Microscopic analysis and seasonality of gemma production in the freshwater red alga Hildenbrandia angolensis (Hildenbrandiales, Rhodophyta)

The development and release of the unique vegetative propagules of the freshwater encrusting alga Hildenbrandia angolensis Welwitsch ex West et West, gemmae, were studied using several different microscopic and histochemical techniques. In addition, the seasonality of gemma production was monitored bimonthly over a 12‐month period in two spring‐fed streams in Texas, USA. Gemmae differentiate within the thallus and are subsequently released from the surface of the crust. Release of the gemmae most likely occurs by digestion of surrounding cells, as suggested by the presence of starch granules and lipid globules in the region between the released gemma and the thallus. The initial separation of the gemmae from the thallus occurs from the sides of the gemma or the bottom, or possibly simultaneously. Contrary to previous studies, we have observed that gemma production occurs endogenously within the thallus of freshwater Hildenbrandia, rather than on the surface of the crust in raised structures. Histochemical tests and electron microscopic examination indicate that the cells of the gemmae contain a large amount of floridean starch. The starch granules frequently form rings surrounding the nuclei of both gemma and thallus cells; a feature infrequently reported for florideophyte red algae. Our seasonality investigations indicate that large fluctuations in gemma production occur over 1 year, but at least some gemma production continues year‐round in the streams examined.     

Apoptotic wing degeneration and formation of an altruism-regulating glandular appendage (gemma) in the ponerine ant <Emphasis Type="Italic">Diacamma</Emphasis> sp. from Japan (Hymenoptera,Formicidae, Ponerinae)

We here show an example of morphological novelties, which have evolved from insect wings into the specific structures controlling social behaviour in an ant species. Most ant colonies consist of winged queen(s) and wingless workers. In the queenless ponerine ant Diacamma sp. from Japan, however, all female workers have a pair of small thoracic appendages, called gemmae, which are homologous to the forewings and acts as an organ regulating altruism expression. Most workers, whose gemmae are clipped off by other colony members, become nonreproductive helpers, while only a single individual with complete gemmae becomes functionally reproductive. We examined histologically the development of gemmae, and compared it with that of functional wings in males. Female larvae had well-developed wing discs for both fore- and hindwings. At pupation, however, the wing discs started to evaginate and later degenerate. The hindwing discs completely degenerated, while the degeneration of forewing discs was incomplete, leading to the formation of gemmae. The degeneration process involved apoptotic cell death as confirmed by TUNEL assay. In addition, glandular cells differentiated from the epithelial cells of the forewing buds after completion of pupation. The mechanism of developmental transition from wing to gemma can be regarded as an evolutionary gain of new function, which can be seen in insect appendages and vertebrate limbs.Edited by P. Simpson     

Development and germination of rhizoidal gemmae of Bryum violaceum

Approximately four to six weeks after transferring gametophores of Bryum violaceum Crundwell & Nyholm to fresh soil, abundant new rhizoids with stalked gemmae were present at the bases of the gametophores. The development of gemmae on rhizoids was followed from single cell initials to multicellular, three-dimensional forms. Mature gemmae, the vegetative diaspores, were pale orange, purple or reddish brown and each had a uniseriate stalk of two to four cells. While remaining on soil, the rhizoidal gemmae showed no in situ germination. However, following the removal of gametophores with rhizoidal gemmae, or rhizoids alone with gemmae, and their placement upon wet filter paper in Petri dishes in light, the gemmae germinated. During germination, the protonemata emerged consistently from mature gemmae at their distal ends, revealing the existence of gemma polarity. In the case of immature gemmae, on the other hand, the protonemata emerged from any surface cell indicating that gemma polarity had not yet been established.     

单芽狗脊蕨的生境观测及其芽胞发育初步研究

初步观察了单芽狗脊蕨的生长环境及其芽胞的发育过程。结果表明,单芽狗脊蕨喜阴湿环境和偏酸性土壤。5月的复叶上开始分化出芽胞,7月成熟的芽胞可在母体上萌发,长出不定须根,芽胞脱离母体后即能在土壤中萌发生长。芽胞越大,其萌芽率和生长速度越大。单芽狗脊蕨一年可产生两代芽胞。进行胎生的单芽狗脊蕨的孢子囊缺失或数目减少。     

Composite bundles,the host/parasite interface in the holoparasitic angiosperms Langsdorffia and Balanophora (Balanophoraceae)

Composite bundles are not simply a type of vascular bundles, but an integrated host/parasite interface. We investigated their structure in tubers of Langsdorffia and Balanophora. Composite bundles in both genera have similar components: 1) a central mass of host vascular tissues among which are located large parasite transfer cells; 2) a sheath of parasite parenchyma surrounding the central host vascular tissues; 3) specialized conducting tissues in the sheath; and 4) apical meristems composed of both host and parasite meristematic cells. Sheath parenchyma is recognizable from parasite tuber matrix by having thinner cell walls, and, especially in Langsdorffia, by the presence of collapsed matrix cells between the bundle sheath and tuber matrix. Sheath-conducting tissues consist of densely cytoplasmic transfer cells and small sieve tube members; in Langsdorffia, tracheary elements are also present. These sheath bundles connect with vascular bundles of the tuber matrix. Direct host/parasite contact only occurs by means of parasite transfer cells in the composite bundles. There is no xylem-xylem contact at the host/parasite interface. Abundance of parasite transfer cells suggests that they play an important role in nutrient absorption and translocation.     

Development and Liberation of Cauline Gemmae in the Moss Aulacomnium androgynum (Hedw.) Schwaegr. (Bryales): An Ultrastructural Study

The leafy shoots of the mossAulacomnium androgynum form clustersof gemmae borne terminally on long pseudopodial axes. The gemmaearise from single initial cells produced by the activity ofa superficial meristem. Mature gemmae comprise an apical anda basal cell with four to seven cells forming two, sometimesthree, tiers in between. The basal cell is connected to thetip of the pseudopodium by a uniseriate filament consistingof an abscission (tmema) cell and a stalk cell. The first divisionof the initial cell produces a proximal cell and a distal cell.The proximal cell elongates without further division formingthe stalk of the gemma; the distal cell gives rise to a lowerand an upper cell by transverse division. The upper cell dividesrepeatedly by oblique septa forming the apical and middle cellsof the gemma; the basal cell and tmema cell arise from a transversedivision of the lower cell. The first two divisions in gemmadevelopment are highly asymmetrical and exogenous, i.e. preceededby cell expansion. A broad interphase cortical band of microtubulesis associated with intercalary cellular growth during this stage.Subsequent gemma development follows an endogenous pattern withcellular expansion following the completion of proliferativedivisions and involving a conventional system of cortical microtubules.While elongating to about four times its original length withoutdeposition of a distinct new wall the tmema cell undergoes cytoplasmicdegeneration and eventually breaks, causing gemma liberation.The stalk cell elongates about eight-fold and its contents alsodegenerate after gemma liberation. Plasmodesmata in the basaland stalk cells are obliterated by the deposition of additionalwall materials. The highly electron-opaque outer walls of themature gemmae and tips of the stalk cells are water-repellant.The gemmae are dispersed either in water films or by air currents. Abscission; asexual reproduction; bryophytes; morphogenesis; microtubules; ultrastructure     

Asexual reproduction in a population of a leafy hepatic species Lophozia silvicola Buch in central Norway

The seasonal pattern and degree of asexual reproduction of a boreal hepatic species, Lophozia silvicola Buch, were studied in a lowland population of the species in central Norway. Asexual reproduction was quantified as numbers of 1–2 -celled gemmae produced per individual shoot, with average production ranging from 1000 to 7000 gemmae per shoot. Despite a slightly increasing trend in early spring, average numbers of gemmae present on shoots were fairly constant during the growing season. Germinability of gemmae was high in early spring, but declined rapidly towards the end of the season, suggesting an increasing proportion of gemmae in the dormant stage in the diaspore bank. Despite repeated sampling, average shoot density remained fairly constant in L. silvicola colonies during the growth season. Gemma production did not correlate with shoot density. Local colonization efficiency was estimated as frequency of occupied substrate patches and proportion of substrate area occupied by L. silvicola in a 10×20 m plot. Colonizable substrates included decaying wood, bare soil and rock surfaces. Colonies of L. silvicola were small, and differences in colony sizes between the substrate types were not significant. Colonies were found in 38% of the substrate patches, but total cover was low, less than 0.1% of all colonizable substrate.     

ROLE OF ASYMMETRIC CELL DIVISION IN PTERIDOPHYTE DIFFERENTIATION. II. EFFECT OF CA2+ ON ASYMMETRIC CELL DIVISION,RHIZOID ELONGATION,AND ANTHERIDIUM DIFFERENTIATION IN VITTARIA GEMMAE

Gametophytes of Vittaria graminifolia reproduce vegetatively by means of gemmae. Each gemma consists of a linear array of six cells: four body cells and a knob-shaped terminal cell at each end. When gemmae are shed from the gametophyte onto Knop's mineral medium, the two terminal cells do not divide, but elongate to form primary rhizoids. The body cells undergo asymmetric cell division, and the smaller daughter cells differentiate into either secondary rhizoids or prothalli. When gibberellic acid is included in the medium, antheridia are formed as a result of asymmetric cell division instead of vegetative structures. We studied the effect of Ca²⁺ on asymmetric cell division, rhizoid elongation, and antheridium formation in gemmae cultured on Knop's mineral medium and variations of Knop's medium. Ca²⁺ inhibited the onset of cell division and rhizoid elongation, but was required for differentiation of antheridia. Treatments which lowered the Ca²⁺ content of gemmae (EGTA and dilute HCl extraction, culture on verapamil-containing and Ca²⁺-deficient medium) caused an early onset of cell division and rhizoid elongation. The stimulation of growth was most pronounced when gemmae were deprived of Ca²⁺ during the first 24 hr of culture. The proportion of cell divisions which differentiated into antheridia in response to GA was greatly reduced when the Ca²⁺ status of gemmae was lowered with verapamil and Ca²⁺-EGTA buffers.     

贯叶连翘的分泌结构及其与金丝桃素积累的关系

贯叶连翘（ＨｙｐｅｒｉｃｕｍｐｅｒｆｏｒａｔｕｍＬ．）地上器官分布着分泌细胞球（黑色腺点）、分泌囊（半透明腺点）和分泌道（半透明腺条）３类内部分泌结构。分泌细胞球在茎、叶和花器官中均有分布,由２层鞘细胞包围多个分泌细胞构成实心的分泌细胞团。分泌囊主要分布于叶片中,分泌道则分布于花器官中,它们都是由１～２层切向扁平细胞围绕圆形或长形腔道构成,腔道的贮存物为精油。利用组织化学方法,结合荧光显微镜观察,证实金丝桃素类物质是由分泌细胞球（黑色腺点）所合成和积累的。通过用戊二醛和锇酸固定样品的显微和超微结构观察,发现金丝桃素类物质积累在成熟腺体分泌细胞的中央大液泡中,细胞周围浓厚的细胞质中分布着大量小液泡和高尔基体、内质网等细胞器。在此基础上对金丝桃素类物质的积累过程进行了初步探讨     

Experimental Investigation of the Mechanism of Reproductive Differentiation in the Queenless Ant,Diacamma sp., from Japan

We test two hypotheses about regulation of the reproductive division of labour in the permanently queenless ponerine ant, Diacamma sp., from Japan. All workers emerge with gemmae (tiny innervated thoracic appendages), but only one individual keeps them in each colony, and she is the only mated reproductive worker (gamergate). The gemmae of all other workers are mutilated by the gamergate soon after their emergence, and they can never mate. In the presence of gamergate, mutilated workers have inactive ovaries and do not behave aggressively. Two possible consequences of mutilation are: 1. olfactory signal — a pheromone inhibiting the oogenesis of mutilated workers is no longer released by the gemmae; and 2. endocrine degeneration of its afferent neuronal connections interferes reproductive physiology of a gamergate. Gemmae of gamergates were coated with shellac (to prevent pheromone emission) or removed, and over three weeks we studied any changes in ovarian activity of the gamergates as well as nestmate workers. Coating of gemmae did not elicit worker oviposition, suggesting that gemmae pheromones do not have a regulatory function. Experimental mutilation of gamergates resulted in a slight increase in both the frequency of dominance interactions and the ovarian activity of mutilated workers, but this effect was much lower than in colonies where the gamergate was removed. This contrasts with the immediate change in the behaviour (aggressive to timid) of newly emerged workers following mutilation.     

The Secretory Structure of Hypericum perforatum and Its Relation to Hypericin Accumulation

Three different kinds of internal secretory structures, secretory cell globules (black dots), secretory cavities (translucent dots) and secretory ducts (translucent streaks) were observed in Hypericum perforatum L. The secretory cell globule, which occurred in flower, leaf and stem, consisted of a core of large secretory cells surrounded by two layers of flattened sheath cells. The secretory cavities were present throughout the lamina and the secretory ducts throughout the flower, both consisted of one or two layers of flattened cells walling an oil chamber or duct. Histochemical and fluorescence microscopy revealed that hypericin was accumulated in the secretory cell globules. Microscopic and ultrastmctural observation further demonstrated that hypericin was accumulated in the large central vacuole of the mature secretory cells. Numerous dictyosome, endoplasmic reticulum and small vacuoles were observed in the dense cytoplasm surrounded the large central vacuole. The process of hypericin accumulation in the secretory cells was preliminarily discussed.     

Clonal dynamics and evolution of dormancy in the leafy hepatic Lophozia silvicola

Dead shoots of colonies of a leafy hepatic species, Lophozia silvicola , are replaced by shoots developing from asexual propagules, the gemmae. Observations of two populations of L. silvicola showed a strong decreasing seasonal trend in germinability of the gemmae. We suggest that the non-germinating gemmae enter dormancy, and that the proportion of gemmae entering dormancy is season-specific. We assume that there are two types of gemmae, dormant and non-dormant and that only the dormant gemmae can survive during winter. Using a stochastic individual-based cellular automaton model, we investigated whether selection on season-specific dormancy fraction would lead to a decreasing proportion of germinating gemmae. Thus the germination schedule is the evolving trait in the model. Parameter estimates for the model were based on data collected from a population of L. silvicola in southern Finland over a three-year study period. In the simulations, the germination schedule shaped by evolutionary change was similar to the observed pattern. Thus the modelling results give support to the dormancy hypothesis. The qualitative pattern of decreasing germinability towards the end of the growing season is robust. Quantitative predictions are influenced by changes in parameters; for example, if winter mortality of shoots increases relative to mortality during the growing season, production of an increased fraction of dormant gemmae is favoured, especially at the end of the season.     

中国藓类植物无性繁殖体的初步观察

无性生殖在苔藓植物的生活史中起着重要的作用,并且常通过各种无性繁殖体来完成。无性繁殖体的形态常被用来辅助鉴定一些不育的藓类植物。本文通过对38种藓类植物的无性繁殖体进行显微观察,结果显示：无性繁殖体在不同的藓类植物之间已经过多次演化;无性繁殖体的形态在种内是相对稳定的,且与其着生位置、配子体的分枝方式密切相关,而与植物的系统位置以及生境并无直接的关系;无性繁殖体的颜色与其胞壁的厚度以及表面纹饰密切相关。此外,无性繁殖体的产生常常与假根、原丝体有共存关系。在研究中发现,藓类植物的无性繁殖体主要包括原丝体芽胞、无性芽胞（叶生芽胞、中肋芽胞、枝生芽胞）、芽体、假根芽胞和假根状块茎;其中原丝体芽胞和无性芽胞最为常见。     

The use of alizarin red S to detect and localize calcium in gametophyte cells of ferns

An aqueous solution of alizarin red S containing chloral hydrate both clears intact chlorophyllous gemma cells of Vittaria graminifolia and stains for protoplasmic calcium. Verification that the stain was protoplasmic rather than in the cell wall was shown by a positive reaction in extruded protoplasm. Similar staining was found in extruded protoplasm of Onoclea sensibilis spores. Differentiating gemma cells show localized protoplasmic accumulations of Ca2+ at sites where asymmetric cell divisions initiate the formation of rhizoids, antheridia or vegetative cells. The staining properties of the dye depend on careful control of pH and the addition of appropriate amounts of KCl to the mixture. Treatment of Onoclea spores and Vittaria gemmae with 100 mM EGTA for 30 min nearly abolishes staining of their extruded protoplasts and also of intact cells of gemmae. The use of alizarin red S with and without chloral hydrate demonstrates different pools of protoplasmic Ca2+. When Onoclea spores are ruptured to extrude the protoplasm, both dye mixtures stain a peripheral, granular protoplasmic component. However, the chloral hydrate-containing dye also reveals Ca2+ associated with small particulate protoplasmic components. Extruded protoplasm of gemma cells stains intensely with alizarin-chloral hydrate, but does not stain with alizarin lacking chloral hydrate.     

Didymodon kunlunensis D.P.Zhao,S.Mamtimin & S.He (Pottiaceae), a new species from Xinjiang,China

Didymodon kunlunensis D.P.Zhao, S.Mamtimin & S.He is described as a new species found in Xinjiang, China. The new species is characterised by a combination of the following characters: ovate leaves with laminal cells always smooth, red colour in KOH, recurved leaf margins from base to apex, a percurrent costa without dorsal and ventral stereid bands in cross section, a distinct stem central strand and the presence of gemmae in the leaf axils. Light microscope photographs of the significant characters are provided and its distinctions from closely related taxa are discussed. A key to Chinese Didymodon species with smooth laminal cell is provided.     

Central projections of the sensory hairs on the gemma of the ant Diacamma: substrate for behavioural modulation?

In the ant genus Diacamma, all workers eclose from their cocoons with little clublike thoracic appendages, called gemmae. Whether these gemmae are mutilated determines individual behaviour, and ultimately reproductive role, in two of the three species examined. The gemmae are covered with sensory hairs, which probably serve a mechanoreceptive function. The sensory afferents arising from these hairs were stained and traced into the central nervous system (CNS). They feature widely distributed collaterals invading all three thoracic ganglia as well as the suboesophageal and the second abdominal ganglia. The multisegmental arborization pattern of the gemma afferents is very similar to that of wing-hair afferents of other ants (queens and males) or other insects in general. This implies that gemmae and wings are homologous structures. We discuss the morphology of the gemma afferents with respect to their possible involvement in the behavioural changes associated with mutilation. The neuronal processing may be modulated by (1) the decrease of sensory input onto interneurons (suggested by the afferents' extensive arborizations); or (2) by the effect of neuromodulatory substances (suggested by the finding that terminals occur within the cell body rind of the ganglion).     

Meristem differentiation in Riella helicophylla (Bory et Mont.) Mont. under the influence of auxin or antiauxin

During the development of the unistratose gemmae of Riella helicophylla, the single intercalary meristem of the very young gemmae is subdivided into two lateral meristems. The duration of the cell reproduction cycle increases from the margin to the median part of the gemmae. This polarization within the meristem disappears after addition of the antiauxin PCIB to the culture medium. PCIB leads to a retardation or blockage of the cell cycle during the light period of the culture. Under the influence of PCIB the amount of starch in the chloroplasts is strikingly increased, probably because of a reduction of starch degradation. Addition of sugars compensates the effect of PCIB on the cell cycle. The effects of PCIB are counteracted by auxin. The results are taken as evidence that auxin plays a role in directing the transport of substances needed for the continuation of the cell reproduction cycle between adjacent cells of the meristem.Abbreviations IAA indole-3-acetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - PCIB p-chlorophenoxyisobutyric acid - G-l-P glucose-l-phosphat - EDTA ethylenediaminetetraacetic acid     

Serial Development of Foliar Gemmae in Tortula (Pottiales, Musci), an Ultrastructural Study

The development and liberation mechanism of foliar gemmae havebeen studied by electron microscopy in two mosses, Tortula latifoliaBruch and Tortula papillosa Wils. The gemmae develop on theadaxial surface of mature leaves from single initial cells onboth the lamina and costa in T. latifolia but only on the costain T. papillosa . Elongation of the initial cell is associatedwith the deposition of a highly extensible new wall whilst theold wall and cuticle in the apical dome rupture. The first divisionis transverse and separates a short basal cell embedded in thefoliar tissue and a distal cell, or gemma primordium, protrudingfrom the leaf surface. Subsequent divisions of the gemma primordiumgive rise to a six-to-eight-celled globose gemma with mucilaginousouter walls. During gemma development the basal cell producesa new wall and elongates again whilst the common wall with thegemma splits apart centripetally along the boundary betweenthe old and new wall in the basal cell; plasmodesmal connectionsare gradually severed and eventually the young gemma remainsconnected to the basal cell only by mucilage. After separationof the first-formed gemma, the basal cell may expand and producea second gemma by the same mechanism. The whole process maybe repeated several times resulting in the formation of a chainof gemmae stuck together by mucilage and which are liberatedonly when the leaves are fully hydrated. Accumulation of abundantlipid deposits in the gemmae after symplasmic isolation reflectsconsiderable photosynthetic autonomy. Abscission; bryophytes; cell wall formation; plasmodesmata; vegetative reproduction     

Neuronal and non-neuronal responses to nerve crush in a pulmonate snail,Melampus bidentatus

Immunohistochemical and ultrastructural techniques were used to study sequelae of nerve injury in the pulmonate snail Melampus bidentatus. Either pedal or tentacle nerves were crushed, severing all axons, and recovery was monitored over 15 days. The axons regenerated from the segment attached to the soma, with no evidence of fusion of proximal and distal segments. The medium to large axons of central neurons, including those monitored with serotonin immunohistochemistry, grow distally across the path of smaller axons extending centrally from peripheral somata. The regions into which the growing axons projected were a focus of phagocytic activity. Cells previously labeled by PKH-26PCL, a fluorescent marker for phagocytic activity, were attracted to the crushed nerve within 6 h and were a consistent feature in the vicinity of the injury for at least 9 days, gradually extending their range as repair progressed in both directions from the crush. Repair proceeded within an intact sheath, and many sheath cells survived the crush, although the nuclear dye Hoechst 33258 revealed an initial distortion of their nuclei. The concentration of cells in the sheath in the crushed region increases after the crush, with the packing of nuclei peaking at 3 days and gradually returning to control conditions; this probably reflects migration of resident sheath cells. Cell division is rare in the sheath of intact nerves, but labeling with bromodeoxyuridine increases at the crush site between 4 and 9 days, indicating that cell replacement also occurs at the site. Electronic Publication