Quantitative short-term uptake of inorganic phosphate by the Chara hispida rhizoid

Abstract The rhizoid of Chara hispida L. made a small contribution to the uptake of inorganic phosphate under laboratory conditions. At 1 mmol m^?3 phosphate the rhizoid contributed about 4% to the uptake of the whole plant over 4 h. Under these conditions about half of the phosphate taken up by the rhizoid was translocated into the shoot. The rates of uptake and translocation increased with increasing external phosphate concentrations. When the shoot was in darkness, ³²P-translocation from the rhizoid was less than half of that found when the shoot was illuminated. When the rhizoid medium was anaerobic the translocation rate was lower than the rate in aerobic conditions and illumination of the shoot had no effect on the uptake or translocation of phosphate. Translocated ³²P accumulated in the apical growing regions of the plant. This was first noted in the secondary apices nearest to the rhizoids.     

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.     

Intercellular Transport and Photosynthetic Differentiation in Chara corallina

Intercellular transport of ¹⁴C-labelled photoassimilates, bothin isolated upper shoots and in isolated internode-branchletcomplexes of Chara corallina, was measured. The isolated uppershoots were composed of a primary apex, two mature internodes,and three branchlet whorls. A 10 min loading of the isolatedupper shoot with H¹⁴CO₃^– resulted in a greater accumulationof ¹⁴C in the apical complex and branchlets than in the internodes,while a subsequent 50 min chase with unlabelled solution inthe light resulted in a greater accumulation of ¹⁴C in internodesthan in other parts of the shoot. In the isolated internode-branchlet complex, when the apex wasnot detached, the amount of ¹⁴C transported from branchletsto internodes was about fives times that transported from internodesto branchlets. Removal of the apex resulted in a decrease intransport from branchlets to internodes and an increase in transportin the opposite direction. In an attempt to explain the mechanism of the polar transportof photosynthetically fixed carbon between branchlets and internodes,photosynthetic activities of both types of cells were investigated.Detached branchlets have higher photosynthetic ¹⁴C-fixationactivities than those of internodes. Chlorophyll contents, measuredin terms of surface area, in internodes and branchlets werealmost identical. The ribulose-l,5-bisphosphate carboxylase(RuBPCase) activity of branchlets was 1.6 times that of internodes,and the rate of ferricyanide-dependent evolution of oxygen inbranchlets was 1.4 times that in internodes. Key words: Chara, internode, branchlet, polar transport, photosynthesis     

Source-sink Characteristic of Photoassimilate Transport in Fertile and Sterile Plants of Chara vulgaris L.*

In cuttings of Chara vulgaris consisting of 3–4 internodal cells with whorls (branchlets) transport of photoassimilates was investigated by ¹⁴C-labelling. The distribution of radioactivity in different component parts of the plant was shown as percentage and calculated in relation to fresh weight. Sucrose was identified as the main transported carbohydrate. In fertile Ch. vulgaris plants, developing oogonia and antheridia were important sinks for transported carbohydrates. Polarity of transport in the internodal cells (nutrients and assimilates) remains constant, as the main characteristic, from base to apex in fertile as well as in sterile plants. In sterile plants, the branchlets being the main source for photoassimilates and the apex the sink, transport from branchlets is mainly in the direction of the internodes. In the case of fertile plants gametangia are additional sinks redirecting transport of assimilates into the branchlets.     

Culturing of stoneworts and submersed angiosperms with phosphate uptake exclusively from an artificial sediment

1. We aimed to demonstrate reproducible nutrition and growth of macrophytes in non‐axenic laboratory cultures preventing growth of phytoplankton and epiphytes. 2. Macrophyte shoot segments were planted in a mixture of commercial acid‐washed silica sand with crystalline tricalcium phosphate, and this artificial sediment was covered with a layer of pure silica sand. The liquid mineral media used did not contain phosphorus but were rich in all other nutrient elements. A CO₂ reservoir provided sustainable CO₂ supply to macrophyte cultures by gas diffusion through a polyethylene membrane. 3. Chara hispida, Chara tomentosa, Chara baltica, Elodea canadensis, Potamogeton pectinatus and Zanichellia palustris could be cultivated for long term without medium exchange and aeration. Microalgae growth was prevented by the absence of phosphate in the water column. Mobilisation of tricalcium phosphate and phosphate uptake by the rhizoids of C. hispida enabled sustainable rapid shoot growth and increased the concentration of inorganic phosphate in the shoot dry weight by five to six times in comparison with plants cultivated on pure silica sand. A significant growth support from tricalcium phosphate was also observed for E. canadensis, but the rate of phosphate uptake by the roots was not sufficient to maintain a storage pool of inorganic phosphate (P_i) in the growing shoots of this plant. 4. Membrane‐controlled CO₂ supply from a reservoir and artificial sediments like the one described provide attractive options for the laboratory culture of macrophytes.     

Intercellular Transport and Cytoplasmic Streaming in Chara hispida

The correlation between the velocities of cytoplasmic streamingand of translocation of ¹⁴C-photosynthate and ³²P-phosphateassociated radioactivity has been investigated in whole plantsof the green freshwater alga Chara hispida L. Tracer was suppliedto the plant's rhizoid system in a split-chamber. The velocityof cytoplasmic streaming of 52±3.3 µm s^–1compares with 57±10 µm s^–1 found for ¹⁴C-transportand 32±20 µm s^–1 found for ³²P-transport.There was no indication of intercellular translocation at avelocity faster than visible streaming. Cytochalasin B inhibitedthe translocation of ³²P and cytoplasmic streaming. CytochalasinB becomes fully effective in inhibiting streaming and transportafter an incubation time of at least 5 h. Key words: Chara hispida, Cytoplasmic streaming, Intercellular transport     

In vitro regeneration of plantlets in Brassica alboglabra

Different vegetative parts of Brassica alboglabra seedlings and mature plants were used as explants in culture.A high frequency (60–100%) of shoot regeneration was obtained from hypocotyl explants, nodal stem segments, internodal segments and shoot apices cultured on Murashige-Skoog basal medium. Addition of 6-benzylaminopurine and kinetin increased the average number of shoots per explant. When detached and transferred to basal medium, the shoots readily developed roots. Regenerated plantlets could be successfully transplanted in soil.     

Distribution of labelled assimilates within a young apple tree after supplying14CO2to a leaf or shoot

Labelled carbon dioxide was supplied for 22 hrs to a leaf of the leader or to the lateral shoot in two-year-old apple seedlings. The distribution of radioactive assimilates within the plant following this treatment was investigated by using radioautography. The transport of labelled assimilates from the young leaf of the leader was very meagre and affected only parts of the stem and the leaves situated in the close vicinity of the treated leaf. The¹⁴C-labelled assimilates from the mature leaf of the leader were transported in a considerable amount to the apex and to the other leaves of the leader. They were also found in an appreciable amount in the stem and the roots, as well as in some lateral shoots. After supplying¹⁴CO₂ to the lateral shoot remarkable transport of labelled assimilates was observed. Radioactivity was detected in the tip and in the youngest leaves of the leader, as well as in the roots. Their path in the stem was studied by dissecting the plant and examining the cross section from each internode. This method revealed that the assimilates from the treated leaf or shoot were transported downward only on one side of the stem in a helical pattern. The lateral shoots situated on the radioactive side of the stem were also labelled, whereas those situated on the opposite (non-radioactive) side were not labelled.     

Characterization of the light reaction in promoting the mobilizing ability of rose shoot tips

Mixed fluorescent and incandescent light increased growth and sink strength of the uppermost young shoot of rose plants (Rosa hybrida cv. Marimba) in comparison to pure fluorescent light. This was manifested by increased apical dominance. Monochromatic low-energy red light, given by means of optic fibers for 24 hours to shoot tips that had been previously darkened for 5 days, increased the transport of ¹⁴C-labeled assimilates to the intact tips and the uptake of [¹⁴C]sucrose by detached tips. Far-red had little or no effect, and blue was not effective at all in these reactions. Red light given directly to detached shoot tips, in vitro, increased the uptake of [¹⁴C]sucrose by the isolated tips. Adding far-red to the red greatly promoted the uptake, whereas blue and blue plus far-red were not active. The main character of the light reaction promoting sink activity in the shoot is that it is perceived by the shoot tip itself. It is operated by red light; far-red promotes the red effect but has little or no effect when alone. Light apparently promotes shoot sink activity by increasing the unloading process.     

Chara hispida beds as a sink of nitrogen: Evidence from growth,nitrogen uptake and decomposition

Chara hispida forms dense beds (0.78–0.95 kg DW m⁻²) in Colgada Lake. The ability of Chara meadows to act as a nitrogen source or sink was evaluated by the following methods: (1) investigating Chara growth, (2) nitrogen incorporation and decomposition laboratory experiments and (3) relating experimental results to field conditions. Sediment oospores were germinated in large aquaria and observed growth rates were 0.001 m day⁻¹ (shoot length) and 0.0002 g day⁻¹ (dry weight). Nitrogen uptake rates were determined by addition of K¹⁵NO₃ during two different periods of Chara growth and the rates were 1.21 and 3.86 μM g DW⁻¹ h⁻¹ when charophytes were 166 days old (not sexually mature) and 323 days old (sexually mature), respectively. After the uptake experiments, the same charophytes were allowed to decompose within two types of litter bags (3 mm-pore litter bags and entire, non-porous plastic litter bags). Decomposition rates of Ch. hispida were 0.016 and 0.009 day⁻¹ in perforated and non-perforated bags, respectively, and fit a negative exponential model. The nitrogen release rate, calculated as the disappearance of N content from Chara tissues, was 0.012 day⁻¹ and there were no statistically significant differences between the values from the two different bag types. The dissolved organic nitrogen concentrations in aquarium and non-perforated litter bags waters increased linearly with time due to the leaching of soluble compounds from Chara. The rate of N loss from Chara tissues, total nitrogen and dissolved organic nitrogen release rates and the decrease in initial dry weight rate were all lower than the daily rate of Chara N uptake. By extrapolating laboratory data to field situations, we determined that approximately 38% of the N taken up by charophytes in Colgada Lake during the growth period is retained. Given the high charophyte biomass in the lake, its ability to incorporate nitrogen, its low decomposition rate and its ability to over-winter, we conclude that Chara beds could be acting as nitrogen sinks in this ecosystem.     

Translocation of some assimilates from the sink to the donor in apple tree

The shoot apex or fruitlets of Jonathan apple trees grafted on M IX rootstock and grown in pots in a greenhouse were exposed to¹⁴CO₂ in an assimilation chamber. The translocation of¹⁴C-labelled assimilates from treated organs to other parts of the plant was studied. It was found that a very small amount of¹⁴C-labelled compounds was translocated from the shoot apex and very young fruitlets to the shoot stem. Preliminary chromatographic studies show that the chemical composition of the labelled substances detected below assimilation chamber differs profoundly from that of those remaining in the supplied leaves. The results support the view that there exists a translocation of some substances, possibly regulators from the sink to the donor.     

The Internode of Sambucus racemosa L. Originates from a Single Cell Layer

The internodes of Sambucus racemosa L. shoot apices originatefrom a single layer of cells in which mononucleate mother tannincells are present. The mother tannin cells increase in lengthand differentiate together with the internodal growth. Sambucus racemosa L., shoot apices, internode, mother tannin cells, tannin coenocytes     

Control of phosphate transport across the plasma membrane of Chara corallina

This paper examines the control of phosphate uptake into Chara corallina. Influxes of inorganic phosphate (Pi) into isolated single internodal cells were measured with ³²Pi. Pretreatment of cells without Pi for up to 10 d increased Pi influx. However, during this starvation the concentrations of Pi in both the cytoplasm and the vacuole remained quite constant. When cells were pre-treated with 0.1 mM Pi, the subsequent influx of Pi was low. Under these conditions the Pi concentrations in the cytoplasm was almost the same as that of Pi-starved cells, but vacuolar Pi increased with time. Transfer of cells from medium containing 0.1 mM Pi to Pi-free medium induced an increase of Pi influx within 3 d irrespective of the concentration of Pi in the vacuole.During Pi starvation, neither the membrane potential nor the cytoplasmic pH changed. Manipulation of the cytoplasmic pH by weak acids or ammonium decreased the Pi influx slightly.Pi efflux was also measured, using cells loaded with ³²Pi. Addition of a low concentration of Pi in the rinsing medium rapidly and temporarily induced an increase in the efflux.The results show that Pi influx is controlled by factors other than simple feedback from cytoplasmic or vacuolar Pi concentrations or thermodynamic driving forces for H⁺-coupled Pi uptake. It is suggested that uptake of Pi is controlled via the concentration of Pi in the external medium through induction or repression of two types of plasma membrane Pi transporters.Key words: Chara corallina, membrane transport, phosphate influx, phosphate starvation      

Decomposition of 14C-labelled lignin and phenols by a Nocardia sp.

A Gram-positive bacterium which was isolated from a Finnish soil and identified as a Nocardia sp., was able to decompose lignin and to assimilate lignin degradation products as a carbon source. It could release ¹⁴CO₂ from ¹⁴C-labelled methoxyl groups, side chains or ring carbons of coniferyl alcohol dehydropolymers (DHP) and from specifically ¹⁴C-labelled lignin of plant material. Furthermore, it could release ¹⁴CO₂ from phenolcarboxylic and cinnamic acids and alcohols labelled in the OCH₃, COOH groups, side chain or aromatic ring carbons.Non-Common Abbreviations Used DHP dehydropolymers of coniferyl alcohol     

Dipeptides and Amino Acids as Nitrogen Sources for the Callus of Atropa belladonna

Morphactin ester applied as a bark band to stems of Monterey pine seedlings (Pinus radiata D. Don) at first stimulated and then inhibited the rate of stem elongation over a 20-day period. The inhibition was caused by a direct action of morphactin on the shoot rather than by indirect action on the root system. ¹⁴C labelled morphactin applied to the bark accumulated in the shoot apices and the amount accumulated could be related to the inhibition of growth. More than 70% of labelled material isolated from roots and shoot apices was in the form of a metabolite similar to the carboxylic acid derivative of the morphactin ester. A preparation of ¹⁴C labelled acid derivative applied to the bark was translocated to the shoot apex and to the roots and inhibited stem elongation.     

Movement of Abscisic Acid in <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> Plants

THE movement of abscisic acid (ABA) in plants seems to have been studied only in isolated segments of tissue^1–4. We have used ¹⁴C-labelled ABA of relatively high specific activity to investigate its movement in a number of plant species, in both isolated tissue segments and whole plants. The movement of 2-¹⁴C-ABA in intact seedlings of Phaseolus vulgaris is described here.     

The response to NaCl of excised fully differentiated and differentiating tissues of the cultivated tomato, Lycopersicon esculentum, and its wild relatives, L. peruvianum and Solanum pennellii

The responses to NaCl of cultured leaf discs and leaflets derived from fully differentiated leaves and of shoot apices excised from the cultivated tomato Lycopersicon esculentum Mill. and its wild salt-tolerant relatives L. peruvianum (L.) Mill, and Solanum pennellii Cor were compared. The results suggest that the tolerance of the whole plant to salt depends largely on the tolerance of plant organs containing meristematic tissues rather than on tissues already differentiated. This suggestion is based on the positive correlation found between the response to NaCl of shoot apices and of the whole plant, i.e. both whole plants and apices of the wild species were more resistant to salt than those of the cultivated species. No difference was found among the species with respect to the responses of the fully differentiated parts. The ion balance (K⁺/Na⁺ and Cl⁻/Na⁺) in detached leaves and apices exposed to salt was different from the balance in the same parts while attached to the salt-treated plant. This difference may be due to the severance of the excised parts from the major sites controlling the balance of ions in the whole plant.     

Effects of shoot bending of apple trees on accumulation and translocation of14C-labelled assimilates

The translocation of¹⁴C-labelled assimilates from a single leaf in bent and intact apple shoots was studied in varying stages of shoot development. In actively growing shoots¹⁴C-labelled assimilates translocated from the treated leaf and accumulated mainly in the shoot apex. In moderately growing apple shoots radioactive assimilates were translocated from the treated leaf in both directions towards and down the shoot. In apple shoots showing only slight growth activity the¹⁴C-labelled assimilates were transported from the treated leaf mainly to the base of the shoot, stem and roots. Bending shoots changed the pattern of distribution of radioactive assimilates. Bending actively and moderately growing shoots resulted in higher concentration of 14-carbon in the shoot apex than in controls. In slowly growing and non-growing apple shoots bending caused a higher accumulation of radioactive assimilates in the bent section than in an equal section of control shoots.     

Short-term Measurements of the Cytoplasmic pH of Chara corallina Derived from the Intracellular Equilibration of 5,5-Dimethyloxazolidine-2,4-Dione (DMO)

Smith, F. A. 1986. Short-term measurements of the cytoplasmicpH of Chara corallina derived from the intracellular equilibrationof 5,5-dimethyloxazolidine-2,4-dione (DMO).—J. exp. Bot.37: 1733–1745. Measurements of the time-course of influx of ¹⁴C-labelled 5,5-dimethyloxazolidine-2,4-dione(DMO) into the cytoplasm and vacuole of internodal cells ofChara corallina, and of efflux of DMO into non-radioactive solutions,have shown that exchange of DMO across the tonoplast is veryrapid compared with exchange across the plasma membrane. Thishas made possible calculations of cytoplasmic pH from distributionof DMO between cytoplasm and vacuole over short periods (5 or10 min) even when intracellular DMO is not at flux equilibriumwith external DMO. Using this new method, estimates have beenmade of the rates and magnitude of: (i) acidification of thecytoplasm caused by acidic growth regulators (IAA and NAA) andby metabolic inhibitors (azide, DNP, CCCP and DCMU), and (ii)alkalinization caused by uptake of ammonium and methylammoniumions. The potential application of the method to future studiesof membrane transport in charophyte cells is assessed. Key words: Charophyles, cytoplasmic pH.