Orthophosphate Absorption by the Sporophyte of Funaria Hygrometrica During Maturation

The absorption of phosphate by the sporophyte of Funaria hygrometrica,during its maturation, has been studied using ³²P. More radioactivitywas found in the capsule when the absorption occured throughthe seta alone than when it look place through the leaves ofthe gametophyte. Each stage of capsule development studied isdescribed from transverse sections and electron micrographs.Phosphate is accumulated very actively by the capsule in theyoungest stage when the spores are being formed. The exchangesbetween the capsular tissues and the spores are low.     

Structure and Function of Transfer Cells in the Sporophyte Haustorium of Funaria hygrometrica Hedw: III. TRANSLOCATION OF ASSIMILATE INTO THE ATTACHED SPOROPHYTE AND ALONG THE SETA OF ATTACHED AND EXCISED SPOROPHYTES

Translocation of products of photosynthesis from gametophyteto sporophyte was examined in the moss Funaria hygrometricaHedw., as an adjunct to companion studies on the ultrastructureof the sporophyte haustorium and its capacity for absorptionof sugars in vitro. Labelled products derived from gametophyticphotosynthesis are transported to the sporophyte at an approximatelylinear rate for up to 12 h after a pulse treatment with ¹⁴CO₂.Large sporophytes receive label at a greater rate than smallerones. Transport is inhibited under conditions of water stress,and by lack of light, though darkening the sporophyte alonehas no effect. Movement of label from the haustorium along theseta occurs at a velocity of 1–3 mm h^–1, and issimilar to the onward movement of label derived from [³H]glucosesupplied to the haustorium in vitro.     

Accumulation of Pb and Zn in Gametophytes and Sporophytes of the Moss Funaria hygrometrica(Funariales)

Accumulation of lead and zinc was studied in the moss Funariahygrometrica Hedw. collected from mine tailings. Heavy metalaccumulation in gametophytes and sporophytes was quantifiedby graphite furnace atomic absorption spectrometry (GFAAS) andinductively coupled plasma-atomic emission spectrometry (ICP-AES).Pb and Zn accumulation in the placental zone was analysed byx-ray scanning electron microscopy (SEM) and transmission electronmicroscopy (TEM) microanalysis. Spectrometry showed that whilemoss gametophytes accumulated considerable concentrations ofheavy metals, sporophytes accumulated only small concentrationsof metals. X-ray SEM and TEM showed that the two metals accumulatedin placental transfer cells on both the gametophytic and sporophyticsides. To investigate the uptake pattern for both metals undercontrolled conditions, F. hygrometrica plants collected froma non-polluted site were treated in the laboratory with separatesolutions of Pb and Zn at two concentrations (10^-2and 10^-4 M)for 24 or 168 h. Metal accumulation was analysed separatelyin gametophytes and sporophytes using GFAAS and ICP–AES.Each generation had a different accumulation quotient for bothmetals, and gametophytes accumulated significantly more metalthan sporophytes. Concentrations of Zn in sporophytes were alwayshigher than concentrations of Pb. The findings are discussedin relation to the role performed by the gametophyte and theplacenta in the accumulation and sequestration of Pb and Zn.Copyright 2001 Annals of Botany Company Atomic spectroscopy, Funaria hygrometrica, gametophyte, Pb and Zn accumulation, sporophyte, x-ray TEM and SEM microanalysis     

African Hepatics: VI. Euosmolejeunea

Abstract

(1) The ratio of ¹⁴CO₂ uptake of fully expanded capsules to that of corresponding gametophytes in the species studied was about 1 : 15 in Mnium hornum, 1 : 35 in Pleurldium acuminatum and around 1 : 1 in Funaria hygrometriea.

(2) Translocation of photosynthate from gametophyte to sporophyte took place within 1 to 4 days of exposure to ¹⁴CO₂; in expanded capsules the final activity was at least equal to that remaining in the gametophyte.

(3) In M hornum there was no evidence of reverse translocation from sporophyte to gametophyte.

(4) Translocation from gametophyte continued at a high level throughout the expansion and differentiation of the capsule in M. hornum. The activity reaching the sporophyte was approximately proportional to its fresh weight.

(5) Both in situ photosynthesis and translocation to the capsule of F. hygrometrica rose to a maximum during the latter part of capsule expansion and ensuing differentiation, subsequently declining as the green tissues of the capsule senesced.

(6) Fully expanded (but premeiotic) capsules of M. hornum contributed approximately 20% of the assiinilate necessary for their growth. The corresponding figure for P. acuminatum was about 10%, and for F. hygrometrica about 50%.     

A hundred-year-old question: is the moss calyptra covered by a cuticle? A case study of Funaria hygrometrica

Background and Aims

The maternal gametophytic calyptra is critical for moss sporophyte development and ultimately sporogenesis. The calyptra has been predicted to protect the sporophyte apex, including the undifferentiated sporogenous region and seta meristem, from desiccation. We investigate the hypothesis that this waterproofing ability is due to a waxy cuticle. The idea that moss calyptrae are covered by a cuticle has been present in the literature for over a century, but, until now, neither the presence nor the absence of a cuticle has been documented for any calyptra.

Methods

The epidermis of the calyptra, leafy gametophyte and sporophyte sporangia of the moss Funaria hygrometrica were examined using scanning and transmission electron microscopy. Thicknesses of individual cuticle layers were quantified and compared statistically. The immunochemistry antibody (LM19) specific for pectins was used to locate cell wall material within the cuticle.

Key Results

A multi-layered cuticle is present on the calyptra of F. hygrometrica, including layers analogous to the cuticular layer, cell wall projections, electron-lucent and electron-dense cuticle proper observed in vascular plants. The calyptra rostrum has a cuticle that is significantly thicker than the other tissues examined and differs by specialized thickenings of the cuticular layer (cuticular pegs) at the regions of the anticlinal cell walls. This is the first documentation of cuticular pegs in a moss.

Conclusions

The calyptra and its associated cuticle represent a unique form of maternal care in embryophytes. This organ has the potential to play a critical role in preventing desiccation of immature sporophytes and thereby may have been essential for the evolution of the moss sporophyte.     

Effect of Lead and Colchicine on Morphogenesis in Protonemata of the Moss Funaria hygrometrica

Protonemata of Funaria hygrometrica grown in artificial mediacontaining different lead concentrations grow more slowly thancontrols and show a disturbance of polar growth, changed arrangementof chloroplasts, alterations of nucleus and septa position.Morphological effects are dose-dependent. At the lowest leadconcentration (10^-6 M), only a delay in development was observed,but no cellular alterations, At 10^-5 M Pb nuclear migration,cellular shape, size and position of plastids, were alteredand a variety of aberrant forms were present. At 10^-4 M, besidesthese alterations, a drastic reduction of the protonemal system,high vacuolation and the growth of protonemal filaments fromleaves were evident. The highest concentration, (10^-3 M), causeddeath. Patterns of protonemal development and cellular arrangementin lead-treated samples showed similarities as well as differences,if compared to alterations induced by colchicine. Indirect immunofluorescencedemonstrated a correlation between lead concentration and alterationof cytoskeletal organization (alterations similar to those inducedby colchicine). Hypotheses are raised to account for effects of lead on microtubulestructure, arrangement and cytoplasm organization.Copyright1995, 1999 Academic Press Funaria hygrometrica, lead, protonemal development, cytomorphogenesis, microtubules     

Effects of Dimethyl Sulphoxide on the Gametophytic and Sporophytic phases of Pteridium aquilinum (L.) Kuhn

Dimethyl sulphoxide (DMSO) at concentrations of up to 10 mlper litre of growth medium was found to have no significantinfluence upon Pteridium gametophyte growth or morphology However,significant effects upon embryo development and sporophyte morphologywere shown The most striking of these was that more than oneembryo developed from each fertilized gametophyte grown on DMSOmedium Two to five embryos regularly developed on each gametophytecultured on media containing this solvent, control gametophytesbore single sporophytes The significance of these findings inrelation to theories concerning polyembryony are discussed Pteridium, dimethyl sulphoxide, polyembryony     

The control of chloroplast division in Funaria hygrometrica II. The effects of kinetin and indole-acetic acid on nucleic acids

Replication of chloroplast DNA is an initial stage in chloroplastdivision during regeneration of isolated moss leaves. Variousconcentrations and combinations of growth substances have beenused to ascertain their effects on initiation of cellular dedifferentiation.Results indicate that during early stages of dedifferentiationof cells of isolated leaves of the moss Funaria hygrometricaa .change in the concentration balance between indole-aceticacid (IAA) and kinetin is effective in controlling the initiationof DNA replication in nuclei and chloroplasts. One of the waysthe balance between growth substances changes in by rapid lossof IAA from the cut leaf surface after isolation of the leaf (Received January 8, 1971; )     

Phytochrome Regulation of Expression of mRNA Encoding the Major Light-Harvesting Chlorophyll a/b-Binding Proteins of Photosystem II in the Haploid Phase of Adiantum capillus-veneris

Light regulates development in the fern Adiantum capillus-venerisduring both the haploid gametophyte and diploid sporophyte phases.In this study, we have investigated the role of the photoreceptorphytochrome in haploid spore germination and regulation of expressionof mRNA encoding the major light-harvesting polypeptides ofphotosystem II {Lhcb mRNAs). Both of these responses appearto be induced by phytochrome and inhibited by blue light, acting.throughseparate photoreceptor(s). Characteristics of phytochrome actionwere similar for both responses, showing no induction in thevery low fluence range and an ‘escape’ from far-redreversal for 50% of the response by 6 h. Overall, these resultsshow that phytochrome regulates both a morphological and a molecularprocess in haploid fern spores, and that these processes mayshare a common signal transduction pathway. (Received February 4, 1998; Accepted April 8, 1998)     

The Effect of the Calyptra on the Plane of Guard Cell Mother Cell Division in Funaria and Physcomitrium Capsules

The calyptra influences the plane of division in guard cellmother cells of Funaria and Physcomitrium. Normally, capsulesexpand while sheathed by the calyptra and the axes of the stomataare parallel to the axis of the capsule in both genera. Removalof the calyptra from an elongating sporophyte leads to setathickening prior to capsule expansion and an essentially randomorientation of stomata. If the calyptra is removed from a sporophyteof Funaria at the time the division of the guard cell mothercells is expected, guard cells of abnormal shape and undividedguard cell mother cells are found in unusually high frequency.     

THE GENETIC STRUCTURE OF SPOROPHYTIC AND GAMETOPHYTIC POPULATIONS OF THE MOSS,FUNARIA HYGROMETRICA HEDW

Patterns of phenotypic and genotypic variability in two populations of the moss, Funaria hygrometrica, were investigated using measurements of gametophytic and sporophytic morphology, sporophytic reproductive output, spore germination, gametophytic growth rates and tolerances of copper, cadmium, and low nutrient conditions, and electrophoretically detectable enzyme variation. The two populations differed in all traits measured, but complete monomorphism within populations at 14 enzyme loci suggested that each represented a single clone. Variability in gametophytic growth rates and responses to different experimental media, however, occurred among haploid sib families (families of meiotic progeny derived from the same sporophyte) and among sibs within families within both populations, suggesting high levels of genetic variability. Low mean reproductive output and a high level of variability among sporophytes in a mine site population probably reflected heavy metal toxicity. Based on this study, in combination with previous work on F. hygrometrica (Shaw, 19906), somatic mutation and/or nongenetic effects appear to contribute significantly to phenotypic variability in natural populations.     

ROLE OF SETTLEMENT DENSITY ON GAMETOPHYTE GROWTH AND REPRODUCTION IN THE KELPS PTERYGOPHORA CALIFORNICA AND MACROCYSTIS PYRIFERA (PHAEOPHYCEAE)1

Laboratory studies were used to examine how variation in the density of spore settlement influences gametophyte growth, reproduction, and subsequent sporophyte production in the kelps Pterygophora californica Ruprecht and Macrocystis pyrifera (L.) C. Ag. In still (non-aerated) cultures, egg maturation in both species was delayed when spores were seeded at densities 300 · mm^?2. Although the density at which this inhibition was first observed was similar for both species, the age at which their eggs matured was not. P. californica females reached sexual maturity an average of 4 days (or ～ 30%) sooner than did M, pyrifera. As observed previously in field experiments, per capita sporophyte production was negatively density dependent for both species when seeded at spore densities of 10 · mm^?2. Total sporophyte production (i.e. number · cm^?2) for both species, however, was greatest at intermediate densities of spore settlement (～ 50 spores · mm^?2). In contrast, total sporophyte production by P. californica steadily increased with increasing spore density in aerated cultures; highest sporophyte density was observed on slides seeded at a density of 1000 spores · mm^?2. Preliminary experiments with P. californica involving manipulation of aeration and nutrients indicate that inhibition of gametophyte growth and reproduction at higher densities of spore settlement in non-aerated cultures was probably caused by nutrient limitation.     

Ion Distribution Across the Cortex of Soybean Nodules: Possible Involvement in Control of Oxygen Diffusion

X-ray microanalysis was used to study ion distribution in nodulesof soybean [Glycine max (L.)] cv. Clarke formed with two strainsof Bradyrhizobium japonicum ; RCR3407 and RCR3442. Previousstudies have shown that the oxygen diffusion barrier in nodulesof the RCR3442 symbiosis has slower responses to acetylene andincreased external oxygen than those of the RCR3407 symbiosisand this variation is correlated with differences in glycoproteincontent. X-ray maps and cross-cortical cell counts of nodulesformed by either strain show very similar zonal distributionsof Mg, K, S and Ca across the cortex. Levels of K appear tobe similar but levels of Mg, S and Ca seem to be lower in nodulesof the RCR3442 symbiosis. These results suggest that the contentand distribution of Mg, S and Ca reflect an involvement in theoperation of the cortical diffusion barrier.Copyright 1994,1999 Academic Press Glycine max, X-ray microanalysis, nodules, oxygen diffusion, ion distribution, Mg, Ca, K, S     

Development of moss plants from isolated and regenerated protoplasts

Proroplasts were enzymatically isolated from protonema of Funaria hygrometrica L. SIBTH., cultured from the spores on Knop's solid medium. The protoplasts were incubated in hanging drops in liquid Knop's medium with 0.3 M mannitol and 0.5% glucose. Isolation of protoplasts was obtained successfully, through the action of Macerozyme®, cellulase and Rohament P® mixed with mannitol. Cultured protoplasts began to regenerate walls within 24 h, followed by the enlargement and divisions within 48 h. These protoplasts started producing active and long strands of protonema within 5 days, while well-differentiated leafy gametophytes were developed just in 20–23 days.     

Structure and Function of Transfer Cells in the Sporophyte Haustorium of Funaria hygrometrica Hedw: I. THE DEVELOPMENT AND ULTRASTRUCTURE OF THE AUSTORIUM

The development of the sporophyte-gametophyte interface in themoss, Funaria hygrometrica Hedw., is described with the aidof light- and electron-microscopy. The outer walls of the cellsthat abut the haustorial cavity in both generations developlabyrinths typical of transfer cells. This feature is more apparentin the epidermal cells of the sporophyte foot (haustorium),where development can be split into three main stages. The primarygrowth stage, which is complete at about the time the calyptradetaches from the ripened archegonium, involves the formationof transfer cells. The secondary stage is characterized by thedeposition of amorphous inclusions in the wall labyrinth ofthe transfer cells. The tertiary stage, which commences as thesporophyte capsule ripens, entails de-differentiation of thetransfer cell wall labyrinth to form a thick, heavily encrusted,outer cell wall. The pattern of development of these cells iscorrelated with changes in gametophyte- sporophyte translocationcapabilities.     

Uptake, transport and metabolism of cytokinin in moss protonema

Uptake, transport and metabolism of cytokinin in the protonemaof Funaria hygrometrica were studied using labelled kinetin(6-furfurylamino [8-¹⁴C]-purine). All cells of the protonema,chloronema and caulonema, were able to take up kinetin, whichwas carried in the symplastic transport system from cell tocell. Radioactivity was especially accumulated in growing cellsof the protonema. Kinetin was metabolized immediately afteruptake. While only very little kinetin (less than 1%) remainedas free kinetin and one part was immobilized in chromatographicseparation [e.g. attached to proteins and incorporated intonucleic acids (17)], most of the remaining kinetin was metabolizedto adenine derivatives. Exogenously supplied adenosine changedthe metabolism of kinetin. In the caulonema, adenosine reducedthe turnover of kinetin to other adenine derivatives and enhancedthe content of labelling in the start fraction. Thus adenosinecan stimulate cytokinin-dependent bud formation in moss protonema. (Received November 24, 1977; )     

Seasonal Changes of Bioelements in the Litter and their Potential Return to Green Leaves in four Species of the Argentine Subtropical Forest

The aim of this work was to analyse the nutrient concentrationsof N, P, Ca, Mg, K and Na in mature leaves, branches and fruitsand to investigate relationships between the contents of thesebioelements in senescent and fresh leaves in four forest species:Gleditsia amorphoides ( Espina Corona), Patagonula americana(Guayaibí),Chlorophora tinctoria ( Mora) and Astroniumbalansae (Urunday). The study site was located in the ColoniaBenítez Estricta Nature Research (Chaco, Argentina).In this subtropical forest, total litter was collected monthlyand was sorted into three groups: (1) leaves; (2) branches andfruit; (3) unidentified. Total dry matter was recorded and analysedfor N, P, Ca, Mg, K and Na. Espina Corona had the highest leafconcentrations of N, while Mora had the highest concentrationsof Ca and Mg. The highest leaf concentrations of P were foundin Espina Corona, Mora and Urunday. No significant differencesin K were found among the different species. Na concentrationswere higher in Espina Corona and Guayaibí than Mora andUrunday. A marked seasonal variability was observed in the concentrationsof N, P and K, with no important differences for Ca and Mg,except in Espina Corona. These variations in nutrient concentrationswere greater in leaves than in branches and fruits. N and Pwere translocated to other tree organs and Ca, Mg and Na wereaccumulated in mature leaves. The bioelement K is the only onethat undergoes leaching and mobilization in all species. ResorbedN and P can be used for the production of new leaf organs inthe following annual cycle. This resorption supports a portionof the production of new foliage, diminishing the demand fromsoil.Copyright 2000 Annals of Botany Company Above-ground production, potential return, nutrient resorption, leaf analysis, tropical forest, Gleditsia amorphoides, Patagonula americana, Chlorophora tinctoria, Astronium balansae, Espina Corona, Guayaibí, Mora, Urunday.     

Forced calcium entry and polarized growth of Funaria spores

We have used both steady electric fields, and gradients of the divalent ionophore, A23187, to control the point at which rhizoids emerge from spores of the common moss Funaria hygrometrica. The spores were grown in a medium containing calcium nitrate as the only major salt. Spores tend to form rhizoids towards the positive electrode, with a half maximal response to a difference of 4–8 mV across each cell. They also tend to form rhizoids towards the end of higher ionophore concentration in response to A23187 gradients. Both of these responses are the same at pH 5.5 and 8.0. Our tentative explanation is that Funaria spores tend to form rhizoids where most calcium enters. However, the point of chloronema emergence is scarcely affected by steady fields of up to 45 mV/cell. Moreover, when steady fields are applied across already developed rhizoids or chloronemata, their subsequent growth is directed towards the negative electrode in both cases, with rhizoids giving a 50% response at only 3—5 mV/cell, and chloronemata being less responsive.From Tsung-Hsien Chen's Ph.D. thesis, Purdue University     

Nuclear and Organelle DNA Replication during Spore Germination in Bryophytes and Equisetum

In contrast to ferns, the spore nuclei of the moss, Funaria hygrometrica, are in the G₂ phase. They do not incorporate the thymidine analogue BUdr before the first mitosis. When spores germinate in the presence of aphidicolin, an inhibitor of the (nuclear) DNA polymerase alpha, they give rise to two-celled, negatively phototropic protonemata, consisting of relatively long and thin cells. Determinations of nuclear DNA content further confirm the presence of G₂-nuclei in Funaria spores. Spores of Pogonatum urnigerum and of Equisetum hyemale likewise contain G₂-nuclei, as shown by BUdr incorporation or experiments with aphidicolin, respectively. In germinating Funaria spores, the organelle DNA is synthesized very early and independent of nuclear DNA. A gradient in replication of organelle DNA is found only in the long tip cells which are formed in the presence of aphidicolin.     

MORE MOSSES WITH CHLOROCHYTRIUM1

The green alga Chlorochytrium is reported intracellular in the mosses Archidium tenerrimum Mitten (Archidiaceae) and Funaria hygrometrica Hedwig (Funariaceae).