The uptake of nitrate and ammonium nitrogen in Chara hispida L.: the contribution of the rhizoid

Abstract The uptake of ammonium and nitrate nitrogen by cultured plants of the green freshwater alga Chara hispida L. has been compared quantitatively with the contribution of its rhizoidal tissue. In the short-term, the rhizoid takes up 7–20% of the ammonium nitrogen, and about 15% of the nitrate that is taken up by whole plants under similar conditions. The uptake was studied over a range of both temperatures and external concentrations. The apparent activation energy for the uptake of NH₄⁺ and NO₃^? by the whole plant was found to be 50 kJ mol^?1 and 30 kJ mol^?1, respectively. For the rhizoid, the values were similar for both nitrogenous ions, 106 kJ mol^?1 and 70–100 kJ mol^?1. The rhizoidal uptake mechanism for ammonium nitrogen operates more efficiently compared to that in the whole plant. Nitrate is taken up by the rhizoid by a mechanism with a substrate affinity higher than in the plant taken as a whole. The possible ecological significance of the results is discussed.     

POLAR GROWTH OF HORMOSIRA BÁNKSII ZYGOTES IN SHAKE CULTURE

Zygotes of the fucalean alga Hormosira banksii initiate rhizoidal outgrowths in stationary culture 15 hr after fertilization and are then recognizably polar. By 24 hr most embryos are two-celled, and a few are four-celled. In a dark-grown population orientation of the developmental axis, as indicated by the direction of the rhizoidal outgrowth, was random around the vertical axis. In a unilaterally illuminated population the rhizoid usually emerged on the shaded side. Zygotes grown in light or darkness in shake culture, where they were continuously reoriented, usually developed as polar embryos, indicating that gradients of environmental factors are not required for initiation of polar growth. Some apolar embryos developed in stationary and shake cultures, but they were most frequent in dark shake cultures.     

THE BIOLOGY OF HARVEYELLA MIRABILIS (CRYPTONEMIALES,RHODOPHYCEAE). VII. STRUCTURE AND PROPOSED FUNCTION OF HOST-PENETRATING CELLS1

The parasitic red alga Harveyella mirabilis (Reinsch) Schmitz & Reinke was examined by light and electron microscopy to determine the structural mechanism involved in nutrient transfer. The host-penetrating rhizoidal cells are unique in possessing an extensive and apparently dynamic endomembrane system as well as other unique cytoplasmic inclusions. The membrane system consists of the plasmalemma, pinocytotic vesicles, multivesicular and concentric bodies, endoplasmic reticulum, dictyosomes, micro-body-like structures and an extensive vacuolar system. It is proposed that this system is active in the uptake and processing of host-derived nutrients. Plasmalemmal extensions (plasmalemmavilli) of Harveyella medullary cells may also function in nutrient uptake.     

TRANSFER OF PRODUCTS BETWEEN EPIPHYTIC MARINE ALGAE AND HOST PLANTS1,2

The red alga Smithora naiadum is normally found only as an epiphyle on the sea grasses Phyllospadix scouleri and Zostera marina. I used ³²P and ¹⁴CO₂ to examine the chemical communication between host and alga. Both ³²P and the product of ¹⁴CO₂ light fixation moved from the host to the alga. Reverse movement between host and epiphyte was also demonstrated. Part of this transfer occurred through the plant and part occurred by leakage from the host into the medium and subsequent uptake by the alga. Although plants were initially labeled in the light, transfer of ¹⁴C was light independent. Transfer of ¹⁴C-labeled products between host and epiphyte was also shown for Punctaria orbiculata and Z. marina; for Microdadia coulteri on Grateloupia doryphora, and between Gonimophyllum skottsbergii and Botryoglossum ruprechtiana. Epiphyte-host associations do not require a penetrating rhizoid for an exchange of the isotopes tested. By their proximity alone, epiphytic flora are apparently capable l exchanging products before these are diluted by the sea.     

SPORE GERMINATION AND DEVELOPMENT OF THE YOUNG GAMETOPHYTE OF THE OSTRICH FERN (MATTEUCCIA STRUTHIOPTERIS)

Light is required for the germination of spores of Matteuccia struthiopteris. Histochemical studies show that dormant spores contain no starch, but have an abundance of storage protein granules. Starch accumulates in the numerous chloroplasts of the spore on exposure to light and becomes gradually more extensive. Protein granules disappear as germination progresses. Following this, the centrally located nucleus migrates toward the proximal spore face. Concomitant with the nuclear migration, an increase of cytoplasmic RNA surrounding the nucleus occurs. An equal nuclear division and unequal cell division give rise to a 2-celled gametophyte consisting of a large prothallial cell and smaller rhizoidal cell. A new peripheral wall forms around the entire protoplast at the time of nuclear migration, while a transverse wall forms after nuclear division. The rhizoid emerges through the split raphe along the proximal spore face; it is rich in cytoplasmic RNA but contains very few chloroplasts and little starch. Electron microscopy of the 2-celled stage revealed a greater concentration of mitochondria, Golgi bodies, and a more extensive endoplasmic reticulum in the rhizoid than was found in the prothallial cell, which, however, was far richer in chloroplasts and lipid bodies. As the rhizoid elongates and becomes more vacuolated, cytoplasmic RNA decreases as cytoplasmic protein increases. The rhizoid undergoes no cell divisions, while the prothallial cell retains the potential for further cell division. The possible significance of the distribution of storage products, cell organelles, and other cell components were considered in relation to the non-equational cell division and differentiation of the 2 cells.     

Orientation ofFucus egg polarity by electric a. c. and d. c. fields

Summary Zygotes of the marine brown alga,Fucus serratus, have been subjected to the different modes of electric fields. 1) The result of a former study with conductive d.c. fields has been confirmed using electrostatic d.c. fields of 0.5 to 4 V/cm: the zygotes develop the cell polarity axis parallel to the imposed field with the rhizoid pole toward the cathode. 2) The frequency response to both, conductive and electrostatic, a.c. fields represents an optimum curve. The response,i.e. rhizoid formation at either or, in rare cases, both cell poles, peaks at square pulse durations,t _E, of 70 to 120 ms. 3) The same frequency response appears if the pulse number is kept constant at 8s^–1 by variation of the interval between the pulses,t _o. Only fort _oo > 200 ms,i.e. a pulse number of 3s^–1 the response declines markedly. The data support our hypothesis that imposed electric fields induce cell polarityvia differential shift of the membrane potential rather than transcellular current flow. Furthermore, the given dose-response curves strikingly resemble those due to the other morphogenetically active signals: percent response consistently approximates the per cent signal intensity gradient which evokes it.     

Rhizome gravitropism precedes gravimorphogenesis after inversion of the green algal coenocyte Caulerpa prolifera (Caulerpales)

Previous investigations reported that the marine alga Caulerpa prolifera, a giant coenocyte, had a fast morphogenetic response to a change in position with regard to gravity: the next rhizoid formed approximately 1 day later on the new underside of the inverted rhizome (Jacobs and Olson, 1980, American Journal of Botany 67: 141–146). Preceding the change in site of rhizoid development was a striking accumulation of amyloplasts at the new prospective rhizoid-initiation site in the rhizome tip. Detailed monitoring with video equipment of inverted Caulerpa plants, growing under controlled conditions, now reveals that after the amyloplast accumulation, but before the gravimorphogenetic effect on rhizoid development, the inverted rhizome tip shows a negative gravitropism that restores it to its normally upturned position. Because the experiments were performed with two-sided illumination, this was clearly not a phototropic response.     

IS THERE AN ECOPHYSIOLOGICAL EXPLANATION FOR THE GAMETOPHYTE–TETRASPOROPHYTE RATIO IN GELIDIUM SESQUIPEDALE (RHODOPHYTA)?1

In the fall, when 61% of the fronds of the Gelidium sesquipedale (Clem.) Born. et Thur. population located in Albufeira (southern Portugal) were reproductive, about 90% of these fronds were tetrasporophytes, whereas an equal percentage of female and male gametophytes was found (5%). The comparison of physiological performances of the reproductive phases (males, females and tetrasporophytes) did not reveal a physiological advantage of tetrasporic fronds. There were no significant differences either in the photosynthesis, nitrogen uptake, nitrate reductase activity, or biochemical composition of adult fronds. On the other hand, vegetative recruitment and spore production in the laboratory were significantly different. The re‐attachment to calcareous substrate and the subsequent rhizoidal growth were faster in tetrasporophytes. Particular levels of temperature, rather than irradiance, had an important effect on the phase differences in the spore release, attachment, and germination rates. Significant results were the higher release of carpospores at all irradiances at 17°C, and the higher attachment percentage of carpospores at 13°C versus tetraspores. Under higher temperatures (21°C), tetraspores showed higher attachment rates while carpospores germinated more. G. sesquipedale cystocarps released carpospores for 2 months, while tetrasporangia stopped shedding tetraspores after 1 month, resulting in a 3‐fold higher production of carpospores than tetraspores. Results showed that vegetative and spore recruitment may explain the low gametophyte–tetrasporophyte ratio of the studied population of G. sesquipedale as opposed to the physiological performance of phases.     

Observations on mechanisms of attachment in the green algaUlva mutabilis Føyn

The development of the rhizoid cells of the green alga Ulva mutabilis was investigated at the ultrastructural level paying special attention to the mechanism of attachment of the plant. Cytochemical data concerning the initial settling of the early zygote are also given. On the basis of histochemical staining and enzyme treatment it is concluded that the adhesive material secreted by the rhizoid cells is chemically different from that secreted by the zygote during the initial settling of the alga.     

Secretion of Lectin-Binding Material in Rhizoid Differentiation of Spirogyra

Some species of Spirogyra (a green alga) anchor to the substratumby differentiating the rhizoid. The transparent material secretedat the tip of the terminal cell or rhizoid is speculated tobe glycoprotein (Nagata 1977). To examine this, we treated Spirogyrafilaments with differentiating rhizoid with fluorescently labeledlectins. Among the nineteen lectins examined, only Bandeiraea(Griffonia) simplicifolia lectin I (BSL-I) strongly stainedthe transparent material, suggesting that the transparent materialcontains     

Identification of two putative adhesive polypeptides in Caulerpa prolifera rhizoids using an adhesion model system

The rhizoid section of the green alga Caulerpa prolifera (Cp) is active in attaching the developing plant to the substratum. A model system for the study of the adhesion of Cp rhizoids has been developed and identification of two putative adhesive polypeptides of Caulerpa (Vn-Cp) was revealed by immunodetection. A method for fast induction of new rhizoids was established using blade-base cutting followed by a few days of incubation. The new rhizoids were gently enclosed between two cover glasses and incubated until firm attachment developed. While analyzing protein extracts, two ∼60–70 kDa polypeptides (Vn-Cp I and Vn-Cp II) were identified by immunodetection with monoclonal antibodies to human vitronectin (Vn). The relative concentration values of the Vn-Cp proteins increased significantly in the ‘cell-wall’ fraction of the attached rhizoids during the incubation period. However, Vn-Cp proteins were not detected in non-attached rhizoids. Furthermore, the Vn-Cp proteins were also detectable on glass substratum subsequent to attached rhizoid removal. The induction and accumulation of Vn-Cp proteins on the ‘cell-wall’ of Caulerpa rhizoids and the firm attachment of the rhizoids to the glass substratum during the incubation period suggest that Vn-Cp proteins play a significant role in adhesion, which may be similar to the function of vitronectin in other adhesion systems. Furthermore, the high accumulation of Vn-Cp proteins on the glass substratum during attachment of new rhizoids suggests that the Vn-Cp proteins are secreted to the extracellular matrix and directly connect rhizoids to the glass substratum as an intermediate compound. These unique properties of Cp make it an excellent model system for the establishment of high amounts of adhesive material for future research. This revised version was published online in August 2006 with corrections to the Cover Date.     

Feeding strategy of the sacoglossan opisthobranch <Emphasis Type="Italic">Oxynoe olivacea</Emphasis> on the tropical green alga <Emphasis Type="Italic">Caulerpa taxifolia</Emphasis>

The feeding behaviour of the shelled sacoglossan Oxynoe olivacea was investigated to better understand the role and importance of this species in influencing encroachments of the alien alga Caulerpa taxifolia in the Mediterranean sea. We tested whether this slug preferred, as preliminary field observations suggested, an aggregative feeding behaviour and which part of the algal thallus, phylloid vs rhizoid, it preferred. Results showed that O. olivacea fed in groups and actively selected phylloid. This outcome poses important questions regarding the possibility that this species, fragmenting the alga thallus, could enhance dispersion and regeneration of C. taxifolia.     

Development of the siphonous green alga Penicillus and the Espera state*

The thallus of Penicillus is composed of two filament types: axial filaments of indeterminate growth and laterals of determinate growth. In vegetative reproduction new plants arise from horizontal rhizoids. Four stages can be distinguished in development. In the primordium stage the tip of a rhizoid swells and forms a primordium. In the germling stage ascending and descending axial filaments arise from the primordium, the former grow into a fascicle and give rise to lateral saccate branches, each of which forms an ascending and a descending arm and branches further into rhizoid-like branchlets. Together these structures constitute the foundation of the stipe. At the same time the descending axial filaments elongate and become main rhizoidal filaments with lateral rhizoidal branchlets. In the early juvenile stage the stipe is formed. The elongating ascending axial filaments form a medulla while their laterals produce a cortex. In the late juvenile stage the axial filaments form the capitulum. The Espera state of Penicillus lacks a stipe since the ascending axial filaments do not join in a fascicle. Espera has been grown in laboratory cultures from Penicillus plants collected in the Caribbean region and also been found uncommonly in nature in this area. This state may be a response to environmental stress. The flattening of thalli and their orientation perpendicular to the direction of waves are discussed. A comparison of Penicillus and Codium indicates that at least two types of development exist in multiaxial Eusiphoniidae.     

ENTRY OF A CHLORELLA-VIRUS INTO ITS HOST CELL1

The endosymbiotic Chlorella sp. (Chlorophyceae) of Paramecium bursaria (Ciliata) can be infected by a double-stranded DNA-containing virus (Chlorella-virus) that has a phagelike entry mechanism. Electron micrographs show that soon after attachment of the virus to the algal cell wall, a hole is formed through which the viral DNA enters the alga. Biochemical studies on a European Chlorella-virus system suggest that digestion of the algal cell wall is caused by glycolytic enzymes, one of which was identified as a β-d -glucosidase. Enzymes are bound to the virus capsid and are activated only after or by the attachment of the virus to its cognate alga or to preparations of the algal cell wall. Common features of viral cell wall-digesting enzymes and algal autolysins are discussed.     

Egg release and germling development in Sargassum horneri (Fucales, Phaeophyceae)

The mature female conceptacle of Sargassum horneri (Turner) C. Agardh has an ostiole filled with a gelatinous plug. The oogonium in the conceptacle has cell walls that can be differentiated into a dense outer and a less dense inner microfibrillar layer. Just prior to egg release, stalk material is produced inside the outer layer and the inner layer disappears. At this stage the gelatinous plug is extruded and mucilage is released through the ostiole. The released eggs are retained on the receptacle by the stalk and are surrounded by a large amount of the mucilage. Three-celled germlings form a primary wall with a polylamellated structure of microfibril layers. In multicellular germlings that have differentiated into thallus and rhizoids, the peripheral thallus cells have an outer cell wall consisting of a microfibril layer under the primary wall, while the cell wall of the rhizoid tip has an amorphous structure. The germlings are released from the stalk and become attached to the substratum by an adhesive substance secreted from rhizoidal cells.     

Changes in Intracellular Distribution of Thiol Groups During the Development of Acetabularia mediterranea

A cytochemical study of intracellular thiol distribution inAcetabularia mediterranea intact cells was performed using thefluorescent thiol-labelling agent monobromobimane (mBBr). Differentdevelopmental stages were examined during the vegetative phaseand generative phase of the algal life cycle up to cyst maturation.Important changes in thiol localization have been found to coincidewith turning-points ofAcetabularia development. During the rapid growth phase, overall thiol content steadilyincreased along the stalk, being maximal shortly before capdifferentiation. At this stage, the thiol distribution patternparalleled that of cap morphogenesis essential processes: thiolsbeing accumulated at the apex where morphogenesis is going tobe expressed. High thiol contents were also present in the rhizoidal partof the alga throughout the vegetative phase. At the onset ofthe generative phase, important alterations in rhizoid thioldistribution coincided with the presumptive time of nucleardivision. Overall thiol content strongly decreased and thiolsbecame highly concentrated in definite zones localized in thecentral area of the rhizoid. Later on, during the sequence of morphogenetic events leadingto cyst differentiation, changes in thiol localization and relativecontent were observed in the cap rays. Positioning of secondarynuclei into the cap coincided with a high increase in thiolcontent in the entire cap. During the process of cyst formation,thiol content slightly decreased and thiols were localized incyst domains. Thiol distribution was also studied during regenerative processesafter merotomy. A spatio-temporal coincidence was shown withcell wall regeneration. Key words: Acetabularia, thiols, development, bromobimanes     

New records of Gelidiella pannosa, Pterocladiella caerulescens and Pterocladiella caloglossoides (Rhodophyta, Gelidiales) from Japan

Three gelidialean species, Gelidiella pannosa (Feld‐mann) Feldmann et Hamel, Pterocladiella caerulescens (Kutzing) Santelices et Hommersand and Pterocladiella caloglossoides (Howe) Santelices, are newly reported from Japan, and their characteristic features are described. Monoecious plants of P. caerulescens produce spermatangial sori on: (i) fertile cystocarpic branchlets; (ii) special spermatangial branchlets on a cystocarpic axis; and (iii) branchlets of a special spermatangial axis. The latter two are newly reported in this species. Gelidiella pannosa has numerous unicellular independent points of attachment, whereas P. caerulescens and P. caloglossoides have the peg type of secondary rhizoidal anchorage. In the molecular phylogenetic study using small subunit ribosomal DNA sequences, G. pannosa is included in the Gelidiella clade with 100% bootstrap support in neighbor‐joining (NJ) analysis and 99% in maximum parsimony (MP) analysis. Pterocladiella caerulescens and P. caloglossoides are included in the Pterocladiella clade with 99.7% bootstrap support in NJ analysis and 100% in MP analysis. Each type of secondary rhizoidal attachment is completely consistent with the respective genus clade, which suggests that this morphological characteristic reflects phylogeny within the order Gelidiales.     

THE BIOLOGY OF HARVEYELLA MIRABILIS (CRYPTONEMIALES,RHODOPHYCEAE). VI. TRANSLOCATION OF PHOTOASSIMILATED 14C122

Pulse-chase labelling experiments demonstrate that photoassimilated ¹⁴C-bicarbonate is translocated from the host red alga Odonthalia floccosa (Esper) Falkenberg to the parasite Harveyella mirabilis (Reinsch) Schmitz & Reinke. The primary path of translocation is from host cortical cells (the site of photoassimilation) to the erumpent parasite pustule via the zone of interdigitation. The latter is a tissue region in which rhizoidal cells of Harveyella grow between, and establish secondary pit plugs with medullary cells of Odonthalia. A secondary translocation pathway occurs from isolated host cells dispersed in the pustule of Harveyella to adjacent parasite cells.     

Material structure, stiffness, and adhesion: why attachment pads of the grasshopper (Tettigonia viridissima) adhere more strongly than those of the locust (Locusta migratoria) (Insecta: Orthoptera)

The morphology, ultrastrucure, effective elastic modulus, and adhesive properties of two different smooth-type attachment pads were studied in two orthopteran species. Tettigonia viridissima (Ensifera) and Locusta migratoria (Caelifera) have a similar structural organization of their attachment pads. They both possess a flexible exocuticle, where the cuticular fibrils are fused into relatively large rods oriented at an angle to the surface. The compliant material of the pad contributes to the contact formation with the substrate. However, the pad material structure was found to be different in these two species. L. migratoria pads bear a thick sub-superficial layer, as well as a higher density of rods. The indentation experiments showed a higher effective elastic modulus and a lower work of adhesion for L. migratoria pads. When the indentations were made at different depths, a higher effective elastic modulus was revealed at lower indentation depths in both species. This effect is explained by the higher stiffness of the superficial pad layer. The obtained results demonstrate a clear correlation between density of the fibres, thickness of the superficial layer, compliance of the pad, and its adhesive properties. Such material structures and properties may be dependent on the preferred environment of each species.     

Sulfation of fucoidan in Fucus embryos. I. Possible role in localization

Zygotes of the brown alga Fucus distichus L. Powell divide into two cells which are structurally and biochemically different from each other. Cytochemical staining and autoradiography indicate that a sulfated polysaccharide is localized in only one of the two cells. Up to 10 hr after fertilization, no localization of sulfated polysaccharides is detectable in zygotes, and little ³⁵S (Na₂³⁵SO₄) is incorporated into an acid-soluble carbohydrate fraction. Between 10 and 16 hr, during rhizoid initiation and several hours before the first cell division, there is a large increase in the amount of ³⁵S incorporated into this fraction. The label is found associated with the sulfated fucose polymer fucoidan. Various extraction techniques and labeling experiments demonstrate that fucoidan is unsulfated at fertilization and undergoes little metabolic activity or turnover during the first 24 hr. Thus, the incorporation of sulfate into this carbohydrate fraction appears to involve a sulfation of a preexisting, unsulfated fucan polymer. The degree of sulfation achieved at this time in vivo is sufficient for migration of fucoidan through an electric field in agarose or acrylamide gels. The possible role of sulfation as a mechanism for the localization of fucoidan in the rhizoid cell by means of an intracellular electrical gradient is discussed.