The Assimilation of Nitrate-nitrogen by Nitrogen-starved Cells of Platymonas (Tetraselmis) striata (Prasinophyceae)

Nitrate refeeding of nitrogen-starved cells of Platymonas striataresulted in approximately a doubling of average cellular nitrogenwithin 24 h. All the nitrate-nitrogen removed from the culturemedium could be accounted for as non-nitrate nitrogen withincells. Thus no significantly sized nitrate pool existed in Platymonasstriata and no assimilated nitrogen was lost from the cellsto the medium over the 48 h period studied. The slight fallin average cellular nitrogen which occurred from 24 to 36 hcould be attributed to cell division. The majority (70–80per cent) of the assimilated nitrate was recovered in the trichloroaceticacid (TCA)-insoluble fraction. There was some increase in thepercentage of nitrogen found in the TCA-soluble fraction duringthe period of most rapid nitrate assimilation (0–24 h).This presumably reflects an inability of the cells to assimilatelow-molecular-weight metabolic intermediates into macromoleculesat the same rate at which they were being formed. The majorityof the TCA-soluble fraction could be accounted for in termsof amino acids, purine and pyrimidine bases and ammonia. Cellswith adequate nitrogen nutrition seemed to maintain amino acidand purine + pyrimidine base nitrogen pools of about 0.8–0.9and 0.3–0.4 pg per average cell respectively. Algal amino acids, algal purine and pyrimidine bases, algal ammonia     

Studies on ammonium-assimilating enzymes of Platymonas striata Butcher (Prasinophyceae)

Platymonas striata Butcher displays significant levels of glutamate synthase (GS) (EC 2.6.1.53) and glutamine synthetase (GOGAT) (EC 6.3.1.2.), but very low levels of glutamate dehydrogenase (GDH) (EC 1.4.1.4). This suggests that the GS/GOGAT pathway is important for nitrogen assimilation. The in vitro rates of enzyme activity can however only account for about 10% of the in vivo rates of nitrogen assimilation. Nitrogen-starvation reduced GS activity to undetectable levels. On nitrate or ammonium ion refeeding the cellular GS activity was rapidly restored, and reached levels of 56% and 91% greater than the unstarved values 24h after refeeding nitrate or ammonium respectively.Abbreviations NAR nitrate reductase - NIR nitrate reductase     

Effect of Source of Nitrogen on the Growth of Fiskeby Soya Bean: the Carbon Economy of Whole Plants

Fiskeby V soya bean was grown from seed germination to seedmaturation with two contrasting patterns of nitrogen metabolism:either wholly dependent on dinitrogen fixation, or with an abundantsupply of nitrate nitrogen, but lacking root nodules. The carbonand nitrogen economies of the plants were assessed at frequentintervals by measurements of photosynthesis, shoot and rootrespiration, and organic and inorganic nitrogen contents. Plantsfixing atmospheric nitrogen assimilated only 25–30 percent as much nitrogen as equivalent plants given nitrate nitrogen:c. 40 per cent of the nitrogen of ‘nitrate’ plantswas assimilated after dinitrogen fixation had ceased in ‘nodulated’plants. The rates of photosynthesis and respiration of the shootsof soya bean were not markedly affected by source of nitrogen;in contrast, the roots of ‘nodulated’ plants respiredtwice as rapidly during intense dinitrogen fixation as thoseof ‘nitrate’ plants. The magnitude of this respiratoryburden was calculated to increase the daily whole-plant respiratory loss of assimilate by 10–15 per cent over thatof plants receiving abundant nitrate. It is concluded that ‘nodulated’plants grew more slowly than ‘nitrate’ plants inthese experiments for at least two reasons: firstly, the symbioticassociation fixed insufficient nitrogen for optimum growth and,secondly, the assimila tion of the nitrogen which was fixedin the root nodules was more energy-demanding in terms of assimilatethan that of plants which assimilated nitrogen by reducing nitratein their leaves.     

The effect of nitrogen refeeding on the carbohydrate content into nitrogen-starved cells of Platymonas striata Butcher

Studies on the mean cellular carbohydrate contents of Platymonas striata Butcher under conditions of nitrogen-starvation, and after refeeding these starved cultures with either nitrate or ammonium ions (growing under continuous illumination or with an alternating light/dark regime) have shown that nitrogen-starved cells accumulated abnormal amounts of cellular carbohydrate and that nitrogen refeeding produced a marked drop in the cellular carbohydrate. Cells grown in a light/dark regime accumulated less carbohydrates than those grown in continuous light. The mean cellular carbohydrate levels 16 h after nitrogen refeeding were still much in excess of those of cells grown with normal nutrition. It was therefore suggested that the differences in nitrogen uptakes in this period — when comparing either the uptake of cells grown in continuous light with that of cells grown in a light/dark regime; or when comparing the uptakes of cells presented with either nitrate or ammonium ions and grown in a light/dark regime —cannot be directly due to shortages of carbohydrate for the provision of carbon skeletons for nitrogen assimilation.     

Carbon and Nitrogen Assimilation by Vicia faba L. at Low Temperature: the Importance of Concentration and Form of Applied-N

Low temperature (6 C) growth was examined in two cultivarsof Vicia faba L. supplied with 4 and 20 mol m^–3 N as nitrateor urea. Both cultivars showed similar growth responses to increasedapplied-N concentration regardless of N-form. Total leaf areaincreased, as did root, stem and leaf dry weight, total carboncontent and total nitrogen content. In contrast to findingsat higher growth temperatures, 20 mol m^–3 urea-N gavesubstantially greater growth (all parameters measured) than20 mol m^–3 nitrate-N. The increased carbon content per plant associated with increasedapplied nitrate or urea concentration, or with urea in comparisonto nitrate, was due to a greater leaf area per plant for CO₂uptake and not an increased CO₂, uptake per unit area, carbon,chlorophyll or dry weight, all of which either remained constantor decreased. Nitrate reductase activity was substantial inplants given nitrate but negligible in plants given urea. Neitherfree nitrate nor free urea contributed greatly to nitrogen levelsin plant tissues. It is concluded that there is no evidence for a restrictionin nitrate reduction at 6 C, and it is likely that urea givesgreater growth than nitrate because of greater rates of uptake. Vicia faba, broad bean, low temperature growth, carbon assimilation, nitrogen assimilation     

Nitrogen Utilization in N-limited Barley During Vegetative and Generative Growth: III. POST-ANTHESIS KINETICS OF NET NITRATE UPTAKE AND THE ROLE OF THE RELATIVE ROOT SIZE IN DETERMINING THE CAPACITY FOR NITRATE ACQUISITION

Barley (Hordeum vulgare L., cvs Golf, Mette, and Laevigatum)was grown under nitrogen limitation in solution culture untilnear maturity. Three different nitrogen addition regimes wereused: in the ‘HN’ culture the relative rate of nitrate-Naddition (RA) was 0·08 d^–1 until day 48 and thendecreased stepwise to, finally, 0·005 d^–1 duringgrain-filling; the ‘LN’ culture received 45% ofthe nitrogen added in HN; the ‘CN’ culture was maintainedat RA 0·0375 d^–1 throughout. Kinetics of net nitrateuptake were measured during ontogeny at 30 to 150 mmol m^–3external nitrate. V_max (which is argued to reflect the maximuminflux rate in these plants) declined with age in both HN andLN cultures. A pronounced transient drop was observed just beforeanthesis, which correlated in time with a peak in root nitrateconcentration. Similar, but less pronounced, trends were observedin CN. The relative V_max (unit nitrogen taken up per unit nitrogenin plants and day) in all three cultures declined from 1·3–2·3d^–1 during vegetative growth to 0·1–0·7d^–1 during generative growth. These values are in HN andLN cultures 15- to more than 100-fold in excess of the demandset by growth rates throughout ontogeny. Predicted balancingnitrate concentrations (defined as the nitrate concentrationrequired to support the observed rate of growth) were below6·0 mmol m^–3 in HN and LN cultures before anthesisand then decreased during ontogeny. In CN cultures the balancingnitrate concentration increased during grain-filling. Apartfrom the transient decline during anthesis, most of the effectof ageing on relative V_max can be explained in terms of reducedcontribution of roots to total biomass (R:T). The loss in uptakeper unit root weight is largely compensated for by the declinewith time in average tissue nitrogen concentrations. The quantitativerelationships between relative V_max and R:T in ageing plantsare similar to those observed for vegetative plants culturedat different RAs. The data support the contention that the capacity for nitrateacquisition in N-limited plants is under general growth control,rather than controlled by specific regulation of the biochemicalpathway of nitrate assimilation. Key words: Barley, nitrogen concentration, root: total plant biomass ratio, V_max     

Uptake of 13C and 15N (ammonium, nitrate and urea) by Microcystis in Lake Kasumigaura

The uptake rate of carbon and nitrogen (ammonium, nitrate andurea) by the Microcystis predominating among phytoplankton wasinvestigated in the summer of 1984 in Takahamaira Bay of LakeKasumigaura. The V_max values of Microcystis for nitrate (0.025–0.046h^–1) and ammonium (0.15–0.17 h^–1) were considerablyhigher than other natural phytoplankton. The ammonium, nitrateand urea uptake by Microcystis was light dependent and was notinhibited with nigh light intensity. The K₁ values were farlower than the I_k values. The carbon uptake was not influencedby nitrogen enrichment. Microcystis accelerated the uptake rateby changing V_max/K _s value when nitrogen versus carbon contentin cells declined. Nitrate was scarcely existent in TakahamairiBay during the summer, when Microcystis usually used ammoniumas the nitrogen source. However, the standing stock of ammoniumin the water was far lower than the daily ammonium uptake rates.Therefore, the ammonium in this water had to be supplied becauseof its rapid turn-over time (–0.7–2.6 h).     

Nitrogenous nutrient uptake by phytoplankton and ammonium regeneration by microbial assemblage in Lake Biwa

In situ rates of nitrate, ammoniwn and urea uptake by the phytoplanktonassemblage, and the regeneration rate of ammonium by the microbialassemblage, in Lake Biwa were measured using the nitrogen 15tracer method from 1985 to 1987. The rate of total nitrogen(sum of ammonium, nitrate and urea) uptake was in the rangeof 62–594 ng N^–1 r^–1 h^–1. The percentagecontribution of ammonium uptake was 41–92%, that of urea4–58% and that of nitrate <1–28% of total uptake.The annual mean new production which was supported by nitrateuptake was 18% of the total production in 1986. The phytoplanktonassemblage in Lake Biwa preferentially utilized regeneratednitrogen, such as ammonium and urea, whose concentration wasmuch lower than that of nitrate throughout the observation penodwithout in summer. The in situ nitrogen uptake rate was almostsufficient to meet the nitrogen requirement of the phytoplanktonassemblage, except in midsummer when the nitrate concentrationwas below the detection limit of 0.3 µg N r^–1. Inthe trophogemc layer, the rate of ammonium regeneration was66–272 ng N 1^–1 h^–1 Although the ambient ammoniumconcentration in the trophogenic layer was maintained at aroundthe half-saturation constant for ammonium uptake kinetics, theammomum uptake rates were always highly correlated with ammoniumregeneration rates. From the size fractionation experimentsand estimates from the literature, it was suggested that themicrobial assemblage <1 µm may have been the most importantagent responsible for the ammonium regeneration processes inthe trophogenic layer.     

Seasonal nitrogen assimilation and carbon fixation in a fjordic sea loch

Carbon (C) fixation and nitrogen (N) assimilation rates havebeen estimated from ¹⁴C and ¹⁵N techniques for a 12 month periodin a Scottish sea loch. The maximum rate of nitrogen assimilated(29.92 mmol N m^–2 day^–1) was in April at the mostseaward station; similar high rates were experienced duringMay at the other stations. Carbon fixation rates were maximal(488–4047 mg C m^–2day^–1) at the time of highphytoplankton biomass (maximum 8.3 mg m^–3 chlorophylla) during May, whilst nitrate concentrations remained >0.7µ.mol l^–1. C:N assimilation ratios suggest nitrogenlimitation only during the peak of the spring bloom, althoughat times nitrogen (nitrate and ammonium) concentration fellto 0.2 µmol l^–1 in the following months. The verticalstability of the water column, influenced by tidal and riverineflushing, varied along the axis of the loch, resulting in markeddifferences between sampling stations. Although ammonium waspreferentially assimilated by phytoplankton, >50% of productionwas supported by nitrate uptake and only during the summer monthswas the assimilation of ammonium quantitatively important.     

Nitrogen Assimilation in Barley (Hordeum vulgare L. cv. Mazurka) in Response to Nitrate and Ammonium Nutrition

Barley plants (Hordeum vulgare L. cv. Mazurka) were grown inaerated solution cultures with 2 mM or 8 mM inorganic nitrogensupplied as nitrate alone, ammonium alone or 1:1 nitrate+ammonium.Activities of the principal inorganic nitrogen assimilatoryenzymes and nitrogen transport were measured. Activities ofnitrate and nitrite reductases, glutamine synthetase and glutamatesynthase were greater in leaves than in roots but glutamatedehydrogenase was most active in roots. Only nitrate and nitritereductases changed notably (4–10 times) in response tothe different nitrogen treatments. Nitrate reductase appearedto be rate-limiting for nitrate assimilation to glutamate inroots and also in leaves, where its total in vitro activitywas closely related to nitrate flux in the xylem sap and wasslightly in excess of that needed to reduce the transportednitrate. Xylem nitrate concentration was 13 times greater thanthat in the nutrient solution. Ammonium nitrogen was assimilatedalmost completely in the roots and the small amount releasedinto the xylem sap was similar for the nitrate and the ammoniumtreatments. The presence of ammonium in the nutrient decreasedboth export of nitrate to the xylem and its accumulation inleaves and roots. Nitrate was stored in stem bases and was releasedto the xylem and thence to the leaves during nitrogen starvation.In these experiments, ammonium was assimilated principally inthe roots and nitrate in the leaves. Any advantage of this divisionof function may depend partly on total conversion of inorganicnitrogen to amino acids when nitrate and ammonium are givenin optimal concentrations. Hordeum vulgare L., barley, nitrate, ammonium, nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, glutamate dehydrogenase, nitrogen transport     

CAM Photosynthesis under Drought Conditions in Kalanchoe blossfeldiana Grown with Nitrate or Ammonium as the Sole Nitrogen Source

K. blossfeldiana Poelln. cv. Hikan was grown in vermiculite,supplied daily with nutrient solution containing 1 mM (or 10mM) nitrate or ammonium as the sole nitrogen source. The nitrate-grownplants had more activity of CAM (Crassulacean acid metabolism)photosynthesis (nocturnal CO₂ uptake in the shoot and nocturnalincreases of titratable acidity and malate content in the leaves)than the ammonium-grown plants. Interruption of the solutionsupply for 5 or more days (drought conditions) increased theactivity of CAM photosynthesis in nitrate- or ammonium-grownplants, and the diurnal CO₂ uptake pattern in the nitrate-grownplants shifted from ‘weak-CAM’ to ‘full-CAM’.The difference in the activity of CAM photosynthesis betweennitrate- and ammonium-grown plants increased under the droughtconditions. When the solution was resupplied, the activity ofCAM photosynthesis rapidly decreased to the levels before theinterruption. The physiological mechanism and ecological significanceof the effect of the nitrogen source on CAM photosynthesis arediscussed (Received January 5, 1988; Accepted April 13, 1988)     

The Nitrogen Content of Plants and the Self-thinning Rule of Plant Ecology: A Test of the Core-skin Hypothesis

The ‘core-skin’ hypothesis postulates that secondarilythickened plants behave energetically as an inert ‘core’covered by an active ‘skin’, the ‘skin’being two-imensional, the ‘core’ three-dimensional.This would explain the ‘self-thinning ‘or‘–3/2’ rule of plant ecology, that is, the tendencyfor log (dry weight per plant) and log (number of plants perunit area) to progress along a straight line relationship, withslope = – 3/2’. The hypothesis was tested as follows. Plant nitrogen contentwas used as an estimate of the mass of ‘skin’ perplant, and dry weight as an estimate of the mass of the ‘core’.As plants mature the slope of the relationship between y = log(mass of nitrogen per plant) and x = log (mass of dry matterper plant) is expected to decline from an initial value of 1.0towards a final value of 0.66. The intercept of the relationshipis expected to reflect the intrinsic content of ‘skin’per unit of ‘core’. Genotypic variation in thisparameter should cause genotypic differences in the maximumattainable yield of biomass per unit area. The expectations were investigated by fitting the function y= p+qx+r exp – x to 30 sets of data on plant nitrogencontent, plant weight and time in 18 different vegetables. Simplelinear regressions of y on x were fitted to more limited setsof data on weights and nitrogen contents of mature trees. Theexpectations were, with some minor exceptions, confirmed. Nitrogen, yield, plant competition, self-thinning     

Heterotrophic growth and nutritional aspects of the diatom Cyclotella cryptica (Bacillariophyceae): effect of nitrogen source and concentration

To investigate the nutritional value of the diatom Cyclotella cryptica Reimann, Lewin, and Guillard as an alternative feed for the use in the aquaculture industry, the heterotrophic growth characteristics, total fatty acids, and the resultant fatty acid profile of the microalga were studied when cultivated with sodium nitrate, ammonium chloride, or urea. All three nitrogen sources supported growth under heterotrophic conditions, and their uptake affected the pH of the cultivation medium, even when buffered. The use of sodium nitrate or urea resulted in a significant increase in the pH of the cultivation media, whereas the use of ammonium chloride caused a minor decrease in the pH of the cultivation media. The maximum specific growth rate was highest when urea and ammonium chloride were supplied at a low concentration; however, the total fatty acid content was not significantly affected (P = 0.101) by the nitrogen source when supplied at 10.7 mM nitrogen. The total fatty acid content and fatty acid profile of C. cryptica was more affected by the growth phase (predominately influenced by the initial nitrogen concentration) than by the source of nitrogen.     

Relation between Nitrogen Status, Carbohydrate Distribution and Subsequent Rooting of Chrysanthemum Cuttings as Affected by Pre-harvest Nitrogen Supply and Cold-storage

This study investigated the relationship between internal nitrogenand carbohydrate distribution in chrysanthemum cuttings of twocultivars (‘Puma’, ‘Cassa’) when affectedby nitrogen supply to stock plants (0.6, 1.5, or 4.0 g N m^-2week^-1)and different periods (2, 3, or 4 weeks) of dark cold-storage(0.5 or 5°C), and adventitious rooting. Concentrations oftotal nitrogen (N_t) and nitrate in cuttings and the levels ofsugars, starch and fructan in different cutting parts (leaves,upper stem, and basal stem) were studied in relation to subsequentadventitious rooting at natural radiation in a greenhouse. Increasingnitrogen supply resulted in substantially lower starch levelsand higher sucrose concentrations in leaves when cuttings wereexcised. Fructan concentrations were low and decreased withincreasing nitrogen levels. Starch completely disappeared fromleaves and to a large extent from stems within the shorteststorage period. A less pronounced decrease in sugar concentrationwas observed, particularly in low-nitrogen cuttings and thecuttings of ‘Puma’. The number and length of adventitiousroots subsequently formed by unstored and stored cuttings waspositively correlated with initial N_t, and to a lesser extentwith initial nitrate concentrations in cuttings. Whereas rootingwas not limited by pre-rooting concentrations of carbohydratesin the different cutting parts, the generally higher rootingcapability of nitrogen-rich cuttings, a stronger nitrogen responseof ‘Cassa’, and increased rooting at a particularharvest date, were associated with higher sucrose:starch ratiosin leaves at harvest. This reflected an increased assimilateexport. By using this characteristic in a linear regressionmodel, total variability of root numbers, ranging from three–35per cutting, could be predicted to 57% for the unstored andto 40% for all cuttings. Increased basipetal transport of carbohydrates,of nitrogen compounds, and of auxins may be causally involvedin these associations. Copyright 2000 Annals of Botany Company Adventitious rooting, nitrogen, sugars, carbohydrates, source-sink, partitioning, quality, storage, cuttings, stock plants, chrysanthemum, Dendranthema grandiflorum     

Active Iron in Plant Leaves

Tomato plants were grown with limited or luxury water supplyand given nitrogen either as the ammonium ion or the nitrateion. The ‘active’ iron fractions of the leaves asextracted by etherized tenth molar hydrochloric acid showedno relationship with the total iron but a very significant linearrelationship was found between the active iron fraction andthe ratio of total phosphorus to total iron. Recalculation of results presented by other investigators formaize plants grown in soil with various levels of moisture,phosphorus and iron also showed a highly significant linearrelationship between the active iron fraction and the ratioof total phosphorus to total iron. Lycopersicon esculentum L., tomato, ‘active iron’, phosphorus, Zea mays     

Influence of Flooding on Net CO2Assimilation, Growth and Stem Anatomy of Annona Species

A series of experiments was conducted to assess net CO₂assimilationand growth responses to waterlogging of grafted and seedlingtrees in the genus Annona. Seedlings of A. glabra, A. muricataandA. squamosa L., and scions of ‘Gefner’ atemoya(A. squamosaxA. cherimola Mill.), ‘49-11’ (‘Gefner’atemoyaxA. reticulata L.), ‘4-5’ (‘Priestley’atemoyaxA. reticulata), A. reticulata grafted onto either A.glabra, A. reticulata orA. squamosa rootstocks were floodedfor up to 60 d. Soil anaerobiosis occurred on the third dayof flooding. Seedlings ofA. glabra and A. muricata, and thescions ‘49-11’, ‘Gefner’ atemoya, andA. reticulata grafted onto A. glabra rootstock were consideredflood tolerant based on their ability to survive and grow inflooded conditions. Scions of the normally flood-sensitive A.reticulata, ‘Gefner’ atemoya, and ‘49-11’tolerated root waterlogging when grafted onto the flood-tolerantspecies, A. glabra. In contrast, flooding of A. squamosa seedlingsand rootstocks, and A. reticulata rootstocks greatly reducedgrowth and net CO₂assimilation rates, and resulted in 20–80%tree mortality. Stem anatomical responses to long-term flooding(12 continuous months) were assessed in seedlings of A. glabraand A. muricata, and trees of ‘49-11’ grafted ontoA. glabra. Flooded trees developed hypertrophied stem lenticels,particularly in A. glabra, and enlarged xylem cells resultingin thicker stems with reduced xylem density. Flooding did notincrease air spaces in pre-existing xylem near the pith or inxylem tissue that was formed during flooding. Thus, flood tolerancedid not involve aerenchyma formation in the stem. Copyright1999 Annals of Botany Company Flood tolerance, net CO₂assimilation, photosynthesis, stem anatomy, shoot growth, anaerobiosis, Annonaceae.     

Leaf and Tiller Production of Prairie Grass (Bromus willdenowii Kunth) and Two Ryegrass (Lolium) Species

A detailed morphological study of three prairie grass cultivars(Bromus willdenowii Kunth) was conducted under ‘vegetative’and ‘reproductive’ growth conditions (short andlong photoperiods) and at different temperatures. Perennialryegrass (Lolium perenne L.) and Westerwolds ryegrass (Loliummuhiflorum Lam.) were compared during vegetative growth. Prairie grass had higher leaf appearance rates (leaves per tillerper day) and lower site filling (tillers per tiller per leafappearance interval) than the ryegrass species. Tillering rates(tillers per tiller per day) were also lower, except under vegetativeconditions at 4C. Low tiller number in prairie grass was notdue to lack of tiller sites but a result of poor filling ofthese sites. Lower site filling occurred because of increaseddelays in appearance of the youngest axillary tiller and lackof axillary tillers emerging from basal tiller buds. In prairiegrass, no tillers came from coleoptile buds while only occasionallydid prophyll buds develop tillers. Low tiller number in prairiegrass was compensated for by greater tiller weight. Prairiegrass had more live leaves per tiller, greater area per leafand a high leaf area per plant. Considerable variation between cultivars was found in prairiegrass. The cultivar ‘Bellegarde’ had high leaf appearance,large leaves and rapid reproductive development, but had lowlevels of site filling, tillering rate, final tiller numberand herbage quality during reproductive growth. ‘Primabel’tended to have the opposite levels for these parameters, while‘Grasslands Matua’ was intermediate and possiblyprovided the best balance of all plant parameters. prairie grass, Bromus willdenowii Kunth, perennial ryegrass, Lolium perenne L., Westerwolds ryegrass, Lolium multiflorum Lam., temperature, photoperiod, leaf appearance, leaf area, tillering, site filling, tillering sites, yield     

Factors Affecting the Abscission of Reproductive Organs in Yellow Lupins (Lupinus luteus L.): III. ENDOGENOUS GROWTH SUBSTANCES IN VIRUS-INFECTED AND HEALTHY PLANTS AND THEIR EFFECT ON ABSCISSION

Extracts of small and mature-size lupin pods yielded four substancesaffecting the growth of wheat-coleoptile sections: one acidpromotor (A), two acid inhibitors(B and X), and one neutralinhibitor(Y). Inhibitor B was extremely active, however, coleoptile sectionsshowed no signs of toxic effects; they resumed growth at a rapidrate after rinsing them and adding ß-indolylaceticand (IAA) to the medium. 1 µg of IAA was required to counteractthe effect of ‘B’ extracted from 230 mg. Of tissue.On an equal fresh weight basis the inhibiting action of ‘B’in lupin pods was 500–1,500 times more potent than thatof ‘inhibitor ß’ in etiolated pea seedlings. Small pods of plants infected with pea-mosaic virus yielded3 times the amount of ‘A’ of healthy plants (equivalentto 1 µg. IAA 0.3 µg. IAA per 25 g. of tissue respectively),and approximately the amount of ‘B’. Mature podsof virus-infected plants again yielded more‘A’,but also 2? times more ‘B’ than pods of healthyplants. Healthy pods yielded more ‘A’ than virus-infectedpods, and there was no difference in ‘X’. A lupin abscission test was developed and the effects of proximaland distal application of -naphthyl acetic acid (NAA) are presented,and discussed with respect to results of other abscission tests. ‘A’ accelerated abscission when applied proximally,and delayed or prevented it when applied distally. ‘B’strongly accelerated abscission when applied in either way.A possible mechanism explaining the abscission-inducing effectof developing pods on later flowers is discussed in terms ofthe substances ‘A’ and ‘B’. The partlyprevented abscission observed on virus-infected plants was foundto agree well with the proposed mechanism.     

The Influence of the Plant Growth-regulator, 2-Methyl-4-Chlorophenoxyacetic Acid, on the Metabolism of Carbohydrate, Nitrogen and Minerals in Solanum lycopersicum (Tomato)

The effects of the foliar application of phytocidal concentrationsof 2-methyl-4-chlorophenoxyacetic acid (MCPA) on change in totaldry weight, and in ‘available carbohydrate’ (starch,‘total’ and ‘reducing’ sugars), totalnitrogen, phosphorus, potassium, calcium, and magnesium of ‘tops’and roots of tomato plants have been followed over a periodof 14 days following spraying. There were two main treatments—‘nutrient’(nutrient supply to roots continued after spraying) and ‘water’(distilled water only supplied to roots after spraying) and‘water’ (distilled water only supplied to rootsafter spraying)—the sub-treatments consisting of ‘MCPA’versus ‘no-MCPA’ for each of the main treatments.Twelve different times of sampling were used. In analysing the present data, the quantity ‘residualdry weight’ (total dry weight less ‘available carbohydrate’),which was originally introduced by Mason and Maskell as a basisof reference for analyses of plant organs in short-period experimentsnot involving appreciable growth, has been used as an estimateof the permanent structure of plant growth. This new use ofthe ‘residual dry weight’ basis has brought outimportant features which were obscured when the data were leftin their primary form (as percentages of total dry weight oramounts per plant). Growth, as measured by increase in ‘residual dry weight’,was greatly inhibited by 2-methyl-4-chlorophenoxyacetic acidshortly after spraying, in both the presence and the absenceof nutrient. In the presence of 2-methyl-4-chlorophenoxyacetic acid, netassimilation rate (estimated as rate of increase in total dryweight per gram ‘residual dry weight’ of the ‘tops’)was greatly diminished while uptake of total nitrogen and ofP₂O₅ (estimated as increase in total nitrogen or of P₂O₅ ofthe whole plant per day per 1 g. ‘residual dry weight’of the roots) appeared to undergo a similar but much smallerdiminution. It seemed probable, however, that in the presenceof MCPA a larger proportion of the carbohydrate actually formedwas utilized for synthesis of aminoacids and protein. In the plant as a whole there was no evidence of actual depletionof ‘available carbohydrate’ as a result of MCPAtreatment, this fraction showing a steady increase in all treatmentsthroughout the experiment. The rate of increase was, however,much reduced by MCPA treatment. The ‘tops’ presentedmuch the same picture as the whole plant, but for the rootsthe situation was quite different. While the roots of the ‘no-MCPA’plants and also of the ‘MCPA-water’ plants showeda steady increase in available carbohydrate, those of the ‘MCPA-nutrient’plants rose only very slightly (from the initial value of 8mg. per plant to about 10 mg.) during the first 2 days, andthen in the next 2 days declined to a value (about 6 mg.) belowthe initial and remained at this low level for the rest of theexperiment. It is suggested that the phytocidal effect of 2-methyl-4-chlorophenoxyaceticacid in the presence of nutrient may be due to depletion ofthe ‘available carbohydrate’ supplies in the roots,which is shown to be brought about, in part, by reduced transportfrom the tops, and partly by the relatively greater utilizationof the carbohydrate present. These results offer an explanationfor the facts that plants showing vigorous growth are more easilykilled by MCPA and that perennial plants, particularly thosewith storage tissues in their roots, are more resistant. Further,they suggest the useful practical application that MCPA treatmentshould be given when the carbohydrate reserves of the rootsare at a minimum. For perennial plants, conditions might beexpected to be optimal for the application of MCPA in late spring,at a time when the first ‘flush’ of growth is slowingdown and before any appreciable new reserves of carbohydratehave been accumulated. It was also shown that 2-methyl-4-chlorophenoxyacetic acid preventedthe net synthesis of starch, but still permitted an appreciablenet formation of sucrose. 2-methyl-4-chlorophenoxyacetic acid appeared to have no effecton the uptake of potassium, calcium, or of magnesium. The lackof effect on potassium is contrasted with the previous observationby Rhodes, Templeman, and Thruston (1950) that sub-lethal concentrationsof MCPA, applied over a relatively long period to the rootsof tomato plants, specifically depressed the uptake of potassium.     

Rapid and slow modulation of nitrate reductase activity in the red macroalga <Emphasis Type="Italic">Gracilaria chilensis</Emphasis> (Gracilariales,Rhodophyta): influence of different nitrogen sources

Nitrate is one of the most important stimuli in nitrate reductase (NR) induction, while ammonium is usually an inhibitor. We evaluated the influence of nitrate, ammonium or urea as nitrogen sources on NR activity of the agarophyte Gracilaria chilensis. The addition of nitrate rapidly (2 min) induced NR activity, suggesting a fast post-translational regulation. In contrast, nitrate addition to starved algae stimulated rapid nitrate uptake without a concomitant induction of NR activity. These results show that in the absence of nitrate, NR activity is negatively affected, while the nitrate uptake system is active and ready to operate as soon as nitrate is available in the external medium, indicating that nitrate uptake and assimilation are differentially regulated. The addition of ammonium or urea as nitrogen sources stimulated NR activity after 24 h, different from that observed for other algae. However, a decrease in NR activity was observed after the third day under ammonium or urea. During the dark phase, G. chilensis NR activity was low when compared to the light phase. A light pulse of 15 min during the dark phase induced NR activity 1.5-fold suggesting also fast post-translational regulation. Nitrate reductase regulation by phosphorylation and dephosphorylation, and by protein synthesis and degradation, were evaluated using inhibitors. The results obtained for G. chilensis show a post-translational regulation as a rapid response mechanism by phosphorylation and dephosphorylation, and a slower mechanism by regulation of RNA synthesis coupled to de novo NR protein synthesis.