In vivo Nitrate Reductase Activity in Barley (Hordeum vulgare L.) during Ear Development

Experiments were conducted during the 1974–75 and 1975–76winter season with the barley (Hordeum vulgare L.) cultivarJyoti. From amongst the various plant parts, the flag leaf bladehad higher in vivo nitrate reductase (NR) activity than thelower two leaf blades, glumes, and grains. However, the potentialof a plant part to reduce NO₃^– is a function of its freshweight and the NR per unit fresh weight. On this basis, thesecond and third leaf blades could reduce more NO₃^– thanthe flag leaf blade. N fertilizer application resulted in enhancementof the activity of the leaf blades alone. N fertilizer appliedduring the reproductive phase was taken up and assimilated bythe various plant parts. The studies suggest that, even whenthe fertilizer is applied at optimum levels for obtaining maximumyields, the upper leaf blades have sub-optimal NR activity andthat there is a likelihood of either a preferential flow ofNO₃^– to the leaf blades or transnational barriers to NO₃^–movement to the ear.     

Response of Melanthera biflora to Salinity and Water Stress

Melanthera biflora (Asteraceae) is a moderately salt-tolerantplant from the Indo-Pacific region. In laboratory studies itsgrowth was inhibited by salt above 50 mol m^–3, but itwas able to survive salinities approaching that of seawater,namely 400 mol m^–3. Shoot potassium concentrations weremaintained over a range of salinities up to 400 mol m^–3,while sodium and chloride accumulation followed closely theincrease in external osmotic pressure. In contrast, the increasein osmotic pressure of the leaf sap of Melanthera biflora, subjectedto water stress, was due mainly to a decrease in the ratio offresh weight/dry weight. 3-dimethylsulphoniopropionate (3-DMSP)and glycinebetaine were identified by fast atom bombardmentmass and ¹H -NMR spectroscopy, with 3-DMSP being the main oniumcompound and glycinebetaine absent in some accessions. Onium(quaternary ammonium and/or tertiary sulphonium) compounds andproline increased during salt and water stress due mainly toa decrease in the fresh weight/dry weight ratio of tissue, althoughpart of the increase in salt-stressed tissue was due to an increasein the accumulation of the onium compound. This salt-inducedincrease in 3-DMSP was inhibited in conditions of low sulphursupply and there was no compensatory increase in proline. Key words: Melanthera biflora, Asteraceae, salinity, glycinebetaine, 3-dimethylsulphonioproprionate     

Salt Tolerance in the Triticeae: Growth and Solute Accumulation in Leaves of Thinopyrum bessarabicum

Gorham, J., McDonnell, E., Budrewicz, E. and Wyn Jones, R. G.1985. Salt tolerance in the Triticeae: growth and solute accumulationin leaves of Thinopyrum bessarabicum.—J. exp. Bot. 36:1021–1031. The diploid wheatgrass Thinopyrum bessarabicum was found towithstand prolonged exposure to 350 mol m^–3 NaCl in hydroponicculture. During the gradual addition of salt to the externalmedium, osmotic adjustment was rapidly achieved by the accumulationof Na and Cl. Following osmotic adjustment constant leaf Naand Cl concentrations were maintained, and K was retained ata high level. Thinopyrum bessarabicum may be described as anosmoconformer, adjusting its internal osmotic pressure to 400–500mOsmol kg^–1 above that of the external medium in hydroponicculture. Both slower shoot initiation and reduced leaf lengthcontributed to the reduced growth rates at higher salinities.Leaf width was not affected. Increasing salinity resulted inincreases in leaf concentrations of phosphate, glycinebetaine,sucrose and proline, and in decreases in the concentrationsof nitrate, sulphate, magnesium, calcium, total amino acidsand organic acids. Thinopyrum bessarabicum exhibits salt tolerancecharacters which may be useful in wheat breeding. Key words: Salt stress, solute accumulation, osmotic adjustment, Thinopyrum     

Effects of elevated CO2 concentrations on three montane grass species: III. Source leaf metabolism and whole plant carbon partitioning

Agrostis capillaris L.⁵, Festuca vivipara L. and Poaalpina L.were grown in outdoor open-top chambers at either ambient (340 3µmol mol^–1) or elevated (6804µmol mol^–1)concentrations of atmospheric carbon dioxide (CO₂) for periodsfrom 79–189 d. Photosynthetic capacity of source leaves of plants grown atboth ambient and elevated CO₂ concentrations was measured atsaturating light and 5% CO₂. Dark respiration of leaves wasmeasured using a liquid phase oxygen electrode with the buffersolution in equilibrium with air (21% O₂, 0.034% CO₂). Photo-syntheticcapacity of P. alpina was reduced by growth at 680 µmolmol^–1 CO₂ by 105 d, and that of F. vivipara was reducedat 65 d and 189 d after CO₂ enrichment began, suggesting down-regulationor acclimation. Dark respiration of successive leaf blades ofall three species was unaltered by growth at 680 relative to340 µmol mol^–1 CO₂. In F. vivipara, leaf respirationrate was markedly lower at 189 d than at either 0 d or 65 d,irrespective of growth CO₂ concentration. There was a significantlylower total non-structural carbohydrate (TNC) concentrationin the leaf blades and leaf sheaths of A. capillaris grown at680µmol mol^–1 CO₂. TNC of roots of A. capillariswas unaltered by CO₂ treatment. TNC concentration was increasedin both leaves and sheaths of P. alpina and F. vivipara after105 d and 65 d growth, respectively. A 4-fold increase in thewater-soluble fraction (fructan) in P. alpina and in all carbohydratefractions in F. vivipara accounted for the increased TNC content. In F. vivipara the relationship between leaf photosyn-theticcapacity and leaf carbohydrate concentration was such that therewas a strong positive correlation between photosynthetic capacityand total leaf N concentration (expressed on a per unit structuraldry weight basis), and total nitrogen concentration of successivemature leaves reduced with time. Multiple regression of leafphotosynthetic capacity upon leaf nitrogen and carbohydrateconcentrations further confirmed that leaf photosynthetic capacitywas mainly determined by leaf N concentration. In P. alpina,leaf photosynthetic capacity was mainly determined by leaf CHOconcentration. Thus there is evidence for down-regulation ofphotosynthetic capacity in P. alpina resulting from increasedcarbohydrate accumulation in source leaves. Leaf dark respiration and total N concentration were positivelycorrelated in P. alpina and F. vivipara. Leaf dark respirationand soluble carbohydrate concentration of source leaves werepositively correlated in A. capillaris. Changes in source leafphotosynthetic capacity and carbohydrate concentration of plantsgrown at ambient or elevated CO₂ are discussed in relation toplant growth, nutrient relations and availability of sinks forcarbon. Key words: Elevated CO₂, Climate change, grasses, carbohydrate partitioning, photosynthesis, respiration     

Relative Importance of Ion Exclusion, Secretion and Accumulation in Spartina alterniflora Loisel.

The extent to which Spartina alterniflora Loisel. excluded,secreted or accumulated the major seawater ions (Cl^-, SO^2-₄,Na⁺, K⁺, Mg²⁺, and Ca²⁺) was investigated under varying salinitytreatments. From a quantitative viewpoint, ion exclusion wasmost prominent and accounted for 91–97% of the theoreticalmaximum ion uptake as a result of transpiration and growth.Of those ions taken up, approximately half was secreted fromthe shoots. Relative to K⁺, a disproportionate amount of Na⁺was excluded at the roots and secreted by the shoots. The concentrationwithin the tissues of S. alterniflora did not change with salinitytreatment for the majority of the ions examined, but Na⁺ wasmore than twice as concentrated at 40 g dm^-3 than at lOgdm^-3.Calculations of the flux of ions from salt marsh sediments tothe flood water via shoot secretion or stem/leaf turnover indicatethat these processes may be important to the ecology of S. alternifloraas mechanisms that limit the accumulation of salt within theroot zone.     

Concentrations and Transport of Solutes in Xylem and Phloem along the Leaf Axis of NaCl-treated Hordeum vulgare

Hordeum vulgare cv. California Mariout was established in sandculture at two different NaCl concentrations (0.5 mol m^–3‘control’ and 100 mol m^–3) in the presenceof 6.5 mol m^–3 K ⁺. Between 16 and 31 d after germination,before stem elongation started, xylem sap was collected by useof a pressure chamber. Collections were made at three differentsites on leaves 1 and 3: at the base of the sheath, at the baseof the blade, i.e. above the ligule, and at the tip of the blade.Phloem sap was collected from leaf 3 at similar sites throughaphid stylets. The concentrations of K ⁺, Na⁺, Mg2⁺ and Ca²⁺were measured. Ion concentrations in xylem sap collected at the base of leaves1 and 3 were identical, indicating there was no preferentialdelivery of specific ions to older leaves. All ion concentrationsin the xylem decreased from the base of the leaf towards thetip; these gradients were remarkably steep for young leaves,indicating high rates of ion uptake from the xylem. The gradientsdecreased with leaf age, but did not disappear completely. In phloem sap, concentrations of K⁺ and total osmolality declinedslightly from the tip to the base of leaves of both controland salt-treated plants. By contrast, Na⁺ concentrations inphloem sap collected from salt-treated plants decreased drasticallyfrom 21 mol m^–3 at the tip to 7.5 mol m^–3 at thebase. Data of K/Na ratios in xylem and phloem sap were used to constructan empirical model of Na⁺ and K⁺ flows within xylem and phloemduring the life cycle of a leaf, indicating recirculation ofNa⁺ within the leaf. Key words: Hordeum vulgare, xylem transport, phloem transport, NaCl-stress     

A Preliminary Study on the Effects of Nitrogen Supply on the Growth In Vitro of Nine Potato Genotypes (Solanum spp)

Cultures of nine potato genotypes (seven Solanum tuberosum oneS. sparsipilum and one S. oplocense genotypes) were examinedfor their response to growing on medium containing either 60mol m^–3, 40 mol m^–3 or 20 mol m^–3 nitrogen.Genotypes differed in their response to nitrogen. Reducing thenitrogen regime tended to produce taller plants with longerinternodes, shoots had larger leaves but contained less chlorophyll.No change in fresh weight or number of nodes was observed. Genotypex nitrogen interactions were significant for chlorophyll content,shoot length and internode length. Results suggest that thechanges observed were as a result of changes in the total nitrogenlevel rather than changes in the ammonium : nitrate ratio. Thisstudy suggests that for certain potato genotypes, nitrogen levelsin MS medium are too high for producing desirable microplantsin terms of leaf area and shoot length Key words: Solanum tuberosum, S. sparsipilum, S. oplocense, micropropagation, morphogenesis     

Soyabean Stem In Vivo Nitrate Reductase Activity

Stem from three- and four-week-old Soyabean [Glycine max (L.)Merr. cv. Tracy] plants reduced from 0.3 to 0.7 µmol nitrateh^–l g^–l f. wt. Leaf activity was 4.7–7.6 µmolnitrate h^–l g^–l f. wt. Outer stem was two to fourtimes more active at reducing nitrate than was inner stem. Plantnitrate nutrition had a strong effect upon the ratio of activitypresent in stem and leaf. More nitrate increased the proportionpresent in leaves. Glycine max L., soyabean, nitrate assimilation, nitrogen metabolism, Rhizobium japonicum     

Salt Tolerance in the Halophyte, Suaeda maritima (L.) Dum.: The Influence of the Salinity of the Culture Solution on the Content of Various Organic Compounds

Plants of the halophyte Suaeda maritima were grown in tap wateror in a culture solution in the presence or absence of sodiumchloride and the levels of sugars, amino acids, organic acidsand quaternary ammonium compounds determined in relation tothe balance between cytoplasmic and vacuolar water potentials.The sugar content (some 7 µmol. g f. wt^–1) was unaffectedby the salinity of the growth medium as was the overall contentof amino acids (about 4 µmol. g f. wt^–1). The organicacid content was maximal in plants kept in tap water alone wherethe dominant acid was malic. Plants grown in culture solutioncontained the same acids, although addition of sodium chlorideto the medium brought about the apparent loss of glycolic acidand the appearance of oxalic acid. Only a single quaternaryammonium compound, glycinebetaine, was apparently present inthe tissues: the content of betaine doubled (to 37·5µrmol. g f. wt^–) when sodium chloride was addedto the culture solution. The content of these various compoundsis discussed in relation to the relative values of the cytoplasmicand vacuolar components of the overall tissue water potential Suaeda maritima, halophyte, salt tolerance, betaine, organic compounds, water potential     

Accumulation of proline and glycinebetaine in Spartina alterniflora Loisel. in response to NaCl and nitrogen in the marsh

Summary The possible interaction of high soil salinity and low soil nitrogen content in affecting the growth of Spartina alterniflora Loisel in the high and low marshes of the Eastern U.S. was explored. Throughout the whole growing season, the short plants growing in the high marsh, where there was a higher soil salinity and lower available soil nitrogen, contained more proline and glycinebetaine and showed a lower leaf water potential than the tall plants growing in the low marsh. In both short and tall plants, the growing season, with the highest content occurring in spring and fall. In contrast, the glycinebetaine content in both short and tall plants remained fairly constant throughout the growing season, and was consistently at least 10 fold higher than the proline content. It is estimated that 19–30% of the total leaf nitrogen was in the form of proline and glycinebetaine in the short plants, and 14–27% in the tall plants. Ammonium nitrate fertilization in the field resulted in increased growth, higher proline and glycinebetaine contents, and lower water potentials in the short plants, but had little effect on these parameters in the tall plants. We suggest that in the low marsh, the plants can obtain sufficient nitrogen for osmoregulation and other metabolism. In the high marsh with higher soil salinity and lower nitrogen content, the plants have to allocate a even greater proportion of the already limited nitrogen supply for osmoregulation. Thus, nitrogen available for osmoregulation and other nitrogen-requiring metabolism is insufficient, resulting in reduced growth.     

Influence of Proline and Glycinebetaine on Salt Tolerance of Cultured Barley Embryos

Lone, M. I., Kueh, J. S. H., Wyn Jones, R. G. and Bright, S.W. J. 1987. Influence of proline and glycinebetaine on salttolerance of cultured barley embryos.—J. exp. Bot. 38:479–490. The addition of exogenous proline and glycinebetaine to culturedbarley (Hordeum vulgare L. cv. Maris Mink) embryos increasedshoot elongation under saline conditions. Inhibition of shootelongation by NaCl was relieved by proline when plantlets weregrown in deep crystallizing dishes but not in Petri dishes whereshoots come into direct contact with the medium. The effectof proline could be related to a decrease in shoot Cl^–and Na⁺ accumulation which was only observed in plantlets grownin crystallizing dishes. Proline but not betaine uptake intocultured plantlets was stimulated by NaCl while each organicsolute inhibited the endogenous synthesis of the other soluteunder salt stress. Comparison of the effects of exogenously supplied proline withenhanced endogenous proline accumulation in the mutant lineR5201 suggested that the increased proline accumulation in themutant is an order of magnitude too low to have a significantphysiological effect. The implications of the effect of prolineon ion transport, discrimination and accumulation are discussed. Key words: Salt tolerance, proline, ion transport, barley embryo culture     

Proline and Polyamine Involvement in Chilling Tolerance of Maize Suspension Cultures

We have assessed the effect of various medium supplements inpromoting the ability of maize (Zea mays L.) inbred FR27rhmsuspension cultures to grow following a period of 4 °C chillingstress. Following a 4 week exposure to 4 °C in culture mediumwithout proline, no cell growth occurred upon subsequent incubationat 28°C for 2 weeks. This inhibition was reversed when 3to 48 mol m^–3 proline or 0.1 mol m^–3 putrescineor 0.01 mol m^–3 spermidine were present in the mediumduring the chilling stress. On the other hand, suspensions weremade more sensitive to 4°C by blocking polyamine biosynthesiswith 1.0 mol m^–3 methylglyoxal bis (guanylhydrazone) (MGBG)or a combination of 1.0 mol m^–3 difluoromethylornithine(DFMO) and 1.0 mol m^–3 difluoromethylarginine (DFMA).The addition of 10 mol m^–3 putrescine to the suspensioncontaining DFMO and DFMA prevented the increased chilling sensitivity.Electrolyte leakage studies conducted to assess membrane integrityafter 4 weeks at 4°C and a 2 week regrowth period showedthat cells treated with no polyamines (control), 0.01 mol m^–3spermidine, 1.0 mol m^–3 putrescine, or 1.0 mol m^–3MGBG lost 43, 32, 14, and 100% of the total electrolyte pool,respectively. These results suggest that proline and polyaminesare beneficial for inducing chilling tolerance in FR27rhm suspension. Key words: Proline, polyamine, chilling stress     

Measurement of the Apoplastic Activity of K+ and Cl- in the Leaf Epidermis of Commelina communis in Relation to Stomatal Activity

Bowling, D. J. F. 1987. Measurement of the apoplastic activityof K⁺ and Cl^– in the leaf epidermis of Commelina communisin relation to stomatal activity.–J. exp. Bot. 38: 1351–1355. Ionic activity of K⁺ and Cl^– in the apoplast of the lowerepidermis of the leaf of Commelina communis was measured usingion selective micro-electrodes. Large gradients across the stomatalcomplex were observed which were related to stomatal aperture.On stomatal closure the activity of K⁺ and Cl^– in theapoplast of the guard cell rose from 3·0 mol m^–3to 100 mol m^–3 and 33 mol m^–3 respectively. It wasconcluded that the apoplast is an important pathway for iontransport between the cells. Key words: Stomata, ionic activity, leaves, apoplast     

Effects of Gibberellic Acid and Naphthaleneacetic Acid on Petiole Senescence and Subtended Peduncle Growth of Cyclamen persicum Mill

Removal of the blade from the leaf subtending the first flowerbud on Cyclamen persicum ‘Swan Lake’ plants causedthe petiole of that leaf to senesce, but had no effect on thegrowth of the flower peduncle in the debladed petiole's axil.A 10 mg NAA l^–1 application generally had no effect onpetiole senescence, peduncle elongation or flowering date whenapplied to the cut end of the petiole after blade removal. A25 mg GA₃ l^–1 application or a combination of 25 mg GA₃l^–1 application or a combination of 25 mg GA₃ l^–1plus 10 mg NAA l^–1 delayed petiole senescence and enhancedpeduncle elongation and subsequent flowering. No treatment significantlyaltered peduncle length at the time of flowering. Cyclamen persicum Mill, ‘Swan Lake’, tissue receptivity, flowering, GA₃, NAA     

Determination of apoplastic K+ in intact leaves by ratio imaging of PBFI fluorescence

The tetraammonium salt of the K⁺ binding fluorescent dye benzofuranisophthalate (PBFI) was used to investigate the influence ofpotassium nutrition (0.1–2.1 mol m^–3) on apoplasticK⁺ inVicia faba leaves by means of ratio imaging. As a referencethe infiltration-centrifugation method was used. Both methodsreflected the influence of K⁺ supply on apoplastic K⁺ concentration.The abaxial leaf side revealed significantly higher K⁺ concentrations(20-25 mol m^–3) than the adaxial side (5–8 mol m^–3).Application of CCCP led to an immediate increase in apoplasticK⁺ demonstrating the reliability of the PBFI method. Key words: Vicia faba, leaf, apoplast, K⁺, PBFI, ratio imaging, ratiometric fluorescence microscopy     

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     

Stomata and Mesophyll Characteristics of Barley Leaf as Affected by Light: Stereological Analysis

The anatomical structure of the second leaf blade of barley{Hordeum vulgare L. cv. Koral) was studied in plants exposedto a photosynthetic photon flux density (PPFD) of 200 µmolm^–2 s^–1 compared with those grown under 25µmolm^–2–11. Design-based stereological methods wereused for the estimation of various leaf anatomical characteristicssuch as mesophyll volume, proportion of intercellular spaces,number of mesophyll cells, mean mesophyll cell volume, and internalleaf surface area. The structure of the mesophyll was more affectedby different levels of PPFD than were the stomatal characteristics.Increased PPFD produced thicker leaves with a larger mesophyllvolume having a higher number of less elongated mesophyll cellsand a larger internal leaf surface area. Key words: Hordeum vulgare, light effect, mesophyll, stereology, stomata     

The Effects of Cytokinins and ABA on Stomatal Behaviour of Maize and Commelina

Epidermal strips and leaf fragments of Commelina and leaf fragmentsof maize were incubated on solutions containing naturally-occurringor synthetic cytokinins and/or ABA. The effects of these treatmentson stomatal behaviour were assessed. Cytokinins alone did notpromote stomatal opening in either species but concentrationsof both zeatin and kinetin from 10^–3 to 10^–1 molm^–3 caused some reversal of ABA-stimulated closure ofmaize stomata. The reversal of the ABA effect increased withincreasing cytokinin concentration. Cytokinins had no effecton ABA-stimulated closure of Commelina stomata. When appliedalone, at high concentration (10^–1 mol m^–3), toCommelina epidermis or leaf pieces both zeatin and kinetin restrictedstomatal opening. Key words: ABA, Cytokinins, Stomata, Maize, Commelina     

Translocation of Nitrogenous Compounds in Symbiotic and Nitrate-Fed Amide-Exporting Legumes

Peoples, M. B., Sudin, M. N. and Herridge, D. F. 1987. Translocationof nitrogenous compounds insymbiotic and nitrate-fed amide-exportinglegumes.–J. exp. Bot. 38: 567–579. The transport of nitrogen from the roots and nodules of chickpea(Cicer anetinum L.), lentil (Lens culinaris Medic), faba bean(Vicia faba L.) and pea (Pisum sativum L.) was examined in glasshouse-grownplants supplied either with nitrate-free nutrients or with nutrientssupplemented with 1,2,4 or 8 mol m^-3153N-nitrate. A sixth treatmentcomprised uninoculated plants supplied with 8–0 mol m^-31513N-nitrate. For each species, more than 75% of the nitrogenwas exported from the nodules as the amides, asparagine andglutamine. In fully symbiotic plants, the amides also dominatednitrogen transport to the shoot When N2 fixation activity wasdecreased by the addition of nitrate to the rooting medium,the N-composition of xylem exudate and stem solutes changedconsiderably. The relative concentrations of asparagine tendedto increase in the xylem whilst those of glutamine were reduced;the levels of nitrate increased in both xylem exudate and thesoluble nitrogen pool of the stem with a rise in nitrate supply.The changes in relative nitrate contents reflected generallythe contributions of root and shoot to overall nitrate reductaseactivity at the different levels of nitrate used. The relationshipsbetween the relative contents of xylary or stem nitrate andamino nitrogen and the plants' reliance on N2 fixation (determinedby the ¹⁵N isotope dilution procedure) were examined. Data suggestthat compositional relationships based on nitrate may be reasonableindicators of symbiotic dependence for all species under studyexcept faba bean when greater than 25% of plant nitrogen wasderived from N2 fixation. Key words: Nitrogen, translocation, legumes