Cellular fatty acid composition of six fastidious,gramnegative, xylem-limited bacteria from plants

Composition of cellular fatty acids was determined for strains of fastidious, Gramnegative, xylem-limited bacteria causing or associated with Pierce's disease of grapevine, phony disease of peach, plum leaf scorch, stunt of ragweed, elm leaf scorch, and periwinkle wilt. The most abundant fatty acids were straight-chain 150, 160, 170, and 180, unsaturated 161, 181, and unsaturated 17-and 19=carbon homologs. Minor fatty acids included straightchain 120, 140, 190, and 200; an unsaturated 15-carbon homolog; hydroxy-substituted 2-OH 120, 3-OH 120, and 3-OH 140; and branched chain iso-140 and iso-200. Cyclopropane acids were not detected. Physiological age had no effect on fatty acid composition. Class analysis of data indicated relative uniformity within the group. Saturated even-carbon chains comprised 31%–42%, unsaturated acids 41%–52%, saturated odd-carbon chains 10%–18%, hydroxysubstituted chains 2%–7%, and branched-chains 1%–4% of total fatty acids. The ratio of saturated-unsaturated acids ranged from 0.8 to 1.2.     

A new type of phosphoglycolipid fromPseudomonas cepacia

A new type of phosphoglycolipid was found inPseudomonas cepacia. A ratio of glucosamine/phosphoric acid/polar fatty acid/nonpolar fatty acid was resolved to be 1 1 1 1, and the molecular weight of the phosphoglycolipid was determined to be 777 by mass spectrometry. An amino group of the glucosamine was free, and both a phosphoric acid and two kinds of fatty acids were bound directly to the glucosamine.     

Isolation of mutants ofLactobacillus bacterias,grown in skim milk,that resist fatty acid inhibition

The effect of saturated fatty acids from 60 to 160 and oleic acid was determined onLactobacillus leichmannii growing in skim milk. The growth of this strain was markedly inhibited by fatty acids from 80 to 120 but not by straight chain fatty acids greater than 130 or less than 70 and oleate. Laurate was the fatty acid with the highest bactericidal effect. Similar results, with little changes depending on strains, were obtained withL. casei, L. plantarum, L. bulgaricus, L. lactis, L. helveticus. Mutants from theseLactobacillus organisms, resistant to fatty acid inhibition, were isolated by a recycling selection procedure. These mutants exhibited high levels of oxidation for laurate. The presence of 2 mM of this compound in the skim milk culture increased the fatty acid oxidation activity four to ten times higher than was exhibited by the parent strains. The practical implications of these observations are discussed.     

Bioassembly of acyl lipids in microspore-derived embryos of Brassica campestris L.

The native lipid composition and the capacity of cell-free extracts to biosynthesize acyl lipids in vitro were determined for the first time using the recently reported microspore-derived (MD) embryo system from the Brassica campestris low erucic acid line BC-2 (Baillie et al. 1992). The total lipid fraction isolated from midcotyledonary stage MD embryos (21 days in culture) was composed primarily of triacylglycerol (76%) with an acyl composition quite similar to that of mature BC-2 seed. When incubated in the presence of glycerol-3-phosphate, ¹⁴C 181-CoA, and reducing equivalents, homogenates prepared from 21-day cultured MD embryos were able to biosynthesize glycerolipids via the Kennedy pathway. The maximum in vitro rate of triacylglycerol biosynthesis could more than account for the known rate of lipid accumulation in vivo. The homogenate catalyzed the desaturation of 181 to 182 and to a lesser extent, 183. The newly-synthesized polyunsaturated fatty acids initially accumulated in the polar lipid fraction (primarily phosphatidic acid and phosphatidylcholine) but began to appear in the triacylglycerol fraction after longer incubation periods. As expected for a low erucic acid cultivar, homogenates of MD embryos from the BC-2 line were incapable of biosynthesizing very long chain monounsaturated fatty acyl moieties (201 and 221) from 181-CoA in vitro. Nonetheless, embryo extracts were still capable of incorporating these fatty acyl moieties into triacylglycerols when supplied with ¹⁴C 201-CoA or ¹⁴C 221-CoA. Collectively, the data suggest that developing BC-2 MD embryos constitute an excellent experimental system for studying pathways for glycerolipid bioassembly and the manipulation of this process in B. campestris.Abbreviations CPT sn-1,2-diacylglycerol cholinephosphotransferase - DAG diacylglycerol - DGAT diacylglycerol acyltransferase - DGDG digalactosyldiacylglycerol - G-3-P glycerol-3-phosphate - G-3-PAT glycerol-3-phosphate acyltransferase - LPA lyso-phosphatidic acid - LPAT lyso-phosphatidic acid acyltransferase - LPC lyso-phosphatidylcholine - LPCAT acyl-CoA: lyso-phosphatidylcholine acyltransferase - LPE lyso-phosphatidylethanolamine - MGDG monogalactosyldiacylglycerol - PA phosphatidic acid - PA Phosphatase, phosphatidic acid phosphatase - PC phosphatidylcholine - PE phosphatidylethanolamine - PG phosphatidylglycerol - TAG triacylglycerol - 181-CoA oleoyl-Coenzyme A - 181 oleic acid, cis-9-octadecenoic acid - 182 linoleic acid, cis-9,12-octadecadienoic acid - 183 -linolenic acid, cis-9,12,15-octadecatrienoic acid - 201 cis-11-eicosenoic acid - 221 erucic acid, cis-13-docosenoic acid; all other fatty acids are designated by number of carbon atoms: number of double bonds National Research Council of Canada Publication No. 35896     

Synthesis of aspartame precursor by solid thermolysin in organic solvent

Summary The enzymatic synthesis of a peptide compound was carried out successfully in homogeneous organic solvent.Solid Thermolysin was found to catalyze the synthetic reaction of N-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester (Z-APM; a precursor of sweetner Aspartame) from N-benzyloxycarbonyl-L-aspartic acid (Z-L-Asp) and L-phenylalanine methyl ester (L-PheOMe) in a 98 percent organic medium (ethylacetatebenzenemethanolwater=5029192). The dissolution of enzyme was not observed. The optimal pH shifted to acidic side by 1.0 pH unit, compared with that in aqueous medium. The enzymatic activity of solid thermolysin with an average size of 3.4×9.5 m was determined to be 0.18 moles-product/(mg-solid)·h under the initial concentrations of L-PheOMe of 0.1M and Z-L-Asp of 0.05M, and at pH 6.0 and 40°C.     

Interaction of free fatty acids with the erythrocyte membrane as affected by hyperthermia and ionizing radiation

The interference of hyperthermia and ionizing radiation, respectively, with the effects of capric (100), lauric (120), myristic (140), oleic (cis-181) and elaidic (trans-181) acids on the osmotic resistance of human erythrocytes was investigated. The results are summarized as follows: (A) not only at 37°, but also at 42° and 47°C lauric acid (120) represents the minimum chain length for the biphasic behaviour of protecting against hypotonic hemolysis at a certain lower concentration range and hemolysis promotion at subsequent higher concentrations; (B) with increasing temperatures the protecting as well as the hemolytic effects occur at lower concentrations of the fatty acids; (C) the increase of temperature promotes the extent of hemolysis and reduces the extent of protection against hypotonic hemolysis; (D) Gamma-irradiation of erythrocytes selectively affects the concentration of oleic acid at which maximum protection against hypotonic hemolysis occurs, without altering the minimum concentration for 100% hemolysis.     

Genetics of fertility restoration in hybrid rice

Summary The cross combination involving 14 male-sterile lines in rice, when crossed with different maintainers, showed fertility restoration in certain combinations. When F₂ segregating populations were classified based on spikelet fertility, fertility restoration was shown to be governed by 31, 9331, and 1231, due to allelic differences. This indicated that the cytosterility of the same group showed monogenic fertility restoration, whereas crossing plants belonging to different cystosterile groups showed a digenic pattern of segregation.     

Fatty acid composition of myelin lipids from developing rat forebrain and spinal cord: Influence of experimental hyperphenylalaninaemia

The fatty acid composition of individual myelin lipids from rat forebrain and spinal cord was analysed at 20 and 30 days p.p. During this phase of rapid myelination the proportions of C 160 and C 180 fatty acids decreased whereas the relative amounts of long chain and unsaturated fatty acids increased in most lipid classes. This developmental shift was more pronounced in the forebrain and was different with respect to both magnitude and direction for each myelin lipid. The experimental induction of chronic hyperphenylalaninaemia (hyper-Phe) from day 3 p.p. lead to alterations in the rate of myelination, which were most pronounced in the forebrain. At 20 days, especially in the forebrain, chain elongation and desaturation of fatty acids were delayed. This delay was maximal in sphingomyelin fatty acids, C 180 and C 241. In hydoxycerebrosides, the shift in the fatty acid composition from C 160 to C 220 was accelerated in hyper-Phe rats. No significant difference in the fatty acid composition of any myelin lipid was found in the more mature myelin (spinal cord, 30 days p.p.). No evidence was found of a primary effect of hyper-Phe on myelination.In memoriam of M. Klapproth who died by a tragic accident.     

Nutrient proportions in foliage of semi-mature loblolly pine

Summary Nitrogen and phosphorus limitations to growth are common in many loblolly pine (Pinus taeda) stands. Interactions of these nutrients may complicate interpretation of foliar nutrient analysis for predicting response to forest fertilization. Proportions of foliar nutrient concentrations (and the changes in these proportions following fertilization) were examined in 36 semi-mature loblolly pine plantations in the southeastern United States. Mean proportions of nutrient concentrations (NPKCaMg) for non-fertilized stands were 1009.336.517.29.2. Potassium and phosphorus were higher. Nitrogen fertilization generally decreased the PN ratio and enhanced growth, indicating a nitrogen deficiency in most stands under study. Additions of nitrogen and phosphorus together yielded a significant increase in the PN ratio. Effects of fertilization effects on other nutrient concentration ratios were also examined.Paper No. 9401 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695.     

Sex determination in Sarotherodon (tilapia)

Summary The simplest possible model of the sex determination process adding autosomal influence to a minimal number of sex chromosomes was developed to explain matings of Tilapia (Sarotherodon) species. Eighteen different genotypes, each having two autosomes (AA, Aa, or aa) and two sex chromosomes (WX, WY, WW, XY, XX or YY) involved in sex determination, are predicted by the theory. Their sex (10 males and 8 females) were determined using a series of directed graphs, showing the relative strength of the chromosome pairs, developed on the basis of Chen's sex ratio results (Chen 1969). This theoretical model predicts eight different sex ratios (01, 13, 35, 11, 97, 53, 31, 10 ); three of them are not predicted by the WXYZ theory. The greatest part of these sex ratios have been obtained experimentally in extensive series of crosses between related species of Tilapia and their hybrids, carried out by several authors. The theory succeeds in explaining all of Chen's results, including those ratios 53 and 01 seen in certain crosses but not predicted by the WXYZ theory. The importance of the autosomes is seen in comparisons of the genotype pairs (AaWY, aaWY), (AaXY, aaXY) and (AAWW, AaWW) in which the first genotype in each case is male while the second is female as proven by the sex ratio results. The members of the pair differ only in the substitution of one autosome for the other. To test the theory, experiments consisting of hormonal sex reversion and a series of crosses are proposed. Finally, theoretical and practical implications of the theory are discussed.     

Characterization of the guanosine 3′∶5′-monophosphate-dependent protein kinase from silkworm eggs and analysis of the endogenous protein substrates

Summary In a previous paper, two types of cyclic nucleotide-dependent protein kinases, namely cGMP dependent G-kinase and cAMP dependent A-kinase, in silkworm eggs has been reported (Takahashi et al. 1975; Takahashi 1976). One of these, G-kinase, has now been purified 2400-fold by means of ammonium sulfate fractionation, chromatography on hydroxylapatite, DEAE cellulose, and gel filtration.Some of the properties of the enzyme are described. The enzyme is highly dependent on cGMP; it is strongly inhibited by GTP in a noncompetitive manner not only for ATP but also for cGMP. GTP was found to be highly inhibitory on G-kinases from various tissues of the silkworm, but did not inhibit the A-kinase.Incubation of the egg extract with [-³²P]ATP and Mg²⁺ led to the formation of three major³²P-labelled proteins, with molecular weights of 42.000, 70.000 and 180.000 as analyzed by SDS polyacrylamide gel electrophoresis. Two of them corresponded to the subunits of vitellin.The silkworm vitellin was effectively phosphorylated both by the highly purified G-kinase and by the A-kinase. It is concluded that the G-kinase is involved in the phosphorylation of vitellin in developing silkworm eggs.Abbreviations cAMP adenosine 35-monophosphate - cGMP guanosine 35-monophosphate - A-kinase adenosine 35-monophosphate-dependent protein kinase - G-kinase guanosine 35-monophosphate-dependent protein kinase - MIX 1-methyl-3-isobutylxanthine     

Bacterial regeneration of ammonium and phosphate as affected by the carbon:nitrogen:phosphorus ratio of organic substrates

The effect of carbonnitrogenphosphorus (CNP) ratio of organic substrates on the regeneration of ammonium and phosphate was investigated by growing natural assemblages of freshwater bacteria in mineral media supplemented with the simple organic C, N, and P sources (glucose, asparagine, and sodium glycerophosphate, respectively) to give 25 different substrate CNP ratios. Both ammonium and phosphate were regenerated when CN and NP atomic ratios of organic substrates were 101 and 161, respectively. Only ammonium was regenerated when CN and NP ratios were 101 and 10–201, respectively. On the other hand, neither ammonium nor phosphate was regenerated when CN and NP ratios were 151 and 51, respectively. In no case was phosphate alone regenerated. As bacteria were able to alter widely the CNP ratio of their biomass, the growth yield of bacteria appeared primarily dependent on the substrate carbon concentration, irrespective of a wide variation in the substrate CNP ratio.     

Glycerolipid-fatty-acid desaturase deficiencies in chloroplasts from fruits of Capsicum annuum L.

Chloroplasts from fruits and leaves of Capsicum annuum cv. Bell Tower were purified on sucrose gradients, and the lipids were separated by column and thin-layer chromatography. The glycerolipids mono- and digalactosyldiacylglycerol (MGDG, DGDG), sulfoquinovosyldiacylglycerol (SQDG), and phosphatidylglycerol (PG) were quantified, and the fatty-acid composition at the 1 and 2 positions of the glycerol moiety (sn-1 and sn-2) was determined after hydrolysis with position-specific lipases. In fruit chloroplasts, ³-trans hexadecenoate (trans-3-161) was absent and replaced by palmitate (160) at sn-2 of PG, and ^7,10,13-hexadecatrienoate (163) at sn-2 of MGDG was greatly reduced and largely replaced by linoleate (182). The ratio of 182 to linolenate (183) was consistently greater in glycerolipids from fruit compared with leaf chloroplasts. The lower percentage of C-16 fatty acids at sn-2 indicated that prokaryotic molecular species were reduced by 15% in DGDG, 40% in SQDG, and 90% in MGDG, in fruit compared with leaf chloroplasts. The MGDGDGDG ratios in fruit and leaf chloroplasts were 1.21 and 2.21, respectively. Taken together, the data indicate that chloroplasts in Capsicum fruit are deficient in three desaturases: those that convert 1) 160 to ³-trans-161 at sn-2 of PG, 2) 160 to ⁷–cis-161 at sn-2 of MGDG, and 3) 182 to 183 at both sn-1 and sn-2 of various chloroplast glycerolipids.Abbreviations Chl chlorophyll - DGDG digalactosyldiacylglycerol - FS free sterol - GL galactolipid - MGDG monogalactosyldiacylglycerol - PE phosphatidyl ethanolamine - PG phosphatidylglycerol - PL phospholipid - SQDG sulfoquinovosyldiacylglycerol We are grateful to Dr. Roger Calza for providing us with the tobacco gt11 cDNA expression library and to Dr. Eric Huttner for his advice throughout the screening procedure. We also wish to thank M. Gosse for his assistance in growing and maintaining our plants. T.W.B. was supported by a BAP research grant from the Commission of the European Communities.     

Das Fettsäurespektrum des Flußkrebses Orconectes limosus und seine Beeinflussung durch die Nahrung

Zusammenfassung Das Fettsäurespektrum von männlichen Flußkrebsen Orconectes limosus wurde gaschromatographisch analysiert. Die Fettsäuren zeigen eine spezifische Verteilung auf die Lipidklassen. Das Gesamtspektrum entspricht dem Schema eines Süßwassertieres im Winter mit den größten Fraktionen: C160, C181, C202 und C204 (zusammen rund 53% der Gesamtfettsäuren). In verschiedenen Fraktionen wurden ungeradzahlige und verzweigte Fettsäuren gefunden.Durch Verfüttern von Kartoffeln und Lebertran kann das Spektrum beeinflußt werden. Die Fettsäuren in der Mitteldarmdrüse, und in weniger starkem Ausmaß im Restkörper, haben sich nach viermonatiger Fütterung mit Lebertran qualitativ und quantitativ dessen Spektrum angepaßt. Nach siebenmonatiger Fütterung mit Kartoffeln zeigen Mitteldarmdrüse und Restkörper ein Spektrum, das dem eines unbehandelten Kontrollkrebses ähnelt und sowohl C182 als auch höher ungesättigte Fettsäuren enthält. Hierfür kommen zwei Deutungen in Frage: Entweder wird das Spektrum der Fettsäuren in Strukturlipiden aufrechterhalten, oder es erfolgt de novo Synthese einer C18:2^6,9-Fettsäure und deren Verlängerung analog dem von Wirbeltieren bekannten Weg.

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The fatty acid composition in the crayfish, orconectes limosus, and the effect of nutrition

Summary The fatty acid composition in the male crayfish, Orconectes limosus, was analysed by gas-liquid chromatography. The fatty acids were found to have a specific distribution in different lipid classes. The composition corresponds to those of a fresh water animal in winter with the most important acids: C160, C181, C202, C204 (including about 53% of the total acids). In several lipid classes oddnumbered and branched chain fatty acids could be detected.Potatoe and fish-liver oil diets influence the fatty acid composition. After four months feeding with fish-liver oil the fatty acids in the hepatopancreas and to a lesser extent in the rest body show a similar spectrum as the fed oil. After feeding for seven months with potatoes hepatopancreas and rest body exhibit a fatty acid composition representative for the untreated animal, which contains C182 as well as higher unsaturated acids. These findings support the hypotheses that either the fatty acid composition in the structure lipids was maintained or that a C182^6,9-fatty acid was de novo synthesized and elongated analogous to the known vertebrate pathway.

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Herrn Prof. Dr. K. Urich bin ich für die Anregung zu der vorliegenden Arbeit und für die Durchsicht des Manuskriptes zu großem Dank verpflichtet.     

On the structure of carminic acid and carmine

Summary The chemical mechanism and histochemical significance of carmine stains are not yet understood. To determine possible effects of dye configuration on staining patterns we built models of dye molecules with the Stuart-Briegleb-type of atomic models. However, steric hindrance prevented construction of carmine according to the formula suggested by Harms. A review of recent chemical literature showed that the widely accepted formula of carminic acid is incorrect; the carboxyl group is not in the 5 but in the 7-position, and the side-chain is not a methylpentose but a hexose. Models based on the revised structural formula could be combined to 211 carminic acid-Al-Ca complexes. But formation of the central Al-O-Ca-O-Al bridge of the conventional 421 carminic acid-Al-Ca formula of carmine was still impossible. It is suggested that carmine may be a 211 compound analogous to the 211 alizarin-Al-Ca complex established by Kiel and Heertjes. Investigations of carmine were rendered difficult by wide variations in the staining properties of dye samples and the lack of data concerning the composition of various batches of carmine.     

Lipid content and related fatty acid composition in fallow and cultivated soil in Southern Italy

Summary Lipid content and fatty acid composition were studied on two soil samples—one fallow and one cultivated with durum wheat. Differences between the cultivated and the fallow subunits in respect of total lipid content and the saponifiable fraction were found to be statistically significant. The same observations were made on some fatty acids showing either accumulation or decrease after three years. The interaction years x cultivation for total lipid content and the saponifiable fraction was significant showing that accumulation and reduction of lipids in the cultivated subunit are in equilibrium. The interaction dates of sampling x cultivation was significant for C150, C160, C180, C240 as their contents increase in the cultivated subunit in July as a result of the decomposition of plant tissues.Results discussed in this paper have been partially presented at the 7th International Symposium Humus et Planta Brno (Czechoslovakia) on August 20–25th 1979. This research has been partially supported by the National Research Council of Italy and carried out at the Experimental Institute of Agronomy — Bari — Italy (Director: F. Lanza).     

Inhibition by n-3 fatty acids of arachidonic acid metabolism in a primary culture of astroglial cells

Docosahexaenoic acid (226 n-3) was present in low concentrations in a primary culture of rat brain astroglial cells, when compared to brain cortex. We have thus supplemented these cells with this fatty acid and investigated the effects of its incorporation in cell phospholipids on the conversion of arachidonic acid, 204 n-6, through the cyclo and lipoxygenase pathways, after cell stimulation. Docosahexaenoic acid-enriched cells produced less thromboxane B₂ and 6-keto-Prostaglandin F₁ and markedly less 12-hydroxyeicosatetraenoic acid than unsupplemented cells, after stimulation with the Ca²⁺-ionophore A23187. The production of 15-hydroxyeicosatetraenoic acid from arachidonic acid was slightly increased in docosahexaenoic acid-supplemented cells. We have also supplemented these cells with eicosapentaenoic acid (205 n-3) and, in addition to accumulation of this fatty acid in cell phospholipids, we found elevation of 225 n-3 and some increment of 226, confirming that glial cells are able to convert eicosapentaenoic acid to the long chain, more unsaturated derivatives. In conclusion, n-3 fatty acids, when supplemented to glial cells, appear to modulate the arachidonic acid cascade and to be converted through the elongation and desaturation pathways.     

Osmotropotaxis inDrosophila melanogaster

Summary Drosophila melanogaster is able to perform osmotropotaxis under open-loop conditions. With an optimal stimulus the average turning tendency to the side of higher concentration corresponds to a circular track with radiusr=0.8 cm. The response amplitude does not decrease within 1 or 2 h. Unilaterally antennectomized flies in an homogeneous odor field show a permanent turning tendency towards their intact side.The smallest concentration ratio to elicit osmotropotaxis in normal flies ranges between 610 and 910 at high and between 210 and 510 at an about 50 times lower odor intensity. No negative tropotaxis (i.e. turning to the side of lower concentration) is observed, even with strong repellents.     

Hexahydroindanone derivatives of steroids formed by Rhodococcus equi

Summary A mutant strain of Rhodococcus equi accumulates three metabolites from the androst-4-ene-3,17-dione or from its degradation intermediate, 3a-H-4(3'-propionic acid)-7a-methylhexahydro-1,5-indanedione (MEPHIP). These three metabolites are: 3a-H-4a(3'-propionic acid)-5-hydroxy-7a-methylhexahydro-1-indanone--lactone (HIL); 3a-H-4(3'-trans acrylic acid)-5-hydroxy-7a-methylhexahydro-1-indanone (^2'-5-hydroxy-MEPHIP); and 3a-H-4(3'-hydroxy-3'-propionic acid)-5-hydroxy-7a-methylhexahydro-1-indanone (3'-hydroxy-HIL). The behaviour of this mutant allows us to propose a pathway for degradation of the intermediates, methylperhydroindanone propionates. However, during this degradation, the side-chain propionate was eliminated by a-oxidation mechanism. Offprint requests to: A. Miclo     

Developmental changes in the fatty acids of synaptic membrane phospholipids: Effect of protein malnutrition

The acyl-linked fatty acid composition of the major phospholipid species in rat cortical synaptic membranes was determined at various stages of development. For most species there was a decrease during development in the short chain saturated fatty acids, 140 and 160, an increase in 180 and 226 (n-3) and an increase in the ratio of 226 (n-3)/225 (n-6). Pups were protein deprived by feeding the dams a 12% casein diet as compared to the 24% casein control diet. Protein malnutrition markedly affected the composition of acyl-linked fatty acids in the synaptic membranes. The increases in the ratio of 226 (n-3)/225 (n-6) fatty acids were especially compromised.Abbreviations used (PC) Phosphatidylcholine - (PE) phosphatidylethanolamine - (EP) ethanolamine plasmalogen - (PS) phosphatidylserine - (PI) phosphatidylinositol - (SP) sphingomyelin - (t.l.c.) thin layer chromatography - (BHT) butylated-hydroxytoluene