ZEITSCHRIFT FÜR NATURFORSCHUNG

SECTION c A EUROPEAN JOURNAL OF

BIOSCIENCES

Council Editorial Board

E. BüNNiNG,Tübingen A. H A G E R , Tübingen A. B U T E N A N D T , München K. H A H L B R O C K , Köln M . E I G E N , Göttingen W. H A S S E L B A C H , Heidelberg

P. K A R L S O N , Marburg F. K A U D E W I T Z , München J. K L E I N , Tübingen J. ST. S C H E L L , Köln E. W E C K E R , Würzburg

Advisory Editorial Board

P. B Ö G E R , Konstanz H . SCHIMASSEK, Heidelberg D. B Ü C K M A N N , Ulm D. S C H U L T E - F R O H L I N D E , Mülheim/R. K. G. G Ö T Z , Tübingen F. F. S E E L I G , Tübingen G. G O T T S C H A L K , Göttingen J. SEELIG, Basel R. J A E N I C K E , Regensburg H . S I M O N , München G. F. M E Y E R , Tübingen W. S T E G L I C H , Bonn M . R A J E W S K Y , Essen A. TREBST, Bochum

EDITED IN CO E L A B O R A T I O N

WITH THE INSTITUTES OF T H E M A X - P L A N C K - G E S E L L S C H A F T

Volume 40 c 1985

V E R L A G D E R Z E I T S C H R I F T F Ü R N A T U R F O R S C H U N G

T Ü B I N G E N

Volume 40c Zeitschrift für Naturforschung 1985

Contents

Conten t s of N u m b e r 1/2

Editorial: Zur Gründung der Zeitschrift für Natur-forschung P. K A R L S O N 1

O r i g i n a l C o m m u n i c a t i o n s

Occurrence of 3,4-Diacetoxybutinylbithiophene in Tagetes p a t u l a and Its Enzymatic Conversion R. PENSL and R. SÜTFELD 3

Isomeric Di-C-glycosylflavones in Fig ( F i c u s c a r i c a L.) F. SIEWEK, K. H E R R M A N N . L. G R O T J A H N , and V. W R A Y 8

Investigations on Callus Cultures of M e l i s s a ofßcina-l i s L , II. Volatile Diterpene Hydrocarbons in Not Differentiated Static Cultures (In German) I. K O C H - H E I T Z M A N N , W . S C H U L T Z E , and F.-C. C Z Y G A N 13

Effect of 5-Azacytidine on the Formation of Sec-ondary Metabolites in C a t h a r a n t h u s roseus Cell Suspension Cultures H.-A. A R F M A N N , W . K O H L , and V. W R A Y 21

Occurrence of uStress"-Proteins in Yeast after Heat-Shock, Acrylonitrile Treatment and during the Stationary Growth Phase U. PFEFFER and B. S C H U L Z - H Ä R D E R 26

Degradation of Glutathione in Plant Cells: Evi-dence against the Participation of a y-Glutamyl-transpeptidase R. STEINKAMP and H. R E N N E N B E R G 29

Purification and Properties of Adenosine Kinase from Rat Liver: Separation from Deoxyadenosine Kinase Activity A. K. D R A B I K O W S K A , L. H A L E C , and D. SHUGAR 34

Amino Acid Composition of Cockroach Hypertreha-losaemic Hormones G . G Ä D E 42

The Scent Substances of Pierid Butterflies ( H e b o -m o i a g l a u c i p p e Linnaeus) and the Volatile Com-ponents of Their Food Plants ( C r a t a e v a r e l i g i o s a Forst.) N . H A Y A S H I , A. NISHI , T. M U R A K A M I , K . M A E S H I M A , H. K O M A E , and T. S A K A O 47

Chemical and Physical Characterization of Four Interfacial-Active Rhamnolipids from Pseudomo-nas Spec. D S M 2874 Grown on n-Alkanes C. S Y L D A T K , S. L A N G , F. W A G N E R , V. W R A Y , and L . W I T T E 51

Production of Four Interfacial Active Rhamnolipids from rt-Alkanes or Glycerol by Resting Cells of Pseudomonas species D S M 2874 C. S Y L D A T K , S. L A N G , U . M A T U L O V I C , and F. W A G N E R 61

Phosphatidylinositol-Specific Phospholipase C from Bovine Blood Platelets. Inhibition by Calmodulin-Inhibitors - Activation by ATP and A D P (In Ger-man) H. BENEDIKTER, G. K N O P K I , and P. R E N Z 68

Phospholipid Transfer from ER to the Peribacteroid Membrane in Soybean Nodules R. B. M E L L O R , T M . I. E. CHRISTENSEN, S. BASSARAB, and D. W E R N E R 73

Higher Cardol Homologues (5-Alkenylresorcinols) from Rye Affect the Red Cell Membrane-Water Transport A. K O Z U B E K 80

Effects of Carbohydrates on the Ion Conductance of the Hemocyanin Channel G. M E N E S T R I N A and F. PASQUALI 85

Kinetics of Calcium-Induced Fusion of Cell-Size Liposomes with Monolayers in Solutions of Dif-ferent Osmolarity N . STOICHEVA, I. TSONEVA, D . S. D l M I T R O V , and I. PANAIOTOV 92

IV Contents

Control of the Activity of Brain Synaptosome-Asso-ciated Acetylcholinesterase by Acidic Phospholi-pids S. TSAKIRIS 97

I n V i t r o Cellular Muscle Calcium Metabolism. Char-acterization of Effects of 1,25-Dihydroxy-Vita-min D 3 and 25-Hydroxy-Vitamin D 3 A. R. DE B O L A N D and R. B O L A N D 102

J. T E N G Ö , E . E N G E L S , W. E N G E L S , R. K I T T M A N N , and D. SCHNEIDER 145

Isolation of Populations of Antipeptide Antibodies Directed against Different Epitopes of the Same Fragment A. CHERSI . C . G R E G E R , and R. A. H O U G H T E N 148

Absorption Spectra of Phytochrome Intermediates P. E I L F E L D and W . R Ü D I G E R 109

Variability of the Light-Harvesting-System during the Cell Cycle of C h l o r e l l a fusca (In German) P. B R A N D T . H. G L E I B S . A. K Ö H N E , and W . WIESSNER 115

Effect of Light lntensity on Photosynthetic 1 4 C 0 2 Fixation of Anabaenaßos-aquae G. D Ö H L E R and A. MÄRTE 122

Photodestruction of P r o p i o n i b a c t e r i u m acnes Por-phyrins T. B . M E L O , G. REISAETER, A. JOHNSSON. and M . JOHNSSON 125

Inhibition of Photosystem II by UV-B-Radiation M . TEVINI and K . PFISTER 129

Notes

Enzymatic Nitrate Assay by a Kinetic Method Em-ploying E s c h e r i c h i a c o l i Nitrate Reductase J. SCHILD and J.-H. K L E M M E 134

Fructose-Stimulated Ferricyanide Reduction by In-tact Blue-Green Algae S. SCHERER and P. B Ö G E R 138

Male Produced Volatiles from Coreid Bug L e p t o -glossus a u s t r a l i s (Heteroptera) A. J. E. G O U G H , D. E. G A M E S , B. W. S T A D D O N , and T. O . O L A G B E M I R O 142

Identification of Cyclic Enolethers from Insects: Alkyldihydropyranes from Bees and Alkyl-dihydro-4H-pyran-4-ones from a Male Moth W. F R A N C K E , W. M A C K E N R O T H , W. SCHRÖDER,

Conten ts of N u m b e r 3/4

O r i g i n a l Communcations

Molecular Pharmacological Investigation of Medici-nal Plant Substances. II. Inhibition of Acetylcho-linesterase by Monoterpene Derivatives i n v i t r o (In German) L . G R A C Z A 151

L-Tyrosine as a Precursor of Flavonoids in Buck-wheat Cotyledons U . M A R G N A . L . LAANEST, E . M A R G N A . and T. VAINJÄRV 154

Two Different Chalcone Synthase Activities from Spinach L . BEERHUES and R. W I E R M A N N 160

Enhanced Production of Cell-Bound and Extracellu-lar Streptolysin S by Hemolytic Streptococci Pre-treated with Proteases A. T A K E T O and Y . T A K E T O 166

Regulation of ß-Galactosidase Synthesis in Wild Type and in a Succinate-Resistant Mutant of Rhi-zobium m e l i l o t i A. P. S I N G H and J. B. S I N G H 170

Do Cyanobacteria Contain a Membrane Bound Cysteine Oxidase? E . L . B A R S K Y , F. D. K A M I L O V A , and V . D. S A M U I L O V 176

Soybean Root Response to Symbiotic Infection Gly-ceollin I Accumulation in an Ineffective Type of Soybean Nodules with an Early Loss of the Peri-bacteroid Membrane D. W E R N E R , R. B. M E L L O R , M . G. H A H N , and H . G R I S E B A C H 179

Contents V

Acetate Concentration and Chloroplast Pyruvate Dehydrogenase Complex in S p i n a c i a o l e r a c e a Leaf Cells B. L I E D V O G E L 182

Isolation of Vacuoles from the Upper Epidermis of P e t u n i a H y b r i d a Petals. I. A Comparison of Isola-tion Procedures J. M . F. G. A E R T S and A. W. SCHRÄM 189

Small-Angle X-Ray and Light Scattering Studies on the Influence of M g 2 + Ions on the Structure of the RNA from Bacteriophage MS 2 G . RIBITSCH, R. D E C L E R C Q , W. F O L K H A R D , P. ZIPPER, J . S C H U R Z , and J . C L A U W A E R T 234

Nucleotide Sequence of Bovine 1.723 Satellite D N A G . P L U C I E N N I C Z A K , J . S K O W R O N S K I , A. P L U C I E N N I C Z A K , and J . J A W O R S K I 242

Isolation of Vacuoles from the Upper Epidermis of P e t u n i a H y b r i d a Petals. II. Vacuolar Localiza-tion of Some Hydrolases J. M. F. G. A E R T S and A . W. SCHRÄM 196

Rate and Rate-Determining Step of Hydrogen-Atom-Induced Strand Breakage in Poly(U) in Aqueous Solution under Anoxic Conditions E. B O T H E and H . S E L B A C H 247

Dicyano-bis(pyridin-2.6-dicarbothioato)-ferrate (II)/ ferrate (III), a Further Fe Containing Redox System from the Culture Medium of Pseudo-monas sp. (In German) U. H I L D E B R A N D , K . T A R A Z , H . B U D Z I K I E W I C Z , H . K U R T H , and G. P U L V E R E R 201

Differential Inhibition by Mevinolin of Prenyllipid Accumulation in Radish Seedlings S. SCHINDLER. T. J . B A C H . and H . K . L I C H T E N T H A L E R 208

On the Chemical Reactivity of the Phytochrome Chromophore in the P r and P f r Form F. T H Ü M M L E R , P. E I L F E L D , W. R Ü D I G E R , D . - K . M O O N , and P.-S. S O N G 215

Physicochemical Characterization of Tetraether Lipids from T h e r m o p l a s m a a c i d o p h i l u m . II. Film Balance Studies on the Monomolecular Organiza-tion of the Main Glycophospholipid in Monofilms C. STROBL, L. SIX, K . H E C K M A N N , B. H E N K E L , and K . R I N G 219

Chemiluminescence in the Coupled Oxidation of Lecithin and Ascorbate K . LICHSZTELD, Z. M A C H O Y , and A. STEPINSKA 223

Properties of Lectins from Snails of the Genus H e l i x Probed by Monoclonal Antibodies H . A. W. SCHNEIDER 254

Sex Pheromone Components in the New Zealand Brownheaded Leafroller Ctenopseustis o b l i q u a n a (Lepidoptera: Tortricidae) H . Y O U N G , R . A. G A L B R E A T H , M . H . B E N N , V A. H O L T , and D . L. STRUBLE 262

Sex Pheromone Components in the New Zealand Greenheaded Leafroller P l a n o t o r t r i x excessana (Lepidoptera: Tortricidae) R. A. G A L B R E A T H , M . H . B E N N , H . Y O U N G , and V. A. H O L T 266

Sex Attractant Blends for Field Trapping of A g r o t i s segetum Males (Lepidoptera: Noctuidae) in Israel and Germany E. D U N K E L B L U M , S. G O T H I L F , H . J. B E S T M A N N , W. K N A U F , and O. VOSTROWSKY 272

Calcium/Sodium Binding Competition in the Gat-ing of Light-Activated Membrane Conductance Studied by Voltage Clamp Technique in L i m u l u s Ventral Nerve Photoreceptor H . STIEVE, M . P F L A U M , J. K L O M F A S S , and H . G A U B E 278

Temperature Dependence of Population Growth in a Green Microalga: Thermodynamic Characteris-tics of Growth Intensity and the Influence of Cell Concentration C. J . SOEDER, E . H E G E W A L D , E . FIOLITAKIS, and J. U . G R O B B E L A A R 227

Notes

Site of H Atom Attack on Uracil and Its Derivatives in Aqueous Solution S. D A S , D. J . D E E B L E , and C. v. S O N N T A G 292

VI Contents

The Effect of Thiol Compounds on the Glutamate Affinity of the Chloroplastic Glutamine Syn-thetase from Mustard Leaves R. M A N D E R S C H E I D and A. W I L D 295

The Influence of Low-Intensity Millimeter-Wave Radiation on the Growth of Cress Roots F. K R E M E R , A . POGLITSCH, L . SANTO, D . SPERBER, and L . G E N Z E L 336

C o n t e n t s o f N u m b e r 5/6

O r i g i n a l C o m m u n i c a t i o n s

Adsorbent Filter - A Tool for the Selection of Plant Suspension Culture Cells Producing Secondary Substances B . KNOOP and R. BEIDERBECK 297

Localization of Chromenes and Benzofurans in the Genus E n c e l i a (Asteraceae) P. PROKSCH, M . PROKSCH, W. W E C K , and E . Ro-DRIGUEZ 301

The Widespread Occurrence in Nature of Antho-cyanins as Zwitterions J. B . H A R B O R N E and M . B O A R D L E Y 305

Studies of the Sterolsynthesis in the Fungus A u r e o -b a s i d i u m ( D e m a t i u m ) p u l l u l a n s (In German) P. F L E S C H and M . SCHAEFER 309

Isolation of the Phytoalexin Medicarpin from Leaf-lets of A r a c h i s h y p o g a e a and Related Species of the Tribe Aeschynomeneae R. N . STRANGE, J. L . I N G H A M , D . L . C O L E , M . E . C A V I L L , C. E D W A R D S , C. J. COOKSEY, and P. J. GARRATT 313

Isoceroptene, a Novel Polyphenol from P i t y r o g r a m -m a t r i a n g u l a r i s K . R. M A R K H A M , C H . V I L A I N , E . W O L L E N W E B E R , V . H . D I E T Z , and G . SCHILLING 317

Flavonoids and Terpenoids from the Leaf Resin of P l u c h e a o d o r a t a E . W O L L E N WEBER, K . M A N N , F. J. A R R I A G A , and G. YATSKIEVYCH 321

Biflavonoids, Quinones and Xanthones as Rare Chemical Markers in the Family Iridaceae C H R . A . W I L L I A M S and J. B . H A R B O R N E 325

Are Spinach Chloroplasts Involved in Flavonoid O-Methylation? K . THRESH and R. K . IBRAHIM 331

Amine-carbamoyl-dihydro-10B-borane, Preparation, and Biological Properties (In German) F. D A L L A C K E R , T. B Ö H M E L , W . M Ü L L N E R S , andH. MÜCKTER 344

Carrierfree Synthesis Products from Irradiated Va-line by Means of Hot 1 1C-Atoms of Giant Reso-nance Reaction (In German) G. G U N D L A C H , E . L . SATTLER, and U . W A G E N -BACH 351

Acylchain Specificity and Kinetic Properties of Phos-pholipase A x and A 2 of Bone Marrow-Derived Macrophages I. F L E S C H , B . SCHMIDT, and E . FERBER 356

A Small-Angle X-Ray Scattering Study on Pre-Irra-diated Malate Synthase. The Influence of For-mate, Superoxide Dismutase, and Catalase on the X-Ray Induced Aggregation of the Enzyme P. ZIPPER, R. W I L F I N G , M . K R I E C H B A U M , and H . D U R C H S C H L A G 364

Physicochemical Properties of Ferredoxin from C h l a m y d o m o n a s r e i n h a r d i i F. G A L V Ä N , A . J. M Ä R Q U E Z , and E . FERNÄNDEZ

373

Fluorescence-ODMR of Light Harvesting Pigments of Photosynthetic Bacteria A . ANGERHOFER, J. U . v. SCHÜTZ, and H . C. W O L F

379

Action of Methylthiopyrimidine Experimental Her-bicids as Diuron-Like Inhibitors of Photosynthesis J . -M. D U C R U E T and R. S C A L L A 388

Inhibition of Photosynthetic Electron Transport by Halogenated 4-Hydroxy-pyridines A . TREBST, B . D E P K A , S. M . R I D L E Y , and A . F. H A W K I N S 391

Proton Uptake by Light Induced Interaction be-tween Rhodopsin and G-Protein A . SCHLEICHER and K . P . H O F M A N N 400

Contents VII

Physiology of Nitrogen Fixation in Two New Strains of A n a b a e n a P E I - C H U N G C H E N 406

Biochemical Oxygen Activation as the Basis for the Physiological Action of Tetrachlorodecaoxide (TCDO) R . J . Y O U N G M A N , G . R . W A G N E R , F. W . K Ü H N E , and E . F. ELSTNER 409

Diversification of Chemoreceptors in E c t o c a r p u s , S p h a c e l a r i a , and A d e n o c y s t i s (Phaeophyceae) D . G . M Ü L L E R , W . B O L A N D , L . JAENICKE, and G . G A S S M A N N 457

Nonconventional Interactions between Photorecep-tor Axons in the Butterfly Lamina Ganglionaris W . C. G O R D O N 460

Isolation of Labeled Lipoprotein from E s c h e r i c h i a c o l i and P r o t e u s m i r a b i l i s after Incubation with [ I 4C]Penicillin G . G R U N E R , M . H . TADROS, and R. P L A P P 415

Studies on Protein Synthesis of Outgrowing Spores of B a c i l l u s s u b t i l i s (In German) M . H E C K E R , A . - M . D U N G E R , G . W A C H L I N , and F. M A C H 421

On the Role of Branched-Chain Amino Acids in Protein Turnover of Skeletal Muscle. Studies i n v i v o with L-Norleucine K . - J . SCHOTT, J. G E H R M A N N , U . PÖTTER, and V . N E U H O F F 427

Regression of the Time-Keeping Ability in Carabid Beetles by Phylogenetic Adaptation to Cave Con-ditions W . M A R T I N and F. W E B E R 438

Notes

y-Radiolysis of 2'-Deoxyguanosine. The Structure of the Malondialdehyde-Like Product D. LANGFINGER and C. v. SONNTAG 446

Prosomes are Involved in the Repression of Viral m R N A A . H O R S C H , K . K Ö H L E R , and H.-P. SCHMID 449

Inhibition of Bluelight-Induced, Flavin-Mediated Membraneous Redoxtransfer by Xenon W . SCHMIDT 451

Photoaffinity Labeling of Spinach Thylakoids and Cytochrome b(Jf-Comp\ex by the Hydrophobie Reagent 3-(Trifluoromethyl)-3-(m-[125I]iodo-phenyl)-diazirine W . OETTMEIER, H.-J . S O L L , andE. OLSCHEWSKI

454

Contents of N u m b e r 7/8

O r i g i n a l C o m m u n i c a t i o n s

Comparative Studies on the Production and Accu-mulation of Essential Oil in the Whole Plant and in the Cell-Culture of P i m p i n e l l a a n i s u m L . (In German) J . REICHLING, H . B E C K E R , R. M A R T I N , and G . BURKHARDT 465

New Flavone Glucoside Malonylesters from B r y u m c a p i l l a r e W. STEIN, S. A N H U T , H . D . ZINSMEISTER, R. M u -ES, W. B A R Z , and J . KÖSTER 469

3-Acetamino-4-hydroxy Benzoic Acid and 2-Acet-amino Phenol from Pseudomonas Cultures (In German) S. W I N K L E R , W. N E U E N H A U S , H . BUDZIKIEWICZ, H . K O R T H , and G . PULVERER 474

Comparison of pH Changes and Elicitor Induced Production of Glyceollin Isomers in Soybean Cotyledons W. F. O S S W A L D , S. Z I E B O L L , and E . F. ELSTNER

477

Isoneorautenol and other Pterocarpan Phytoalexins from C a l o p o g o n i u m mueunoides J. L . INGHAM and S. T A H A R A 482

Natural Inhibitors of Germination and Growth, III. New ct-Pyrones from Seeds of Rosa c a n i n a E . L O H A U S , C. Z E N G E R , W. RÜDIGER, and E . C M I E L 490

The Regulation of Acetyl Coenzyme A Synthesis in Chloroplasts H.-J . T R E E D E and K . - P . H E I S E 496

VIII Contents

Isomerase and Decarboxylase Activities in the 4-Hy-droxyphenylacetate Catabolic Pathway of Pseudo-monas p u t i d a M . L . M A R T I N and A . GARRIDO-PERTIERRA 503

The Presence of a Specialized-ß-Glucosidase: Lina-marase, in the Leaves of T r i f o l i u m repens is Con-trolled by the Gene L i P. K A K E S and H . E E L T I N K 509

Biosynthesis of Trichothecene Mycotoxins in F u s a -r i u m c u l m o r u m Cultures N . C. P. B A L D W I N , B . W . BYCROFT, P. M . D E -WICK, J . GILBERT, and I. H O L D E N 514

2-Hydroxy-Swietenin, a New Limonoid from Swiete-n i a m a h a g o n i D C . (In German) A . D A I L Y , O. S E L I G M A N N , H . LOTTER, and H . W A G N E R 519

The Action of S a r a c a asoca Roxb. de Wilde Bark on the P G H 2 Synthetase Enzyme Complex of the Sheep Vesicular Gland T. B . M I D D E L K O O P and R. P. L A B A D I E 523

Effect of Salinity on Photosynthetic i 4 C 0 2 Fixation and Amino Acid Pools of B e l l e r o c h e a y u c a t a n e n s i s (v. Stosch) and T h a l a s s i o s i r a r o t u l a (Meunier) G. D Ö H L E R and J . Z I N K 527

ESR and E N D O R Investigations of Adrenochrome Semiquinone and Related Amino-l,2-Benzosemi-quinone Radicals H . B . STEGMANN, H . D A O - B A , K . SCHEFFLER, and M . G . PETER 531

Three-Dimensional Models of the Carbohydrate Moieties of Murein and Pseudomurein H . F O R M A N E K 555

Composition and Distribution of Lipids in Tissues of Bogue ( B o o p s b o o p s ) V . M . K A P O U L A S and S. MINIADIS-MEIMAROGLOU

562

X-Ray Studies on Phospholipid Bilayers. V . Inter-actions with D D T M . S U W A L S K Y , N . B U G U E N O , J . T A P I A , and F . NEIRA 566

Invariance of Stoichiometry of the Sarcoplasmic Ret-iculum Calcium Pump at Physiological Calcium Concentrations — a Reevaluation W . H A S S E L B A C H and A . M I G A L A 571

Nature of Yeast Cells Immobilized by Radiation Polymerization Activity Dependence on the Mo-lecular Motion of Polymer Carriers T. FUJIMURA and I. K A E T S U 576

Notes

Fast Atom Bombardment Mass Spectrometry of Coumaric Acids C H . G . D E KOSTER, G . J . N I E M A N N , and W . H E E R -MA 580

Detection of Semiquinone Radicals of N-Acylcate-cholamines in Aqueous Solution M . G . PETER, H . B . S T E G M A N N , H . D A O - B A , and K. SCHEFFLER 535

A Comparison of Decoration Techniques for the Demonstration of Immunocomplexes by Scanning Electron Microscopy: Labeling of a Protein Anti-gen on the Surface of the Yeast C a n d i d a a l b i c a n s (In German) M . BORG 539

Isotopic Effect on the Kinetic of Thermal Denatura-tion of Ceruloplasmin L . SPORTELLI, A . DESIDERI , and A . C A M P A N I E L L O

551

Flavonoid Glycosides from Thymus membranaceus F. T O M A S , L . H E R N Ä N D E Z , F. A . T . B A R B E R Ä N , and F. FERRERES 583

Chichoric Acid and Its Derivatives from E c h i n a c e a Species (In German) H . B E C K E R and W . C H . H S I E H 585

Pattern Formation and Marangoni Convection du-ring Oscillating Glycolysis S. C. M Ü L L E R , T . PLESSER, A . B O I T E U X , and B . H E S S 588

Hyperglycemia in Freshwater Field Crab ( O z i o t e l -p h u s a senex senex) Produced by Pesticides K . S. J . R A O 592

Contents IX

Conten t s of N u m b e r 9/10

O r i g i n a l C o m m u n i c a t i o n s

Synthesis of l,2-Dipalmitoyloxypropyl-3-(2-ammo-niumethyl) Phosphinate M . C. MOSCHIDIS 595

Comparative Analysis of Epicuticular Waxes from Some High Alpine Plant Species C. L Ü T Z and P . -G. G Ü L Z 599

Physicochemical Characterization of Tetraether-lipids from T h e r m o p l a s m a a c i d o p h i l u m . IV. Calo-rimetric Studies on the Miscibility of Tetraether-lipids with Dipalmitoyl Phosphatidylcholine and Dipalmitoyl Phosphatidylglycerol D. BLÖCHER, R. G U T E R M A N N , B . H E N K E L , and K . R I N G 606

Occurrence and Characteristics of Amino Alcohols and Cyclohexenone. Components of Fungal Mycosporines F. L E M O Y N E , J . B E R N I L L O N , J . F A V R E - B O N V I N , M . L . B O U I L L A N T , and N . A R P I N 612

Isoflavonoids from B o l u s a n t h u s speciosus (Bolus) Harms Leguminosae K . A S R E S , P. M A S C A G N I , M . J . O ' N E I L L , and J . D . PHILLIPSON 617

Studies on Sporopollenin Biosynthesis: The Effect of Inhibitors of Carotenoid Biosynthesis on Sporopollenin Accumulation A . - K . P R A H L , H . SPRINGSTUBBE, K . G R U M B A C H , and R. W I E R M A N N 621

Origin of Cardenolides in Rhizomes and Roots of C o n v a l l a r i a m a j a l i s L . . Biogenesis i n s i t u or Trans-port from Leaves? R. SCHRUTKA-RECHTENSTAMM, B . K O P P , and W. LÖFFELHARDT 627

On the Active Principles of the Spurge Family (Euphorbiaceae). X I . [1] The Skin Irritant and Tumor Promoting Diterpene Esters of E u p h o r b i a t i r u c a l l i L . Originating from South Africa G. FÜRSTENBERGER and E . H E C K E R 631

Effects of Norfloxacin and Rifampicin on Growth and Streptolysin S Production in Hemolytic Strep-tococci A . TAKETO and Y . T A K E T O 647

Acetohydroxyacid Synthase Inhibitors: N-Phthalyl-L-valine Anilide and Related Compounds J . L . H U P P A T Z and J . E . C A S I D A 652

Volatile Secretions in Three Species of D u f o u r e a (Hymenoptera: Halictidae) Bees: Chemical Com-position and Phylogeny J . T E N G Ö , I. G R O T H , G . BERGSTRÖM, W. SCHRÖ-DER, S. K R O H N , and W. F R A N C K E 657

Volatile Constituents of the Red Wood Ant F o r m i c a rufa L . (Hymenoptera: Formicidae) W. F R A N C K E , J . BORCHERT, and D . K L I M E T Z E K 661

Mandibular Gland Secretions as Alarm Pheromones in Two Species of the Desert Ant C a t a g l y p h i s A . H E F E T Z and H . A . L L O Y D 665

(Z)-9-Dodecenyl Acetate, a Component of the Sex Pheromone of C n e p h a s i a l o n g a n a Haworth (Lepidoptera: Tortricidae) H . - J . B E S T M A N N , A . A T T Y G A L L E , H . P L A T Z , O. VOSTROWSKY, and M . G L A S 667

Mode of Action of the Hypertrehalosaemic Peptides from the American Cockroach G. G Ä D E 670

The Kinetics of the Reaction of Thiocyanate and Nitrite Ions with Orconectes l i m o s u s Oxyhemo-cyanin R: G O N D K O , E . SERAFIN, J . M A Z U R , B . JEZOWSKA-TRZEBIATOWSKA and A . JEZIERSKI 677

Regioselective O-Demethylation of Scoparone: Dif-ferentiation between Rat Liver Cytochrome P-450 Isozymes D . M Ü L L E R - E N O C H , E . B Ü T T G E N , and A . N O N N E N -MACHER 682

Degradation of Bacteriochlorophyll a in Rhodo-pseudomonas sphaeroides R 26 H . H A I D L , K . K N Ö D L M A Y R , W. RÜDIGER, H . SCHEER, S. SCHOCH, and J . U L L R I C H 685

A Photoreversible 39 kDalton Fragment from the P f r Form of 124 kDalton Oat Phytochrome U . R E I F F , P. E I L F E L D , and W. RÜDIGER 693

On the Correlation between Oxygen Uptake in Plastids of Greening Etiolated Oat Leaves and Pigment Photooxidation F. F R A N C K and G . H . SCHMID 699

X Contents

Separation of Chloroplast Thylakoid Membrane Polypeptides by Electrophoresis on Polyacryl-amide Gradient Gels with a Length of 60 cm H . O . SCHMIDT and K . - H . Süss 705

Phosphorylation of Coformycin and 2'-Deoxycofor-mycin, and Substrate and Inhibitor Properties of the Nucleosides and Nucleotides in Several En-zyme Systems A . B Z O W S K A , P. LASSOTA, and D . SHUGAR 710

Stereospecific Synthesis by the Sodium Salt Glyco-sylation Method of Halogeno Benzimidazole 2'-Deoxyribose Analogues of the Inhibitor of hnRNA Synthesis, 5,6-Dichloro-l-(ß-D-ribo-furanosyl)benzimidazole (DRB) Z . K A Z I M I E R C Z U K , R. STOLARSKI, and D . SHUGAR

715

Potassium and Rubidium Effluxes in Saccharomyces cerevisiae M . D . ORTEGA and A . RODRIGUEZ-NAVARRO 721

Thyroxine Induced Transformation in Sarcoplasmic Reticulum of Rabbit Soleus and Psoas Muscles M . T. N U N E S , A . C. B I A N C O , A . M I G A L A , B . AGOSTINI, and W. H A S S E L B A C H 726

Protoplast Dielectrophoresis in Axisymmetric Fields N . STOICHEVA, I . T S O N E V A , and D . S. DIMITROV735

Conten t s of N u m b e r 11/12

O r i g i n a l Communications

Studies on the Triple Helical Structure of ß-D-1,3 Xylan M . A . H A L E E M and S. Q A M R U D D I N 751

A-ct-hydroxymesobiliverdin, a New Bile Pigment E. BENEDIKT and H. -P . KÖST 755

Alteration of Tocochromanol Pattern in Some Plant Oils (Soybean, Lupine, Sunflower and Wheat) during Germination and Growth (In German) H . S C H U L Z , G . L A U S C H , and W. F E L D H E I M 760

Quinolizidine Alkaloids as Nitrogen Source for Lu-pin Seedlings and Cell Cultures M . W I N K and L . WITTE 767

Cellular Differentiation of Sucrose Metabolism in Anabaena v a r i a b i l i s N . SCHILLING and K . EHRNSPERGER 776

The Crystal Structure of Ectoine, a Novel Amino Acid of Potential Osmoregulatory Function (In German) W. S C H U H , H . P U F F , E . A . G A L I N S K I , and H . G . TRÜPER 780

Notes

Biotransformation of Tryptophan to Indole-3-acetic Acid by Xanthomonas campestris pv. campestris D. J . ROBESON and D . R. C O O K 740

A Fungal Elicitor of the Resistance Response in Wheat K . - H . K O G E L , B . H E C K , G . K O G E L , B . MOERSCHBACHER, and H . - J . REISENER 743

Water Relations in Young Growing Wheat Leaves after Application of (2-Chloroethyl)trimethyl-ammoniumchloride (CCC) to the Roots of Wheat Seedlings J . B O D E and A . W I L D 745

Associations of Mitochondria with Desmosome-Like Structures in the Oocytes of R a i l l i e t i e l l a aegypti (Pentastomida, Cephalobaenida) R. R I E H L and V . W A L L D O R F 748

Elimination of Ammonium Ion from the ot-Hy-droxyalkyl Radicals of Serine and Threonine in Aqueous Solution and the Difference in the Reac-tion Mechanism G. B E H R E N S and G . K O L T Z E N B U R G 785

A Disappearance of a 24-kDa Acid-Soluble Protein from Liver Chromatin of Normal and Starved Hens Following D-Galactosamine Administration J . P A L Y G A 798

Further Evidence for a Functional Relationship be-tween L-Amino Ac id Oxidase Activity and Photo-synthetic Oxygen Evolution in Anacystis n i d u l a n s . Effect of Chloride on the Two Reactions E. K . PISTORIUS 806

Antiserum Against the 33-kDa Herbicide-Binding Protein G. H E R R M A N N , A . T H I E L , and P. BÖGER 814

Contents

Preparation of an Enzyme Associated with Carth-amin Formation in C a r t h a m u s t i n c t o r i u s L . K . SAITO, Y . T A K A H A S H I , and M . W A D A 819

Binary Oscillation of Delayed Luminescence: Evi-dence of the Participation of Q B in the Charge Recombination E . H I D E G and S. DEMETER 827

The Role of N A D P H in the Reversible Phototrans-formation of Chlorophyllide P 6 8 2 into Chlorophyl-lide P 6 7 8 in Etioplasts of Oat F . F R A N C K and G . H . SCHMID 832

Biochemical Characterization of Propylglyoxal Bis(guanylhydrazone). Facile Synthesis of Mono-alkylglyoxal Bis(guanylhydrazones) H . E L O , R. L A I N E , L . A L H O N E N - H O N G I S T O , J . JÄN-N E , I. M U T I K A I N E N , and P. L U M M E 839

Tumor Inhibition by Ferricenium Complexes: Sys-temic Effect i n v i v o and Cell Growth Inhibition i n v i t r o P. K Ö P F - M A I E R 843

Chemical Composition of the Wing Gland and A b -dominal Hair Pencil Secretions of the Male Afri-can Sugarcane Borer, E i d a n a s a c c h a r i n a (Lepidoptera: Pyralidae) B . V . B U R G E R , W. M . M A C K E N R O T H , D . SMITH, H . S. C. SPIES, A N D P. R. A T K I N S O N 847

Presumed Sex Pheromone from Androconial Glands of Male Cotton Harlequin Bug T e c t o c o r i s d i o -phthalmus (Heteroptera; Scutelleridae) Identified as 3,5-Dihydroxy-4-pyrone D. W. K N I G H T , B . W. S T A D D O N , and M . J . T H O R N E 851

Further Observations on Periodicities of Nucleotide Occurrences in Natural D N A ' s N . B U R R F U R L O N G and K . M A R I E N 854

M T D Approach to Quantitative Structure-Activity Relationships for Cardiotonic Steroids M . B O H L and Z . SIMON 858

Reaction of Fluorescein Isothiocyanate with Thiol and Amino Groups of Sarcoplasmic ATPase G. SWOBODA and W. H A S S E L B A C H 863

XI

Formation and Decay of the Vanadate Complex of the Sarcoplasmic Reticulum Calcium Transport Protein P. M E D D A and W . H A S S E L B A C H 876

Linear Dichroism Spectroscopy of Retinal with Picosecond Time Resolution M . E . LIPPITSCH, M . R I E G L E R , F. R. A U S S E N E G G , N . F R I E D M A N , M . SHEVES, Y . M A Z U R , and L . M A R -GULIES 880

A Patch Clamp Study of Tonoplast Electrical Prop-erties in Vacuoles Isolated from Chenopodium r u b r u m Suspension Cells F. - W . B E N T R U P , B . H O F F M A N N , M . G O G A R T E N -B O E K E L S , J . P. G O G A R T E N . and C H . B A U M A N N 886

Reculturing of Cells from Primary C F U - C Colonies G . E . H Ü B N E R , F. A L I - O S M A N , M . K A S T N E R , C. PAPADIMITRIOU, and H . R. M A U R E R 891

Mammalian Spermatogenesis as a Biological Indi-cator for Ionizing Radiation (In German) U . H A C K E R - K L O M , E . - M . M E I E R , and W . G Ö H D E

898

Notes

Analytical Ultracentrifugation of Proteins in Solu-tions of Triton X - 1 0 0 Using Absorption Optics D. SCHUBERT and K . Boss 908

Calcium, Iron and Cobalt Accumulation in Root Hairs of Soybean ( G l y c i n e max) D. W E R N E R , K . - P . K U H L M A N N , F. GLOYSTEIN, and F . - W . RICHTER 912

Seasonal Variation of Free Flavone Aglycones from S i d e r i t i s l e u c a n t h a (Lamiaceae) F. A . T. B A R B E R Ä N , F. T O M A S , and J . M . N Ü N E Z

914

On the Specificity of the Herbicide Chlorsulfuron in Intact Spinach Chloroplasts U . H O M E Y E R , D . SCHULZE-SIEBERT, and G . SCHULTZ 917

Bleomycin-Iron Complex and Oxygen Activate A l -gal Ribonucleotide Reductase R. H O F M A N N and H . F O L L M A N N 919

XII Contents

Growth Response of Mouse Lymphoma Cells to Low Concentrations of Mercuric Chloride ST. P. JOST, J . C O L E , and B . C . GOODWIN 922

The U V Absorption Spectra of the C(5) and C(6) O H Adduct Radicals of Uracil and Thymine De-rivatives. A Pulse Radiolysis Study D. J . D E E B L E and C. VON SONNTAG 925

High Yields of DNA-Transfer into Moyse L-.Cells by Electropermeabilization H . STOPPER, U . Z I M M E R M A N N , and E . W E C K E R 929

Female Urinary Chemosignals Influence Scent-Marking Behavior in Male Mongolian Gerbils ( M e r i o n e s u n g u i c u l a t u s ) B . PROBST 936

Specificity of Sexual Attractants in X e s t i a t r i a n g u l u m Hufn. and X . d i t r a p e z i u m Schiff. (Lepidoptera: Noctuidae) E . PRIESNER 939

Inhibitors of Sexual Attraction in the Moth A g r o t i s e x c l a m a t i o n i s E . PRIESNER 943

D N A in the Nucleomorph of C r y p t o m o n a s Demon-trated by D A P I Fluorescence P. H A N S M A N N , H . F A L K , and P. SITTE 933

Subject Index

Authors Index

947

969

Degradation of Bacteriochlorophyll a in Rhodopseudomonas sphaeroides R26 H . H a i d l , K . K n ö d l m a y r , W . R ü d i g e r , H . Scheer, S. Schoch

Botanisches Institut der Universität, Menzinger Str. 67, D-8000 München 19

and

J. U l l r i c h

Physikalisches Institut der Universität, Pfaffenwaldring 57, D-7000 Stuttgart

Z . Naturforsch. 40c, 685-692 (1985); received May 24, 1985 Rhodopseudomonas s p h a e r o i d e s , Bacteriochlorophyll a , Bacteriopheophytin a , Bacterio-pheophorbide a , Pigment aggregation

A series of pigments of the bacteriopheophytin a spectral type have been isolated from ageing cultures of Rhodopseudomonas s p h a e r o i d e s strain R26. These pigments are formed in varying amounts, and can be most readily analyzed i n v i v o by their absorption in the 530 nm spectral ränge. They are enriched in sedimenting cells, but their formation is not affected by light. By Chromatographie comparison with authentic pigments and chemical correlation, the following pigments have been identified: bacteriopheophytin a , bacteriopheophorbide a (which is the pre-dominant produet), pyrobacteriopheophorbide a and a fourth, very polar bacteriopheophytin a -type produet of unknown structure. The major portion of these newly formed pigments is present in the cells in a State, in which the near-infrared absorption band is shifted to longer wavelengths. As shown by low temperature fluorescence spectroscopy, these forms are very similar to bac-teriopheophorbide a aggregates.

Introduction

One of the most widely aeeepted indicators for plant senescence is the disappearance of Chloro-phylls. Nonetheless is the biochemistry of this proc-ess — in contrast to Chlorophyll biosynthesis [1 — 3] and to the degradation of the structurally related hemes [4] — at present only poorly understood. In senescing green plants, produets have been identified which arise from demetalation [5], hydrolysis of the phytylester [5, 6] and/or oxidation at the isoeyclic ring [7, 8]. Loss of the [132]carbomethoxy group has been reported for E u g l e n a [9]. Beyond these rela-tively early stages, a phytol containing pyrrol derived from ring D has been reported by Park et a l . [10], but most of the phytol is assumed to be seavenged as fatty acid esters [11]. There is even less known about the degradation of bacterial Chlorophylls.

The major handicap in Chlorophyll breakdown studies is probably the lack of suitable Systems,

A b b r e v i a t i o n s : behl, bacteriochlorophyll a \ bphe, bac-teriopheophytin a ; bpheid, bacteriopheophorbide a ; (see also formulas); Rp. Rhodopseudomonas; tlc, thin layer chromatography; H P L C , high Performance liquid chromatography.

Reprint requests to Dr. S. Schoch.

Verlag der Zeitschrift für Naturforschung, D-7400 Tübingen 0341 -0382/85/0009-0685 S 01.30/0

which accumulate the early produets sufficiently. Düring a Screening for suitable Systems, we had ob-served, that cultures of the carotenoid - less mutant R26 from the purple photosynthetic bacterium, Rhodopseudomonas s p h e r o i d e s , often change color if they are kept over times extending the common growth periods of approx. 7 days. Here, we wish to report the results of a spectroscopic and chemical analysis of the major newly formed pigments, which indicate a rather similar breakdown pattern for bac-teriochlorophyll a than has previously been reported for Chlorophyll a.

Materials and Methods

R p . s p h a e r o i d e s (strains 2.4.1 and R26) were grown anaerobically in the Light (> 1500 Lux from incandescent lamps) in 0.7 1 bottles in Hutner's medium [12]. The cultures were not stirred. The cells were harvested by centrifugation at 10,000 x g .

Pigments were extracted with methanol aecording to Strain and Svec [13]. Reference pigments (see Scheme 1 for formulas and abbreviations) were pre-pared by the following Standard procedures: Bac-teriopheophytin a: Demetalation with dilute hydro-chloric acid [14]; bacteriopheophorbide a: Hydrolysis with cold hydrochloric acid (18%) [15]; bacterio-pyropheophytin a: Refluxing of bacteriopheophytin

686 H . Haidl et a l . • Degradation of Bacteriochlorophyll a in Rhodopseudomonas s p h a e r o i d e s R 26

COOR

M R. R2

bchl a Mg phytyl C O O C H , bphe a 2H phytyl C O O C H , pyro-bphe a 2H phytyl H bpheid a 2H H C O O C H , pyro-bpheid a 2H H H methyl-bpheid a 2H C H , C O O C H , pyromethyl-bpheid a 2H C H , H

a in pyridine [16]. Methyl esters ( m e t h y l - b p h e i d and p y r o m e t h y l - b p h e i d ) were prepared by refluxing in methanol containing 5% (v/v) sulfuric acid (modified from Fischer and Stern [17]). A l l pigment handling was done under subdued light, and all reactions were carried out under nitrogen.

Absorption spectra were recorded with a PE320 (Perkin-EImer, Ueberlingen), a Unicam SP1800 (Philips), or a ZWS II spectrometer (Sigma, Berlin) equipped with a model 8000 "intelligent recorder" (Bryans, Mitcham).

Fluorescence spectra were recorded with a home-built apparatus (Fig. 1). The light of a Xenon arc lamp (Osram X B O 900) was focused on the entrance slit of a double monochromator (Spex 1672). Part of the light leaving the monochromator was reflected by a quartz plate onto a photodiode (PD). The signal of the photodiode was used as a reference for the slit control (SC). It controled a variable slit at the en-trance of the monochromator to obtain a constant excitation intensity over the entire spectral ränge. The sample was cooled to 4K in a liquid helium cryo-state. The light emited by the sample passed a double monochromator (Spex 1680) and was detected by a photomultiplier (PM) (Sl photocathode, type E M I 9684 for near infrared detection; S20 photocathode, type E M I 9558 for the visible region). In order to compensate for the intensity minima in the spectral Output of the xenon lamp, which cannot be ade-quately regulated by the slit control, the amplified signal from the photomultiplier was divided by the signal from the photodiode.

Analytical thin layer chromatography was done on reverse phase H P T L C plates (Merck, Darmstadt, RP-8, F254S) and methanol or mixtures of methanol/ water (95:5, v/v) as eluents. H P L C was performed on a System consisting of 2 model 6000A pumps and a model 660A solvent programer (Waters, König-stein), a LiChrosorb RP-18 column 250x4.6 mm (Merck, Darmstadt) and two absorption detectors model spectro monitor II (Latek, Heidelberg) set at 525 and 595 nm. Flow: 1.5 ml/min, solvent A : methanol—water = 80:20 (v/v), solvent B: ethyl ace-tate, program #8 from 0 to 50% B in 30 min.

Fig. 1. Apparatus for fluorescence measurements. See materials and methods for details.

Results

I n v i v o a b s o r p t i o n s p e c t r a

Ageing cultures of R p . s p h e r o i d e s R26 change gradually their color from a clear blueish-green to a violet hue. The time evolution of this color change is rather different for different cultures and culture conditions. Two extreme situations are illustrated in Fig. 2. The slowly degrading culture (Fig. 2a) shows over a period of 46 days a decrease of the bchl Qx-band at 592 nm by about 50%. At the same time, there is an increase of a broad and structured band around 530 nm. This region is typical for the Qx-band of demetalated bchl, e.g. bacteriopheophytin (bphe) type pigments. The changes in the Qy region

H . Haidl et a l . • Degradation of Bacteriochlorophyll a in Rhodopseudomonas s p h a e r o i d e s R 26 687

500 600 (nm) 500 600 (nm)

Fig. 2. I n v i v o absorption spectra (Qx region of the tetrapyrrolic pigments) of ageing cultures of R h o d o -pseudomonas s p h a e r o i d e s R26. A : Slowly degrading cul-ture after 6 (a), 14 (b), 22 (c) and 46 days (d) of inocula-tion; B: Rapidly degrading culture after 4 (a), 14 (b), 20 (c) and 27 days.

are less significant with a uniform decrease and a minor blue-shift (see Fig. 3).

The fast degrading culture shows qualitatively the same changes, but both the decrease around 590 nm and in particular the increase around 530 nm are much more pronounced and rapid (Fig. 2b). In this culture, the 530 nm region shows two well resolved maxima at 522 and 538 nm. The difference between

500 600 700 800 900 (nm)

Fig. 3. I n v i v o absorption spectra of a slowly degrading culture of Rhodopseudomonas s p h a e r o i d e s R26 after 6 (a) and 46 days (b) of inoculation, and sedimented cells after 30 days of inoculation (c).

the two cultures is as yet only poorly defined. It is noteworthy, however, that light is not an important factor. Parallel cultures grown under otherwise iden-tical conditions in the dark or under continuous il-lumination (< 2000 lux), respectively, yield the same spectra.

The occurrence of a prominent absorption around 530 nm is accompanied by the formation of a pur-plish Sediment, which in light grown cultures may also extend to the walls of the culture vessel facing the light source, where generally a blanket of cells develops in cultures which are more than 10 days old. The Sediments show the aforementioned spectral changes in the Qx band region much more clearly than the still suspended cells. Pronounced changes are visible even in the Sediments of slowly degrading cultures. Moreover, the sedimented cells show also distinct changes in the Qy region which was obscured in the absorption spectra of the whole culture (Fig. 3, trace c). The maximum of this band is shifted to 860 nm, and second band arises at its short-wavelength Shoulder (?^max = 805 nm). In the fast de-grading culture, the Sediment is completely free of bchl if judged from the absence of the Qx band around 600 nm, and the 805 nm band becomes a distinct absorption band in the near infrared region (data not shown).

I d e n t i f i c a t i o n of t h e p i g m e n t s

The spectra of the methanolic extracts of aged cul-tures are typical for pigment mixtures of the bchl and bphe spectral type, which can be analyzed most read-ily by their Qx absorptions around 600 and 530 nm, respectively. In contrast to the i n vivo spectra, the extracts have only a Single, unstructured absorption around 530 nm (data not shown). Tic and H P L C analysis of the extracts yielded besides bchl several fractions which all have the same absorption spec-trum identical to bphe. The following pigments have been found in the extracts: Bacteriopheophytin a (bphe), bacteriopheophorbide a (bpheid), bac-teriopyropheophorbide a (pyro-bpheid) and a fourth, unidentified pigment. The latter is the most polar produet in the extract and is only found in the Sediment of fast degrading cultures. The identifica-tion of these pigments is based on (a) their absorp-tion spectra, (b) Chromatographie comparison (tlc, H P L C ) with authentic samples (s. Table I) and (c) conversion to the methyl esters and subsequent

688 H . Haidl et a l . • Degradation of Bacteriochlorophyll a in Rhodopseudomonas s p h a e r o i d e s R 26

Pigment bchl a bphe a pyro-bphe a bpheid a pyro-bpheid a

Retention 32 34 36 6 9 time [min]

Table I. Retention times of bac-teriochlorophyll a and its degradation produets (see materials and methods for details of the H P L C System).

Chromatographie comparison with the authentic methyl- and pyromethyl-bacteriopheophorbides a . The quantitative analysis of a fast degrading culture at different times, as well as of the Sediments after prolonged standing, is given in Table II. The bchl content decreases continuously in all cultures, and the major new produets are the unesterified, demeta-lated pigments, e.g. bpheid and pyro-bpheid. The re-lative amounts of these pigments varies, but bpheid is generally formed in larger amounts than is pyro-bpheid (the latter is absent in slowly degrading cul-tures). The esterified, demetalated pigment, bphe, never amounts to more than 5% of the total pigment.

F l u o r e s c e n c e s p e c t r a

Fresh cultures of R p . s p h e r o i d e s R26 have a Single major emission maximum at about 910 nm, with a typical bchl excitation profile [18]. Other pigments have been reported to be present in minor amounts, most likely as biosynthetic precursors of bchl. A slowly degrading culture of this bacterium shows essentially the same major fluorescence pattern (Fig. 4a, excitation at 600 or 400 nm, Fig. 4b, emis-sion at 925 nm). However, a distinct second near-

Table II. Pigment composition in ageing cultures of Rhodopseudomonas s p h a e r o i d e s R26. The data are taken from the H P L C analysis of the samples shown in Fig. 1B, all values are given in (%) of the total tetrapyrroles (bac-teriochlorophyll + bacteriopheophytin type pigments). " X " is the unknown polar Compound with bphe type ab-sorption spectrum (see text). The same extinetion coeffi-cients have been assumed for all the pheophytin-type pig-ments in the analysis. See Fig. 2 for the absolute decrease in pigment contents, which is always

CD O c d> o

o 3

9]1 exc i t a t ion

600 nm

600 700 800 900 X/nm -

525 nm

400 nm

1000

emission

765 nm

865 nm

925 nm

— i 1 1 1 1 1— 400 500 600 700 800 900

Ä/nm o » Fig. 4. Low temperature (4K) fluorescence emission (A , left) and excitation spectra (B, right) of a 28 day old slowly degrading culture of Rhodopseudomonas s p h a e r o i d e s .

O c O 00 (L> O D

exc i t a t ion

520 nm

600 700 800 900 1000 X/nm o

ü c a> o 00 0J \_ o 3

emission

754 nm

875 nm

909 nm

400 500 600 700 800 900 X/nm o

Fig. 5. Low temperature (4K) fluorescence emission (A, left) and excitation spectra (B, right) of the sedimented cells of a fast degrading culture.

690 H . Haidl et a l . • Degradation of Bacteriochlorophyll a in Rhodopseudomonas s p h a e r o i d e s R 26

Discussion

A l l the degradation produets investigated are of the bacteriopheophytin a spectral type. The degrada-tion is thus most easily detectable in the 530 nm

spectral ränge, where bacteriopheophytins but not the related bacteriochlorophylls possess a moderately intense absorption band. This finding is in agreement with the reports on degradation of Chlorophyll a in higher plants, where the demetalation to pheophytin

H . Haidl e t a t . • Degradation of Bacteriochlorophyll a in R h o d o p s e u d o m o n a s s p h a e r o i d e s R 26 691

692 H . Haidl et a l . • Degradation of Bacteriochlorophyll a in Rhodopseudomonas s p h a e r o i d e s R 26

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