Purification and Properties of Manganese Superoxide Dismutase from Norway Spruce (Picea abies L. Karst)

A manganese-containing superoxide dismutase (SOD; EC 1.15.1.1 [EC] )was purified to electrophoretic homogeneity from seeds of Norwayspruce (Picea abies L.). The apparent molecular mass of thepurified enzyme was 86 kDa, as determined by gel filtration.The subunit molecular mass, estimated by SDS-polyacrylamidegel electrophoresis, was 22 kDa both in the presence and inthe absence of 2-mercaptoethanol. Thus, the native enzyme isa homotetramer with subunits that were not linked by disulfidebonds. The isoelectric point of this Mn-SOD was 5.5. The specificactivity of the Mn-SOD was strongly pH-dependent and was 400units per nmol SOD at pH 7.8 and 30 units per nmol SOD at pH10.4. The first 25 amino acid residues in the amino terminalregion of spruce Mn-SOD exhibited a high degree of sequencehomology to those of Mn-SODs from other organisms. In Mn-deficientneedles the activity of Mn-SOD was only half of that in non-deficientneedles, whereas the activity of CuZn-SOD was doubled. (Received May 20, 1994; Accepted October 31, 1994)     

Effects of Chilling, Light and Nitrogen-containing Compounds on Germination, Rate of Germination and Seed Imbibition of Clematis vitalba L.

Effects of chilling (5 °C) period, light and applied nitrogen(N) on germination (%), rate of germination (d to 50% of totalgermination; T_50%) and seed imbibition were examined inClematisvitalba L. In the absence of chilling, light and N, germinationwas minimal (3%). When applied alone, both chilling and N increasedgermination. Chilling for 12 weeks increased germination to64%, and 2.5 mM NO^-₃or NH⁺₄increased germination to 10–12%.Light did not increase germination when applied alone, but didwhen applied in combination with chilling and/or N. Half theseed germinated when light was combined with 2.5 mM NO^-₃or NH⁺₄.The influence of chilling, light and/or N on germination wasgreater when combined, than when either factor was applied alone.Both oxidized (NO^-₃) and reduced (NH⁺₄) forms of N increasedgermination, but non-N-containing compounds did not, suggestingthe response was due to N and not ionic or osmotic effects. Without additional N, T_50%decreased from 16–20 d at zerochilling, to around 5 d at 8 and 12 weeks chilling. AlthoughT_50%was not influenced by an increase in NO^-₃or NH⁺₄from 0.5to 5.0 mM , it did increase with additional applied N thereafter.However, the magnitude of the N effect was small compared tothat of chilling. Like germination, seed imbibition increasedwith a longer chilling period, but in contrast imbibition decreasedslightly with increased applied NO^-₃or NH⁺₄. It is argued thatincreased imbibition is not directly related to an increasein total germination, but that it may be related to the rateof germination. Possible mechanisms involved in the reductionin dormancy ofC. vitalba seed are discussed. Clematis vitalba L.; germination; dormancy; imbibition; rate of germination; chilling; light; nitrate; ammonium; nitrogen; phytochrome     

Conformational modulation of slow skeletal muscle troponin T by an NH(2)-terminal metal-binding extension

TroponinT (TnT) is an essential element in the thin filamentCa²⁺-regulatory system controlling striated musclecontraction. Alternative RNA splicing generates developmental andmuscle type-specific TnT isoforms differing in the hypervariableNH₂-terminal region. Using avian fast skeletal muscle TnTcontaining a metal-binding segment, we have demonstrated a role of theNH₂-terminal domain in modulating the conformation of TnT(Wang J and Jin JP. Biochemistry 37: 14519-14528,1998). To further investigate the structure-function relationship ofTnT, the present study constructed and characterized a recombinantprotein in which the metal-binding peptide present in avian fastskeletal muscle TnT was fused to the NH₂ terminus of mouseslow skeletal muscle TnT. Metal ion or monoclonal antibody binding tothe NH₂-terminal extension induced conformational changes in other domains of the model TnT molecule. This was shown by thealtered affinity to a monoclonal antibody against the COOH-terminal region and a polyclonal antiserum recognizing multiple epitopes. Protein binding assays showed that metal binding to theNH₂-terminal extension had effects on the interaction ofTnT with troponin I, troponin C, and most significantly, tropomyosin.The data indicate that the NH₂-terminal Tx [4-7repeats of a sequence motif His-(Glu/Ala)-Glu-Ala-His] extension confers a specific conformational modulation in the slowskeletal muscle TnT.

     

Topological analysis of NHE1, the ubiquitous Na+/H+ exchanger using chymotryptic cleavage

Proteases,glycosidases, and impermeant biotin derivatives were used incombination with antibodies to analyze the subcellular distribution andtransmembrane disposition of theNa⁺/H⁺exchanger NHE1. Both native human NHE1 in platelets and epitope-tagged rat NHE1 transfected into antiport-deficient cells were used for thesestudies. The results indicated that1) the entire population ofexchangers is present on the surface membrane of unstimulated platelets, ruling out regulation by recruitment of internal stores ofNHE1; 2) the putative extracellularloops near the NH₂ terminus areexposed to the medium and contain all the N- andO-linked carbohydrates;3) by contrast, the putativeextracellular loops between transmembrane domains 9-10 and11-12 are not readily accessible from the outside and may befolded within the protein, perhaps contributing to an aqueous iontransport pathway; 4) the extreme COOH terminus of the protein was found to be inaccessible toextracellular proteases, antibodies, and other impermeant reagents,consistent with a cytosolic localization; and5) detachment of ~150 amino acidsfrom the NH₂-terminal end of theprotein had little effect on the transport activity of NHE1.

     

Response of Glutamine Synthetase and Glutamate Synthase Isoforms to Nitrogen Sources in Rice Cell Cultures

As a model system with no photorespiration and no long distancetransport, rice cell cultures (Oryza saliva L. cv Sasanishiki)were used to investigate the effect of nitrogen sources on thelevels of isoforms of glutamine synthetase (GS) and glutamatesynthase (GOGAT). Isoforms of GS and GOGAT were analyzed byimmunoblotting methods and their activities in early growthphase of the cells. Cytosolic type GS (41 kDa subunit) and NADH-GOGATwere the major isoforms in the rice cells grown in normal R-2medium. However, contents of plastid type GS (44 kDa subunit)and Fd-GOGAT increased in response to NO_–³ supply. NADH-GOGATactivity also increased following the supply of NO_–³.In vitro translated products from poly(A)⁺RNA prepared fromthe cells showed that the precursor of plastid type GS (49 kDa)was detected at 48 h after the inoculation. Supply of NH₊⁴ resultedin an increase in NADH-GOGAT activity but had no effect on thelevels of Fd-GOGAT, of polypeptides of the plastid type GS orof the corresponding mRNAs. (Received May 30, 1990; Accepted August 23, 1990)     

The Influence of Nitrate and Ammonium Nutrition on the Growth of Wheat (Triticum aestivum) and Maize (Zea mays) Plants

The effects of NO^-₃ and NH⁺₄ nutrition on hydroponically grownwheat (Triticum aestivum L.) and maize (Zea mays L.) were assessedfrom measurements of growth, gas exchange and xylem sap nitrogencontents. Biomass accumulation and shoot moisture contents ofwheat and maize were lower with NH⁺₄ than with NO^-₃ nutrition.The shoot:root ratios of wheat plants were increased with NH⁺₄compared to NO^-₃ nutrition, while those of maize were unaffectedby the nitrogen source. Differences between NO^-₃ and NH⁺₄-fedplant biomasses were apparent soon after introduction of thenitrogen into the root medium of both wheat and maize, and thesedifferences were compounded during growth. Photosynthetic rates of 4 mM N-fed wheat were unaffected bythe form of nitrogen supplied whereas those of 12 mM NH⁺₄-fedwheat plants were reduced to 85% of those 12 mM NO^-₃-fed wheatplants. In maize supplied with 4 and 12 mM NH⁺₄ the photosyntheticrates were 87 and 82% respectively of those of NO^-₃-fed plants.Reduced photosynthetic rates of NH⁺₄ compared to NO^-₃-fed wheatand maize plants may thus partially explain reduced biomassaccumulation in plants supplied with NH⁺₄ compared to NO^-₃ nutrition.Differences in the partitioning of biomass between the shootsand roots of NO^-₃-and NH⁺₄-fed plants may also, however, arisefrom xylem translocation of carbon from the root to the shootin the form of amino compounds. The organic nitrogen contentof xylem sap was found to be considerably higher in NH⁺₄- thanin NO^-₃-fed plants. This may result in depletion of root carbohydrateresources through translocation of amino compounds to the shootin NH⁺₄-fed wheat plants. The concentration of carbon associatedwith organic nitrogen in the xylem sap of maize was considerablyhigher than that in wheat. This may indicate that the shootand root components of maize share a common carbon pool andthus differences induced by different forms of inorganic nitrogenare manifested as altered overall growth rather than changesin the shoot:root ratios.Copyright 1993, 1999 Academic Press Triticum aestivum, wheat, Zea mays, maize, nitrogen, growth, photosynthesis, amino acids, xylem     

Control of Glutamate Dehydrogenase from Green Tobacco Callus Mitochondria by Ca2$ and pH

Glutamate dehydrogenase (GDH) (EC 1.4.1.3 [EC] .) purified from greentobacco callus mitochondria was activated markedly by Ca^2$ inthe amination reaction. This activation was detectable evenat concentrations below 5 µM Ca^2$. Saturation curves for the three substrates of the aminationreaction showed normal Michaelis-Menten kinetics in the presenceof 1 mM of Ca^2$, but pronounced substrate inhibition occurredwithout Ca^2$. The effect of Ca^2$ was chiefly on the maximalvelocity. The saturation curve for NH₄Cl in the presence of Ca^2$ was modulatedby a change in pH. The apparent K_m value for NH₄Cl markedlydecreased whereas that for -ketoglutarate increased slightlywhen the pH was raised from 7.3 to 9.0. In contrast, the K_mfor NADH was little affected by raising the pH. The characteristicof GDH which increases its affinity for NH₄Cl when the pH israised may be compatible with the detoxification of ammonia. ¹ Present address: Mochida Pharmaceutical Co., Ltd. (Received August 24, 1981; Accepted November 28, 1981)     

Regulation of AE2 anion exchanger by intracellular pH: critical regions of the NH(2)-terminal cytoplasmic domain

The role ofintracellular pH (pH_i) in regulation of AE2 function inXenopus oocytes remains unclear. We therefore compared AE2-mediated ³⁶Cl efflux fromXenopus oocytes during imposed variation of extracellular pH(pH_o) or variation of pH_i at constantpH_o. Wild-type AE2-mediated ³⁶Clefflux displayed a steep pH_o vs. activity curve, withpH_o(50) = 6.91 ± 0.04. SequentialNH₂-terminal deletion of amino acid residues in tworegions, between amino acids 328 and 347 or between amino acids 391 and510, shifted pH_o(50) to more acidic values by nearly 0.6 units. Permeant weak acids were then used to alter oocytepH_i at constant pH_o and were shown to beneither substrates nor inhibitors of AE2-mediated Cltransport. At constant pH_o, AE2 was inhibited byintracellular acidification and activated by intracellularalkalinization. Our data define structure-function relationships withinthe AE2 NH₂-terminal cytoplasmic domain, which demonstratesdistinct structural requirements for AE2 regulation by intracellularand extracellular protons.

     

Biochemical properties of 1-aminocyclopropane-1-carboxylate N-malonyltransferase activity from early growing embryonic axes of chick-pea (Cicer arietinum L.) seeds

In the present work, certain biochemical characteristics ofthe enzyme 1-aminocyclopropane-1-carboxylate N-malonyltransferase(ACC N-MTase) which is responsible for the malonylation of 1-aminocyclopropane-1-carboxylate(ACC) in chickpea (Cicer arietinum) are described. Phosphatebuffer was the most appropriate buffer with regard to enzymestability and, therefore, ACC N-MTase was extracted, assayedand purified in the presence of this buffer. ACC N-MTase waspartially purified approximately 900-fold from embryonic axesof chick-pea seeds using ammonium sulphate precipitation, hydrophobicinteraction and molecular filtration chromatography. By gelfiltration chromatography on Superose-12, the molecular massof the enzyme was estimated to be 54 4 kDa. ACC N-MTase hadan optimal pH and temperature of 7.5 and 40C, respectively,as well as a K_m for ACC and malonyl-CoA of 400 M and 90 M,respectively. D-Phenylalanine was a competitive inhibitor ofACC N-MTase with respect to ACC (K_i of 720 M), whereas co-enzymeA was a competitive product inhibitor with respect to malonyl-CoA(K_i of 300 M) and a non-competitive inhibitor with respectto ACC (K_i of 600 M). Under optimal assay conditions, ACC N-MTasewas strongly inhibited by (a)divalent [Zn²⁺>Mg²⁺>>Co²⁺>Co²⁺>(NH₄)²⁺>Fe²⁺]and monovalent metal cations (Li⁺>Na⁺>K⁺), without activitybeing detected in the presence of Hg²⁺, and (b) PCMB or mersalicacid, suggesting that sulphydryl group(s) are involved at theactive site of the enzyme. Key words: ACC-N-malonyltransferase, Cicer arietinum, embryonic axes, ethylene, germination, seeds     

cDNA Cloning and Expression of the Plastidic Copper/Zinc-Superoxide Dismutase from Spinach (Spinacia oleracea L.) Leaves

A cDNA clone for copper/zinc-superoxide dismutase (Cu/Zn-SOD)was isolated from spinach (Spinacia oleracea L.) leaves. Itsnucleotide sequence showed that it codes for a precursor polypeptideof 222 amino acids, including the NH₂-terminal 68-residue extensionwhich corresponds to a plastidic transit peptide. Northern hybridization,using plastidic and cytosolic Cu/Zn-SOD cDNAs as the probes,revealed that these two genes are differentially expressed inthe roots and leaves of spinach. ¹Present address: Department of Biochemistry and Microbiology,Cook College, Rutgers University New Brunswick, NJ 08903-0231,U.S.A.     

Cell Cycle Timing of Replication of the Nuclear Gene Encoding Chloroplastic NADP-Specific Glutamate Dehydrogenase Isoenzymes in Chlorella sorokiniana

In synchronized Chlorella sorokiniana cells, the NH₄⁺ inducibleNADP-specific glutamate dehydrogenase enzyme (NADP-GDH) accumulatedin a linear manner throughout the first cell cycle. Early inthe following second cell cycle, an increase in its rate ofaccumulation occurred that was proportional to the increasein total cellular DNA in the previous cell cycle. In synchronizedbacterial cells, increases in rate of linear accumulation ofinducible enzymes coincide with the time of replication of theirstructural genes. To determine whether the rate change in NADPGDHaccumulation resulted from a delay in replication of its nuclearstructural gene (gdhN) in fully induced C. sorokiniana cells,the cell cycle timing of replication of this gene was comparedto that of another nuclear gene, nitrate reductase (nia), andof a chloroplast gene, ribulose bisphosphate carboxylase large-subunit(rbcL), in synchronized cells cultured in NH₄⁺ or NO₃^–(uninduced) medium. The gdhN and nia genes replicated withinthe period of nDNA synthesis and rbcL within the period of ctDNAsynthesis in cells growing in either nitrogen source. Therefore,the delayed rate change in enzyme accumulation results froma process that regulates expression of the gdhN gene after itsreplication. (Received July 16, 1994; Accepted November 28, 1994)     

Control of the germination of secondary dormant cocklebur seeds by various germination stimulants

The responsiveness of non-dormant, upper cocklebur (Xanthiumpennsylvanicum Wallr.) seeds to various germination stimulants,such as CO₂ C₂H₄ CS(NH₂)₂, BA and enriched O₂, decreased withincreasing periods of water imbibition and was completely lostin the state of secondary dormancy. Unlike CO₂ BA and CS(NH₂)₂however, C₂H₂ and enriched O₂ effectively prevented the developmentof secondary dormancy, and their combination was the most effectivefor stimulating the germination of seeds which had undergoneimbibition for a long time. CS(NH₂)₂ and BA were effective,not by themselves but either under anaerobiosis or elevatedO₂ tension. Growth of the axial and cotyledonary segments excisedfrom aged seeds remained responsive to these germination stimulantsand could be further stimulated by exogenous C₂H₂. With imbibitionat a lower temperature, the seeds maintained high germinationin response to various stimulants and a high rate of C₂H₂ andCO₂ production during a long period of water imbibition. Theseresults are discussed in terms of the two possible causes forthe loss of responsiveness or induction of the secondary dormancy. (Received June 27, 1978; )     

Effect of Ammonia on Dark CO2 Fixation by Anabaena Cells Treated with Methionine Sulfoximine

Dark CO₂ fixation by Anabaena cylindrica was stimulated aboutthree-fold by the addition of NH₄Cl to the cells. The ¹⁴CO₂incorporation experiments showed that ¹⁴C is most rapidly incorporatedinto aspartate and then glutamine by adding NH4CI. Glutamineaccumulated predominantly after the addition of NH₄Cl showingthat NH₄ is incorporated into glutamine by glutamine synthetase.The stimulating effect of NH4Cl on CO₂ fixation and amino acidsynthesis was suppressed by methionine sulfoximine, an inhibitorof glutamine synthetase. It was suggested that dark CO₂ fixationwas stimulated by the action of glutamine synthesis which isenhanced by ammonia. (Received February 10, 1981; Accepted April 2, 1981)     

Assimilation of gaseous ammonia and the transport of its products in barley and spinach leaves

The interactions between the assimilation and transport of nitrogenand carbon were investigated in barley and spinach leaves. Bothplants were fumigated with NH₃ (1 mg m^–3 and the contentof amino acids, sucrose and carbon intermediates of amino acidmetabolism were analysed in the leaves, apoplast and phloemsap. The following changes took place in the C- and N-metabolismof barley leaves during 5 h of fumigation with NH₃ (a) The contentsof amino acids, especially glutamine, largely increased andthe contents of sucrose, 2-oxoglutarate, phosphoenolpyruvate,and glycerate-3-phosphate declined. (b) A decrease in the phophoenolpyruvatecontent was accompanied by an increased activity of phosphoenolpyruvatecarboxylase. (c) The altered cytosolic concentrations of aminoacids and sucrose during NH₃ fumigation correlated with similarchanges in the apoplast and phloem sap. The altered percentageof each amino acid relative to the total amino acid concentrationin the cytosol, caused by NH₃ fumigation, is reflected in theapoplast and the phloem sap. The results indicate that the concentrations of amino acids in the cytosol determine their concentrationsin the phloem. Key words: Amino acids, ammonia fumigation, barley leaves, C: N partitioning, phosphoenolpyruvate carboxylase, phloem sap, spinach leaves     

Primary Structure and Properties of Glutathione Reductase from Arabidopsis thaliana

A cDNA encoding glutathione reductase (EC 1.6.4.2 [EC] ) from Arabidopsisthaliana was cloned by immunoscreening. The amino acid sequencededuced from the nucleotide sequence agrees with the N-terminalamino acid sequence of the major isozyme (GR II) purified fromleaves of A. thaliana. The predicted polypeptide comprises anN-terminal leader sequence of 74 amino acids, which has featuresof chloroplast-targeting peptides, and a mature polypeptideof 491 residues with a molecular mass of 52.7 kDa, which showshomology with glutathione reductases from other species. TheK_m for GSSG was 44 µM and that for NADPH was 5.0 µMfor GR II at 25C. The pH optimum for GR II was 7.5 to 8.0.The native molecular mass of GR II was 110 kDa, indicating thatGR II is a homodimer. GR II had an isoelectric point of 4.8.The cDNA hybridizes with a 2.1-kb poly(A)⁺RNA from leaves ofA. thaliana. Genomic Southern analysis indicates that the genecorresponding to the cDNA is likely a single-copy gene. (Received July 1, 1993; Accepted September 8, 1993)     

Structural and Kinetic Properties of NADP-Malic Enzyme from the Inducible Crassulacean Acid Metabolism Plant Mesembryanthemum crystallinum L.

NADP-malic enzyme (EC 1.1.1.40 [EC] ), which is involved in Crassulaceanacid metabolism (CAM), was purified to electrophoretic homogeneityfrom the leaves of the inducible CAM plant Mesembryanthemumcrystallinum. The NADP-malic enzyme, which was purified 1,146-fold,has a specific activity of 68.8 µmol (mg protein)^–1min^–1. The molecular weight of the subunits of the enzymewas 64 kDa. The native molecular weight of the enzyme was determinedby gel-filtration to be 390 kDa, indicating that the purifiedNADP-malic enzyme is a hexamer of identical subunits. The optimalpH for activity of the enzyme was around 7.2. Double-reciprocalplots of the enzymatic activity as a function of the concentrationof L-malate yielded straight lines both at pH 7.2 and at pH7.8 and did not reveal any evidence for cooperativity of bindingof L-malate. The K_m value for L-malate was 0.35 mM. Hill plotsof the activity as a function of the concentration of NADP⁺indicated positive cooperativity in the binding of NADP⁺ tothe enzyme with a Hill coefficient (nH) of 2.0. An S_0.5 value(the concentration giving half-maximal activity) of 9.9 µMfor NADP⁺ was obtained. Oxaloacetate inhibited the activityof the NADP-malic enzyme. Effects of succinate and NaHCO₃ onthe activity of NADP-malic enzyme were small. (Received October 30, 1991; Accepted May 1, 1992)     

Effect of Nitrate, Ammonium and Some Amino Acids on Growth and Nitrate Reductase Activity in Suspension Cultures of Atropa belladonna

Growth and nitrate reductase activity (NRA) of Atropa belladonnacells were studied in medium supplemented with NaNO₃, NH₄NO₃,and amino acid precursors to tropane alkaloids. Growth and NRAwere stimulated by NH₄⁺ and by proline, by proline plus ornithine,but not by glutamate, in NO₃^–-containing medium. Testedamino acids inhibited neither utilization of inorganic nitrogennor growth. (Received September 30, 1988; Accepted August 28, 1989)     

Potassium-Ammonium Uptake Interactions in Tobacco Seedlings

Short-term (< 12 h) uptake experiments were conducted with6–7-week-old tobacco (Nicotiana tabacum L. cv. Ky 14)seedlings to determine absorption interactions between K⁺ andNH₄⁺. At equal solution concentrations (0.5 mol m^–3) netK⁺ uptake was inhibited 30–35% by NH₄⁺ and NH₄⁺ uptakewas decreased 9–24%. Removal of NH₄⁺ resulted in completerecovery in K⁺ uptake rate, but NH⁴⁺ uptake rate did not recoverwhen K⁺ was removed. In both cases, inhibition of the uptakerate of one cation saturated as the concentration of the othercation was increased up to 0.5 mol m^–3. The relative effectof K⁺-NH₄⁺ interactions was not altered when Cl- was replacedwith SO₄^2–, but the magnitudes of the uptake rates wereless in the absence of Cl-. The V_max for NH₄⁺ uptake was reducedfrom 128 to 105 µmol g^–1 dry wt. h^–1 in thepresence of 0.5 mol m^–3 K⁺ and the K_m for NH₄⁺ doubledfrom 12 to 27 mmol m^–3 in the presence of K⁺. The resultsof these K⁺-NH₄⁺ experiments are interpreted as mixed-noncompetitiveinteractions. However, an enhanced efflux of K⁺ coupled to NH₄⁺influx via an antiporter cannot be ruled out as contributingto the decrease in net K⁺ uptake. Key words: Nicotiana tabacum, K⁺, NH₄⁺, Uptake interactions     

Relationship between Coleoptile Elongation and Alcoholic Fermentation in Rice Exposed to Anoxia. II. Cultivar Differences

The relationship between coleoptile elongation and survivalvs. alcoholic fermentation of rice under anoxia is examinedusing eight cultivars differing in submergence tolerance. Anoxiawas imposed on either 1 or 4 d aerated seeds either by N₂ flushingsubmerged tissues or by incubating tissues in stagnant deoxygenatedagar at 0·1% w/v; the latter simulated the stagnant conditionsof waterlogged soil. Two cultivars that were most tolerant tosubmergence also had the greatest tolerance to anoxia, whilea submergence intolerant cultivar was also intolerant to anoxia. Coleoptile growth under anoxia was related to rates of ethanolsynthesis (R_E), however differences between growth during anoxiaand survival after anoxia indicated that post-anoxic injurymay also be important in rice seeds exposed to anoxia. The correlationbetween coleoptile growth and R_E measured on a tissue basisusing intact seeds was r² = 0·67 among six varietiesover 0-3 d anoxia. This correlation improved to about r² = 0·85using R_E of (embryos plus coleoptiles) over 0-3 d, or coleoptilesat 3 d after anoxia. Coleoptile growth of individual seeds wasusually poorly correlated to R_E in these cultivars at 2-3 dafter anoxia. When coleoptiles of similar lengths were obtainedfrom different cultivars using 4 d aerated seeds, there weredifferences in R_E and coleoptile growth which were related tocoleoptile growth during 3 or 5 d anoxia, either on a tissue(r² = 0·85) or a fresh weight basis (r² = 0·70-0·97respectively). Results are discussed in relation to factorswhich may limit ethanol synthesis in rice exposed to anoxiaand the importance of growth to the survival of seeds and matureplants during submergence in the natural environment.Copyright1994, 1999 Academic Press Anoxia, ethanol, alcoholic fermentation, Oryza sativa L., rice, submergence     

Changes in the growth rates of saline-lake diatoms in response to variation in salinity, brine type and nitrogen form

The growth rates of four saline-lake diatom taxa were measuredunder varying conditions of salinity (5, 8 and 11), brine type(sulfate- versus bicarbonate-dominated) and nitrogen form (NH₄⁺versus NO₃^–), using a full factorial design. With NO₃^–as the nitrogen source, Cyclotella quillensis, Cymbella pusillaand Anomoeoneis costata exhibited lower growth rates in thesulfate versus bicarbonate media. The strain of Chaetoceroselmorei used in these experiments, isolated from a sulfate-dominatedlake, was unable to grow on NO₃^– alone. In the NH₄⁺ treatments,neither salinity nor brine type affected the growth rates ofC.quillensis or C.elmorei. When supplied with NH₄⁺, C.pusillaand A.costata had higher growth rates in the bicarbonate versussulfate media, although for C.pusilla the difference on NH₄⁺was not as great as on NO₃^–. The impact of brine typeon NO₃^– use is consistent with the theory that sulfateinhibits molybdate uptake, as molybdenum is required for NO₃^–use but not NH₄⁺. Cymbella pusilla was the only taxon affectedby changes in salinity. The four taxa used in these experimentsare frequently found in saline lakes and saline-lake sediments,hence they are used in paleoclimate reconstructions; the resultspresented here provide additional information that may enhancethese diatom-based reconstructions.