Base-Specific Endo-Exonucleolytic Activity of Chlamydomonas Nuclease C1&2

The reaction kinetics of nuclease C1&2 from Chlamydomonasreinhardtii were studied. It showed endo-exonucleolytic activitywith sugar non-specificity. The relative rates of RNA breakdownwere in order of poly(U) > poly(A) > yeast sRNA. In contrast,poly(G) and poly(C) released almost no acid-soluble materialsafter reacting with nuclease C1&2. The major products ofa 100% limit digest of synthetic RNA homopolymers were mononucleotideswith 3'-phosphate termini. Large oligonucleotides produced duringendo-exonucleolytic degradation also appeared carrying 3'-phosphatetermini. Nuclease C1&2 hydrolyzed single stranded DNA 20times faster than double stranded DNA by endo-exonucleolyticaction, releasing acid-soluble materials. High performance liquidchromatography of a 100% limit digest of salmon testes DNA demonstratedthat the major products were deoxymononucleotides with phosphateat 3'-position. Furthermore, the level of 3'-dCMP among themwas found to be extremely low. Poly(dC) and poly(me⁵dC) werehydrolyzed much more slowly than single stranded (or denatured)DNA, releasing acid-soluble materials. The present results suggestthat nuclease C1&2 is a base-specific nucleate 3'-oligonucleotidohydrolasedifferent from the restriction enzymes. (Received January 13, 1986; Accepted March 25, 1986)     

Purification of Major Isozymes of Nuclease C and Production of Active Fragments by Trypsin

Most nucleases from gametes of Chlamydomonas reinhardtii needCa^2$ for full activation. They have been named nuclease C andat least six species of isozymes have been found in the femalegamete (Ogawa and Kuroiwa 1985a). Nuclease C1&2 and C3 were purified from the vegetative cellsof this organism. Nuclease C1&2 exhibited a sharp pH optimumat 9.5, while nuclease C3 preferred a more neutral pH at 7.0–8.5.Use of the Ca^2$-EGTA [ethylene-glycol-bis-(2-aminoethyl ether)-N,N,N',N'-tetraaceticacid] buffer in the reaction mixture made it possible to determinetheir activity at the physiological Ca^2$ concentration. NucleaseC3 was not activated at low Ca^2$ concentration and exhibiteda sharp optimum at 10^–3{small tilde}10^–4 M. NucleaseC1&2 were activated at a physiological concentration of10^–6 M; increasing the Ca^2$ concentration did not affectthe activity. Nuclease C gave active fragments upon trypsin digestion. Trypticfragments of nuclease C1&2 and C3 had molecular weightsof 21,000 (referred as C6T) and 16,000 (C4T), respectively.Upon regulating the digestion, a few fragments were identifiedas intermediates of nuclease C6T by the in situ nuclease assay.These tryptic fragments were similar in molecular size to theminor components of nuclease C found in the cell lysates ofgametes and early zygotes. This finding suggests that a minorspecies of nuclease C may be produced from the major nucleaseC during gametogenesis. (Received June 26, 1985; Accepted August 28, 1985)     

Nuclease C Polymorphism of Calcium-Dependent Nucleases in Chlamydomonas reinhardtii

DNases were extracted from cells of Chlamydomonas reinhardtii.Their polymorphism and metal ion requirements were investigated.Most of the nucleases from this organism required Ca²⁺ for fullactivation; therefore, the name nuclease C has been given tothem. Zn²⁺ and Mn²⁺ restored activation, but their respectivepotencies were 1/8 and 1/32 that of Ca²⁺. An in situ nucleaseassay revealed that there are at least 6 species of nucleaseC. The molecular weights of the components were 26,000 (C1),25,000 (C2), 22,000 (C5), 21,000 (C6), 18,000 (C3) and 17,000(C4) by SDS PAGE. Ca²⁺-dependent nuclease activity was slightlyhigher in the female gamete than in the male gamete. There wasno marked change in the apparent nuclease activity during thefirst 3 h after mating. An in situ assay, however, showed thatnuclease C6 and C4 are present in smaller amounts in the malegamete in comparison to the female gamete, and that the contentsor activities of nuclease C5 and C6 diminish during the first30–60 min after mating. This evidence is discussed interms of the possible participation of nuclease C in the maternalinheritance of chloroplast genes in this organism. (Received October 8, 1984; Accepted January 21, 1985)     

Thermal Properties of Membrane Lipids from two Cyanobacteria, Anacystis nidulans and Synechococcus sp.

The composition and positional distribution of fatty acids inmonogalactosyldiacylglycerol, digalactosyldiacylglycerol, phosphatidylglyceroland sulphoquinovosyldiacylglycerol from two cyanobacteria, Anacystisnidulans and Synechococcus sp. grown at 25°C have been determinedand compared with measurements of the phase separation temperaturesof the lipids. Only monogalactosyldiacylglycerol in Anacystisand sulphoquinovosyldiacylglycerol in Synechococcus showed phaseseparation temperatures above 0°C. The phase transitiontemperature of a sample of sulphoquinovosyldiacylglycerol containingover 90% of the dihexadecanoyl molecular species has been determinedto be 43°C for the Na⁺ salt and 38°C for the Mg⁺⁺ salt. ^*Deceased. September 14, 1986. (Received June 25, 1986; Accepted August 25, 1986)     

Quantal Response of Seed Germination in Seven Genera of Cruciferae to White Light of Varying Photon Flux Density and Photoperiod

The response of the germination of seeds of Barbarea vema (Mill.)Aschers, Brassica chinensis L., Brassica juncea (L.) Czern.& Coss., Brassica oleracea L. var. gongylodes L., Camelinasaliva (L.) Crantz, Eruca saliva Mill., Lepidium sativum L.,Nasturtium officinale R. Br., and Rorippa palustris (L.) Besserto white fluorescent light of different photon flux densitiesapplied for different daily durations in a diurnal alternatingtemperature regime of 20 °C/30 °C (16 h/8 h) was quantifiedby linear relations between probit percentage germination andthe logarithm of photon dose, the product of photon flux densityand duration. The low energy reaction, in which increasing dosepromotes germination, was detected in all the seed populationsbut in Barbarea vema and Brassica Juncea the lowest photon doseapplied (10^–5–2 and 10^{–5 7} mol m^–2 d^–1,respectively) was sufficient to saturate the response. Comparisons,where possible, between photoperiods demonstrated reciprocity,i.e. germination was proportional to photon dose irrespectiveof photoperiod, for the low energy reaction in Brassica oleracea(1 min d^–1 to 1 h d^–1), Camelina saliva (1 min d^–1to 8 h d^–1), Eruca saliva (1 min d^–1 to 24 h d^–1),Lepidium sativum (I min d^–1 to 8 h d^–1) and Rorippapalustris (1 min d^–1 to 8 h d^–1), but not in Brassicachinensis and Nasturtium officinale. The high irradiance reaction,in which increasing dose inhibits germination, was detectedin Barbarea vema, Brassica chinensis, Brassica juncea, Brassicaoleracea, and Camelina saliva. The minimum dose at which inhibitionwas detected was lO^–0–3 mol m^–2 d^–1.These results are discussed in the context of devising optimallight regimes for laboratory tests intended to maximize germination The response of germination to photon dose was also quantifiedwith 3 x 10^–4 M GA₂, co-applied (Brassica chinensis, Camelinasaliva, and Lepidium sativum) and with 2 x 10^–2 M potassiumnitrate co-applied (Brassica chinensis). In the latter casepotassium nitrate had no effect in the dark and inhibited germinationin the light, but GA₂, promoted germination substantially inall three species. Variation amongst seeds in the minimum photondose required to stimulate germination was not affected by co-applicationof GA₂, in Brassica chinensis and Camelina saliva, whereas seedsof Lepidium salivum showed a narrower distribution of sensitivitiesto the low energy reaction in the presence of GA₂ Barbarea vema (Mill.) Aschers, Brassica chinensis L., Brassica juncea (L.) Czern. & Coss., Brassica oleracea L. var. gongylodes L., Camelina saliva (L.) Crantz, Eruca saliva Mill., Lepidium satiaum L., Nasturtium officinale R. Br., Rorippa palustris (L.) Besser, Cruciferae, light, gibberellic acid, seed germination, seed dormancy     

The Potential of Two Perennial C4 Grasses and a Perennial C4 Sedge as Ligno-cellulosic Fuel Crops in N.W. Europe. Crop Establishment and Yields in E. England

Three perennial C₄ rhizomatous species, Cyperus longus L., Spartinacynosuroides and Spartina pectinata Link, were examined as potentialrenewable energy crops. These species are unusual among C₄ plantsin showing natural distributions which extend into cool temperateregions. This study examined whether these species could beestablished in the cool temperate climate of eastern Englandand whether they could consistently attain the relatively highdry matter yields associated with C₄ plants of warmer regions.Clonally produced material was planted in 1986, on two siteswith contrasting soil types in Essex, eastern England. Plantingwas within a randomized-block design incorporating replicatedplots of each species, both with and without fertilizer. Survivorshipand stem demography were monitored at monthly intervals from1986 to Jun. 1989 for stem recruitment and to Dec. 1991 forstem density. Yields were determined from 1987 (the year followingestablishment) to 1993. Survivorship of the planted propagules over the first 12 monthswas 92% for S. pectinata , 96% for S. cynosuroides and 100%for C. longus. Recruitment of new stems peaked in Apr. of mostyears, although a significant number of new stems appeared asearly as Feb. Stem death peaked in Sep. or early Oct. and allabove-ground stems had died by mid-Nov. Stem density trendsindicated that 2-4 years were required to reach a steady-statedensity, depending upon species. The stem density of the twoSpartina species had reached more than 1000 m^-2 in 1989 althoughthat of S. pectinata fluctuated considerably in the subsequentyears. C. longus stem density rose to approx. 600 m^-2 by 1988and did not change significantly in the subsequent years. In the 6 years following establishment, annual yields averagedacross all fertilizer treatments and both sites were 1·0,1·1 and 1·3 kg m^-2 for C. longus, S. cynosuroidesand S. pectinata, respectively. The average annual yield ofall three species at the site with the heavier soil was 1·3kg m^-2. This was significantly greater than the 1·0 kgm^-2 on the lighter soil. Nitrogen addition did not significantlyincrease yield. Even in the absence of any nitrogen addition,the annual yield of S. pectinata averaged 1·2 kg m^-2over the 6 years, with no evidence of any decline with the increasingage of the stands.Copyright 1995, 1999 Academic Press Cyperus longus, Spartina cynosuroides, Spartina pectinata, energy crop, dry matter yield     

Inhibition of Cl- Channel Activation in Chara corallina Membrane by Lanthanum Ion

We examined a role of Ca²⁺ in the activation of the two majorion channels, i.e., Cl^– and K⁺ channels at the excitationof the characean plasmalemma. The current-voltage relation (I-Vcurve) of the Chara membrane was compared under the ramp voltageclamp condition before and after external application of 20µMof La³⁺ (a Ca²⁺ channel blocker). The transient inward currentcomponent, which is carried mainly by the efflux of Cl^–,disappeared almost completely in about 30 min with La³⁺ treatment.On the other hand, no effect was observed on the late largeoutward current, which is mainly carried by the efflux of K⁺in a large depolarization region (less negative than –50mV). These results suggest that the Cl^– channel in theChara plasmalemma is activated by Ca²⁺ influx, while the K⁺channel is simply activated by depolarization. (Received April 7, 1986; Accepted June 6, 1986)     

The Genetic Basis of Preference for Sweet Substances Among Inbred Strains of Mice: Preference Ratio Phenotypes and the Alleles of the Sac and dpa Loci

Five inbred strains (129/J, BALB/cByJ, C3HeB/FeJ, C57BI/6J andDBA/2J) were examined with two-bottle (48 h) preference ratiotesting across concentrations of sodium saccharin (3 x 10^–4M, 10^–3 M, 3 x 10^–3 M and 10^–2 M), d-phenylalanine(10^–3 M, 10^–2 M and 10^–1 M), and l-glutamine(10^–2 M, 3 x 10^–2 M, 10^–1 M and 3 x 10^–1M). Three consistent groupings of strains were observed acrosssubstances and concentrations:

1. C57BI/6J (preference at low andhigh concentrations);
2. BALB/cByJ and C3HeB/FeJ (preferenceat high concentrations);
3. 129/J and DBA/2J (preference at highconcentration for sodiumsaccharin and indifference to d-phenylalanineand l-glutamine).

If a single locus (presumably dpa or Sac) determines these phenotypes,there are likely to be three alleles. If two independent loci(presumably dpa and Sac) determine these phenotypes, an allelicassignment of Sac^b/dpa^+s for the C57BI/6J strain, Sac^b/dpa^–sfor the BALB/cByJ and C3HeB/FeJ strains, and either Sac^d/dpa^+sor Sac^d/dpa^–s for the 129/J and DBA/2J strains is suggested.Chem. Senses 20: 291–298, 1995.     

Transport and Fixation of Inorganic Carbon in the Cells of Chlorella vulgaris 11h Grown under Ordinary Air

Intracellular accumulation of inorganic carbon (C_i) and itsfixation in photosynthesis were investigated using siliconeoil layer filtering centrifugation technique with the cellsof Chlorella vulgaris 11h grown under ordinary air. Both CO₂and HCO₃^– were transported into the cells from the reactionmedium and accumulated in the cells, but the rate of transportwas much faster for the former than the latter. ¹⁴C-fixationfrom the total transported C_i was much more efficient when CO₂was added in the external medium than when HCO₃^– was added.This indicates that CO₂ and HCO₃^– were not converted tothe common compound in the cells during the initial period ofphotosynthesis. Accumulation of Cⁱ into the cells was much lesssusceptible to low temperature than its fixation. Accumulationof Cⁱ was also observed in the dark. Ethoxyzolamide, an inhibitorof carbonic anhydrase (CA), inhibited the fixation of accumulatedCO₂ in the cells, suggesting that CA enhanced the supply ofCO₂ to the reaction site of ribulose bisphosphate carboxylasein the stroma. Mechanism for transport and fixation of Cⁱ duringphotosynthesis in low-CO₂ cells of C. vulgaris 1lh was proposedfrom these results. (Received March 19, 1986; Accepted June 26, 1986)     

Ion Effluxes during Excitation of Characeae

Ion effluxes during excitation of Chara and Nitellopsis measuredby conductometry method were compared with results obtainedby two other analytical methods, Ag-AgCl method for Cl^–and ion chromatography method for K⁺. In both species, the averageefflux measured by the conductometry method was very close tothose of K⁺ and Cl^–. (Received May 12, 1986; Accepted June 25, 1986)     

Utilization Modes of Inorganic Carbon for Photosynthesis in Various Species of Chlorella

Rates of CO₂ and HCC₃^– fixation in cells of various Chlorellaspecies in suspension were compared from the amounts of ¹⁴Cfixed during the 5 s after the injection of a solution containingonly ¹⁴CO₂ or H¹⁴CO₃^–. Results indicated that irrespectiveof the CO₂ concentration during growth, Chlorella vulgaris 11h and C. miniata mainly utilized CO₂, whereas C. vulgaris C-3,C. sp. K. and C. ellipsoidea took up HCO₃^– in additionto CO₂. Cells of C. pyrenoidosa that had been grown with 1.5%CO₂ (high-CO₂ cells) mainly utilized CO₂, whereas those grownwith air (low-CO₂ cells) utilized HCO₃^– in addition toCO₂. Cells that utilized HCO₃^– had carbonic anhydrase(CA) on their surfaces. The effects of Diamox and CA on the rates of CO₂ and HCO₃^–fixation are in accord with the inference that HCO₃^– wasutilized after conversion to CO₂ via the CA located on the cellsurface. CA was found in both the soluble and insoluble fractions;the CA on the cell surface was insoluble. Independent of the modes of utilization, the apparent K_m (NaHCO₃)for photosynthesis was much lower in low-CO₂ cells than in high-CO₂ones. The fact that the CA in the soluble fraction in C. vulgarisC-3 was closely correlated with the K_m(NaHCO₃) indicates thatsoluble CA lowers the K_m. ¹ Dedicated to the late Professor Joji Ashida, one of the foundersand first president of the Japanese Society of Plant Physiologists. ⁴ On leave from Research and Production Laboratory of Algology,Bulgarian Academy of Sciences, Sofia. (Received September 14, 1982; Accepted March 1, 1983)     

Estimating the Bioenergetic Cost of a Developing Kiwifruit Berry and its Growth and Maintenance Respiration Components

A response surface was developed by regression analysis to quantifythe seasonal respiratory losses by a kiwifruit [Actinidia deliciosa(A. Chev.) C. F. Liang et A. R. Ferguson var. deliciosa cv.Hayward] berry growing in Fresno, CA. The equation of the surfacewas LNRESP = 1·622 + 0·0697 x TEMP –0·0472x DAY + 0·000165 x DAYSQ, where LNRESP is the naturallogarithm of the respiration rate (nmol CO₂ g d. wt^–1s^–1), TEMP is fruit temperature (°C), DAY is the numberof days after flowering, and DAYSQ is the square of the numberof days after flowering. Respiratory losses for a fruit witha final dry mass of 18·5 g were calculated to be 5·57and 5·92 g glucose per fruit per season in 1985 and 1986,respectively. Maintenance respiration was estimated to be 2·84and 3·19 g glucose per fruit per season for 1985 and1986, respectively. The total calculated bioenergetic cost ofkiwifruit berry growth and respiration was 25·25 and25·60 g glucose per fruit per season for 1985 and 1986,respectively. Respiratory losses, expressed as a proportionof the total carbohydrate required for fruit growth, were significant(mean 22·6%). The cost of fruit growth was estimatedto be very similar for two cooler sites (Davis and Watsonville)but estimates of maintenance respiration based on Fresno fruitrespiration data were unrealistically low for the Watsonvillesite. Actinidia deliciosa (A. Chev.) C. F. Liang et A. R. Ferguson var. deliciosa cv. Hayward, kiwifruit, growth respiration, maintenance respiration, bioenergetic costs, model     

Vanadate Sensitive ATPase and Phosphatase Activity in Guard Cell Protoplasts of Commelina

Flicker, M. D. and Willmer, C. M. 1986. Vanadate sensitive ATPaseand phosphatase activity in guard cell protoplasts of Commelina.—J.exp. Bot. 38: 642–648. Phosphatase activity was measured in extracts of guard cellprotoplasts of Commelina communis L. using the artificial substratep-nitrophenylphosphate. A pH optimum of 5.8 to 6.3 was determined.Ammonium molybdate (Ol mol m^–3) and sodium vanadate (1–0mol m^–3) gave almost complete inhibition of phosphataseactivity at pH 60. ATPase assays were, therefore, conductedin the presence of 0–2 mol m ^–3 molybdate and vanadatewas used as a specific inhibitor of plasmamembrane ATPase activity.Vanadate sensitive ATPase activity showed a pH optimum of 6.6and activity was stimulated by KC1. These properties are characteristicof plasmamembrane proton pumping ATPases in other systems andsuggest that proton extrusion in guard cells could be mediatedby a similar enzyme. The maximum ATPase activity is sufficientto account for all the proton flux observed during the stomatalopening response. Key words: ATPase, Commelina, guard cell protoplasts, phosphatase, vanadate     

Changes in Steady-State Photosynthetic 14C-Incorporation into cellular Metabolites of Chlorellax pyrenoidosa during Transitions from High to Low Irradiance

Changes in the levels of ¹⁴C-labelled metabolites were monitoredin Chlorella pyrenoidosa cells during a transition from highto low irradiance, i.e., from 700 to 430 µmol quanta (400–700nm) m^–2 s^–1. Chlorella cells assimilated ¹⁴CO₂ photosynthetically(steady-state ¹⁴C-labelling) for 12 min at the high irradianceand then 10 min at the low irradiance. With the transition tolow light, the level of ¹⁴C-labelled ribulose 1,5- bisphosphate(RuBP) did not decrease, even though the rate of total ¹⁴C-incorporationdecreased by 80%. The data suggest that RuBP carboxylase deactivatesrapidly (within 1 or 2 min) on exposure to low light, causingRuBP pool sizes to be maintained (or even increased) in spiteof a decreased rate of RuBP regeneration. There was also evidenceof light modulation of other enzymes, including some enzymesinvolved in sucrose synthesis. The rate of sucrose synthesisdecreased with decrease in light intensity while the level ofuridine diphosphoglucose increased, but within a few minutes,both returned to their former levels. ¹Present address: Chemical Biodynamics, Lawrence Berkeley Laboratory,Building 3, 1 Cyclotron Road, Berkeley, CA 94720, U.S.A. (Received March 8, 1986; Accepted June 25, 1986)     

Induction of Low Molecular Weight Cadmium-Binding Compound in Soybean Roots

Blakeley, S. D., Robaglia, C, Brzezinski, R. and Thirion, J-P.1986. Induction oflow molecular weight cadmium-binding compoundin soybean roots.—J. exp. Bot. 37: 956–964. A Cd-binding compound has been identified in roots of Cd-treatedsoybean plants (Glycine max L.). Induction of synthesis of thiscompound after treatment with CdCl₂ was monitored by labellingwith either ¹⁰⁹Cd in vitro or with ³⁵S-cysteine or ³⁵S-methioninein vivo. The apparent molecular weight as determined by gelfiltration was about 14000 daltons. However, after carboxymethylationand electrophoresis under denaturing conditions its molecularweight was less than 3400 daltons. Simultaneous labelling with¹⁰⁹ and ³⁵S-cysteine demonstrated that the compound containscysteine residues. Gel electrophoresis of ³⁵S-cysteine or ³⁵S-methioninelabelled protein samples showed the compound to be rich in cysteinebut not in methionine, and that it was induced about 60-foldby 1?0 mol m^–3 CdCl₂ after 7 d. Key words: Soybean, cadmium, induction, root     

Isocitrate Lyase from Germinating Soybean Cotyledons: Purification and Characterization

Ruchti, M. and Widmer, F. 1986. Isocitrate lyase from germinatingsoybean cotyledons: purification and characterization.—J.exp. Bot. 37: 1685–1690. Isocitrate lyase (E.C. 4.1.3.1 [EC] ) was purified from the cotyledonsof 7-d-old soybean seedlings. Three molecular forms were detectedwith pi values of 6·46, 6·25 and 6·0. Themain form (pl = 6·46) had an approximate Mr of 130000,a pH optimum of 8·0, a K_m (isocitrate) close to 2·0mol m^–3 and a molecular activity of 615 min ^–1 at25 °C. The purified enzyme is not a glycoprotein and isheat labile. Key words: Isocitrate lyase, soybean     

Laboratory and field observations on the scuticociliate Histiobalantium from the pelagic zone of Lake Constance, FRG

Histiobalantium sp. was found regularly in the pelagic zoneof Lake Constance, FRG, over five annual cycles. Maxima of upto 6400 cells l^–1 were recorded in late summer, with similarnumbers in the 0–8 and 8–20 m depth intervals. Onan annual average, the population accounted for 10–17%of the total biomass of planktonic ciliates. In the laboratory,Histiobalantium grew well on a diet of the cryptophyte Rhodomonassp. Maximum growth rates obtained in batch cultures were 0.21and 0.33 day^–11 at 9 and 18°C, respectively. In situexperiments using diffusion chambers yielded positive growthrates in autumn and winter. The highest values recorded at theambient temperatures 5, 14 and 17°C were 0.17, 0.32 and0.40 day^–1, respectively. Comparing these results withthe different seasonal distributions and higher measured growthrates of other ciliates from Lake Constance, we conclude thatHistiobalantium is a superior competitor at relatively low algalfood concentrations. ²Present address: Fisheries & Oceans Canada, 4160 MarineDrive, West Vancouver, BC, V7V 1N6, Canada     

Plasma Membrane H+-ATPase in Guard-Cell Protoplasts from Vicia faba L.

The activity of plasma membrane H⁺-ATPase was measured withmembrane fragments of guard-cell protoplasts isolated from Viciafaba L. ATP hydrolytic activity was slightly inhibited by oligomycinand ammonium molybdate, and markedly inhibited by NO₃^–and vanadate. In the presence of oligomycin, ammonium molybdateand NO₃^–, the ATP-hydrolyzing activity was strongly inhibitedby vanadate. It was also inhibited by diethylstilbestrol (DES),p-chloromercuribenzoic acid (PCMB) and Ca²⁺, but slightly stimulatedby carbonyl cyanide m-chlorophenylhydrazone (CCCP). The acitivityhad higher specificity for ATP as a substrate than other phosphoricesters such as ADP, AMP, GTP and p-nitrophenylphosphate; theK_m was 0.5 mM for ATP. The activity required Mg²⁺ but was notaffected by K⁺, and it was maximal around pH 6.8. When guard-cellprotoplasts were used instead of membrane fragments, the ATPaseactivity reached up to 800µmol Pi.(mg Chl)^–1.h^–1in the presence of lysolecithin. These results indicate thatthe guard cell has a high plasma membrane H⁺-ATPase activity. (Received December 23, 1986; Accepted April 28, 1987)     

Inhibition of Sterol Biosynthesis and Stem Elongation of Tobacco Seedlings Induced by Some Hypocholesterolemic Agents

Incorporation of DL-[2-¹⁴C]mevalonic acid ([2-¹⁴C]MVA) into4-desmethylsterols in Nicotiana tabacum cv. Turkish Samson seedlingswas inhibited by SK&F 7997-A₃,¹ SK&F 7732-A₃, AY 9944,and the plant growth retardant, Amo 1618. Reductions in 4-desmethylsterol levels resulted from treatmentwith AY 9944 and Amo 1618, but not the SK&F compounds. Amo1618 and SK&F 7997-A₃ both significantly reduced the specificactivity of each of the four major 4-desmethylsterols examined.Although SK&F 7732-A₃ reduced the specific activity of campesterol,and AY 9944 reduced the specific activity of stigmasterol, neitherhad an effect on the specific activity of ß-sitosterol. Stem elongation of tobacco seedlings was retarded by SK&F7997-A₃, AY 9944, and SK&F 7732-A₃, particularly the former,and the retarded plants thus produced were morphologically indistinguishablefrom the Amo 1618-treated plants. Application of exogenous stigmasterol,or GA₃, to the chemically-retarded plants resulted in a reversalof stem growth retardation.     

Carbon and nitrogen content of Noctiluca scintillans in the Seto Inland Sea, Japan

The carbon and nitrogen content of Noctiluca scintillans cellsfrom the Seto Inland Sea, Japan was investigated in order toestimate its biomass in natural samples. The carbon contentof N.scintillans ranged from 123 to 627 ng C cell^–1 witha mean value of 353 ng C cell^–1, or 1.12 to 2.67 fg Cµm^–3 with a mean value of 1.98 fg C µm^–3.The nitrogen content ranged from 36.0 to 232 ng N cell^–1with a mean value of 131 ng N cell^–1, or 0.499 to 0.910fg N µm^–3 with a mean value of 0.694 fg N µm^–3.Total cell carbon and nitrogen increased but the carbon andnitrogen per cell volume decreased with increasing cell volume.The C/N ratio of the cells ranged from 2.3 to 4.4, which wasrelatively low compared with the Redfield ratio. The carbonand nitrogen content was extremely low (91.2 ng C cell^–1,41.8 ng N cell^–1) for starved cells, whereas it was extremelyhigh (528 ng C cell^–1, 205 ng N cell^–1) for cellswhich had ingested the large diatom, Coscinodiscus wailesii.Our results suggest that the carbon and nitrogen content ofN.scintillans varies depending on its physiological conditionand the type of food that it has recently consumed.