Some characteristics of the carbon compounds released by Daphnia

The Daphnia species studied released 18–100% of the algalcarbon ingested as dissolved and particulate carbon compounds,presumably mainly as feces. The particulate fraction constitutedon average 79 5% of the total released compounds, leaving21% as dissolved compounds. The particles released were verysmall and transparent, not visible by light microscopy Moreover,they contained significant amounts of chlorophyll derivatives.The dissolved compounds consisted mainly of small molecules(mol. wt >10³ daltons), and were shown to be utilized byplanktonic bacteria. Our results show that particulate organiccarbon and chlorophyll a should not be used as measures foralgal carbon in grazing experiments with Daphnia. Both theseparameters were influenced by the animals' fecal particles,yielding lowered clearance rates compared with those obtainedby using cell numbers as a measure for algal carbon.     

Seasonal Dynamics of Nitrogenous Compounds in a Nitrophilic Weed II. The Role of Free Amino Acids and Proteins as Nitrogen Store in Urtica dioica

The free amino acid fraction, representing the most importantnitrogen store in the overwintering below ground plant partsof Urtica dioica, consisted mainly (up to 80%) of asparagineand arginine. While asparagine was dominant in rhizomes, upto one year old, argrnine was specifically accumulated in theolder rhizomes and roots. In spring first the nitrogen storedin asparagine and with a delay of about three weeks that inarginine was mobilized and translocated to the rapidly growingshoots. Proteins may function as nitrogen store too, in particularin the case of the seeds. In the below ground organs, however,a special storage protein, rich in amides and/or arginine, whichwould correspond to the large quantities of these amino acidsin the overwintering organs, could not be detected. (Received September 3, 1984; Accepted November 2, 1984)     

Auxin-Induced Changes in the Molecular Weight Distribution of Cell Wall Xyloglucans in Avena Coleoptiles

The effect of auxin on the molecular weight (Mw) distributionof cell wall xyloglucans was investigated by gel permeationchromatography using coleoptile segments of Avena sativa L.cv. Victory, and the following results were obtained.

1. The water-insoluble hemicellulose (HC-A) mainly consisted ofxyloglucans. Iodine staining method revealed that relativelylarge amounts of xyloglucans were present in the water-solublehemicellulose (HC-B) and water-soluble polysaccharide (WS) fractions.
2. IAA did not cause remarkable changes in xyloglucan contentsin the hemicellulose, but significantly increased the xyloglucancontent in the WS fraction.
3. IAA substantially decreased theweight-average Mw of HC-A. Thiseffect became apparent within30 min of the incubation period,and was not affected by the0.15 M mannitol or 2% sucrose applied.Hydrogen ions also causeda decrease in the weight-average Mwof HC-A; its effect beingreversible.
4. Neither IAA nor hydrogen ions caused any remarkablechangesin the weightaverage Mw of water-soluble xyloglucansin theHC-B.

These results suggest that cell wall xyloglucans have an importantrole in auxininduced cell wall loosening in oat coleoptile cells. (Received May 10, 1984; Accepted August 20, 1984)     

The Period of Circadian Rhythm in Lemna gibba G3 is Influenced by the Substitution of Rubidium for Potassium

Substitution of Rb⁺ for 70% or more of the K⁺ in Lemna gibbaG3 extends the period of the uptake rhythm under a fluence ratelower than 2.3 W/m². The extension is less, the higher the fluencerate and the lower the temperature (2O–33?C). Under 12.4W/m² light at 20?C, the substitution somewhat shortens the lengthof the period. (Received April 23, 1984; Accepted July 14, 1984)     

Separation and Properties of the Inner and Outer Membranes of a Psychrophilic Bacterium, Vibrio sp. Strain ABE-1

The inner and outer membranes from a marine psychrophilic bacterium,Vibrio sp. ABE-1, were separated and purified by desalting,lysozyme treatment, and ultracentrifugation. Enzyme assay, electrophoresis,and chemical analysis showed that they had been highly purified.The isolated outer membrane had no 3-deoxy-D-mannooctulosonate(KDO) and chemical analysis revealed that it contained aboutthree times as much total lipid as the inner membrane. Consequently,the density of the inner membrane was higher than that of theouter membrane (1.19–1.23 gµcm³ for IM; 1.16–1.20g–cm³ for OM). The absolute majority of the lipids inthese membranes were phospholipids, more than half of whichwas identified as phosphatidylethanolamine. Virtually all cardiolipinwas found exclusively in the inner membrane, whereas an unidentifiedphospholipid containing hexosamine was only in the outer membrane.The ratios of unsaturated to total fatty acids in the innerand outer membranes were 76.0% and 69.6%, respectively. (Received February 23, 1984; Accepted August 8, 1984)     

Xyloglucan in the Cell Walls of Suspension-Cultured Rice Cells

The xyloglucan present in the 24% KOH extract of the cell wallsof suspension-cultured rice cells was characterized by fragmentationanalysis with Trichoderma viride cellulase and Aspergillus oryzaeß-D-glucosidase. The xyloglucan is composed mainlyof the following oligosaccharide units: Results showed that the xyloglucan of suspension-cultured ricecells is more extensively branched than is that of rice seedlings.Another structural characteristic of the former xyloglucan isthe presence of D-galactosyl-D-xylosyl side chains that arenot found in the latter. (Received June 15, 1984; Accepted January 11, 1985)     

Purification and Further Characterization of Pyrenoid Proteins and Ribulose-1,5-bisphosphate Carboxylase-Oxygenase from the Green Alga Bryopsis maxima

Pyrenoid proteins and ribulose-1,5-bisphosphate carboxylase-oxygenase(RuBisCO) in the green alga Bryopsis maxima were purified tohigh degrees and their peptide compositions were studied bySDS-polyacrylamide gel electrophoresis. RuBisCO had a largesubunit of 50 kDa and a small one of 16 kDa. The apparent molecularweight of the purified RuBisCO was estimated as 460 kDa by gelfiltration. Pyrenoid proteins had two major polypeptides: 52kDa and 17 kDa. The peptide map of the 52 kDa pyrenoid polypeptidecoincided well with that of the large subunit of RuBisCO, stronglysuggesting that the major component of the pyrenoid of thisalga was RuBisCO. We attempted to survey the distribution ofRuBisCO in the chloroplasts. The results suggested that muchof the RuBisCO of Bryopsis maxima was localized in the pyrenoid.The pyrenoid also contained more than 10 minor polypeptidesnot found in the RuBisCO fraction. The minor polypeptides comprisedabout 15% of the total pyrenoid protein and differed from thepolypeptides of the thylakoid membranes and from those foundin the starch grains surrounding the pyrenoid. (Received February 3, 1984; Accepted July 21, 1984)     

Non-Autotrophic CO2 Fixation and Drought Tolerance in Mosses

Cratoneuron filicinum, a drought-sensitive moss, and Tortularuralis, a drought-tolerant moss, fix CO₂ non-autotrophicallyat a rate of about 1.2 and 2.2 µmol h^–1 g^–1dry wt. respectively. During drying, T. ruralis fixes CO₂ atan undiminished rate until the tissue loses about 60% of theinitial fresh weight. Thereafter, CO₂ fixation rapidly declinesto zero. Dark CO₂ fixation by C.filicinum declines steadilyduring the dehydration period. On rehydration, dark CO₂ fixationis resumed immediately in T. ruralis but not in C.filicinum.When dried T. ruralis is equilibrated with an atmosphere ofnearly 100% relative humidity, its weight increases to about40% of the original fresh weight and dark CO₂ fixation resumesat a rate about 60% of the fresh moss. In C.filicinum thereis only a small increase in weight and little CO₂ fixation inthe dark. The non-autotrophically fixed carbon, in both mossesstudied, is incorporated into amino acids (more than 60% ofthe total, mainly into aspartate, alanine and glutamate) andorganic acids (less than 40% of the total, mainly into malate).It is suggested that on rehydration immediate availability ofNADPH, known to be produced by transhydrogenation from NADHduring dark CO₂ fixation, may be an important factor in therepair of drought-induced cellular damage by reductive biosynthesisof membrane components and other cellular constituents. Key words: Mosses, Dehydration, Rehydration, Dark CO₂ fixation, Amino acids, Organic acids, NADPH, Drought tolerance.     

The Influence of Nitrogen Nutrition on the Accumulation of Free Amino Acids in Root Tissue of Urtica dioica and their Apical Transport in Xylem Sap

Urtica dioica plants were grown on a nitrogen supply of 3, 15and 22 mM with nitrate and ammonium as nitrogen source. In contrastto nitrate reductions amino acid synthesis occurred in roottissue. At 3 mM ammonium obviously the amino acids were rathertransported via xylem upwards to the shoots than stored in theroots. Particularly increased ammonium supply led to stimulatedstorage of free amino acids in the roots, mainly as asparagineand arginine. In xylem asparagine was the dominant nitrogentransporting compound, while arginine was hardly translocated.With the enhancement of nitrogen supply, the second amide, glutamine,became more and more important with respect to the transportof nitrogen. (Received September 3, 1984; Accepted November 2, 1984)     

Comparison of Glycolipids and Plastids in Callus Cells and Leaves of Alnus and Betula

The fine structure of plastids and fatty acid composition ofglycolipids (e.g. monogalactosyl diacylglycerides, MGDG; digalactosyldiacylglycerides, DGDG) in callus cells of Alnus glutinosa,A. incana and Betula pendula cultured in light was comparedwith that in intact leaves. The tissues were qualitatively verysimilar but a rather high amount oflignoceric acid (24:0) wascharacteristic for the callus of A. incana. This fatty acidwas found only in trace amount in other tissues. Linolenic (18:3)and palmitic (16:0) acids are the most abundant (25–65%and 17–27% respectively) fatty acids in all tissues studied.The proportion of 18:1 and 18:2 was much higher in the calluscompared with corresponding intact leaves, which are especiallyrich (48–65%) in 18:3. In callus cultures a higher proportion(17–19%) of linoleic acid (18:2) is found in both Alnusspecies than in the two callus strains of Betula (9–12%). All leaf and callus samples contained esterified steryl glycosidesand two cerebrosidelike spots in thin-layer chromatography,but they were more prominent in callus cultures than in leaves.The callus cells have plastids with rather well developed thylakoidswhich explains the similarity of the main glycolipid components(MGDG and DGDG) to that of leaves. (Received April 23, 1984; Accepted August 17, 1984)     

Location of the Electron Accepting Site of a b-Type Cytochrome Oxidase in Purified Chromatophore Membranes and Spheroplast Membrane Vesicles from Photosynthetically Grown Rhodopseudomonas sphaeroides

Purified chromatophore membranes and spheroplast membrane vesicleswere prepared from Rhodopseudomonas sphaeroides by the methodof Michels and Konings (1978). The cytochrome c oxidase activityof the spheroplast membrane vesicles was about 15-fold higheron a bacteriochlorophyll basis than that of the chromatophoremembranes. The cytochrome c oxidase activity of the chromatophoremembranes was greatly stimulated (about 10-fold) by an additionof Triton X-100 (0.1%), but there was less stimulated (about1.5-fold) in the case of spheroplasts. The pH optimum of theoxidase activities of the spheroplast membrane vesicles andof chromatophore membranes treated with 0.1% Triton X-100, andthe salt dependencies of the activity of both preparations werethe same. These results show that the membrane-bound b-type cytochromeoxidase of this bacterium accepts electrons from ferrocytochromec on the periplasmic side of the membrane (on the outer surfaceof the spheroplast membrane vesicles). These results also areconsistent with the fact that cytochrome c₂ is located in theperiplasmic space in this bacterium. (Received January 24, 1984; Accepted April 24, 1984)     

Feeding of Sagitta enflata and vertical distribution of chaetognaths in relation to low oxygen concentrations

Zooplankton samples were collected in Mejillones Bay, northernChile (23°00'15'S, 70°26'43'W ). Sampling was conductedat 4 h intervals, for 24 h during three seasons, austral spring(October 2000), summer ( January 2001) and winter (August 2001)at three different strata (0–25, 25–50 and 50–100m). Five species of chaetognaths were collected. Sagitta enflatawas the most abundant species, representing up to 65% of allchaetognaths in total numbers, followed by Sagitta bierii, makingup 34% of the total abundance of chaetognaths. S. enflata wasdistributed mainly above the Oxygen Minimum Zone, while S. bieriiremained below this zone. Feeding rates were relatively constantwithin the upper layer (0–25 m depth), for each samplingdate, averaging 1.2 prey S. enflata day^–1, and decreasingwith depth. Gut content analyses demonstrated that predationwas principally focused on small copepods (<1500 µm),with greatest feeding activity occurring at night. The dailypredation impact on the total standing stock of small copepodsvaried seasonally between 6% in spring and 0.4% in winter. Thispercentage may represent a negligible impact on the entire copepodcommunity, but it is relevant at the species or genus level,since S. enflata removed more than 20% of the standing stockof Centropages brachiatus and Corycaeus sp. Thus, during someperiods of the year, chaetognaths may strongly influence theabundance and size distribution of copepods in coastal upwellingecosystems.     

Embryogenic Callus Induction from Coffea arabica Leaf Explants by Benzyladenine

Coffea arabica leaf explants cultured on medium with 5 µM6-benzyladenine (BA) as the sole plant growth regulator producedwhite friable calluses that formed somatic embryos. These calluseshave been subcultured on the same medium for more than 2 yearsand maintain the ability to produce somatic embryos. (Received October 3, 1984; Accepted January 18, 1985)     

Specific Labeling of Cell Wall Polysaccharides with myo-[2-3H] Inositol during Germination and Growth of Phaseolus vulgaris L.

myo-[2-³H]Inositol was fed to bean seeds by imbibition and itsmetabolic fate was studied during germination and seedling growth.The largest amount of myo-inositol was taken up from a 500 HIMsupply (8 mg/seed) and the highest percentage was from 1 HIM(29%). myo-Inositol was incorporated to new cell wall polysaccharidesof hypocotyl and roots, mostly as uronic acid and pentose residues.In the 80% ethanolinsoluble cell walls of hypocotyls at 3, 4and 5 days after imbibition, 47 to 52% of ³H was detected asuronic acids, 20 to 24% as arabinose and 11 to 19% as xylose.Glucogenesis from myo-inositol was low: less than 6% was recoveredas hexoses. The ³H in uronic acid and arabinose residues decreasedwith increasing age (i.e. 0 to 6 cm from cotyledons) and increasedin older segments (further than 6 cm from cotyledons). In theoldest segment of 5-day-old hypocotyl (> 10 cm), ³H in thesugar residues was more than that in the youngest part (0–2cm). On the other hand, ³H in xylose residues increased steadilyin the older part, but did not exceed that in arabinose. The results show that the myo-inositol oxidation pathway functionsin growing hypocotyls and roots of bean seedlings to provideexclusively uronic acid and pentose units for cell wall synthesis.Results also show that incorporation of arabinose and uronicacids derived from myo-[2-³H]inositol to cell wall polysaccharidesis active in two regions of the hypocotyl; first, for the constructionof the primary walls in the young, growing region of the hypocotyl,and second, for thickening of the walls after completion ofelongation growth. ¹Supported by NSERC of Canada. (Received April 10, 1984; Accepted June 12, 1984)     

A Novel Gene Isolated from Human Placenta Located in Down Syndrome Critical Region on Chromosome 21

Down syndrome is the most common birth defect, which is causedby trisomy 21. We identified a novel gene in the so-called Downsyndrome critical region by EST mapping to genomic DNA and followingcDNA cloning. The gene, designated DCRB (Down syndrome CriticalRegion gene B), consisted ofthree exons of1095 bp in total andencoded a large open reading frame of118 amino acid residues.The amino acids sequence ofDCRB showed no significant homologyto any known protein. Northern blot analysis showed that DCRBis mainly expressed in the placenta, in which a major 1.1-kbband and a minor 2.0-kb band were detected. Minor bands of 1.4kb and 2.2 kb were also detected in adult heart and skeletalmuscle.     

Column Chromatographic Separation of Chlorophyllide {alpha} and Pheophorbide {alpha}

Chlorophyllide a, pheophorbide a, chlorophyll a and pheophytina were separated in a short time by anion-exchange chromatographywith a short column of DEAE-Sepharose CL-6B. (Received February 16, 1984; Accepted April 13, 1984)     

Digitoxin Biosynthesis in Isolated Mesophyll Cells and Cultured Cells of Digitalis

In the laminae of Digitalis, most of the digitoxin present isfound in the mesophyll. A new method for determining the amountof digitoxin biosynthesis using a digitoxin antibody was devisedto estimate this activity in isolated mesophyll cells and culturedcells. Isolated mesophyll cells showed significant activity,which suggests that the site of biosynthesis and the accumulationof cardenolides in a lamina of Digitalis is mainly in the mesophyllcells. Of five liquid cultures of D. purpurea; green shoot-formingcultures, white shoot-forming cultures, root-forming cultures,undifferentiated green cells and undifferentiated white cells,the green shoot-forming cultures had the highest activity. Thewhite shoot-forming cultures had about one-third the activityof the green shoot-forming cultures, and the other three cultureshad very low activity. No stimulatory effect of light was foundduring the 48-h incubation. (Received January 19, 1984; Accepted June 8, 1984)     

Mixed Nitrogen Nutrition and Productivity of Wheat Grown in Hydroponics

The objective of this study was to study the effects of nitrogen(N) supplied as either mixtures of NO₃ and NH₄ or as all NO₃on the final yield of spring wheat. Two separate greenhouseexperiments evaluated the durum spring wheat (Triticum durumL.) cultivar ‘Inbar’ in 1986, and the hard red springwheat (Triticum aestmum L.) cultivar ‘Len’ in 1987.Nitrogen treatments consisted of all NO, or mixtures (75/25or 50/50) of NO₃ and NH₄. At maturity, plants were harvested,separated into leaves, stems, roots, and grain, and each partanalysed for dry matter and chemical composition Compared to plants receiving only NO₃ as the source of N, mixedN nutrition resulted in greater accumulation of whole plantreduced-N (49 to 108% more), phosphorus (38 to 69% more), andpotassium (25% more) for both cultivars. In all cases, plantsproduced higher grain yields (28% for Len to 78% for Inbar)when grown with mixed N nutrition than with only NO₃. The yieldincrease was not associated with heavier grains or more grainsper ear, but rather with an increase in the number of ear-bearingtillers per plant. For both cultivars, the higher yields withmixed N resulted from the production of more total biomass (36to 76%) as the partitioning of dry matter between plant partswas not altered by N treatment. Under the hydroponic conditionsof this experiment, the utilization of both NO₃ and NH₄ resultedin greater growth, nutrient absorption, and yield than NO₃ alone,which was primarily associated with an enhancement in tillerdevelopment Triticum aestivum L., Triticum durum L., spring wheat, hydroponics, ammonium nutrition, nitrate nutrition, tillering, yield components, partitioning     

Messenger RNA in the Ungerminated Pollen Grain: A Direct Demonstration of its Presence

The presence of presynthesized messenger RNAs in the mature,dehydrated pollen grains of Tradescantia paludosa L. has beendemonstrated by translation of total RNA and poly (A)⁺ RNA ina wheat germ cell-free system, and a comparison of in vitroand in vivo synthesized proteins by SDS-polyacrylamide gel electrophoresis.The mRNAs are capped at their 5'-termini with a guanosine 5'phosphate moiety which is methylated. messenger RNAs, guanosine 5' phosphate, pollen, Tradescantia paludosa L     

Different Intracellular Localization of Two Cytochrome P-450 Systems, Ipomeamarone 15-Hydroxylase and Cinnamic Acid 4-Hydroxylase, in Sweet Potato Root Tissue Infected with Ceratocystis fimbriata

Ipomeamarone 15-hydroxylase activity was mainly recovered inthe pellet fraction between centrifugations at 10,000 and 100,000?gfrom a crude extract of Ceratocystis fimbriata-infected sweetpotato root tissue, whereas cinnamic acid 4-hydroxylase activitywas found between centrifugations at 300 and 10,000?g. Whenparticles in the crude extract were fractionated by sucrosedensity gradient centrifugation, the rough-surfaced microsomeswere distributed over a wide density range from 1.09 to 1.14g cm^–3, judging from the distributions of protein, RNAand NADPH-cytochrome c reductase activity. Phosphorylcholine-glyceridetransferase activity was only in the lighter half of the microsomalfraction (density: 1.09–1.11 g cm^–3). Ipomeamarone15-hydroxylase activity was found in heavier half of the microsomalfraction (density: 1.10–1.14 g cm^–3). We proposethat this tissue has two rough-surfaced endoplasmic reticulumspecies, only one of which carries phosphorylcholine-glyceridetransferase, and that the cytochrome P-450 system is localizedon the species lacking the enzyme. Cinnamic acid 4-hydroxylaseactivity was mainly found in a fraction that had densities of1.17–1.19 g cm^–3 and contained vesicular particlesof various sizes. ¹ Present address: Laboratory of Food Hygienics, Faculty ofAgriculture, Kagawa University, Miki-cho, Kida-gun, Kagawa 761-07,Japan. (Received September 6, 1984; Accepted December 27, 1984)