Influence of photoperiod on seed development in the genetic line of peas G2 and its relation to changes in endogenous gibberellins measured by combined gas chromatography — Mass spectrometry

When apical senescence in the genetic line of peas G2 was prevented by short days fruit development was also found to be retarded. The levels of GA₂₀ and GA₂₉ in cotyledons and pods grown under long or short days were measured by gas chromatography — mass spectrometry multiple ion monitoring using extracts derivatised with deuterated trimethylsilyl groups as internal standards. The levels of GA₂₀ but not GA₂₉, were increased by short days. Conventional gas chromatography — mass spectrometry showed that relative to GA₂₉ the levels of GA₁₉, the other GA identified in G2 cotyledons, were also increased in short days. The levels of GA₂₀ in the pods were highest during the main phase of pod growth early in fruit development.Abbreviations GA_n gibberellin A_n - GC/MS gas chromatography — mass spectrometry - MIM multiple ion monitoring - Me methyl ester - SIM single ion monitoring - TIC total ion current - TMS trimethylsilyl ether - TLC thin layer chromatography - TTLC instant thin layer chromatography     

Metabolic products of microorganisms. 185. The anthraquinones of the Aspergillus glaucus group. I. Occurrence,isolation, identification and antimicrobial activity

The occurrence of emodin, erythroglaucin, physcion, physcion-9-anthrone, questin, catenarin, and catenarin-8-methyl ether in different species of the Aspergillus glaucus group (genus Eurotium) was investigated. So far catenarin-8-methyl ether (1, 4, 6-trihydroxy-8-methoxy-3-methylanthraquinone) has not been described as a natural product; it was therefore given the name rubrocristin. The chemical and physical properties of rubrocristin are reported. In addition a new violet pigment (C₁₆H₁₂O₅) was isolated and characterized by its MS-, IR- and UV-spectra.The antimicrobial properties of all substances were examined in the agar diffusion assay. Gram-positive bacteria were the most sensitive organisms and catenarin was the most active naturally occurring substance. Synthetically obtained 1, 4, 6, 8-tetrahydroxy-anthraquinone was slightly more active than catenarin, whereas rubrocristin showed no antibacterial activity.Abbreviations MIC Minimal inhibitory concentration - TLC Thin layer chromatography - PTLC Preparative thin layer chromatography Metabolic products of microorganisms. 184. H. Anke: On the mode of action of cladosporin. J. Antibiotics 32, 952–958 (1979)     

Mass Spectroscopy and Chromatography of the Trichloromethyl Radical Adduct of Phenyl Ter T-Butyl Nitrone

Positive structural identification of the PBN-trichloromethyl spin adduct in vim was accomplished with the use of high pressure liquid chromatography and/or gas chromatography coupled with mass spectrome-try. Both thin layer and liquid chromatography were used to separate a complex mixture of compounds from rat liver extracts treated with CCI, in vitro and in vivo. Deuterated PBN's (PBN-d, text-butyl deuteration, or PBN-d₁₄; both phenyl and tert-butyl deuteration) were also used to aid in the mass spectral analysis of spin adducts from liver extracts of CCI, exposed rat livers, since the rerr-butyl group fragment ion, C₄D₉+ (m/z = 66) is always present for PBN and PBN spin adducts. In addition, the masses of the ion peaks increase by the amount of deuteration, i.e. an increase of 9 for PBN-d, or PBN-d₁₄ in comparison to normally synthesized PBN.     

Nuclear mutations affecting plastoquinone accumulation in maize

We have isolated and characterized two nuclear mutations which affect plastoquinone accumulation in maize. The mutations, hcf103 and hcf114, modify the same genetic locus. Plants homozygous for either mutant allele exhibit reduced PS II electron transport activity, reduced variable chlorophyll fluorescence and reduced delayed fluorescence yield. In these ways, hcf103 and hcf114 resemble previously described PS II mutants which lack stably assembled PS II reaction center complexes. However, unlike most previously described PS II mutants, hcf103 and hcf114 possess stable membrane-associated PS II complexes. Plastoquinone (PQ-9), which performs a variety of redox functions essential to normal non-cyclic electron transport, is severely depleted in the mutants. The lack of PS II electron transport activity is attributed to the absence of PQ-9. This is the first report of mutants deficient in PQ which do not also suffer serious pleiotropic defects.Abbreviations PS II Photosystem II - PQ plastoquinone - Q_A and Q_B primary and secondary stable electron acceptors of PS II - HPLC high pressure liquid chromatography - LDS-PAGE lithium dodecyl sulfate polyacrylamide gel electrophoresis - TLC thin layer chromatography     

A simple system for determining the phloem mobility of compounds using excised pods of lupin (Lupinus albus L.)

Lupin pods were isolated from the plant and the pedicels placed in solutions of either radiolabeled sucrose, glucose, abscisic acid, or gibberellin A₉ (GA₉). Phloem exudate was collected from the cut stylar ends of the pods and both quantitative and qualitative data were obtained on the activity in the sap. Fed sucrose, abscisic acid, and GA₉ were found in phloem exudate, but labeled glucose was immobile as the monosaccharide and activity in exudate appeared in sucrose.Abbreviations GA gibberellic acid - ABA abscisic acid - TLC thin layer chromatography     

In vitro cultures of Solanum malacoxylon Sendt.: nutritional requirements and sterol production

Callus cultures were established from hypocotyl, root and leaf explants of Solanum malacoxylon. The growth rate of calli was evaluated on Murashige and Skoog medium as well as Gamborg's B5 medium. Sterols were isolated from calli grown on both media however the B5 proved to be more effective in promoting the production of these metabolites. No calcitriol was found in the cultures. Feeding experiments with vitamin D₃ were scarcely effective in promoting the production of vitamin D₃ hydroxylated metabolites.Abbreviations GC/MS gas-liquid chromatographic-mass spectrometric analysis - CHCL₃ chloroform - GLC gas liquid chromatography - PCV packed cell volume - TLC thin layer chromatography     

Structure, gas chromatographic measurement, and function of suberin synthesized by potato tuber tissue slices

The polymeric material (suberin) of the wound periderm of potato tuber slices was analyzed after depolymerization with LiAIH₄ in tetrahydrofuran or BF₃ in methanol with the use of thin layer chromatography, chemical modification, and combined gas-liquid chromatography and mass spectrometry. Fatty acids (C₁₆ to C₂₆), fatty alcohols (C₁₆ to C₂₆), octadec-9-ene-1, 18-dioic acid, and 18-hydroxy-octadec-9-enoic acid were identified to be the major components. Based on the structural information that the two bifunctional C₁₈ molecules constituted a major portion of suberin, a gas chromatographic method of measuring suberization was developed. This method consisted of hydrogenolysis of powdered tissue followed by thin layer chromatography and gas chromatographic measurement of octadecene-1, 18-diol as the trimethylsilyl ether. With this assay it was shown that the development of resistance to water loss by the tissue slices was directly proportional to the quantity of the bifunctional C₁₈ molecules, thus providing evidence that a function of suberin is prevention of water loss.     

Arginine catabolism in Aphanocapsa 6308

The catabolic products of arginine metabolism were observed in Aphanocapsa 6308, a unicellular cyanobacterium, by thin layer chromatography of growth media, by limiting growth conditions, and by enzymatic analysis. Of the organic, nitrogenous compounds examined, only arginine supported growth in CO₂-free media. The excretion of ornithine at a concentration level greater than citrulline suggested the existence in Aphanocapsa 6308 of the arginine dihydrolase pathway which produced ornithine, CO₂, NH₄, + adenosine 5-triphosphate. Its existence was confirmed by enzymatic analysis. Although cells could not grow on urea as a sole carbon source a very active urease and subsequently an arginase were also demonstrated, indicating that Aphanocapsa can metabolize arginine via the arginase pathway. The level of enzymes for both pathways indicates a lack of genetic control. It is suggested that the arginase pathway provides only nitrogen for the cells whereas the arginine dihydrolase pathway provides not only nitrogen, but also CO₂ and adenosine 5-triphosphate.Nonstandard Abbreviations CCCP carbonylcyanide mchlorophenyl hydrazone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethyl urea - CGP cyanophycin granule protein - PS II photosystem II - PSI photosystem I - TLC thin layer chromatography - TCA trichloroacetic acid - DPM disintegrations per min     

Synthesis of salicyl-peptides and their effect on human platelet aggregationin vitro

Summary A series of leucine dipeptide amides containing at their N-terminal amino group the salicyl-residue [(o)-RO-C₆H₄-CO-, where R=H or CH₃CO] have been synthesized by conventional solution techniques and tested for their inhibitory activity on human platelet aggregationin vitro induced by collagen, ADP or adrenaline. The salicyl-peptide (o)-HO-C₆H₄-CO-Leu-Asp(OBzl)-NH₂ was found to exert strong inhibitory activity on platelet aggregation induced by collagen with an IC₅₀ value 4.5 mM. The corresponding dipetide H₂N-Leu-Asp(OVzl)-NH₂ was also examined and was found to be less active, indicating that the presence of the lipophilic-benzyl ester group in combination with the salicyl group enhance the inhibitory activity. All the other salicyl-peptides examined either didn't show any inhibitory or aggregatory activity or a slight inhibition at the concentration of 9–10 mm.The abbreviations used are in accordance with the rules of IUPAC-IUB Joint Commission on Biochemical Nomenclature: Eur J Biochem (1984) 138: 9–37; J Biol Chem (1989) 264: 663–673. Additional abbreviations are: ADP adenosine diphosphate - Boc tert-butyloxycarbonyl - Bzl benzyl - DIEA N,N-diisopropylethylamine - DMSO dimethyl sulfoxide - DMF N,N-dimethylformamide - EtOAc ethyl acetate - FCC flash column chromatography - IC₅₀ molar concentration of salicyl-peptide for 50% inhibition of platelet aggregation - iso-BCF isobutyl chloroformate - NMM N-methylmorpholine - PyBOP (benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate - TLC thin layer chromatography - TMS tetramethylsilane     

Relationship between gibberellin and cytokinin activity and flowering in Rosa damascena Mill.

Kinetin at 10 mg l^–1 increased the number of flowers produced on Rosa damascena plants while GA₃ inhibited flowering. In the leaves of non-flowering plants GA-like activity was high while specific cytokinin activity (fraction-II) was significantly higher in flowering plants. A novel compound 10- methyldihydrozeatin riboside and isopentenyl-adenine were identified from TLC fraction-II while TLC fraction-I yielded zeatin and 2-hydroxy-6-methylaminopurine.Abbreviations TLC thin layer chromatography - BA N⁶-benzyladenine - GA₃ gibberellic acid CIMAP communication No. 92-40J     

The identification of gibberellins in immature seeds of Vicia faba,and some chemotaxonomic considerations

GA₁₇, GA₁₉, GA₂₀, GA₂₉, GA₄₄ and 13-hydroxy-GA₁₂, now named GA₅₃, were identified by GC-MS in immature seeds of Vicia faba (broad bean). Also identified were a GA catabolite, two polyhydroxykauranoic acids, and abscisic, phaseic and dihydrophaseic acids. The GAs of Vicia are hydroxylated at C-13, in common with those of other legumes. However the GAs of Vicia are not hydroxylated at C-3, nor do they appear to be readily conjugated. In these respects Vicia resembles Pisum, another member of the tribe Viciae. Vicia differs from Phaseolus and Vigna, of the tribe Phaseoleae, in both these respects.Abbreviations ABA abscisic acid - DPA dihydrophaseic acid - GA_n gibberellin A_n - GC gas chromatography - GC-MS gas chromatography mass spectrometry - KA kauranoic acid - PA phaseic acid - TLC thin layer chromatography     

Vanillate hydroxylase from the white rot basidiomycete Phanerochaete chrysosporium

A soluble enzyme fraction from Phanerochaete chrysosporium catalyzed the oxidative decarboxylation of vanillic acid to methoxy-p-hydroquinone. The enzyme, partially purified by ammonium sulfate precipitation, required NADPH and molecular oxygen for activity. NADH was not effective. Optimal activity was displayed between pH 7.5–8.5. Neither EDTA, KCN, NaN₃, nor o-phenanthroline (5 mM) were inhibitory. The enzyme was inducible with maximal activity displayed after incubation of previously grown cells with 0.1% vanillate for 30h.Abbreviations MHQ Methoxy-p-hydroquinone - GLC gas liquid chromatography - TMSi trimethylsilane - TLC thin layer chromatography     

Ein Beitrag zur Analyse der Gibberelline in Samen von Pisum sativum L.

Summary Unripe seeds of Pisum sativum were analyses for gibberellins and gibberellin-like substances. Forty kg of seeds were extracted with methanol, the methanol was evaporated and the residue distributed between water and ethylacetate, and between water and butanol at different pH-values. The acidic ethylacetate and the butanol fractions were separated by thin layer chromatography. In the acidic ethylacetate fraction a new substance tentatively named gibberellin A_x was isolated by preparative thin layer chromatography. In different solvent mixtures this substance migrates closely behind gibberellin A₃. It is biological active in a dwarf pea and in dwarf maize mutant bioassays. IR and fluorescense spectra point to a gibban structure. Furthermore gibberellin A₅ was found to be the main gibberellin in pea seeds. It was identified by its R_f-values and the R_f-values of its methylester in different solvent systems as well as by its characteristic relative biological activity in three biological test systems. Most probably gibberellin A₆ is also present in the pea seeds, but because the amounts detected were small, no clear evidence can be presented.In the butanol phase two gibberellin-like substances could be separated by thin layer chromatography.

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Mit Hilfe der Deutschen Forschungsgemeinschaft.     

Zeatin-9-glucoside,a major endogenous cytokinin of Vinca rosea L. crown gall tissue

Cultured crown gall tissue of Vinca rosea L. was found to contain, in addition to the previously reported cytokinins zeatin, zeatin riboside, and the 0-glucosides of these two compounds, relatively high levels of zeatin-9-D-glucopyranoside. This is the first conclusive identification of an endogenous cytokinin 9-glucoside.Abbreviations GC gas chromatography - HPLC high-performance liquid chromatography - I.D. internal diameter - RFE rotary film evaporation - TLC thin layer chromatography - TMS trimethylsilyl - UV ultraviolet - Z zeatin - Z7G zeatin-7-glucoside - Z9G zeatin-9-glucoside - Z0G zeatin-0-glucoside - ZR zeatin riboside - ZR0G zeatin riboside-0-glucoside     

Transport properties of N-acyl derivatives of the coprogen and ferrichrysin classes of siderophores inNeurospora crassa

Several derivatives of the coprogen and ferrichrysin classes of siderophores were synthesized as potential affinity labels of the iron uptake system inNeurospora crassa. While only one of these compounds has proved useful as an affinity label, all were recognized and transported byNeurospora crassa. One derivative, chloroacetyl-ferrichrysin, proved to be an unexpectedly potent reversible inhibitor (K ₁=0.4 M) of both ferrichrysin and coprogen uptake, similar to the natural siderophore, ferrirubin. The reported results provide further understanding of the steric and electronic requirements of siderophores for the iron uptake system inNeurospora crassa.Abbreviations amu atomic mass units - DMF dimethylformamide - FAB tast atom bombardment - NMR nuclear magnetic resonance - ppm parts per million - tlc thin layer chromatography     

Suberin production by isolated tomato fruit protoplasts

The multilamellar wall secreted by protoplasts isolated from locule tissue of tomato (Lycopersicon esculentum L.) fruit was purified, and an extract was obtained after depolymerization with BF₃-methanol. Analysis of this extract using thin layer chromatography demonstrated the presence of fatty acid methyl esters, fatty alcohols, dicarboxylic acid dimethyl esters, and ω-hydroxy acid methyl esters. These components were quantified using an Iatroscan thin layer chromatography-flame ionization detection system. The different chain lengths in each group were identified and quantified using gas chromatography. The results clearly indicated the presence of suberin.     

Establishment of callus,cell suspension and shoot cultures of Leonurus cardiaca L. and diterpene analysis

Summary Callus cultures, cell suspension cultures and shoot cultures of Leonurus cardiaca L. (Motherwort) were established and growth conditions optimized. Shoot cultures showed constant growth whether in the dark or under continuous light, accumulating varying amounts of the furanic labdane diterpenes leosibiricin, preleosibirin, leosibirin and isoballotenol acetate, which are also present in the soil-grown plants. Only traces of leosibiricin were detected in callus cultures, while cell suspension cultures did not produce any furanic diterpenes. A small amount of furanic labdane diterpenes was found in the medium of shoot cultures. Callus and shoot culture induction of several other Lamiaceae species is also described.Abbreviations 2 4-D 2,4-dichlorophenoxyacetic acid - BAP benzylaminopurine - GA₃ gibberellic acid - HPLC high pressure liquid chromatography - NAA naphthyl acetic acid - TLC thin layer chromatography     

A new bacteriochlorophyll from brown-colored chlorobiaceae

A new bacteriochlorophyll has been isolated by thin layer chromatography from all strains of the brown-colored Chlorobiaceae Chlorobium phaeobacteroides and Chlorobium phaeovibrioides. The new bacteriochlorophyll e —like the bacteriochlorophylls c and d—represents the major amount of bacteriochlorophyll in the cells in addition to small amounts of bacteriochlorophyll a. Bacteriochlorophyll e can be differentiated from the bacteriochlorophylls c and d by its absorption maxima in aceton and its different R _f -value in the thin layer chromatogram. The structure of the new bacteriochlorophyll e has been elucidated on the basis of mass spectra, ¹H- and ¹³C-NMR-spectra, the UV/VIS-spectrum as well as IR-, ORD-, and CD-spectra. The new bacteriochlorophyll has the same relationship to bacteriochlorophyll c as chlorophyll b from green plants to chlorophyll a; therefore, bacteriochlorophyll e represents the first formyl-substituted chlorophyll from bacteria. Similar to the bacteriochlorophylls c and d, the new bacteriochlorophyll e consists of a mixture of at least three homologues which differ from each other by different substituents on the pyrrol rings II and III.Abbreviations Used DSM Deutsche Sammlung von Mikroorganismen, Göttingen - Bchl. bacteriochlorophyll - Bph. bacteriopheophytin - P phytol - Gg geranylgeraniol - F farnesol - C Chlorobium This work was made possible by the technology program of the Bundesministerium für Forschung und Technologie.     

Separation of dolichylpyrophosphoryloligosaccharides by liquid chromatography

Fourteen dolichylpyrophosphoryloligosaccharides, precursors of the asparagine-linked oligosaccharides of glycoproteins, have been separated by liquid chromatography on silica gel. The dolichylpyrophosphoryl-N-acetylglucosamine and the dolichylpyrophosphoryl-(N-acetylglucosamine)₂-(mannose)₉(glucose)_2,3 thus resolved were shown to retain their activity as substrates in enzyme catalyzed reactions. The chromatography procedure for the first time makes available many of these single intermediates for further study.     

Establishment of an effective TLC bioautographic method for the detection of Mycobacterium tuberculosis H37Ra phosphoglucose isomerase inhibition by phosphoenolpyruvate

A bioautographic assay based on thin layer chromatography was developed for phosphoenolpyruvate (PEP) detecting as a known but rarely studied inhibitor of phosphoglucose isomerase (PGI). The protocol with NADP⁺/NBT/PMS (β-nicotinamide adenine dinucleotide phosphate/nitrotetrazolium blue chloride/phenazine methosulfate) staining was capable of detecting Mycobacterium tuberculosis H₃₇Ra PGI inhibition using PEP. According to this method, visibly brighter spots (zones) against purple background are observed in the area of inhibition of the above-mentioned enzyme activity. The detection limit for PEP as an inhibitor of Mycobacterium tuberculosis H₃₇Ra PGI was 226?μg per spot/zone. Noteworthy is that we are the first authors to have successfully used a bioautographic assay to detect Mycobacterium tuberculosis H₃₇Ra PGI inhibition by PEP.