not belong in the Vicieae but has much more in common with the Ononideae.     

Cytogenetics of species of Chamaecrista (Leguminosae – Caesalpinioideae) native to southern Brazil

Chromosome numbers, karyotypes, meiotic behaviour and pollen analysis are presented for species of Chamaecrista Moench (Leguminosae, Caesalpinioideae, Cassieae ) native to southern Brazil: C. nictitans ssp. patellaria , C. nictitans ssp. disadena , C. repens , C. rotundifolia , C. flexuosa , C. vestita and C. desvauxii . Meiotic behaviour is reported for the first time for all the taxa and was very regular; only bivalents were formed at diakinesis and metaphase I, chromosome disjunction and segregation were regular at anaphases I and II, meiotic indexes were over 99% and pollen fertility was over 92%. Pollen grains were subprolate in C. flexuosa and C. vestita and prolate–spheroidal in the other taxa. Karyotypes were symmetrical in all six species and the data are original, except for C. nictitans ssp. p atellaria. Chromosome number is presented for the first time for C. repens (2 n  = 16) and has been confirmed for the other taxa: 2 n  = 14 for C. desvauxii , 2 n  = 32 for the tetraploid C. nictitans ssp. patellaria and C. nictitans ssp. disadena , and 2 n  = 16 for the other species. These two basic numbers found in the genus, x  = 7 and x  = 8, point to chromosome evolution by dysploidy, which has also been accompanied by polyploidy.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 150 , 429–439.     

Seed of Cercis siliquastrum L (Leguminosae,Caesalpinioideae): Reserves and their Utilization after Germination

An ultrastructural study on the reserves stored in cotyledons and endosperm and on their utilization after germination has been carried out in Cercis siliquastrum seeds. The observations have shown that all the cotyledonal reserves of proteins and lipids are used during seedling growth. However, inside the endosperm cells only the protein reserves are depleted, and large quantities of lipids remain undigested. In both cell walls and intercellular spaces of the endosperm cells huge masses of polysaccharide reserves are stored. However, only the polysaccharides inserted in the cell walls are dismantled, while those located in the intercellular spaces are not attacked by the lytic enzymes. Some hypotheses regarding the possible roles of those endospermic non-nutritional materials in the strategies of seed germination and seedling growth are suggested.     

Apical pattern of fruit production in the racemes of Ceratonia siliqua (Leguminosae: Caesalpinioideae): role of pollinators

Fruit production and arrangement within the raceme were studied in two dioecious populations of Ceratonia siliqua (Leguminosae: Caesalpinioideae), an arboreal species that produces caulogenous racemes (emerging only from the old branches) with numerous flowers. Fruit production per raceme was low and similar between years and populations and even between individuals. During flowering, there were considerable flower losses from predation and lack of pollination. A mean of nine flowers per raceme began the transformation into fruits, of which 77% aborted. The final fruit production per raceme increased significantly following hand pollination, but was always very much lower than the availability of flowers in the raceme. The results suggest that fruit production of each raceme is limited by both availability of resources and a deficient pollination. In racemes setting fruit arrangement follows a definite pattern that remains constant between years and populations: fruit production was significantly higher in the apical zone of the raceme and lower in the basal zone. The pollinators of C. siliqua (flies and wasps) showed a clear preference for beginning their visits at the apex of a raceme. As a result, the pollen load deposited on the stigmas decreased from apex to base of the raceme. In most of the flowers situated in the central and basal zone of the raceme, the number of pollen grains deposited on their stigmas was lower than the number of their ovules. The high number of seeds in developed fruits suggests that the plant selectively aborts flowers that receive a smaller pollen load. The results indicate that the final pattern of fruit arrangement within the raceme is a direct result of pollinator activity.     

Direct observation of spectral differences between normal and basal cell carcinoma (BCC) tissues using confocal Raman microscopy

Raman spectroscopy has strong potential for providing noninvasive dermatological diagnosis of skin cancer. In this study, confocal Raman microscopy was applied to the dermatological diagnosis for one of the most common skin cancers, basal cell carcinoma (BCC). BCC tissues were obtained from 10 BCC patients using a routine biopsy and used for confocal Raman measurements. Autofluorescence signals from tissues, which interfere with the Raman signals, were greatly reduced using a confocal slit adjustment. Distinct Raman band differences between normal and BCC tissues for the amide I mode and the PO2- symmetric stretching mode showed that this technique has strong potential for use as a dermatological diagnostic tool without the need for statistical treatment of spectral data. It was also possible to precisely differentiate BCC tissue from surrounding noncancerous tissue using the confocal Raman depth profiling technique. We propose that confocal Raman microscopy provides a novel method for dermatological diagnosis since direct observations of spectral differences between normal and BCC tissues are possible.     

Con A conjugated to Europium(III) cryptate as a new histological tool for prostate cancer investigation using confocal microscopy

Lectins are carbohydrate recognition proteins that can be used as probes to reveal the glycosylation state of cells. They frequently have been used for diagnostic and prognostic cancer studies. For fluorescence based analysis, lectins commonly are conjugated to fluorescein isothiocyanate (Con A-FITC); however, this molecule loses its fluorescence quickly. We conjugated Europium cryptate to Con A (Con A-cryp-Eu) for use as a histochemical luminescent probe to recognize glucose/mannose residues in benign prostatic hyperplasia and prostatic carcinoma tissues, and used confocal microscopy instead of commercial Con A-FITC. Tissues were treated with Evans blue to suppress intrinsic tissue fluorescence before incubation with Con A-cryp-Eu or Con A-FITC. Con A-cryp-Eu exhibited hemagglutinating activity. Con A-cryp-Eu showed the same binding pattern as Con A-FITC in prostate stroma and gland cells. Staining was strong in benign prostate hyperplasia and prostate carcinoma tissues. Con A-cryp-Eu probe stained glucose/mannose residues in prostatic carcinoma more intensely than Con A-FITC. Furthermore, staining with Con A-cryp-Eu showed greater fluorescence intensity than Con A-FITC and the emission of Con A-cryp-Eu was more stable than the Con A-FITC for seven days under the same storage conditions. Maintenance of the luminescent properties and the binding pattern of Con A-cryp-Eu favor its use as an auxiliary histochemistry probe for prostatic tissue studies.